text:
Great location
"Inadequate rooms with subpar service. Vibe is not too good."
Hotel description
First Class Hotel is located in the coastal town of Alanya. It features a private beach, spa facilities and free Wi-fi. The contemporary rooms have air conditioning and a balcony. The hotel has a variety of leisure facilities available on site including sauna, massage, gym and traditional Turkish hammam. The complex also has an indoor and outdoor pool with a bar. First Class Hotel has a private beach with sun loungers which is a 5 minute walk from the hotel.
All requested cribs and extra beds are subject to the hotel's confirmation.
Pets are not allowed.
Food & Drink
Health & Fitness
Room service
Total price for 1 night, 2 adults, 1 room


text:
Humans are a unique and interesting kind of creature. They champion both conformity and individuality. Teamwork and self-reliance. Independence and codependence. These seemingly opposing concepts are equally a part of our nature, and guide the very way we function as a species. So how is this possible?
There is an important value that ties these things together and allows humankind to be successful. This value is Loyalty. Loyalty gives us the ability to come together, and work towards a common goal. Humans, for all of our existence, have been the most successful when we work in packs. Whether we are hunting, farming, building, renovating, brainstorming, inventing- everything is a team effort. Those who turn against, or stray from their groups, are lost. However, in every group individuality is important. Every member must have different sets of skills and attributes that contribute to the group as a whole.
Loyalty is found when your personal identity is congruent with the group you are a part of. If that is not the case, then maybe it is time to find a new group. One that you can identify with, remain loyal to, and reach a common goal with. Some people are part of a group that does not share the same vision. Neither the group, not this person is benefited by this situation. Some people believe they don't need a group of like-minded individuals. This person is willingly limiting himself, and will not last long in the real world. But those who are truly effective find the group with a consistent goal, that lines up with their own, and remain actively loyal until that goal changes or is achieved.
Before you make the decision of Loyalty, make the decision of who you are loyal to.
See you on the mat,
Coach Monkey
Best group of martial artists I've ever been around. Youngsters are great and the instructors are out of this world. Could not recommend any more highly!
Great coaches and great opportunity for kids to gain confidence in their own abilitiies
Pride MMA has been a wonderful experience from the moment that my family walked in the door. Great staff! Great instructors! Great facility! Great atmosphere! Everyone welcomes you in like family. There have been no egos and everyone truly wants to help you acheive your goals. My biggest regret is not starting sooner. Don't wait. You won't be disappointed.

subset: commoncrawl


text:
Crest Hill Real Estate - Find Your Perfect Home For Sale!
Crest Hill - Town vs. County Stats
Avg Price in Crest Hill: $186,900 / County Avg $338,300
Avg Taxes in Crest Hill: $3,900 / County Avg $6,300
Avg Sq. Ft. in Crest Hill: 1,499 / County Avg 2,480
Avg Price per/ft2 in Crest Hill: $125 / County Avg $136
Avg Walkscore in Crest Hill: 28 / County Avg 24
Avg Year Built in Crest Hill: 1990 / County Avg 1989
Avg Days on Website in Crest Hill: 67 / County Avg 88
Crest Hill Real Estate Market Health
|Areas in Crest Hill IL
|Crest Hill
|Neighborhoods in Crest Hill IL
|Carillon Lakes
|Remington Lakes
|Rockdale Junction


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Unlike THC, which primarily binds to CB-1 receptors located in the brain, CBD works in the body by manipulating receptors throughout organ tissues, the immune system, the pain response system, the hormonal system, and other whole-body regulatory systems. Basically, since its receptors have been found to exist in virtually every cell and tissue type in the body, CBD is believed to work on every aspect of human health and behavior – from the subcellular level to the whole-body leve and beyond.
This means that CBD oil products that are derived from these "not marihuana" parts of imported hemp plants are not federally banned, and the natural cannabinoids in hemp-derived products are exempt from DEA enforcement. While hemp cannot be legally cultivated in the U.S. except under state-regulated programs, hemp-derived oil has been a legal import in the U.S. for decades.
CBD oil is prohibited for sale by terms of service on the major online marketplaces, including Amazon, eBay, and Groupon. Most of the products sold as CBD oil on those websites are hemp seed oil and don't contain any CBD. We've also heard many reports of counterfeit and potentially dangerous products being sold as CBD oil on those websites. Your best bet is to always buy directly from the CBD company's official website.
Our bodies' ECS is responsible for producing natural cannabinoids that contribute to our feeling pleasure, pain, alleviating anxiety, regulating sleep patterns, and controlling appetite. When one consumes CBD oil, his/her body's cannabinoids gets supplemented with cannabinoids. As a result, the body's natural cannabinoids production increases. In doing so, your ECS system's natural healing ability gets enhanced.
The unflavored oil is derived from organic hemp seed oil. It is considered both vegan and gluten-free, and does not have any artificial sweeteners or preservatives. The oil contains 0.3% THC, meaning those who face the possibility of drug tests may want to avoid taking it, but this relatively small amount produces little, if any, psychoactive effect. The price per mg is $0.11, making the Spruce Lab Grade CBD Oil less expensive than average.
All this talk about THC lands us nicely in the whole "Full Spectrum vs. Pure Isolate" debate. Once you begin shopping for CBD products, you'll notice a lot of jargon that gets thrown around without much explanation. Now that we've introduced THC into the conversation, we can talk about the difference between, and relative benefits of, Full Spectrum CBD and CBD Isolate (and the lesser-known contender: Broad Spectrum).
This article provides a wealth of practical information for the individual considering CBD as an adjunctive or alternative treatment for pain and or anxiety. CBD works in the body by manipulating receptors throughout organ tissues, the immune system, the pain response system, the hormonal system, and other systemic regulatory systems. While CBD oils have not been reviewed or approved by the FDA for the treatment of these conditions, a wealth of literature, both anecdotal and research-related now exists to help describe both their safety and effectiveness. As discussed in this article, the potential new adopter must be mindful of several important items. First, only those products that are sourced from Industrial Hemp will be considered legal in all states. One must be careful if the product you choose is sourced from the marijuana plant, as those products may contain THC levels above the legal limit in your given state. Secondly, all products are NOT created equal – they differ significantly in strength, absorption, and elimination by the body and in the manner in which they are formulated. One should be mindful of the differences in doses available for each of these products, starting at a low or moderate dose and increasing as needed in order to find the lowest dose that provides the desired relief. In this way, one can individualize usage to maximize effectiveness, while minimizing risk, a proper goal for the use of all medicinals.


