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Earlier today, I made my first blog post. After a few hours, I was excited to see that a couple of people had commented their thoughts and opinions. When I went to read their comments, however, I got extremely confused. They both referred to me as Anna.
pic found here
My name is Rachel.
At first, I assumed that I somehow made a mistake and displayed my name as Anna. I went to check, and surely enough my name said Rachel Sara Anton. I came to the conclusion that the first person must have seen my last name, Anton, and confused it with Anna. But how does that explain the second comment? I called my friend who is also in Science 200 and told her leave a comment that started off with "Hi RACHEL" in all caps so that no one else would get confused. I check back an hour later, and get this: a fourth person left a comment: "Anna, this blog was very interesting."
COME ON GUYS. How could three people make the mistake of calling me Anna after my friend commented my real name in all caps, and my blog has my name clearly displayed as Rachel?
Then it hit me—social psychology.
Social psychology is the umbrella term that includes informational social influence and conformity, two of the concepts that I suspect caused my name to be Anna for the day.
Let's start with informational social influence. This is defined as the phenomenon where people adopt the actions of others in an effort to perform a behavior correctly for a given situation. In my blog situation, the second person most likely assumed that the first person was correct and therefore adopted the behavior of calling me Anna. In this article, splits social influence into two types: private acceptance and public compliance. In this situation, public compliance was used. Public compliance is when we copy our peers because we fear rejection if we behave otherwise. Most people would do just as the second person did in this situation. We as people often assume that if our peers do something, it is okay for us to do it as well. Think about your mom's favorite quote to keep you out of trouble: "If all your friends jumped off a bridge, would you do it too?" This is especially prevalent in situations where the mission at hand is significant to us, such as a homework assignment this early in the school year.
Conformity also plays a role in my blog dilemma. Conformity is when we alter our own way of thinking in order to fit in with the group. Think about the fourth person who commented. He or she most likely saw two people call me Anna and only one person call me Rachel. In this case, the person most likely conformed to the majority, and the majority thought my name was Anna. Even if this person may have questioned if my name could be Rachel, he or she felt more comfortable calling me Anna because that's what just about everyone else did. Although my blog post is a simple example, conformity is actually very complex and can be traced back to our brain (check out this article for a cool explanation).
I think this could all be related to the idea that science is not derived from eminent people, a concept we learned in class. Just as science is its own entity and cannot be claimed by "special" people, my name cannot be decided by my classmates. In the same way that we cannot simply trust observations and opinions about science, we cannot trust our peers just because they all trust each other.
This post was brought to you by RACHEL 🙂


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Tips for Selling Your Home | Process of Selling a House – Buy & Sell. Most Popular The Everything Guide to Selling Your First Home. Buy;. How to Find the Right Person to Sell Your House.. How to Get Your home show ready. In Preparing Your Home to Sell. How to Get Your Home Show Ready. In Buy.
6-month homebuyer countdown | Bankrate.com – 6-month homebuyer countdown. If one of your New Year's resolutions is to buy a home this summer, early in the year is the perfect time to get started.
How Do You Know When You're Ready To Buy A Home? – Buying a house is one of the largest financial decisions of your life; but how do you know when you're ready to buy a home? 5 questions to help you find out.
The "Am I Ready to Buy a House" Checklist – The Allstate Blog – October 9, 2012; By Quicken Loans Guest Blogger; https://www.allstate.com/blog/the-am-i-ready-to-buy-a-house-checklist/ If you're considering buying your first home.
3 signs you aren't ready to buy a home – CNBC – 3 signs you aren't ready to buy a home.. size of house (45 percent) and whether or not to buy a house in need of. it could be over a decade before typical millennials are ready to buy,
How Much Of A Mortgage Can We Afford How Much House Can I Afford? — The Motley Fool – We license calculators from CalcXML, who estimates how much house you can afford based on a few important items, including income, amount of money saved for a down payment, and monthly obligations.
How to Buy a House: 8 Steps (with Pictures) – wikiHow – · How to Buy a House. Purchasing a house is an incredible accomplishment and one of the biggest steps you'll take during your life. Though the process may seem overwhelming, breaking everything down will help you keep your head straight.
Knock.com | Learn how we buy or sell your house in 8 weeks. – You just keep paying your old mortgage and Knock will settle with you for any costs we covered on the new home when your old house is sold.
Under-40s now prepared to buy homes "with a stranger" to escape. – Which is why the news that an astonishing one in seven young Brits are prepared to buy a house with a total stranger to get on the property.
New Home Buyer Credits First-time homebuyer credit texas grants for first time home buyers | Internal Revenue Service – The home is destroyed or condemned and you do not replace it by rebuilding or buying a new main home within two years of the event You must repay the full amount of the credit. You must complete Form 5405 and attach it to your federal tax return for the year of the destruction or condemnation of the home to report the destruction or condemnation.
6 things every kid should know how to cook before leaving home – Sometimes I assisted my mom as she prepared blintzes. less and be healthier than if they buy it in a package. A base level confidence – which comes with a base level of comfort in the kitchen – is.
5 ways to prepare to buy your first house – WorkWell – Buying your first house is exciting. You get to go out and find a place to call your own and create memories. While this is a fun time, the reality is that buying a house is the most expensive investment you'll ever make.
17 things to know about closing your house deal | The Star – Closing day in a house deal is a milestone for both the seller and the buyer. To make it go smoothly, both buyer and seller must be prepared.

subset: commoncrawl


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IT & Software is now considered a key industry within the UK's fast-growing digital sector. Skilled candidates can find a range of information technology (IT) careers on offer, within IT Services, Software and Technology organisations.
In Britain, according to an industry organisation, the UK technology sector is growing more than two-and-a-half times faster than the overall economy, with employment in the tech category having increased by over 50% in the last decade.
The sector needs professionals at every level. Fields vary from Software, SaaS, to IT services and Technology.
Our job listings show the variety of vacancies available in the sector, such as:
Sales Director
Sales Manager
Head of Customer Success
Business Development Manager
Account Manager
Customer Success Manager
Account Executive
Inside Sales Manager
Customer Success Executive
Cambridge is one of the key hubs for world leading Software and Technology companies in the UK. There are also a key number of Software companies throughout the South East including Milton Keynes, Hitchin, Watford, Hemel Hempstead and throughout the home counties. With salaries ranging from £22k for entry level jobs to six-figure packages, this is a good time to make your next career move.
For the latest vacancies visit our jobs page.
According to Buckinghamshire's Business Secretary Greg Clark, "Buckinghamshire is already the fourth most productive area in England and has the highest employment rate in the Oxford-Cambridge Arc area". With many firms choosing Milton Keynes or Aylesbury as their base,
Over the last six years, Buckinghamshire-based firms received £30m of innovation funding from the UK Government, via Innovate UK, and £9m of innovation funding, via the Horizon 2020 programme. 60% of Innovate UK funding coming into Buckinghamshire was awarded to small and medium-sized firms. The bulk of this funding has been used to undertake innovation in the manufacturing, materials and mobility sector.


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- kwikfixAsked on April 01, 2012 at 08:33 AM
i have set up conditions to show a different additional field when different radio buttons are checked but they don't appear when these options are selected? Have I set something up incorrectly?
- JotForm SupportmlizAnswered on April 01, 2012 at 08:45 AM
Have you tried clearing the browser's cache and cookies or tried using a different browser? I pulled up the form online and the conditions you have set are working fine, the corresponding fields for the options Service, Sales and ITV were displayed when option is selected.
Let us know if this resolves the issue. Thanks!
- kwikfixAnswered on April 01, 2012 at 08:53 AM
i can see them working in IE but not Chrome, even after clearig cookies etc.
Is there a known issue with conditional fields in Chrome? Another thing I noticed, is that if I just copy the forms jotform url to test it, those conditional fields will show once but when I select another option and then back to the same option, it will not show again.
- JotForm SupportmlizAnswered on April 01, 2012 at 10:14 AM
I was able to replicate the issue with the odd behaviour of Conditions in Chrome for this reason I have created a ticket for our developers to look into what could be causing this.
We will keep you posted for whatever updates we have on this.
- webministerAnswered on April 05, 2012 at 03:20 PM
I too am having similar problem... The form works great with IE and Firefox, but I had a problem with layout redering in Chrome. All conditions looks and functions well except for Child1.
Form url https://secure.jotformpro.com/form/20948103134951
Please let me know of the solution
- pjsullivan75Answered on July 27, 2012 at 01:36 PM
It appears both Chrome and Safari have issues with conditional fields. Is this still being worked on by JotForm developers?
- pinoytechAnswered on July 27, 2012 at 01:51 PM
Hi pjsullivan75,
Yes, our developer is still working on this bug. Don't worry, as soon as our developer fixed this, we will inform you immediately via this thread.
Thank you!
- jamesozAnswered on December 22, 2013 at 05:48 PM
Did this issue get resolved? I have similar issues.
- JotForm SupportjonathanAnswered on December 22, 2013 at 09:23 PM
@ jamesoz
This issue was resolved already.
But please create a new separate thread regarding your issue.
Use this link http://www.jotform.com/contact/
Provide as much details as you can and we will attend to it as soon as we can.
- jamesozAnswered on December 22, 2013 at 09:35 PMHi,
The issue was not resolved, that's why I replied.
Please confirm,
- JotForm SupportEltonCrisAnswered on December 23, 2013 at 12:44 AM
Please stick to this thread http://www.jotform.com/answers/308261 to get further updates about your question. Thank you!


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Great Yarmouth charter celebrations
PUBLISHED: 13:13 24 February 2009 | UPDATED: 13:07 03 July 2010
THE end of the 800th anniversary year for the signing of the King John Charter will be marked at the weekend.
Celebrations to mark the end of the charter year will be held on Saturday where Royal Navy ship HMS Hurworth will host a service on the quayside.
THE end of the 800th anniversary year for the signing of the King John Charter will be marked at the weekend.
Celebrations to mark the end of the charter year will be held on Saturday where Royal Navy ship HMS Hurworth will host a service on the quayside.
Winterton Marine Cadets TS Fearless will lead a procession, including mayor and mayoress Terry and Jenny Easter, from the Town Hall to the quay shortly before 2pm with residents being asked to line the quayside.
The service will be led by canon Michael Woods in his last civic engagement before his retirement and will include readings from the vice lieutenant of Norfolk, Lady Knowles and deputy council leader Barry Stone.
At the end of the service sirens from boats in the harbour, including lifeboats from Gorleston, Caister and Hemsby, will sound along with the bells at St Nicholas' Church in Yarmouth.
The 750 tonne vessel will arrive in Yarmouth on Friday and commanding officer Lieutenant Commander Charles Maynard will host a reception attended by the mayor and mayoress and other invited guests.
Lt Cmdr Maynard said: "We are very much looking forward to spending time in Yarmouth and fell honoured to have including in this important celebration. I hope as many Yarmouth as possible will be able to attend the event."
Councillor Bert Collins said: "All through the year we have celebrated the achievements of the borough without paying due attention to the wealth which has come from the sea and the river."
HMS Hurworth, which is named after a village in County Durham, is making the first of two potential visits to Yarmouth. The Hunt class mine counter measure vessel is also pencilled in to return in September for the Maritime Festival. The ship carries a crew of 45 many of them clearance divers whose job it is to swim to mines in order to place explosives.
The charter was granted to Yarmouth in 1208 by King John bringing freedom to the borough forever. Celebrations have included a visit by The Princess Royal in June, a medieval banquet, and a painting by local artist Ernie Childs using faces of today's residents depicted in 13th century dress and exhibitions.
If you value what this story gives you, please consider supporting the Great Yarmouth Mercury. Click the link in the orange box above for details.


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Minnesota Wild defeat Nashville Predators, 5-4
At the end in Chicago, Andrew Ebbett only broke a sweat chomping on pressbox popcorn.
At the end in Montreal, Guillaume Latendresse broke a bigger sweat getting out of his car than on the ice.
But Ebbett and Latendresse have come to Minnesota, discovered new life, and in exchange have helped bring new life to a team that was in need of resuscitation.
One game after Latendresse scored the tying goal and Ebbett scored his first career shootout goal at Colorado, the Wil...


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Trusted, Effective Treatment for Fear Of Achievement.
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Because people are coming over for a holiday dinner, Aki will get a bath today. I hide this from her. The little dog has a love/hate relationship with soap and water. She hates the bath but loves to dash around the house when freshly clean. She also loves to explore tidal meadows—a place she is unlikely to leave without mud and muck imbedded in her poodle fur.
We drop down from a well-used trail to the meadow and hear a series of "rock dropping into a well" sounds. Ducks start migrating over our heads, at first in one and twos, and then in dozens. I blame the Labrador retriever that went down the trail ahead of us for flushing the birds until I spot him heading back to the trailhead with his owner. Do the math Dan. The sound of something being plunked into water followed by ducks in frantic flight equals crab pot placements. Someone just dropped a line of crab pots into a chunk of Smuggler's Cove covered with waterfowl. We won't have much to watch when we reach the cove overlook so I snap pictures of fleeing mallards as they pass into front of the Mendenhall Towers and Mount McGinnis.
It's a day for rain forest dwellers to cash in on an early spring day with sun and temperatures in the 50's. Some with boats spend this warm Easter Morning dropping crab pots. Others troll for feeder king salmon off False Outer Point. Many, like Aki and I, just look at things.
We run into a group of serious bird watchers with serious spotting scopes and serious tripods for mounting them. Understandably, they are not happy to receive Aki's happy, if loud greeting. I would have told them where the Smuggler Cove ducks now feed but did not want to intrude.
The little dog and I, we spend this holiday morning with sun on our faces listening to eagle song and duck complaints. I wonder at the beauty so accessible to one with eyes and interest; the little dog rolls in something rich in sea smells that last night's tide left in the meadow grass. Thank God for the day and that Aki is scheduled for a bath.


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Are you one if the millions, who love ironstone china? Count me in!
I brought home my modest collection from the shop this week, because it just wasn't selling. I attribute that to two things. 1) I'm not be buying it right (price wise) or, 2) people don't know it's history, or appreciate its longevity and simplicity.
One thing for sure is that you can't buy it on eBay, pay for shipping and still have room for profit! I have made some good buys, here in town, though.
Since buying my first piece, I've learned a couple of things. By no means am I an expert!
I'll avoid pieces that are from China. I understand that there are lots of reproductions on the market that are being sold as originals. This wasn't one of them.
U. S. companies, like Homer Laughlin, have been making ironstone in the country for years. But it's been made recently as well. It's easy to sell a contemporary piece as something old.
I've been told to look for wear that would be natural for a piece that's over 100 years old. If it looks too good to be true, it probably is!
These two Johnson Brothers pieces look like they are from different eras.
I've seen plates displayed over headboards for a while now. I love the look, although, it doesn't make a lot of sense to me. What are you supposed to do? Have breakfast in bed, with plates off the wall? Oh well, I've jumped on the bandwagon, here in my daughter's room.
What about you? Do you love ironstone? Have a special place to buy your stash?
Ready to spill the beans?
Do tell!
Have you entered my wax brush give away? Sure hope so!


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Alexander Biggs Still You Sharpen Your Teeth (Vinyl)
Still You Sharpen Your Teeth (Vinyl)
'Still You Sharpen Your Teeth is the debut EP from 23-year-old Melbourne artist, Alexander Biggs. Alexander has a clear vision for his new music; emotionally tuned, melodically captivating, and philosophically grounded. With the release of his new single 'Figure It Out', the singer-songwriter emerges from a place of fear and uncertainty with hope and self-acceptance. The past year has seen Alexander hone his live talents, with a myriad of support slots for artists including Evan Dando, Frightened Rabbit and Gretta Ray. It's clear that Alexander's journey up until now has been one of constantly learning, questioning and, more recently, coming to terms with his rapidly changing environment. Still You Sharpen Your Teeth, showcases Alexander's ability to set a scene and sustain a mood. "The themes all came from the same place," he says. "It's about how everything will pass, the ever-changing world, nothing sticks for too long."
|DISC 1 - LP - Side A
1 I'll Go My Way
2 Now I Leave You Here
3 Figure It Out
DISC 1 - LP - Side B
1 It's
2 New York - Explicit Content
3 Gone Again - Explicit Content


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- Description
Brand New & Sealed, Official UK Stock, Next Working Day Delivery (M-F.)
1 Year
Forza Motorsport 5
Forza Motorsport 5 is the latest edition of the highest-rated racing franchise of the past 10 years.
Built from the ground up to take advantage of Xbox One and the infinite power of the cloud, prepare yourself for the amazing sensation of being behind the wheel. Forza Motorsport 5 will be available exclusively for Xbox One at launch.
- Specification
SKU V5D-00010-Feed
- Delivery
Delivery information
To cater the needs of all customers, we offer two delivery options at checkout so you can choose one which best suits your requirements.
Free UK Delivery
In this option we will deliver your product free of charge but it will take 3 to 5 working days for the product to be delivered.
Next Working Day Delivery
If you want your desired product to be delivered next working day you will have to place your order before 3pm. The delivery charge is £6.95 at the time of placing order. This option does not include delivery on Saturday or Sunday.
- Reviews