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Search our database of 173,466 films.
|Matching films
Faliyu, 2019, 11 min
Translation: Green Courtyard
Faliyu: a boy in his mid 20s traces inspiration in his childhood to clean a public place.
Asian Documentary Educational Experimental Independent National
Whispered voices, 2019, 5 min
Doug Duncan Lisa Genovese Lisa Genovese
London, United Kingdom
Engaging and compassionate portrait of a woman's mental health.
Drama Experimental Foreign Language Independent Mystery Romance
Everybody Shut Up, 2019, 13 min
George O'Reilly Michelle Milanko
London, United Kingdom
Short documentary about the importance of social interaction.
Documentary Educational Political
داين تدان, 2019, 12 min
Translation: Karma Of Life
Karim El Shafei Basem Fouad
Cairo, Egypt
KARMA never loses an address
Drama Independent Short
Wind Humans, 2019, 15 min
Doidy Florian, Florian Doidy Doidy Florian
Migrants are wind humans. Wind is music.
Documentary Experimental Political
The Wildlings Trap, 2019, 9 min
Jane Sanger Jane Hamlet
Sevenoaks, International
A wild girl living in the woods is not as innocent as she looks.
Crime Drama Murder Mystery Teen Thriller
Hello Darkness, 2019, 9 min
Christian Skibinski, Sönke Hünding Christian Skibinski
A confrontation at a train station with a tramp.
Art-house Horror Independent Short
Phombies, 2019, 5 min
Manoj Anand AJ. H., Manoj Anand Dale Anthony Church, Manoj Anand
Leicester, United Kingdom
A Mobile phone virus turns people into Phombies
Horror Short
Ed Asner: On Stage and Off, 2019, 87 min
Kurt Jacobsen, Warren Leming Kurt Jacobsen, Warren Leming
Chicago, Illinois, United States
A portrait of Ed Asner as actor and activist
Documentary Historical Independent
Alone, 2019, 7 min
Glen Mitku Glen Mitku Glen Mitku
New York City, United States
After leaving work a woman goes home unaware of a sinister presence following her.
Drama Independent Mystery Short Thriller
Pravaas, 2019, 4 min
Translation: Migration
Sharad Uikey Sharad Uikey
Kolkata, India
A boy who has to migrate.
Adaptation Animation Children Educational Family Short
Upama, 2019, 77 min
Translation: The Simile
S S Jishnu Dev Nithin Noble S S Jishnu Dev
Trivandrum, India
The simile shot on natural light
Low Frequencies, 2019, 4 min
Gemma Ghelardi Gemma Ghelardi Gemma Ghelardi
A frustrated writer discovers he can listen to the stories the city walls have witnessed.
Ken e il coniglio., 2019, 14 min
Translation: Ken and the rabbit
geofrey king geofrey king
Kampala, Uganda
my father is a real nightmare
L'eau est la Vie: From Standing Rock ..., 2019, 25 min
Translation: Water is Life
Sam Vinal Melissa Cox
Los Angeles, United States
On the banks of Louisiana, fierce Indigenous women are ready to fight-to stop the corporate blacksnake and preserve their way of life. They are risking everything to protect Mother Earth from the predatory fossil fuel companies that seek to poison it.
Black Disaster Documentary Independent Political Women
Eyes to See, 2019, 7 min
Ranchana Butler Ranchana Butler
Bangkok, Thailand
A Day in a Thai Rain forest
Asian Natural History
Mazhachillukal, 2019, 15 min
Translation: Rain Crystals
Naseer Badarudeen Pradeep Joseph
THIRUVANANTHAPURAM, India
Love and Relationship Between a Father and his 10-year old son
Asian Children Drama Family Short
An Act of Betrayal, 2019, 16 min
Nick Lazar Graham Campbell Nick Lazar
Brighton, United Kingdom
John Bower thinks he has all his affairs under control. His wife disagrees.
Black Comedy Crime Independent Noir Short
Sunflower, 2019, 4 min
Tessa Kennedy Tessa Kennedy Tessa Kennedy
London, United Kingdom
A short lesbian film
Mockingbird, 2019, 4 min
Renata Gheller Renata Gheller
The lives of two sisters are forever changed on a late summer evening.
Art-house Drama Independent Short Silent Thriller


text:
Selecting either option below will execute a search of our records for any previous licenses. Licenses purchased online are subject to a $2.50 (minimum) service .
Digital Counties Survey Winner 2017. Government Experience Winner 2017. NACO Winner 2018. Automation Alley. Advantage Oakland. Destination Oakland.
All dogs over the age of four months require a license in Oakland County. In order to purchase a license, a valid rabies certificate must be presented.
Greetings,. Each summer, the Oakland County Animal Control Division conducts a census to ensure compliance with the. Michigan State Dog Law of 1919 that .
Oakland County Animal Control 1200 N. Telegraph Rd, Bldg. 42 East Pontiac, MI 48341, 248-858-1070. 1-Year Licenses, Waterford Township Treasurer's Office
Dog licenses may also be purchased online through the Oakland County website at: https://land.oakgov.com/opls/ . Follow the directions given on the website.
Where do I purchase a dog license? Oakland County dog licenses can be purchased at Walled Lake City Hall. All dogs over the age of four months require a .
Rochester Hills Treasury Department (Make check payable to City of Rochester Hills); Some Local Veterinarians; Oakland County Animal Control and Pet .


text:
The Time Has Come to KonMari Your Cleaning Supplies
Less really is more.
I Never Brought Lunch to Work Until I Got This Screw-Top Bowl
No schlepping around bulky plastic here.
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Sponsored By marvin
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text:
Dumpster Rental in Harpersville AL
Rent a Dumpster in Harpersville Now!
Quick 1-minute quote in Harpersville
Dumpster rental specialists are standing by to give you a quick, no-hassle quote. Let us know what dumpster size you are looking for, when you need it, and what zip code your roll-off is going to.
We are always available to help
Simply give us a call and we will answer any questions you may have about the Harpersville dumpster rental procedure, allowed materials, size you may need, etc.
Next-Day delivery of your dumpster
Our roll off containers can usually be delivered in Harpersville the day after you place your order. Make sure to call us early to ensure timely delivery.
Get a Quote! 888-880-4457
Garbage Dumpster Rentals Tips
The Appeal of Garbage Dumpster Rentals
It's contingent on which trash removal company that you hire. What's more, it is very important to explain how much waste is going to be deposited. Then you have to locate a respectable waste disposal facility for the Lorain region.
So there you've got it, a very simple procedure for gathering the information that you want to provide to the rental businesses. After somebody has each one of the essential information on hand, it's wise to generate a list of rental businesses with a positive reputation. Don't hesitate to call at any certain time to talk about problems, logistics or changes otherwise be assured your dumpster is going to be delivered as requested.
What the In-Crowd Won't Tell You About Garbage Dumpster Rentals
Dumpster rentals from such services will guarantee that the rest of the debris is correctly treated and processed in a means that has minimal negative and maximum beneficial effect on the surroundings. The reach of rentals utilized in Illinois makes certain you will have the ability to find the unit that you would like for your own personal project. Portland residents don't need to be concerned about hidden charges.
The Secret to Garbage Dumpster Rentals
Many sorts of garbage are accepted, but there are a number of things that aren't accepted. After you figure out who you would like to rent your dumpster from, there is going to be a couple of factors that impact the charge to lease a dumpster. A company has to know what sort of waste will be set inside the dumpster.
The entire process is simple to do. If you're trying to find a big, yet compact it is possible to look at a 15 yard dumpster. The typical cost to lease a dumpster is $421.
Roll Off Dumpster vs. Dumpster Tote
Dumpster bags may offer an option to roll of dumpsters. Whether this option works well for you, however, will depend on your endeavor. Consider these pros and cons before you decide on a disposal alternative that works for you.Roll Off Dumpsters
It's difficult to overcome a roll off dumpster when you've got a big job that'll create plenty of debris. Most rental companies include dropping off and picking up the dumpster in the prices, in order to avoid additional fees. Roll off dumpsters typically have time restrictions because companies need to get them back for other customers. This is a possible downside if you aren't good at meeting deadlines.Dumpster Bags
Dumpster bags are often convenient for small jobs with free deadlines. If you do not want lots of room for debris, then the bags could work well for you. Many businesses are also pleased to let you maintain the bags for so long as you need. That makes them useful for longer jobs.
Call Now! 888-880-4457
Dumpster Rental Companies Near Harpersville
|Company Name
|Alabama Dumpster
|(334) 288-1500
|(256) 757-9900
|(800) 661-6307
|(334) 265-7326
|Mark Dunning Industries
|(334) 983-1506
|Jameson Inn
|(205) 345-5018
|Med-Waste Inc
|(334) 262-5770
|David Jordan & Co Waste Removal
|(334) 834-3484
|Truck Trailers of Alabama Inc
|(205) 808-0042
|Joe Jackson Environmental Services
|(205) 646-3070
Our Service Area
Zip Codes in Harpersville35078
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Dumpster Sizes Available Harpersville
Get started in less than 60 seconds. Pick up the phone and get a quote now!
10 Yard
Great for small projects
- 12 feet long
- 8 feet wide
- 4 feet tall
20 Yard
For moderate sized cleanouts
- 22 feet long
- 8 feet wide
- 4.5 feet tall
30 Yard
All around good dumpster size
- 22 foot long
- 8 feet wide
- 6 feet tall
40 Yard
For the largest projects
- 22 foot long
- 8 feet wide
- 8 feet tall
*Dimensions & availability may vary by location. Call to confirm.
Thousands of satisfied customers
We take pride in providing our community with great prices and excellent customer service on dumpster rentals. Join thousands of others and get the dumpster you need, for the price you deserve!
We accept
All major credit cards accepted for dumpster rentals in Harpersville.