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Event description
Have you ever considered being an entrepreneur but have hesitated about taking the next steps? This session will provide you with all of the critical considerations before making the leap.
event ended
Me Inc Workshop - September 13, 2016 - Nelson, BC
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|Ethan on a Rock in Danby, NY
Sometimes I make mountains out of mole hills.
Sometimes I look at a problem and think it is much bigger than it really is and wonder how I am ever going to be successful.
Sometimes I look at the schedule for the week ahead and wonder how I am ever going to make it to Saturday.
Every so often though, the rock is really big!
And I still get through it. Sometimes it just takes a little help.
-- Bob Doan, writing from Danby, NY


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As President and CEO of Florida Hospital Memorial Medical Center, Daryl Tol has a gameplan for keeping Ormond Beach healthy.
If one were to ask Daryl Tol's father how his son can successfully navigate the unpredictable waters of healthcare to lead multiple patient-care facilities within Flagler and Volusia counties, he would probably say that, for his son, it's all somewhat of a game.
"He loves to play strategy games and, I think life, for him, has always been a strategy game," says Bill Tol, Daryl Tol's father and development officer at Florida Hospital Flagler.
Born in small-town Tennessee, Daryl Tol was only a few months old when his parents relocated to Indonesia to work as missionaries. After spending six years overseas, his family relocated to California and, eventually, Tol moved to Volusia County to work for the Adventist Health System.
Now, as President and CEO of Florida Hospital for Volusia and Flagler markets and Memorial Medical Center, Tol's job is to ensure the hospital system runs efficiently, and effectively.
The strategy for Tol is to take on big tasks in small doses, and to surround himself with the right teammates.
"You really have to focus on being balanced and being calm in the face of difficulty," Tol says. "If you don't manage your schedule it will manage you."
Tol says the hospital system must be ready for change and have the ability to reinvent itself.
"We are right in the midst of healthcare reform," Tol says. "Many changes have already happened, and in 2013 and 2014, more changes will come."
The biggest challenge locally, he added, will be broadening the focus on the patient. Through the strengthening of health plan partnerships, new programs will be created that emphasize wellness and prevention. The hope is to confront illnesses throughout all all stages of life before they become illnesses. And that'll take planning.
But for Tol, he takes it all one day at a time. It's about staying true to the fundamentals of Florida Hospital.
"We focus on what we do as sacred work, very meaningful, faith-based care," Tol says.
Regardless of one's faith, though, Tol says his objective is to maintain quality healthcare while providing "an exceptional patient experience." And that's easier than some might think, given the advanced facilities at his disposal. The technological capabilities of his hospital extend beyond the size of the community, he says. But he's still not satisfied.
Expected to be completed by summer, a $6 million project is in the works at Florida Hospital Oceanside and Peninsula Rehabilitation Center to make private 40 patient rooms and improve rehabilitation areas. Renovations will also include new visitor amenities and gathering areas.
For any size project, though, it's important for Tol to have the right teammates in place. Much of his time is spent meeting not just with other hospital leaders but also with community members and physicians.
"Every day is really about working with a large team of people to get the job done, and that's the fun part of it," Tol says.
But fun comes in other forms, as well. Family time is important to Tol, and he is careful to let work stay at the workplace. Although they each work for the hospital, Bill and Daryl Tol rarely talk of work when they're together.
Tol also uses his spare time to exercise, do work outside and cook.
"When you're used to working with your head all the time, it's nice to go do yard work or go work in the kitchen," Tol says.
As healthcare continues to change, Tol is tasked with making sure the hospitals and patient-care facilities he oversees can adapt. And as long as he keeps committed to his strategy, for Tol, the clear winner of this game is the people of Flagler and Volusia counties.
Age: 39
Family: He and his wife, Stacey, have three kids, one 13-year-old and two 11-year-old twins.
Position: President and CEO of Florida Hospital for Volusia and Flagler markets, and Memorial Medical Center
Quirky fact: He plays piano and French horn.


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My daughter, Steph, and I tackled a little project the other evening.
This is a great way to reuse baby food or other small jars and scrap wood.
We followed this great tutorial, so I'm not going to go into a ton of detail here.
If you want to make your own and have questions, follow the link.
We laid out all our supplies on the dining room table and gave the baby snacks and toys.
This was Steph's first time handling a drill.
Measuring to make sure we had the clamps centered and evenly spaced was a bit tedious.
The little clamps are ready for jars!
Although it can be hung, I opted to prop my new votive holder / bud vase against the wall.
Isn't the little kitty teapot so cute? She's a recent antiquing treasure.
Doesn't it cast some pretty shadows?
I think at some point I will hang it on the back porch, but for now…
…I like it in the kitchen where I can see it most of the day.
My new wall decor is making me smile.
Thank you for stopping by today!!


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Tips on How to Choose an Incredible Painting Company
Painting is very important in determining how good or bad a house or office looks like at the end of construction or renovation or designing. Paints of varying designs and colors exists and it is very crucial to choose a design and color that matches perfectly with the surrounding objects and designs too. Due to this, it is of uttermost importance that you choose a painting company that is good at its work and has the necessary experience and expertise in painting to do the painting work for you. There are many painting companies available to do painting works and here are some tips on how to choose a good Painting company.
Licensing is a crucial factor of consideration when choosing a painting company and this should be the fact thing to look out for so as to ensure you do not hire the services of quacks ad amateurs. Any incredible painting company has to have legitimate papers and certification from the relevant authorities to carry out contracts with clients. Any painting company that is certified to carry on contracts is safe to work on your premises in the safest way possible and with a lot of expertise. Take some time to look at the previous painting contracts a painting company has taken on before choosing them.
The track record of any painting company worth working with should be ravishing and one that motivates you to choose to hire their services. You might want to ask for photos of the previous contracts that they may have worked on to ascertain that their work is impressive and of the highest quality attainable. If the painting company has got an online platform where clients can give their feedback on the services provided to them, this would be the best place to check on their track record by reading the comments and feedback posted by previous clients.
Any incredible painting company has an impressive track record that is satisfying enough to make you want to work with them. Besides the track record, make inquiries on the grade and quality of the paint the painting company uses for their work. There are paints that are of a good grade and of very high quality which are meant to last for quite a long time, water proof, looks good and does not cause any harm to your health and this is the kind of paint that is mostly used by incredible painting companies. Those are the guideline that should help you identify the quality of the paint the contractor uses.
When it comes to contracts, cost and charges is always an important aspect of consideration. Go through the charges of various painting companies that are available in the market. Quality painting works, of course, come at a higher cost than that of lower quality and that is why it is recommended to avoid painting companies that have got very low quotations.
What I Can Teach You About Services


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Redesigning an FKBP–ligand interface to generate chemical dimerizers with novel specificity
Edited by Christopher T. Walsh, Harvard Medical School, Boston, MA, and approved June 18, 1998 (received for review May 1, 1998)
FKBP ligand homodimers can be used to activate signaling events inside cells and animals that have been engineered to express fusions between appropriate signaling domains and FKBP. However, use of these dimerizers in vivo is potentially limited by ligand binding to endogenous FKBP. We have designed ligands that bind specifically to a mutated FKBP over the wild-type protein by remodeling an FKBP-ligand interface to introduce a specificity binding pocket. A compound bearing an ethyl substituent in place of a carbonyl group exhibited sub-nanomolar affinity and 1,000-fold selectivity for a mutant FKBP with a compensating truncation of a phenylalanine residue. Structural and functional analysis of the new pocket showed that recognition is surprisingly relaxed, with the modified ligand only partially filling the engineered cavity. We incorporated the specificity pocket into a fusion protein containing FKBP and the intracellular domain of the Fas receptor. Cells expressing this modified chimeric protein potently underwent apoptosis in response to AP1903, a homodimer of the modified ligand, both in culture and when implanted into mice. Remodeled dimerizers such as AP1903 are ideal reagents for controlling the activities of cells that have been modified by gene therapy procedures, without interference from endogenous FKBP.
A number of natural and synthetic compounds are known that bind with high affinity to the human protein FKBP12 (1–5). Bivalent versions of these compounds—chemical "dimerizers"—recently have been developed as cell-permeant reagents to control intracellular signaling events that are naturally regulated by protein–protein interactions (6, 7). In these applications, cells are engineered to express a chimeric protein comprising a signaling domain fused to one or more FKBPs; treatment with dimerizer crosslinks the proteins and initiates signaling. Because genes for such fusion proteins can be delivered via gene therapy strategies, dimerizers might be used to control a wide range of proliferation and differentiation events for therapeutic purposes (8, 9).
Use of FKBP dimerizers in vivo is complicated by their potential interactions with endogenous FKBPs, which are ubiquitous and highly expressed in mammals. These interactions could blunt potency by allowing nonproductive dimerization events and compound sequestration. High concentrations of ligand also might interfere with the physiological functions of FKBP12, such as calcium channel modulation and regulation of cardiac development (10–12). The ideal dimerizer therefore would interact minimally with endogenous FKBP; but any modifications that reduce binding would have equivalent effects on the affinity for the chimeric target protein.
We set out to solve this specificity problem by engineering a unique pocket into the target FKBP that differentiates it from the endogenous protein. This approach is feasible because dimerizer applications involve the delivery of the target as well as the drug. Our aim was to add "bumps" to ligands to block sterically binding to wild-type FKBP and then generate compensating "holes" by mutagenesis that could be installed into target fusion proteins. We report here the design of such a remodeled ligand–FKBP pair and evaluation of the resulting bumped dimerizer as a signaling reagent in vivo.
Synthetic Chemistry.
The synthesis of compound 1 has been described (13). Isomers of compound 5 were prepared as shown in Fig. 3. Compounds 2–4 were prepared analogously by coupling amine 7 (Fig. 3) with 3,3-dimethyl-2-methoxy-pentanoic acid, 3,3-dimethyl-2-ethyl-pentanoic acid, or 2-phenyl-butanoic acid, respectively. FK506 was coupled to 4′-(aminomethyl)-fluorescein (4′-AF; Molecular Probes) analogously to the synthesis of FK1012 (6). Fluoro-5S was prepared by coupling 5S to 4′-AF by using 1,3-dicyclohexylcarbodiimide and 4-dimethylaminopyridine in dichloromethane.
Protein Engineering and Expression.
FKBP and its mutants were expressed with an N-terminal hexa-histidine purification sequence and an influenza hemaglutinin (HA) epitope tag. Expression vector pET-H6HA-FKBP was assembled by cloning an NdeI–NdeI oligonucleotide pair encoding the amino acid sequence MHHHHHHYPYDVPDYAAMAHM and an NdeI–BamHI PCR product encoding human FKBP12 into NdeI/BamHI-digested pET20b(+) (Novagen). Mutants were engineered as described (14). Proteins were expressed in Escherichia coli BL21(DE3) and released in ≥50% purity by a single freeze-thaw step as described (15) and were purified further on Ni-NTA agarose (Qiagen) under native conditions. For structural studies, an expression vector for untagged F36V-FKBP was generated by excising the NdeI fragment from pET-H6HA-F36V-FKBP followed by recircularization. Protein for crystallography was expressed and extracted as before and purified to homogeneity by DEAE ion-exchange chromatography followed by gel filtration on S-100 Sepharose.
FKBP Binding Assay.
Fluorescence polarization competition assays were performed as described (C.T.R., E. Laborde, D.A.H., and T.C., unpublished work). In brief, subsaturating concentrations of protein were incubated for 30 min with 2.5 nM fluoro-FK506 and serial dilutions of competitor ligand in the wells of a Dynatech microfluor plate. Polarization was read on a Jolley FPM-2 (Jolley Consulting and Research, Grayslake, IL). The increase in polarization on binding was used as a direct readout of percentage of bound probe, compared with controls containing no competitor (100%) or no protein (0%). The concentration of competitor resulting in a 50% probe displacement (IC50) was determined by a nonlinear least square fit by using the four-parameter algorithm where Pmin, Pmax, and P represent the minimal, maximal, and measured polarization values, respectively. Direct binding experiments with fluoro-5S used 10 pM ligand, and polarization was read on a Jolley FPM-1 reader.
X-Ray Crystallography.
Crystals of the complex of F36V-FKBP with 5S were obtained by vapor diffusion in hanging drops containing 40 mg/ml complex in 1.2 M ammonium sulfate, 0.1 M sodium phosphate (pH 6.0), and 20 mM DTT over reservoirs of 2.4 M ammonium sulfate and were triclinic (a = 32.73, b = 41.85, c = 43.59 Å, α = 62.61, β = 82.75, γ = 85.29°) with two complexes in the asymmetric unit. Data were collected at room temperature with a Rigaku R-AXIS II area detector with graphite monochromated Cu Kα x-rays. Data were collected as 2° oscillation images, reduced to integrated intensities with the program denzo (16), and scaled with rotavata, and agrovata (17). The structure was solved by molecular replacement with amore (17) and refined by using x-plor (18). The current R value for reflections with F > 2σ from 10.0 to 1.9 Å is 0.21 and Rfree is 0.26; rms deviations are 0.009 Å for bonds and 1.78° for angles. Coordinates have been deposited with the Brookhaven Protein Data Bank (PDB ID code 1bl4).
Cell Line for in Vitro Fas Studies.
Construction of the retroviral plasmid pSRα-myristoylation-2xFKBP-Fas-E has been described (13). An equivalent vector incorporating the F36V mutation in both FKBPs was prepared analogously. Cloned HT1080 cell lines (ATCC CCL-121) retrovirally transduced with Fas constructs were prepared as described (13). Cell viability after overnight incubation with AP1903 was measured by Alamar Blue assay (13).
Cell Line for in Vivo Fas Studies.
pSRα-LNGFR-2x(F36V-FKBP)-Fas-E encodes the extracellular and transmembrane portions (amino acids 1–274) of the human low affinity nerve growth factor receptor (19), derived by PCR from a human cerebellum cDNA library, in place of a myristoylation sequence. The constitutive human growth hormone (hGH) expression plasmid pCEP4-hGH was made by subcloning a genomic hGH clone (20) into the episomal plasmid pCEP4 (Invitrogen). To prepare cells that constitutively express both F36V-FKBP-Fas and hGH, a G418-selected population of HT1080 cells retrovirally transduced with pSRα-LNGFR-2FKBP F36V-Fas-E was transfected with pCEP4-hGH by lipofection, and clones were selected in hygromycin B. Clone HTFasGH-3 was selected for its high response to AP1903 in the absence of actinomycin D, and high hGH production.
Animal Experiments.
Male nu/nu mice were obtained from Charles River Laboratories (Wilmington, MA). For injection, HTFasGH-3 cells were harvested from tissue culture dishes in PBS/0.1% glucose/10 mM EDTA, washed, and resuspended in PBS/0.1% BSA/0.1% glucose at a concentration of 2 × 107 cells/ml. Between 2 and 4 × 106 cells were implanted into two i.m. sites. After 24 h, mice were administered i.v. AP1903 formulated in [50% N,N-dimethylacetamide/50% (90% PEG-400/10% Tween 80)] at 2 ml/kg. After a further 24 h mice were killed and serum hGH concentrations were determined by ELISA (Boehringer Mannheim 1-585-878).
Molecular Design.
The natural product FK506 and synthetic derivatives exemplified by compound 1 (Fig. 1A) share a conserved α-keto-pipecolylamide core that binds in a complementary hydrophobic pocket on FKBP, as revealed by x-ray crystallography (Fig. 1B) (2, 5, 21). We focused on the C9 carbonyl oxygen, which packs tightly against Tyr26, Phe36, and Phe99, all making unusual edge-on ɛ-hydrogen contacts. Inspection of crystal structures suggested that replacing this carbonyl with larger substituents would dramatically reduce FKBP binding through steric clashes with either Phe36 or Tyr26 (depending on the stereochemistry at a tetrahedral C9 carbon; Fig. 1A). This in turn suggested that mutations that truncate these side chains might provide compensatory holes that would restore binding.
Interface Remodelling.
We engineered a set of FKBP mutants in which Phe36 was replaced with smaller hydrophobic residues and measured their affinities for C9-alkylated analogs of compound 1 by using a competitive fluorescence polarization assay (Table 1). This assay reports IC50 values that are proportional to dissociation constants (Kd). Compound 1 bound to wild-type FKBP with an IC50 of 86 nM [corresponding to a Kd of ≈20 nM (2)], and binding was largely unchanged for the Phe36 mutants. Even modest enlargements of the C9 substituent to methoxy (compound 2) or ethyl (compound 3) reduced affinity to undetectable levels (IC50 > 10,000 nM), as expected. However, binding could be rescued by any of the Phe36 mutations, and although affinities were fairly low, the compounds showed apparent selectivity of up to 50-fold for the mutants over wild type. Two further trends were apparent. First, compound 3 bound more tightly than 2, indicating that nonpolar substituents may be preferred. Second, the mutants clearly favored the S-isomer of compounds over the R. This is the preference predicted by modeling, so it suggested that the modifications were acting as specificity determinants as envisaged by our design.
To try to improve the affinities of these specific pairings, we retained the ethyl bump of 3 and varied the tert-pentyl "bottom" C9 substituent (2, 4, 5). Installing a phenyl group (compound 4) improved affinities for each of the mutants by 3- to 5-fold while maintaining the lack of detectable binding to wild-type protein, and additional 5- to 20-fold increases were observed when a trimethoxyphenyl group was incorporated (compound 5). As the affinities improved, so did the preference for the S-isomer over R, suggesting that the changes were not altering the binding of the engineered bump. Strikingly, none of the bumped compounds showed any major preference for a particular mutant; instead, affinities were largely insensitive to the size of the new side chain. This implied that, rather than a precise "lock-and-key" fit, molecular recognition in the new pocket was rather loose and flexible.
The best combination was the pairing of 5S with the Phe->Val mutant (F36V-FKBP), with an IC50 (1.8 nM) close to the resolution limit of the assay. To assess the true affinity of the compound, we coupled it to fluorescein and performed direct fluorescence polarization titration. The labeled compound, fluoro-5S, had a Kd of 0.094 nM for the F36V mutant, compared with 67 nM for the wild-type protein (data not shown). Thus, although binding to the wild-type protein was still detectable, specificity for F36V-FKBP was almost three orders of magnitude. Moreover, the affinity of fluoro-5S for F36V-FKBP exceeded that of any known ligand for the wild-type protein (1, 5).
Crystallographic Analysis of Remodeled Interface.
To determine the structural basis for these binding characteristics, we crystallized the complex between 5S and F36V-FKBP and determined its structure to 1.9-Å resolution (Fig. 2). Overall, except for the mutation, the structure of the protein is essentially superimposable with wild-type FKBPs from unmodified complexes (rms deviation for main chain atoms 0.45 Å). The F36V substitution opens up a cavity of ≈90 Å3 (Fig. 2A), and surprisingly—apart from a 0.2-Å movement of the Val36 main chain—the protein does not relax to fill it. As a result, the pocket is lined by the predominantly hydrophobic side chains that usually pack against Phe36, and the Gly28 mainchain is exposed at the base of the pocket (Fig. 2B). The S-ethyl bump of the ligand inserts into this cavity exactly as predicted, penetrating what would be the surface of Phe36 in the wild-type protein and making multiple van der Waals contacts (Fig. 2 C and D). Compared with a complex between wild-type FKBP and a conventional ligand (2), the atoms of the core motif move by only 0.2–0.4 Å (Fig. 2D). Thus the modifications lead to a largely "surgical" deletion and insertion of contacts.
One remarkable consequence of this is that the bump only occupies ≈40% of the engineered hole, so that a large, fully enclosed cavity of ≈60 Å3 remains within which no ordered water molecules are visible. Improving the fit by extending the ethyl bump of 5S to an allyl group did increase affinity but only ≈3-fold (data not shown). Thus, affinity is largely insensitive to the quality of packing in the engineered pocket, possibly because recognition occurs mainly through nondirectional hydrophobic interactions (5). These structural observations correlate with our binding data showing that many Phe36 substitutions confer good binding affinity and that nonpolar bumps are preferred (Table 1).
The remaining peripheral substituents of 5S pack into hydrophobic channels, essentially as observed for unbumped compounds (2). The conformations of these groups can vary widely between ligands (2, 4, 5), and because we do not have the structure of unbumped 5S for direct comparison, we cannot assess effects of the modification—although the C9-trimethoxyphenyl group would have to shift because of the altered C9 stereochemistry (Fig. 2D). The substantial binding energy this group confers to 5S (Table 1) probably stems from two good hydrogen bonds to Arg41 and His87 and also extensive intramolecular contacts (Fig. 2 A and B) that may preorganize the ligand and decrease the entropic cost of binding (2, 5).
Activation of Fas Signaling with a Remodeled Dimerizer.
Having identified 5S as an appropriately specific ligand, we coupled together two molecules through a previously identified linker (13) to create the bumped dimerizer AP1903 (Fig. 3). The binding affinity and specificity characteristics of AP1903 mirrored those of its monomeric precursor (Fig. 4), confirming that AP1903 should interact minimally with endogenous FKBP inside cells. As a first test of the potency of AP1903, we determined its ability to activate signaling through a chimeric Fas receptor. Previous work showed that cells expressing membrane-tethered fusions between FKBP and the intracellular domain of the Fas receptor undergo apoptosis in response to appropriate dimeric FKBP ligands (8, 13). This approach is a promising general method for eliminating engineered cells following gene therapy procedures, but clinical application may require ligands that can operate independently of endogenous FKBP.
The human fibrosarcoma line HT1080 was engineered to express stably a fusion protein comprising a myristoylation sequence, two copies of F36V-FKBP, and the human Fas intracellular domain. AP1903 elicited potent and dose-dependent apoptotic death of these engineered cells in culture, with an EC50 of ≈0.1 nM (Fig. 5A). This potency corresponds directly to the binding affinity and represents a 60-fold improvement over the previous best unbumped compound (AP1510, a synthetic dimer of 1; Fig. 1B) (13). By contrast, activity was reduced by over two orders of magnitude on an analogous cell line expressing a wild-type FKBP Fas fusion protein. Of interest, the level of discrimination (200-fold) was lower than that observed for binding in vitro (1,000-fold). We speculate that this reflects stabilization of the weak interactions between AP1903 and membrane-tethered tandem wild-type FKBP fusion proteins (the avidity effect). No effect was observed on unmodified HT1080 cells at concentrations up to 1 μM (data not shown). Thus, the affinity and specificity of 5S were reflected in dimerizer potency in cells.
Activation of Fas Signaling in Vivo by AP1903.
To test whether AP1903 can function in vivo, we determined its efficacy in an mouse model of conditional cell ablation. We engineered a second HT1080 cell line expressing a Fas-F36V-FKBP construct that also constitutively secretes hGH. Constitutive hGH secretion provides a convenient and accurate way to monitor the number of viable cells in vivo because hGH has a serum half-life of only ≈3 min in mice (22, 23). Over 3 consecutive days, we implanted these cells i.m. into nude mice, treated the animals i.v. with various doses of AP1903, and then determined serum hGH levels as a measure of the number of surviving cells. Fig. 5B shows that AP1903 elicited a dose-dependent decrease in serum hGH levels, with a half-maximal effective dose of 0.4 ± 0.1 mg/kg. Thus, the bumped dimerizer retains potent activity in vivo, indicating that the remodeled interface is functional in a whole animal context.
General Implications for Drug Design.
In this study, we redesigned a molecular interface to generate compounds that can differentiate 1,000-fold between two proteins differing by a single point mutation. This problem resembles a common challenge in conventional drug discovery, where efficacy can depend on discrimination between closely related homologs. For example, anti-inflammatory drugs that specifically inhibit human cyclooxygenase-2 (COX-2) while sparing COX-1 are expected to show significantly reduced toxicity (24). A cavity caused by the amino acid substitution Ile523Val is the major differentiating feature of the COX-2 binding site (25). Therefore, lessons learned in our work may be generally relevant to drug design.
In particular, we found that our designed ligand has subnanomolar affinity for its target despite the presence of an enclosed hole in the binding interface. Cavities at interfaces previously have been considered incompatible with high affinity ligand binding, which is invariably characterized by excellent shape complementarity (5). Furthermore, holes engineered into protein cores are destabilizing (26). Our observations suggest surprisingly that such holes need not be detrimental to affinity. Selectivity of conventional drugs might therefore be attained by relatively imprecise recognition of surface cavities in target proteins—particularly when those cavities are largely nonpolar—and binding affinity then improved by secondary modifications, as in this work.
Comparison to Other Remodeled Interfaces.
We found that our interface modifications caused only local structural changes. Redesigned interfaces also have been engineered between cyclophilin and cyclosporin (27, 28) and between Src tyrosine kinase and ATP (29). Of interest, molecular modelling generally suggests that, in these cases, holes would remain unless rearrangements occurred (30), but no direct structural data are yet available. However, our results are in striking contrast to a recent report of a remodeled complex between two proteins, where major structural adjustments were propagated across the entire interface (31). Additional studies will be required to determine whether this reflects an inherent difference in "plasticity" between the two types of complex.
Future Applications of AP1903.
By engineering out binding to endogenous FKBP, we have eliminated an important potential obstacle to the therapeutic use of chemical dimerizers. Unexpectedly, our design efforts also produced a ligand with higher affinity than any previous dimerizer. AP1903 should therefore be effective for controlling a wide range of cellular events inside cells and whole organisms, at low concentrations and independently of endogenous FKBP.
Inducible apoptosis may be an important early clinical application because the ability to eliminate engineered cells could be broadly useful—for example, to abort a graft-versus-host response after allogeneic bone marrow transplantation (32) or as a general failsafe for gene therapy procedures. Current approaches involve expressing nonhuman enzymes that can metabolize pro-drugs to lethal products (for example, the combination of herpesvirus thymidine kinase and gancyclovir), but immune recognition of the foreign protein is a serious problem (32, 33). AP1903-inducible Fas activation is an attractive alternative because apoptosis is a cell-autonomous and noninflammatory process that occurs rapidly and in the absence of cell division, and Fas-FKBP-F36V is built from human proteins and should be minimally immunogenic.
We thank Terry Keenan and Edgardo Laborde for reagents, Brian Gladstone and Stuart Schreiber for early discussions and advice, and Dale Talbot and Rebecca Ward for helpful comments on the manuscript.
↵* To whom reprint requests should be addressed. e-mail: clackson{at}ariad.com.
↵† Present address: MGH Cancer Center, 13th Street, Charlestown, MA 02129.
↵‡ Present address: Harvard University School of Public Health, Huntington Avenue, Boston, MA 02115.
This paper was submitted directly (Track II) to the Proceedings Office.
Data deposition: The atomic coordinates have been deposited in the Protein Data Bank, Biology Department, Brookhaven National Laboratory, Upton, NY 11973 (PDB ID code 1bl4).
- human FK506-binding protein 12;
- FKBP mutant with Phe36 replaced by Val;
- hGH,
- human growth hormone
- Received May 1, 1998.
- Copyright © 1998, The National Academy of Sciences
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Torii Hunter is wondering why Boston police officer Steve Horgan didn't try to help him when he dived into the Red Sox bullpen in the 8th inning last night.
The Detroit Tigers pitching had totally dominated Boston up to that point over two games, and the Tigers seemed on their way to an nearly insurmountable 2-0 lead in the American League Championship series. The Sox had mustered just one hit in game one, and were also no-hit by Max Scherzer for 5-2/3 innings in game two. As the Boston Herald points out, "Through the series' first 16 innings, the Sox were 3-for-49 (.061) with a whopping 30 strikeouts."
In the 8th inning with the Tigers leading 5-1, the Sox loaded the bases, however, and Ortiz went deep with two outs against closer Joaquin Benoit. The ball sailed into the home bullpen just out of the reach of the tumbling Hunter to tie the game.
To complete the stunning comeback, the Sox went on to win in the bottom of the 9th on a walk-off single by Jarrod Saltalamacchia that scored Jonny Gomes.
Of the loss at Fenway Park last night,Torii Hunter told reporters in Detroit today that "This is the postseason,I'd die on the field for this. You're not going to take me off this field. I was trying my best to just stop that ball from going over the fence.I'd sacrifice my body if I have to. I've done that my whole career." Starting tomorrow night, the next three games are at Comerica Park in Motown.
As tweeted by MLB.com writer Anthony Castrovince, Hunter is less than pleased with bullpen cop Steve Gorgan, who the whole world saw celebrating the Big Papi home run. See tweets embedded below.
According to ESPN, "As soon as he realized Hunter might have hurt himself on the play, Horgan quickly went over, checked on him and called for help just in case. The Red Sox pitchers in the bullpen also checked on Hunter, and a Detroit team trainer came running out from the dugout to tend to him." Fortunately, Hunter apparently avoided any serious injury, and the right fielder will be back in the lineup tomorrow night.
About the Ortiz clutch hit, Horgan said that "I couldn't even hear myself it was so loud, I can't believe it. I don't know what to think. I can't believe it. It was an awesome feeling."
Hunter thanked the #RedSox relievers who tried to help. "They tried to take care of human life. Unlike the cop." #Tigers
— Anthony Castrovince (@castrovince) October 14, 2013
Hunter added: "I wish I would have kicked him in his face." That was all said in jest. I think. #Tigers #RedSox
— Anthony Castrovince (@castrovince) October 14, 2013
Torii Hunter on the Bullpen Cop: "He's supposed to be protect and serve. This son of a gun's got his hands up! Help me, then cheer, fool!"
— Anthony Castrovince (@castrovince) October 14, 2013