text:
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One of the most exciting new developments within the area of global change science has arisen from the growing body of paleoclimatological evidence for rapid changes in the climate system. Until recently it would have been hard to imagine that the global climate, together with other aspects of the Earth system to which it is intimately linked, could have undergone repeated oscillations on millennial timescales as described earlier, let alone that the most dramatic of these "switches" occurred within a matter of decades, with major consequences for temperatures, hydrological regimes, plants, and animals that were at the very least hemispheric in their extent. The evidence now available provides a fascinating insight into the climatic backdrop to the development of human societies in the past. It also highlights one of the major challenges for the future. If the potentially disastrous impacts of climatic "surprises" like the ones that have already been well documented from ice core and sediment records are to be avoided in the future, research devoted to deepening our understanding of abrupt changes in the past is of paramount importance.
Any definition of "abrupt" climate changes is necessarily somewhat subjective, as it depends in large measure on the sample interval used in a particular study and the pattern of longer-term variation within which the sudden shift is embedded. Here, I make no attempt at a general definition but focus attention on two examples of rapid transition found in the paleorecord from the geologically recent past, glacial terminations and the Younger Dryas.
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We are the professional manufacturer and exporter in producing series of dyestuff. our main products are top grade direct dyestuff used for dyeing paper, cotton, wool, nylon and leather, also produce dyestuff intermediates (J Acid, Tobias Acid, etc.; along with liquid dyes used for paper, neutral dyes, salt-free dyes, and special chemicals, with an annual output of fifty thousand Metric Tons
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Acid dye
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Malinda Cao
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If you haven't had a Krispy Kreme doughnut, you don't know what you're missing. But a lot of folks in Minnesota along the route between Minneapolis and Clive, Iowa do know how scrumpdeleicious they are and are getting their sugary fix thanks to the efforts of a young enterprising entrepreneur according to the St. Paul Pioneer Press.
This young entrepreneur is Jayson, Gonzalez, who is 21 and a college student at Metropolitan State University in St. Paul, MN. He got the idea of picking up boxes of Krispy Kreme doughnuts in Clive, Iowa and then delivering and selling them to Minnesotans while on a trip to Iowa to coach in a soccer tournament.
Krispy Kreme doughnuts were no longer available in Minnesota ever since the company closed its last store there in 2008.
Gonzalez posted an ad in the Facebook Marketplace just to see if anyone would be interested in buying some Krispy Kreme doughnuts. And sure enough in just a few days over 300 people responded. Today his Facebook page called 'Krispy Kreme Run Minnesota' has over 5,000 followers.
Gonzalez uses his Facebook page to take orders as well as to communicate with customers updating them on quantities and drop-off/pick-up locations. He uses a lot of local Target parking lot locations.
Krispy Kreme sells their famous original glazed doughnuts for $9.49 at a dozen per box and Gonzalez turns around and sells them for around $17 to $20 per box depending on the variety.
Gonzalez usually operates his business on Saturdays by getting up at 2 a.m. in the morning to drive over 250 miles one way from the Twin Cities to Clive, IA to pick up the doughnuts.
Gonzalez has a Ford Focus and he says he can fit 100 boxes of Krispy Kreme doughnuts in it. Once he's loaded up, he makes his way back to Minneapolis stopping along his route dropping off boxes of Krispy Kreme doughnuts to happy fans of the sugary treats.
At one time, after Gonzalez had 19 successful trips under his belt, he posted on Facebook that Krispy Kreme had told him that he needed to stop his operation because the company was concerned about the quality standard of their product not being met due to the long dive back to Minnesota.
There was such a tremendous response in support of Gonzalez's operations, according to CNN that the company worked out a deal for Gonzalez to do business as an independent operator/contractor.
Krispy Kreme says it is happy to help Gonzalez reach his goals of graduating from college without any debt in 2021.


text:
Construction Company — I applied for od
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9-4-134/76, Hyderabad, Andhra Pradesh, India
ShShaik Nader on Nov 14, 2019
I have applied for b. Tech od (2018 passedout)
Online convocation services through online url:"http://epayments.jntuh.ac.in/onlineconvocationservices/postconv", but i haven't received any acknowledgment of that please help me on this. Transactionid. 2a70377 [protected] for the hall ticket no : 11q81a01a1, email id : [protected]@gmail.com, and all the required documents and photo are uploaded. Please let me know the status so that i can proceed for further.
I am not able to find the status no where in any site. But amount transferred to jntu hyd. Can anyone help me on this please.
Thanks and regards,
Shaik nader
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About Sonic
Welcome To sonar and sonar comics Guest Book Photos About Sonar made Comics About Sonic Favorite Links Other comics Sprites! My Photos Photo
All about sonic
In the below you will find out about sonic, not tails, knuckles, amy, or shadow.
About Sonic
Sonic is a fast paced hedgehog who had his first big debut in 1991 in the game "Sonic the hedgehog". He is the mascot of SEGA who replaced Alex Kidd as the mascot. He also can run at the speed of sound, use the power of the chaos emeralds and 50 rings to turn super sonic and he is the fastest thing alive.