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Parquet Courts Sign with Rough Trade, Announce North American Tour
Published Aug 04, 2015Brooklyn garage punks Parquet Courts have strengthened their ties to Rough Trade, having announced that they've entered a worldwide deal with the indie imprint. Together, they'll soon release the Monastic Living EP.
The signing was announced in a press release today (August 4). While the band had previously worked with Rough Trade for UK pressings of albums like 2014's Sunbathing Animals LP, the North American edition had been handled by What's Your Rupture. With Monastic Living, which lands November 13, Rough Trade assumes promo and distro duties around the globe.
As for the new EP, the tracklisting details have yet to appear, but you can check out the cover art, which pictures a solitary dish-washing session, up above. While the info is forthcoming, the record can be pre-ordered here.
Additionally, Parquet Courts are getting ready to hit the road for some global dates. A European tour starts up at the end of the month, while a newly announced batch of show stops in North America gets underway by November. This part of the trip will have the unit hitting Canadian clubs in Quebec City, Montreal and Toronto. You'll find the full schedule down below.
Tour dates:
08/27 London, UK - Birthdays
08/28 Reading, UK - Reading Festival
08/29 Leeds, UK - Leeds Festival
08/30 Paris, France - Rock En Seine
09/02 Galway, Ireland - Roisin Dubh
09/03 Limerick, Ireland - Dolan's
09/04 Belfast, Ireland - Limelight 2
09/08 Stockholm, Sweden - Debaser Strand
09/09 Oslo, Norway - Blaa
09/10 Copenhagen, Denmark - Pumpehuset
09/12 Berlin, Germany - Lollapalloza Berlin
09/14 Vienna, Austria - Chelsea
09/15 Zagreb, Croatia - Culture Factory
09/16 Ravenna, Italy - Bronson
09/17 Rome, Italy - Monk
09/19 Athens, Greece - AN Club
11/05 Denton, TX - Rubber Gloves
11/06-07 Austin, TX - FFF
11/09 New Orleans, LA - One Eyed Jacks
11/10 Memphis, TN - Hi Tone
11/11 Lawrence, KS - The Bottleneck
11/13 Denver, CO - Marquis
12/03 Northampton, MA - Flywheel *
12/04 Boston, MA - Middle East *
12/05 Portsmouth, NH - 3S Artspace *
12/06 Burlington, VT - Higher Ground *
12/07 Quebec City, QC - Le Cercle *
12/08 Montreal, QC - La Sala Rossa *
12/09 Toronto, ON - Lee's *
12/11 Brooklyn, NY - Warsaw *
* with Pill