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Hot Girl Meg
12 days ago (2019-12-01 20:41:16)
Post text language: EN
Lmao when I was little I wanted to be a pussycat doll so bad 😂😂😂Download video
i lmao when was little wanted to be a pussycat doll so bad
This buttons will redirect you to instagram.com.
Top Comments:
Im fucking weeeeaaak
❤❤❤❤❤ @pussycatdolls
" don't chaaa😭"
Meeeeee tooooo! Its alll THE ENERGY😂😂😂😂😂
Never in my life have i seen an Aquarius have this much fun 🤔
i wanna be your bestie so baaaaad😩
That hair thooo😍🔥
Lmao this was us in your living room all over the place jumping on the couch and everything dancing with them to their video 😭😭😭 @k_woodburn
Beautiful ass
😂😂😂😂😂😂👌Enjoy your life!! I love this!!
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Give a bald BLACK BITCH a wig n she all a sudden a pussycatdoll
You low-key look like Nicole though
I freaking luv you ❤❤❤
@ace_3426 ok but why does this remind me of you? 🤣


text:
Zooming in on the VISTA view of the Tarantula Nebula
This zoom sequence starts with a broad panorama of one of our neighbouring galaxies, the Large Magellanic Cloud, and we gradually close in on the Tarantula Nebula, a very active star-forming region. First we see a detailed visible light image obtained with Wide Field Imager on the MPG/ESO 2.2-metre telescope at ESO's La Silla Observatory in Chile and the sequence ends with a new near-infrared VISTA image of the region around the Tarantula Nebula that was obtained as part of the VISTA Magellanic Cloud Survey.Autorska prava:
ESO/M.-R. Cioni/VISTA Magellanic Cloud Survey and Eckhard Slawik (firstname.lastname@example.com). Music: John Dyson (from the album "Moonwind"). Acknowledgment: Cambridge Astronomical Survey Unit.
O video klipu
|Datum objavljivanja:
|11. avgust 2010. 12:00
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|56 s
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O objektu
|30 Doradus


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604 Melody Lane 604 Melody Lane Cameron, NC#5613722
- 4 Bed
- 2.5 Bath
- $1,650
NEW LISTING located in Harnett County- Theatre Room and Swimming POOL! - DON'T MISS THIS GEM! Located in the Quiet Neighborhood Forest Ridge. This 4 bedroom, 2.5 bath features a Bonus Room that has been converted into a Theatre Room with a prjector and projector screen. Home features an open floor plan that flows room to room. Enjoy family dinners in your formal dining room, and quiet time in your office located downstairs. The master bedroom is located on the main floor and has a master bathroo
18 Old Montague Way 18 Old Montague Way Cameron, NC#4957267
- 4 Bed
- 2.5 Bath
- $1,600
New Home in Lexington - Beautiful newly built home nestled in Lexington Plantation. This home boasts granite counter-tops, walk in closets, stainless steel appliances, and fenced in yard. Plenty of space with four bedrooms and 2.5 baths, with Formal living and dining room! Home includes amenities access. (RLNE4322493)
242 Checkmate Ct 242 Checkmate Ct Cameron, NC#5324925
- 3 Bed
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- $1,420
Bonus Room W/ Wet Bar. +/- ½ Acre Lot - 3 Bedroom 2 1/2 Bath located in Yorkshire Plantation. Short Drive to Shopping Center. House features Formal Living Room and Dining Room. Great Room has a Fireplace and open area. Wet Bar and Separate Office Area in Bonus Room, Great if You Work from Home. Enjoy Cool Nights on the Back Deck and Protect your Cars with this Two Car Garage. Home sits on a Cul-de-sac. Hardwood floors throughout Dining Room and Foyer. Master Bathroom has Double Vanity, Garden T
73 Blue Bay Lane 73 Blue Bay Lane Cameron, NC#5347563
- 3 Bed
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- No Cats Allowed (RLNE3821393) Amenities Small Dogs Allowed No Cats Allowed Large Dogs Allowed
930 Dalrymple Rd. 930 Dalrymple Rd. Cameron, NC#5439229
- 3 Bed
- 2 Bath
- $1,150
Available Now! - Available for immediate occupancy. Renovated, and freshly painted, this 3 bedrooms home is located on a large private, lot. Split bedroom floor plan. No carpet, engineered hardwood in main living area and bedrooms. Large closets in bedrooms and a fireplace in the living room. The private location gives you that country living feel while only 5 minutes from town. Easy commute to Sanford, Ft Bragg, and Pinehurst. Pets will be considered on a case by case basis and are subject to
82 Knollwood 82 Knollwood Cameron, NC#5365678
- 3 Bed
- 2 Bath
- $1,050
Beautiful Home Located in Carolina Seasons- Minutes from Shopping and Fort Bragg! - Well maintained home with hardwood floors in Vaulted ceiling living rm, dining rm. & kitchen. Also hardwood in master BR & Bath. Neutral color paint scheme. Golf Course community with pool & club house. Located on cul-de-sac on .55 acre lot with easy to maintain landscaping. Directions: via NC-210 N/NC-24/NC-87, Turn left onto Milton Welch Rd, Turn left onto Ponderosa Rd, Turn right onto Ponderosa Trail, and th
319 Lakeridge Drive 319 Lakeridge Drive Cameron, NC#5629354
- 3 Bed
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- $950
Harnett Co. Country Living Close to Dining, Shopping & Ft Bragg! - Lovely brick ranch home with a large living room with fire place, Kitchen with large dining area, large master bedroom suite. Large privacy fenced back yard and double garage. Convenient to Ft Bragg, Sanford & Southern Pines. Pets Upon Approval - Pet Fee $150 Per Pet Contact Harnett County Schools to Confirm School Assignments Utilities: Central Electric Membership Corp., Harnett County Water No Cats Allowed (RLNE4692267)
1121 Heritage Way 1121 Heritage Way Cameron, NC#5624778
- 3 Bed
- 2 Bath
- $700
1121 HERITAGE WAY ***COMING SOON!!*** - 3 bedroom 2 bath home. Open kitchen/living room. Spacious master bedroom and master bath. Cozy pot belly style heater for additional warmth. Huge backyard. Located in the Heritage Way subdivision. (RLNE4575775)
91 Eunice Lane 91 Eunice Lane Cameron, NC#5497526
- 2 Bed
- 1 Bath
- $575
- Beautiful 3 Bed 2 Bath Located in the rural Cameron area located 25 minutes from downtown Sanford! No Pets Allowed (RLNE5301612) Amenities No Dogs Allowed No Cats Allowed


text: On Monday 07 November 2005 11:47, Charles Swiger wrote: > Yes, but most of the tunables are on the server side. If the server > is running Samba, look for the client caching tuning option in > smb.conf ("csc policy") and the various oplocks options. You want to > have level-2 oplocks working for best performance... Nuts. The server is a Windows XP (or 2003) machine. I'll see what our Windows guy can come up with. -- Kirk Strauser -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/pgp-signature Size: 155 bytes Desc: not available Url : http://lists.freebsd.org/pipermail/freebsd-questions/attachments/20051107/6d31b335/attachment.bin