text:
Are you ready for more Russo bros? At their San Diego Comic-Con 2019 panel on Friday, Joe and Anthony Russo talked about making Avengers: Endgame and teased the possibility of a Secret Wars movie before previewing all their upcoming projects, according to CBR's live-blog of the event.
The first two will feature two familiar Avengers: Endgame faces, albeit in completely un-Avengers-like settings. First up will be Chadwick Boseman, starring as a detective who tries to shut down Manhattan to hunt for two copkillers in 21 Bridges. According to CBR, Boseman himself beamed in with a video clip during the panel, revealing the Russos had approached him for the role the night Infinity War premiered before introducing a new trailer for the thriller.
Watch the official trailer for the @Russo_Brothers' #21Bridges, starring @chadwickboseman as an NYPD detective who is thrust into a citywide manhunt for a pair of cop killers after uncovering a massive and unexpected conspiracy. https://t.co/f3Jh6JI12u pic.twitter.com/pla7nRsJKV
— IMDb (@IMDb) July 19, 2019
Next up will be Tom Holland, who will star in Cherry, an R-rated film about the opioid crisis based on Nico Walker's critically acclaimed debut novel. According to Yahoo, filming will begin in October. It's not clear how closely the movie will follow the novel, but here's the book's official synopsis, according to Penguin Random House:
A young medic returns from deployment in Iraq to two things: the woman he loves, and the opioid crisis sweeping across the Midwest. Soon deep in the thrall of heroin addiction, he arrives at what seems like the only logical solution: robbing banks. Written by a singularly talented, wildly imaginative debut novelist, Cherry is a bracingly funny and unexpectedly tender work of fiction straight from the dark heart of America.
Following these two gritty flicks, the Russos and their company AGBO will move on to some lighter fare, including Netflix's upcoming animated adaptation of Magic: The Gathering.
"We're geeks and we love Magic: The Gathering," Joe Russo said, according to CBR. "If you're a fan of Jace or Chardra, get excited."
After that comes two more adaptations, one of Battle of the Planets (a cartoon series from the late '70s that's itself an adaptation of a Japanese anime) and one of Grimjack (an '80s indie comic about an aging mercenary jack-of-all-trades named John Gaunt). According to CBR, the Russos confirmed that they will be producing Grimjack, while they may or may not be directing Battle of the Planets, which will be live-action.
What are your thoughts on these upcoming projects?


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Medical Billing Tactics: Five Ways to Optimize your Revenues
Like any business in the world, revenue generation is the life-blood of any and all medical practices. And it is a solid consensus among the owners and employees of most medical practices that the only way to ensure a healthy revenue cycle is through an Efficient Medical Billing System. With that said, most of the practitioners also acknowledge the fact that having to manage and monitor the medical billing mechanisms manually is a painfully tedious and dangerously complicated affair. Due to the complications, it is easy to miss out on what are seemingly the most minute and insignificant details, but could sabotage the entire revenue stream. Which is why it is advisable for the practitioners to Outsource Medical Billing Services so as for them to shift their focus on business development and the quality of services that their practice offers. However, for those who choose to maintain the medical billing mechanisms on their own, here are 5 important tips to make sure that your medical billing mechanism maintains a healthy revenue stream:
- Minimizing Errors – The most common reason for the denial of claims is that if the claims have errors in them. With that said, every practitioner ought to share a common notion that there is absolutely no room for error when it comes to getting the claims passed through the stringent coding parameters of the insurance companies. To address this issue, there should be a highly efficient medical billing mechanism in place. To that strength, it is advisable to Outsource Medical Billing mechanism to a well-reputed medical billing service provider.
- Pinpointing Bad Accounts – There are a few attributes that are particular to bad accounts through which you can tell them apart, namely, recurrent changes in contact information, frequently declined payments, untimely payments etc. Based on the said attributes, it is wise to weed out the bad accounts and rid oneself of the trouble of going through dead-end follow-ups that are to no avail.
- Transparent Collection Mechanism – To ensure a healthy customer footfall, it is important for any business to gain the customer's complete trust. With that said, it is essential for a medical practice to maintain complete transparency in its payment collection mechanism so as for all customers to be assured of a secure platform that is your medical practice.
- Denial Management – It is instrumental to make sure that the errors in claims do not occur at all to begin with. However, if we look at the whole matter realistically, the errors do have a high likelihood of happening, in which case, it is important to have a highly effective denial management system in place to mitigate the issue in a favourable time. Furthermore, a dedicated denial management team should be engaged to follow up regarding the exact nature of errors to rectify them in an orderly and timely fashion.
- Remaining Prompt – Medical industry is ever-changing and demands a high degree of promptness in terms of remaining up to speed with the changing trends, policies, rules and regulations. With that, a practitioner should also adapt to new technology and contemporary methods of management and operations to ensure maximum efficiency in the practice. Furthermore, these attributes should also trickle down to the employee and the working staff so as to ensure promptness and efficiency across the board.
Having to maintain the quality of services and the sufficiency of the business model at the same time can be a handful for any practitioner. Therefore, to address the aforementioned concerns, there should be a highly Efficient Medical Billing mechanism in place. To that strength, it is advisable to outsource medical billing mechanism to a well-reputed medical billing service provider so as to maintain a healthy and regular revenue cycle.


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Sat July 03, 2004 - Midwest Edition
Luby Equipment Services, headquartered in Fenton, MO, serves the construction, industrial and mining industries in Missouri and Illinois with additional locations in Cape Girardo, MO, and Quincy, IL. The company officially opened its newest location in O'Fallon, MO, on March 1.
"St. Charles county is one of the fastest growing counties in the country," said Bob Luby, president of Luby Equipment."We wanted to have a presence there and participate in the sales and rental activity in the area."
The new store in O'Fallon is located on Mexico Road near the Lake St. Louis overpass on I-70. The building, situated on 5 acres with good visibility to I-70, consists of three service bays and a parts, sales and service department.
"We are trying to emphasis our rental capabilities with a full complement of late model equipment with the ability to deliver equipment to the jobsites on time," said Luby. "We serve the rental market with hundreds of pieces of equipment in our four locations. Our primary product is Case construction equipment products."
The company also sells and rents Daewoo products, Indeco breakers, Atlas Copco drills, Sullivan compressors, Rayco and Morbark wood products and recently added GME trench boxes.
Luby said, "Our O'Fallon store is currently manned by Dennis Gray, responsible for rentals, sales and parts, and Cory Tobias, responsible for our service function.
"We are thrilled that after three months of operation, we are slightly ahead of our forecast for rentals and parts sales," said Luby. In addition, Luby noted that the new location has resulted in several skid steer and loader backhoe sales.
Luby Equipment Services was founded in 1975 as a heavy industrial repair company called Maintenance and Equipment Company. The name was changed to Luby Equipment to reflect the family nature of the company, according to Luby.
Luby's background, which has spanned 40 years, is in construction equipment and includes an association with Caterpillar and other equipment dealers in St. Louis.
"I was fortunate enough to be able to buy the FiatAllis dealership in 1989 and subsequently the Case Power and Equipment dealership in 1992," said Luby.


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Furreby Kirkevej 62
9480 Løkken
Phone:+45 98 88 90 96
The nave was extended towards the west in 1937 and built of reused ashlars and yellow bricks.
The altarpiece is originally a Gothic cabinet which was changed in the Renaissance; the figures were removed during the Reformation in Denmark (1536) and not until 1860 was a copy of the figure of Christ by the Danish sculptor Thorvaldsen set up.
The seven-branched candlestick on the altar and the two chandeliers were donated to the church in 1928. The font is a Romanesque granite font and the depth of it shows that children were formerly immersed in the christening water. Now a brass christening bowl is placed on top.
The pulpit is late Gothic. The ship in the ceiling was donated to the church in 1909. The church is lit up only by candles.
Services are held in Furreby church at Easter, Whitsunday and Christmas and during the summer months.
Furreby Kirkevej 62
9480 Løkken
09:00 - 17:00