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text:
One of the most common concerns we come across is the issue of low back pain. We hear it all the time: "I'd really love to be able to work out to lose weight, but my back bothers me too much and I don't want to hurt it more by lifting weights." This thought process, while somewhat understandable, becomes problematic because the individual will typically become sedentary for fear of injury, which will only exacerbate their pre-existing condition. The key to alleviating back pain, is first to understand how back pain, in most cases, is caused, and then devising an educated plan of attack to eliminate these causes safely and effectively through exercise.
According to their website, The Mayo Clinic identifies five (5) main risk factors that are typically linked to chronic low back pain: lack of exercise, excess weight, improper lifting/poor mobility, diseases (like osteoporosis, certain kinds of cancer, and arthritis), and smoking. While all of these factors is important to watch out for, we are going to focus on the issue of lack of exercise as it relates to back pain, and what can be done about it.
Lack of exercise is arguably one of the biggest causes of back pain because it applies to a vast number of people, and when we take a closer look at the anatomy of the spine, especially the lumbar spine (lower back), it's easy to understand why. This portion of the spine is incredibly susceptible to being affected by pain due to its capacity for a wide range of motion, and the fact that it carries all of the weight of the torso. As a result, it is vital that the muscles surrounding the spine are both properly strengthened and flexible enough to support the demands that are constantly placed on the spine. If any one of these muscle groups becomes weakened or too tight due to lack of proper exercise, a number of injuries could occur such as a slipped disc, or nerve impingement (sciatica).
Therefore, one of the most important things that can be done to prevent this type of back pain is regular and specific exercise. In particular, some of the best exercises and stretches to work the extensor muscles include the deadlift, supermans, and cobra pose stretch. For the flexor group, you should incorporate planks, traditional crunches (with feet on the floor) and the kneeling lunge stretch. Finally, to work the oblique muscles, you can incorporate the bicycle crunch and cable trunk-rotations. Additionally, it also advisable to do all your other exercises with freeweights (either dumbbells, barbells, or kettlebells). This will force your body to use the muscles surrounding your lumbar spine to activate while you are completing an exercise that might be focused on completely different muscle group like your chest or legs. This way, every exercise becomes an ab exercise!!
As you can see, when taking into consideration action steps to prevent low back pain from occurring in your future, an enormous amount of focus needs to be placed on regular, and properly performed exercise – specifically strength training. Who knows? After spending enough time working out lifting weights, you may find the only source of back pain you experience is the kind only brought on by your spouse and kids!!
Don't forget we have new Sentinel swag for you to sport, so make sure to pick yours up TODAY! email@example.com email@example.com firstname.lastname@example.com!


text:
Osteogenesis imperfecta (OI) comprises a group of inherited disorders characterized by bone fragility and increased susceptibility to fractures. Historically, the laboratory confirmation of the diagnosis OI rested on cultured dermal fibroblasts to identify decreased or abnormal production of abnormal type I (pro)collagen molecules, measured by gel electrophoresis. With the discovery of COL1A1 and COL1A2 gene variants as a cause of OI, sequence analysis of these genes was added to the diagnostic process. Nowadays, OI is known to be genetically heterogeneous. About 90% of individuals with OI are heterozygous for causative variants in the COL1A1 and COL1A2 genes. The majority of remaining affected individuals have recessively inherited forms of OI with the causative variants in the more recently discovered genes CRTAP, FKBP10, LEPRE1,PLOD2, PPIB, SERPINF1, SERPINH1 and SP7, or in other yet undiscovered genes. These advances in the molecular genetic diagnosis of OI prompted us to develop new guidelines for molecular testing and reporting of results in which we take into account that testing is also used to 'exclude' OI when there is suspicion of non-accidental injury. Diagnostic flow, methods and reporting scenarios were discussed during an international workshop with 17 clinicians and scientists from 11 countries and converged in these best practice guidelines for the laboratory diagnosis of OI.
Classification of osteogenesis imperfecta
Osteogenesis imperfecta (OI) comprises a heterogeneous group of disorders characterized by susceptibility to bone fractures with severity that ranges from death in the perinatal period to subtle increase in fracture frequency and in almost all cases presumed or proven defects in type I (pro)collagen biosynthesis.1 In 1979, David Sillence developed a four-type classification, which is still in use for classification according to clinical/radiological features: OI type I (mild OI with bone fragility and blue sclerae), II (perinatal lethal), III (progressive deforming) and IV (normal sclerae and mild deformity).2, 3 Dominant COL1A1/2 variants appeared to be causative in the majority of OI types. In 2004, OI types V and VI were added to this classification because of specific clinical/radiological and/or histological features, absence of abnormalities of type I (pro)collagen synthesis or structure on gel electrophoresis and absence of causative variants in the COL1A1/2 genes.4, 5 With the discovery of rare recessive genetic causes of OI (see Table 1), it was proposed to extend the classification with OI types VII and VIII.6 However, the classification and subdivision into different types of OI is still under discussion1 because the phenotypic spectrum that results from mutations in some of these genes is almost as broad as that with mutations in type I collagen genes. There is also debate as to whether Bruck syndrome type I7 and II,8 characterized clinically by bone fragility and congenital contractures of the large joints, should be classified as a type of OI.
Biosynthesis of type I collagen
Most individuals affected with OI are heterozygous for a causative variant in either of the two genes, COL1A1 or COL1A2, which encode the proα1(I) and proα2(I) chains of type I procollagen, respectively. Type I collagen is the major structural protein in bone, tendon and ligament. Recently, rare recessive genetic causes of OI have been described and, in all but one, the identified genes encode proteins involved in the biosynthesis of type I procollagen (Figure 1, Tables 1 and 2).9, 10
Genes, proteins and causative variants
Autosomal dominant OI
The most common form of OI in most populations is OI type I. Cultured dermal fibroblasts from affected individuals produce about half the normal amount of type I procollagen molecules, which have normal structure. OI type I usually results from variants in one COL1A1 allele (frameshift, nonsense and splice-site alterations) that lead to mRNA instability and haploinsufficiency. In a small subset of individuals with OI type I, substitutions for glycine by small amino acids (cysteine, alanine and serine) near the amino terminal ends of the triple-helical domains of either COL1A1 or COL1A2 are found. In contrast, OI types II–IV are usually caused by sequence variants in either COL1A1 or COL1A2 that result in substitutions for glycine residues in the uninterrupted Gly-X-Y triplet repeat of the 1014-residue triple-helical domains encoded by each gene. Less common causative variants include splice-site alterations, variants in the carboxyl-terminal propeptide coding-domains or insertion/deletion events that lead to in-frame sequence alterations. Most of these variants result in synthesis of abnormal type I procollagen molecules characterized by post-translational over-modification of the triple-helical domain, which results in alterations visible by SDS-polyacrylamide gel electrophoresis. Loss of expression of either gene because of large-scale deletions appears to be rare.11, 12, 13, 14, 15 More than 1000 distinct variants in the COL1A1 and COL1A2 genes have been identified that cause OI (R Dalgleish: Osteogenesis Imperfecta Variant Database (https://oi.gene.le.ac.uk, accessed 3 May 2011).16, 17
Autosomal recessive OI including Bruck syndrome
In the last 5 years, candidate–gene approaches have identified several loci in which causative variants have been identified which result in the long-sought causes for recessively inherited forms of OI (see Figure 1, Table 2). We classify these genes in four groups—(i) one in which the genes encode proteins that contribute to the initial phase of chain recognition and propagation of molecular folding (CRTAP, LEPRE1 and PPIB)18, 19, 20, 21, 22, 23, 24, 25, 26 the second in which the genes encode proteins involved in the final quality control of type I procollagen (FKBP1027, 28, 29 and SERPINH130), the third, which involves a gene encoding proteins involved in late modification and crosslink formation (PLOD28, 31) and the fourth group, which involves recently recognized genes encoding proteins involved in bone cell differentiation (SP732 and possibly SERPINF133).
Reasons for referral
Prenatal ultrasounds showing abnormalities suggestive of OI (diminished mineralization of skull, platyspondyly or bowing, shortening and/or fractures of long bones) are an important reason for referral for molecular diagnostics of OI.
Radiographs at birth suggestive of OI, recurrent and/or unexplained fractures with or without suspicion of non-accidental injury (NAI), primary (idiopathic) low bone mass, preferably with exclusion of secondary causes, a family history of OI and request for confirmation of the clinical diagnosis are all reasons for referral for molecular diagnostics of OI. However, in some cases dentinogenesis imperfecta or even blue sclerae might be the only reason for referral. As such, physicians from many specialties (gynaecologist, paediatrician, orthopaedic surgeon, clinical geneticist, general practitioner, dentist, ear nose throat specialist, endocrinologist, internist and ophthalmologist) might refer a patient for molecular analysis. Depending on the age of presentation, OI can be difficult to distinguish from some other genetic conditions, for example, Ehlers–Danlos syndrome arthrochalasis type (former EDS VIIA and B), isolated dentinogenesis imperfecta, blue sclerae and corneal fragility, hypophosphatasia, and non-genetic causes of fractures including NAI and idiopathic juvenile osteoporosis.2, 34, 35, 36
Non-accidental injury
Many referrals for OI diagnostics occur in the context of suspected NAI in an attempt to exclude a genetic cause of fractures. Fractures resulting from NAI occur in 24 per 10 000 children under 3 years of age whereas the OI prevalence is 1 per 10 000–20 000.34 The incidence of OI among children evaluated for NAI is 2–5%.34 Differentiation is aided by an experienced clinician and radiologist familiar with OI36 as the nature and localization of fractures in OI and NAI can often be distinguished.37 However, because injuries may involve infants before many of the clinical features of OI are apparent, laboratory diagnostics for OI can certainly be helpful in this differentiation.
Materials and methods
A group of clinicians and scientists involved in OI diagnostics met on 28–29 June 2010 at a workshop in Amsterdam, the Netherlands, to formulate Best Practice Guidelines for the molecular genetic diagnosis of OI supported by the European Molecular Genetics Quality Network (EMQN).
Discussions focussed on diagnostic flow, methodologies, interpretation of results and reporting. Consensus guidelines were established.
The diagnostic flow
The 'traditional' way to establish or confirm the diagnosis of OI is based on studies of collagens synthesized by cultured dermal fibroblasts. Previous studies35 demonstrated that either quantitative or qualitative alterations can be identified in about 90%38 of individuals with clinically confirmed OI. Biochemical analysis will separate individuals with quantitative defects (OI type I), from those with qualitative defects (OI types II–IV) because of causative variants in the COL1A1/2, CRTAP, LEPRE1, PPIB genes and those with no abnormalities detected. Electrophoretic analysis of type I procollagen may also identify other disorders that can mimic some aspects of OI: EDS kyphoscoliotic type (type VIA), EDS arthrochalasia type (types VIIA and VIIB) and EDS dermatosparaxis type (type VIIC).11 Also, cultured fibroblasts provides a resource for the analysis of RNA splicing and unclassified variants. However, biochemical analysis will not identify some quantitative defects of type I procollagen, certain causative variants that alter sequences in some coding regions of the COL1A1/COL1A2 genes and recessive forms of OI (including Bruck syndrome) that result from variants in FKBP10, PLOD2, SERPINF1, SERPINH1 or SP7. Furthermore, prenatal diagnosis by biochemical analysis is only possible in chorionic villus cells and is effective with qualitative alterations9 but unsuitable for quantitative defects and delays time of diagnosis by 2–4 weeks compared with genetic analysis.
In contrast, direct genomic analysis (sequencing) of the known genes should identify causative variants in >95% of affected individuals in most populations.39, 40 Next generation sequencing has gained importance in the laboratory diagnosis of OI – in part because of shorter time to diagnosis and in part because of the added value of the information as well as reduced costs, which is important for the precise determination of recurrence risk (autosomal dominant versus recessive), prenatal diagnosis and pre-implantation genetic diagnosis.
Given these considerations and current facilities, the consensus of the EMQN Best Practice in OI meeting was to initiate laboratory-based diagnostic studies with direct genomic sequencing of the type I procollagen genes, COL1A1 and COL1A2. The approach to diagnosis is detailed in Figure 2. It was agreed that the 'traditional' approach can still be used in the context in which genomic DNA sequencing is unavailable or the financial barriers are high. Moreover, analysis of proteins and mRNA/cDNA from cultured fibroblasts remain valuable tools for follow-up studies.
Explanation of diagnostic work flow