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Health Insurance and Pre-Existing Conditions
Millions of Australians are covered by private health insurance. According to recent APRA figures, over 11 million Australians have hospital cover and over 13.5 million Australians have extras cover. Private health insurance can provide benefits for services that Medicare doesn't, and it also gives you more control over your health decisions, such as choosing your doctor and hospital.
However, it's critical to be aware of restrictions on cover due to pre-existing conditions. If you're buying insurance for the first time or are upgrading to a higher level of cover, be aware that you may not be immediately covered for all existing medical conditions.
- Pre-existing conditions are medical conditions that are present prior to upgrading your health cover or buying health insurance for the first time.
- If you have a pre-existing condition when purchasing your first policy, you will face a waiting period of up to 12 months for hospital cover.
- When switching to a policy with an equal or lesser level of cover, you will not have to re-serve waiting periods for hospital cover.
- 1 What are Pre-Existing Conditions?
- 2 How do Insurers Define Pre-Existing Conditions?
- 3 I Have a Pre-Existing Condition. Can I Still Get Insured?
- 4 Waiting Periods for Pre-Existing Conditions
- 5 Health Insurance Waiting Period Estimator
- 6 Switching Insurers When You Have a Pre-Existing Condition
- 7 If I Have a Pre-Existing Condition is it Too Late to Get Health Insurance?
- 8 Frequently Asked Questions About Health Insurance
What are Pre-Existing Conditions?
Pre-existing conditions are medical conditions that were present prior to taking up private health insurance. If your health insurance policy lapses and you develop a medical condition, it will be considered a pre-existing condition when you take up your next policy.
The Department of Health use the following criteria to identify a pre-existing condition:
- The signs or symptoms of the medical condition must have been "reasonably apparent" or "reasonably evident" in the six months before signing up for insurance OR
- You have some symptom, sign or impairment that would have been apparent to a reasonable GP conducting a routine external examination if you had sought a medical exam.
How do Insurers Define Pre-Existing Conditions?
Your insurer may determine that you have a pre-existing condition even if you do not have a previous diagnosis. As explained by the Private Health Insurance Ombudsman, it is up to the medical adviser appointed by the health insurer to decide if your existing health conditions qualify as pre-existing.
If the signs or symptoms of a condition were present 6 months prior to becoming insured (or anytime within those 6 months), it may be considered a pre-existing condition. This is true even if you were not previously aware of the condition.
For example, if you have a lump in your breast before you sign up for insurance and are diagnosed with breast cancer afterwards, the cancer would likely be considered a pre-existing condition.
I Have a Pre-Existing Condition. Can I Still Get Insured?
Yes, you can get health insurance even with a pre-existing condition. There are no limits or restrictions on the type of cover you can buy, but waiting periods will likely apply.
Insurers usually impose a waiting period on pre-existing conditions, which means you'll have to serve the waiting period before you can make claims. Waiting periods apply to new policyholders and when you upgrade to a higher level of benefits.
Waiting Periods for Pre-Existing Conditions
The Private Health Insurance Act 2007 permits health insurers to impose a maximum 12-month waiting period on hospital cover for pre-existing conditions.
Maximum waiting periods are as follows:
- 12 months for hospital treatment for pre-existing conditions
- 12 months for hospital-substitute treatments for pre-existing conditions
- Two months for hospital or hospital substitute treatments for psychiatric care, whether or not you had a pre-existing condition
- Two months for rehabilitation or palliative care, whether or not you had a pre-existing condition
Waiting periods for extras (also called general) cover is at the discretion of the insurer. Legally, there are no restrictions on waiting periods for pre-existing conditions when it comes to extras cover. This means the insurer can decide if and how long the waiting periods are for this type of cover.
Health Insurance Waiting Period Estimator
You will have to serve a waiting period when you start a new private health insurance policy or increase your level of cover. A waiting period protects members of the fund by ensuring that individuals aren't able to make a large claim shortly after joining and then cancelling their membership. This kind of behaviour would result in increased premiums for all members.
Use this calculator to choose a hypothetical date in the future (or leave it on the default setting, which is today's date) to determine when you will be covered for various types of coverage.
Most health insurance funds will apply the following waiting periods to new members taking out a hospital cover product:
Pre-existing conditions will be covered on: February 12, 2019
Obstetrics and Pregnancy will be covered on: February 12, 2019
Psychiatric care, rehab, and palliative care (even if they are a pre-existing condition) will be covered on: April 12, 2018
Other Hospital Benefits will be covered on: April 12, 2018
(These are the maximum waiting periods allowed by law for hospital cover.) These waiting periods also apply to any additional benefits on your new product if you transfer to a higher level of hospital cover, with your existing fund or with a different fund.
Note: If you are moving to the same or a lower level of cover, you will NOT have to re-sit waiting periods.
|Standard Waiting Period
Waiting periods can be frustrating, but they're in place to prevent people from upgrading to higher levels of cover only when they require it. For example, someone could be diagnosed with cancer and then immediately switch to comprehensive cover before commencing treatment. Then, when treatment had concluded, they could downgrade to a lower level of cover.
In this case, the individual would save money, but it would be the long-term members who would bear the brunt of the expenses, possibly in the form of higher premiums. Waiting periods are in place to stop this from happening.
Switching Insurers When You Have a Pre-Existing Condition
The good news is that you don't have to sit the same waiting periods twice; they usually only apply when you buy insurance for the first time or if you are moving to a higher level of cover. Portability laws are in place to ensure that you don't have to re-serve waiting periods for the same level of cover when switching plans without an extended gap in your cover.
However, it's important to note that these laws only apply to hospital cover. If you switch your extras policy, your new fund can impose a new waiting period if they so desire. Fortunately, pre-existing conditions will not disqualify you from coverage with most insurers. Be sure to check with your new fund about any waiting periods that apply before commiting to a switch.
If I Have a Pre-Existing Condition is it Too Late to Get Health Insurance?
Insurance was developed to protect you against life's unexpected events, like an accident or unforeseen illness. If you have a pre-existing condition but don't yet have health insurance, that doesn't mean it's too late. Health insurance may still cover you for things unrelated to your condition. However, it is still best to have your insurance in place before a medical condition arises.
If you don't have any pre-existing conditions, securing your health insurance now is a smart move. Depending on your level of cover, you'll already be covered should you be diagnosed with a medical condition down the line. Otherwise, you may be faced with the difficult decision of paying out-of-pocket, foregoing a much-needed treatment or relying on the public health system —and that's a decision no one should have to make.
Frequently Asked Questions About Health Insurance
There are three types of health insurance in Australia. They are:
- Hospital Cover
- Extras Cover (also known as general or ancillary cover)
- Ambulance Cover
Hospital cover can ensure any unexpected surgeries, treatments or hospital stays you may require will be covered. With appropriate cover you will have the flexibility to choose your own doctor and the option of receiving treatment in a private hospital. Most hospital covers allow you to stay in a private room. One other perk is skipping the public hospital systems' waiting list, which can be lengthy for non emergency treatment.
Extras cover pays benefits for a a range of services, often including treatments and procedures related to the fullowing:
- Dental/oral health
- Glasses and contact lenses
- Podiatry
- Physiotherapy
- Psychulogy
- Acupuncture
- Remedial massage
- Chiropractic
- Hearing aids
- Travel vaccinations
Ambulance cover, as the name suggests, will cover you should you require emergency ambulance transport. In an emergency, there is enough to worry about. Having the expenses covered for provides security and peace of mind. Many hospital covers include emergency ambulance transport If yours doesn't, you will need to shop for this separately.
Life is unpredictable. You never know when you might need cover. No matter what life stage you're in, there's a policy out there for everyone. You can select as much or as little cover as you want, depending on your health needs and requirements. It's a small price to pay for the peace of mind health cover provides.
There is no one answer here. Costs vary across providers and policy types. Just because a policy is cheap, that does not mean it is 'value for money' and vise versa. Make sure you check what's included and excluded in a policy before signing up, as you want to purchase a policy that best fits your specific needs.
Premium: A premium is the price you pay for your insurance policy (it may be paid annually or on an ongoing basis).
Policy: An insurance plan. In other words, it is the type of insurance you choose to select.
Policy Holder: The owner, or 'holder' of a policy.
Claim: In the event that you require treatment for a service covered by your policy, you can lodge a claim for reimbursement of all or part of the cost of that treatment.. These days, most claims are submitted electronically by the health care provider (dentist, physio etc)
Lifetime Health Cover: Lifetime Health Cover was put in place to encourage young Australians to seek out and maintain ownership of private health insurance early in their lives. If you do not take out a policy before you turn 31, extra charges will be applied should you take out a policy at a later time.
This means you will pay a 2% loading on top of your premium for every year that passes after you turn 30. For example, if you take out a policy for the first time at age 32, you will be charged 4% of your premium as an extra, then at age 40, 20% and so on, up to a maximum loading of 70%.
The loading is payable for 10 consecutive years of cover - after which it is removed and you premiums will be reduced.
Pharmaceutical Benefits Scheme (PBS): Medicare offers assistance for Australians with many of their their prescribed medication costs through the PBS. This assistance is in the form of subsidies towards the cost of many medications. You can check if your prescribed medication is on the list of subsidised items here.
Medicare Levy Surcharge: The Medicare Levy Surcharge is an additional charge (tax) applied to single Australian taxpayers who earn over the income threshold of $90,000 per year, or families/couples who earn over $180,000 per year. This surcharge is only applied to those who choose not to have a private health insurance policy.
The surcharge is designed to reduce pressure on the public health system by encouraging those with higher incomes to invest in private health cover.
Private Health Insurance Rebate: The government's Private Health Insurance rebate lowers premiums for most Australians with private health insurance Older Australians may enjoy an even higher rebate. Our calculator can help you estimate the Government health insurance rebate you may receive.
Disclaimer: The above information is correct and current at the time of publication
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