Sequencing of all type I procollagen gene exons
Procollagen type I gene sequencing should identify causative variants in 90% of affected individuals,39, 40 provided that the clinical diagnosis of OI is accurate. In some cases a follow-up study is needed to determine whether a variant is causative. This will often include genetic studies in the parents with correlation to the phenotype observed in the parent, or analysis of mRNA splicing and protein-based biochemical studies on cultured dermal fibroblasts.
If no causative variant in the COL1A1/2 genes is identified by sequence analysis, the next step is to determine if a deletion or duplication of some or all of the coding regions of either gene has occurred. Strategies such as array-based analysis, MLPA or qPCR if properly validated are considered equivalent by the working group in their detection of such alterations. From currently available data in the represented laboratories, the added causative variants expected from this approach should be about 1–2%. In case of OI types II–IV, after identifying the causative variant in the index patient, determination of parental mosaicism by sequence analysis of DNA from blood from each parent will provide data for genetic counselling with respect to recurrence risk and care in subsequent pregnancies.
Re-review of clinical data and the question of NAI
When no causative COL1A1/2 variant is found, clinical priorities should be re-considered. Failure to find a causative variant occurs if there is technical failure, or if no causative variant is present in these genes (because of causative variants in other (un)known genes or because the patient does not have OI). Therefore, referring physicians should be encouraged to provide a completed clinical checklist (Supplementary Table 1) and X-rays of the patient. This clinical/radiological information should be reviewed by a clinical geneticist or a clinician with experience in OI. This can help to determine the likelihood of OI in the particular patient. With clear evidence of OI, further analysis should proceed (Figure 2).
However, if the primary reason for genetic study is to identify children with OI among the larger group suspected for NAI, analysis can reasonably stop after COL1A1/2 sequencing unless there is a strong suggestion of consanguinity or recessive inheritance. Justification of this decision rests on the following considerations. First, previous studies indicate that fewer than 5% of infants studied for suspicion of NAI are found to have OI by biochemical or DNA-based studies.34 Second, DNA-based analysis will identify a causative variant in >90% of all individuals with OI39, 40 so that the remaining risk that an infant has OI, will be about 0.5%. Third, at present all infants with recessive OI have obvious radiographic abnormalities fitting a diagnosis of OI and not of NAI.
Identification and characterization of causative variants in autosomal recessive OI
Variants in the genes causing recessive OI are estimated to account for about 5 or 6% of individuals with OI. This represents the pooled estimates from all the laboratories represented at the workshop. Although laboratory context and clinical criteria may in some cases require otherwise, the preferred strategy is to analyse all genes at the same time to minimize turnaround time. If no, or only one, causative variant is identified by sequence analysis, the next step is to determine by array-based analysis, MLPA or qPCR if a deletion or duplication of some or all of the coding regions of either allele has occurred. Once recessive causative variants are identified, parental confirmation of carrier status should be completed, first to confirm that the causative variants in the index case are inherited in trans and, second, to exclude the possibility of uniparental disomy in the case of homozygosity in the infant.
Analysis of proteins and mRNA/cDNA from cultured fibroblasts (functional analysis)
By following the guidelines to this point, virtually all causative variants in type I procollagen genes and in the recessive OI-related genes will have been identified. However, in some cases analysis of proteins and mRNA/cDNA from cultured fibroblasts can have an additive value. First of all, mRNA/cDNA analysis provides a tool for studying the effect of unclassified variants suspected to alter splicing. Moreover, this approach can unravel the effect of deep intronic variants that alter the mRNA sequences usually by conversion of cryptic exons to active exons and which would not be detected by gDNA sequencing. In these cases, biochemical analysis of type I procollagen chains can detect quantitative defects if the new exon results in introduction of a premature stop codon and nonsense-mediated mRNA decay, or qualitative defects if the mRNA is stable and translated into an abnormal protein. When OI type I is suspected, but genomic DNA sequencing is unavailable or too costly, COL1A1 null-allele14 testing on cDNA can be used to confirm the diagnosis. However, this approach relies on the availability of informative markers and will not lead to the identification of the disease-causing variant.
Analysis of remaining samples
Some diagnostic laboratories maintain research arms and the identification of the additional OI-related genes has profited from the repositories of cells and DNA samples from individuals with OI because of unknown genetic causes. This practice should continue and cells and DNA samples should be banked for future analysis. If the diagnostic laboratory does not maintain such a resource, banking with ones that do, should be considered.
Laboratory diagnosis of OI
Molecular analysis for OI
Molecular analysis for OI includes sequencing or mutation scanning and deletion/duplication testing for COL1A1 and COL1A2, followed by sequencing of recessive OI-related genes (see Table 1). Preferred material for testing is genomic DNA from blood, but other sources can be used as well (see Tables 3a and b). For general issues regarding sample labelling and identification, sequence analysis and interpretation see 'Practice guidelines for Sanger Sequencing Analysis and Interpretation' (http://www.cmgs.org/BPGs/pdfs%20current%20bpgs/Sequencingv2.pdf).
gDNA and cDNA sequencing
Current practices in sequencing of the type I collagen genes differ among laboratories in that some use exon-by-exon amplification followed by sequence determination (Supplementary Appendix Tables 3A and B) and others use a multi-exon substrate for analysis. If properly validated, both will provide the same mutation capture probability and their use should be determined by local preferences. To identify splice variants, primer sequences for PCR used for direct sequencing should be designed far enough from intron–exon boundaries to allow reading of the branch sites, consensus splice donor and acceptor sites or UTR sequences of acceptable quality. With regard to sequence analysis of cDNA, large fragments should be analysed to allow detection of multi-exon deletions, because these could be missed when primers are located close to the exon junctions at multiple sites. The analysis of cDNA from fibroblasts is warranted when an unknown variant is found that might affect splicing. In addition, cDNA sequence analysis may detect null-alleles without a known cause (eg, an unknown variant in the promoter region) if care is taken to analyse regions in which coding sequence heterozygosity is known.
Quantitative analysis
Quantitative analysis by MLPA (kits for COL1A1 and COL1A2 have been designed by MRC-Holland, available at http://www.mlpa.com) and qPCR is particularly important in cases without a detected molecular cause in which only genomic DNA is available and without heterozygous polymorphic variants detected by sequencing. Recently, complete COL1A1 allele deletions have been reported to cause OI type I.13 For reliable quantitative testing (MLPA/qPCR) patient and control materials from the same source (eg, blood or fibroblasts) should be used for comparison and the same DNA purification protocol should be used for all samples within a test series.
Protein analysis
Protein analysis of type I (pro)collagen (Supplementary Appendix Figures 3A–D) is used to detect quantitative and qualitative changes. The method used for this analysis is based on in vitro labelling of collagen in cultured fibroblasts, followed by electrophoresis on 2 M urea SDS-PAGE gels (the 2 M urea enhances chain separation) and autoradiography.41 The analysis of procollagens is achieved by omitting the pepsin digestion step from the collagen protocol. Procollagen electrophoresis enhances detection sensitivity of quantitative type I (pro)collagen defects (OI type I), visualizes defects in type I (pro)collagen located in the N-terminal region of the type I collagen triple helix more clearly and distinguishes between EDS VIIC and EDS VIIA and B (if samples are analysed after labelling in the presence and absence of dextran sulphate to enhance proteolytic processing).42
Prenatal diagnosis
Prenatal diagnosis is possible in case of identification of known disease-causing variant(s) both on genomic DNA extracted from chorionic villus sample (CVS) cells and amniocytes. 'Testing for Maternal Cell Contamination (MCC) in Prenatal Samples for Molecular Studies' (http://www.cmgs.org/BPGs/pdfs%20current%20bpgs/MCC_08.pdf) should be applied. In those rare instances when post-translational over-modification of (pro)collagen type I is visible in, for example, cells from an affected sibling but no disease-causing variant(s) have been detected in the genes involved in OI, electrophoresis of type I (pro)collagen from cultured CVS cells can be used for prenatal diagnosis. Amniocytes are good sources of DNA but they do not make type I procollagen in sufficient abundance to permit prenatal diagnosis.
Interpretation of performed diagnostics
For the interpretation of an observed sequence variant it is essential to establish the causal role of the variant in the pathogenesis of the disease. Textboxes 1 and 2 include descriptions of variants in the genes involved in OI that are likely to be pathogenic or that should be considered as unclassified variants.
General information on requirements for variant reporting can be found in the OECD Guidelines for Quality Assurance in Molecular Genetic Testing (http://www.OECD.org/dataoecd/43/6/38839788.pdf) and in the guidelines issued by the Swiss Medical Genetics Society (http://sgmg.ch/user_files/images/SGMG_Reporting_Guidelines.pdf).
Reporting scenarios
Finding a causative variant in the COL1A1/COL1A2 genes in an affected index case
The report should state that a causative variant has been detected and that this confirms the clinical diagnosis. The report should include a description of the reason why a particular variant is considered causative. Determination of the parental origin of the detected variant(s) should be advised. Testing of relatives at risk should be offered in conjunction with appropriate counselling.
In the case of severe/lethal OI because of a causative variant in COL1A1 or COL1A2, the measured recurrence risk is 2% after the birth of one affected child. The recurrence risk is increased after the birth of two affected individuals, presumably because the proportion of germ cells that carry the mutation is higher.43 This information may be mentioned in the results letter. If parental mosaicism can be demonstrated in the father, the referring clinician could be encouraged to request a sperm sample from the father to clarify the risk.
When the referring physician is not a clinical geneticist, it is recommended that the patient be referred to a clinical genetics centre for counselling. Referral to OI centres for therapy can be proposed if available.
Not finding a causative variant in COL1A1 and COL1A2 genes in an affected index case
If no causative variant is detected in the COL1A1 and COL1A2 genes after gDNA analysis and screening for large gene deletions (eg, MLPA), an interim report can be made with the recommendation for review of the original patient diagnosis (detailed clinical information, eg, with use of the clinical checklist, radiographs, contact with the referring physician (see Supplementary Appendix Table 1)). This review of the clinical diagnosis should involve a clinician with expertise in diagnosis and classification of OI. The outcome of this review might indicate the desirability for testing of additional OI genes and cDNA and protein analysis of type I (pro)collagen. If applicable, a new request for additional testing should be sent by the referring physician to the laboratory. Laboratories that do not offer analysis of the recessive genes should suggest further analysis in another laboratory.
In the case of doubtful pathogenicity of the identified variant, the 'Practice guidelines for the Interpretation and Reporting of Unclassified Variants (UV's) in Clinical Molecular Genetics' (http://www.cmgs.org/BPGs/pdfs%20current%20bpgs/UV%20GUIDELINES%20\ratified.pdf) should be applied. If the genetic testing procedure has identified an unclassified variant as the only sequence change, this should be reported as such. However, the report should clearly state that the clinical significance of the variant is unknown and that its identification does not provide an explanation for the clinical phenotype of the patient.
Finding causative variants in the CRTAP, LEPRE1, PPIB, FKBP10, SERPINF1, SERPINH1, PLOD2, SP7 genes in an affected index case
The final report should state that causative variants have been detected. It is necessary to investigate the carrier status of the parents and siblings in order to determine whether a variant is causative. Testing of relatives at risk may be offered in conjunction with appropriate counselling. Such testing might be useful for prenatal diagnosis.
Finding one causative variant in the CRTAP, LEPRE1, PPIB, FKBP10, SERPINF1, SERPINH1, PLOD2, SP7 genes in an affected index case
When one causative recessive variant has been detected, it cannot be excluded that a second causative variant has been missed, for example when it concerns a deep intronic variant. Dependent on the laboratory, a second interim report can be made indicating the desirability for cDNA and protein analysis of type I (pro)collagen in the case of one causative variant in the CRTAP, LEPRE1 or PPIB genes. When these additional tests are not informative or cannot be performed in the laboratory, the final report should state that it is not possible to confirm the clinical diagnosis OI by biochemical and/or molecular testing.
Not finding causative variants in the CRTAP, LEPRE1, PPIB, FKBP10, SERPINF1, SERPINH1, PLOD2, SP7 genes in an affected index case
In the case of doubtful pathogenicity of the identified variants, the same procedure as described above for the COL1A1 and COL1A2 genes should be followed. In the case where no causative variants in the genes involved in recessive OI have been found, the final report should state that it is not possible to confirm the clinical diagnosis OI by biochemical and/or molecular testing.
Not finding causative variant(s) in the genes known for autosomal dominant and autosomal recessive OI: type I (pro)collagen electrophoresis
Reports of these studies are necessarily more descriptive than the DNA-based results. They should list the types of genetic alterations that could not be identified. When low production or post-translational over-modification of type I (pro)collagen is observed, the diagnosis of OI can be confirmed and in the case of over-modification of type I (pro)collagen, prenatal diagnosis on chorionic villus cells is possible. In reports concerning cells and samples with no molecular and/or biochemical diagnosis of OI, it should be stated that it is not possible to confirm the clinical diagnosis OI.
Prenatal or preimplantation genetic diagnosis
Prenatal or preimplantation genetic diagnosis with the intention of terminating a pregnancy or not selecting embryos carrying the causative variant(s) is possible in the case of identification of known disease-causing variant(s). This service should only be offered in a clinical genetics service and must be accompanied by appropriate genetic counselling. Requests for prenatal or preimplantation diagnosis should always be referred and announced in advance to a clinical genetics service. In addition, the original result letter of the laboratory that identified the causative variant should accompany the request.
Best practice guidelines have been established for the molecular genetic diagnosis of OI. The most noteworthy issue is that molecular analysis of the COL1A1/2 genes is recommended as the starting point in the diagnostic flow as opposed to protein analysis. It is to be expected that molecular genetic testing in OI will only gain in importance as new OI-genes are discovered and with the development of new technologies. However, in certain selected cases protein analysis will remain important.
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Funding for the Best Practice Meeting was provided by the European Molecular Genetics Quality Network (http://www.emqn.org).
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Competing interests
The authors declare no conflict of interest.
Additional information
This paper was presented at an EMQN Best Practice Meeting, 28–29 June 2010, Amsterdam, The Netherlands. Other participants at the EMQN Best Practice Meeting were Javier Garcia-Planells, Filomena Valentina Gentile, Anne-Sophie Lebre, Shirley McQuaid, Rebecca Pollitt, Agnieszka Rusinska, Thomas Schwarzbraun and Janneke Weiss. Affiliations can be found in Supplementary Appendix Table 2.
Supplementary Information accompanies the paper on European Journal of Human Genetics website
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This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
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Cite this article
van Dijk, F., Byers, P., Dalgleish, R. et al. EMQN best practice guidelines for the laboratory diagnosis of osteogenesis imperfecta. Eur J Hum Genet 20, 11–19 (2012) doi:10.1038/ejhg.2011.141
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- best practice
- osteogenesis imperfecta
- type I (pro)collagen
- diagnosis
- reporting
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Massive parallel sequencing and osteogenesis imperfecta: An essential tool for forensic investigation over child abuse
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Osteogenesis imperfecta in Brazilian patients
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DefinitionIn the early Feudal System, different courts dealt with different types of offences. Minor matters such as a nagging wife was dealt by village courts. More serious matters such as heresy were dealt by the church courts. Other offences such as rebellion were dealt by the Kings court. The courts had regular intervals to law breakers. There were three places, the parish church, the building owned by the lord or by the oldest tree in the kingdom. The Kingdoms in the middle ages had small villages all around them. All villages had a list of local by-laws. All the laws were listed in a document called the custumal. If a villager saw someone break any by-laws they then became responsible to raise the alarm, chase after them and if possible detain the law breaker. In some cases, if a villager did not attempt to chase he/she would be charged as well. When crimes were committed in the middle ages the court would either execute the law breakers or send them to the torture chambers. Torture chambers were found in many castles usually in the lowest room in the castle. There were no fixed set of rules for treatment. Some tortures used were; hung, drawn and quartered, rat torture, head crusher, saw torture, foot roasting and flaying torture.
SourcesThere have been many historians in the past, however not many historians have wrote about medieval punishments. When I was searching this topic not many historians names have come up. I did though find a few major historians from the time period. They include; Gildas, Bede, Asser, Ethelweard, William and John of Worcester. As shown, there are not many names from the time period. I believe it was because peasants and villagers were too scared of the punishments that would be inflicted on them. Some historians from our time period though have written about medieval punishment based on fossils and primary sources they have found. They include; Michael Antonio, Lucy Inglis, Stephen Copper and Richard Almound.
BackgroundWhat factors lead to the punishments?Different punishments varied on different offences. Offences such as a nagging wife were dealt with punishments such as a scodal brace or the offender would be dumped in a lake or river. Theft and treason were punishable by death such as hanging and thieves could also
get their hands chopped off. Rare cases such as rebellion were dealt by the kings court and were punishable by death, any torture and the torture used regularly to rebellion was the rat torture.
EffectsWhat was the impact of individuals in society?The punishments in the middle ages effected society deeply. Citizens were too scared to question the way of life. The punishments impacted their beliefs, jobs and day to day life. People of the Kingdom knew they could not go against anything the King said or they could go to the torture chambers. No-on ever questioned church as they knew they would be declared a heretic or a witch. Woman would never quarrel with other woman and no-body could work on Sundays. Citizens were always under pressure to do their jobs correctly or they could be put on a stock for public embarrassment and they always lived in fear and uncertainty.
ReflectionHow does Middle Ages punishments compare to todays?H 523 words.


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About Blog My husband recently died after a long struggle with alcoholism and I am making the slow climb back to normality.
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Since May 2008
Blog alcoholicdaze.blogspot.com
Domain Authority 17ⓘ


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Group's bond sale will be useful but improving advertising sales would be even better?
Church deeply divided as Holy See accused of not wanting to offend Beijing
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Corruption allegations linked to Israel's $2bn contract for German vessels
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