text:
foolsbabe41 Review
I just love this piece. My husband and I have been searching for a cabinet to put in our master bathroom. The measurement of this was perfect and the price was excellent for this quality. My father was a furniture representative for many years. I am familiar with the Pulaski brand and new this would be a quality piece of furniture at this price point. The color is not showing up correctly on Overstock's website. But you can go to Pulaski's website and it is more accurate. The piece is a dark walnut and the drawer & cabinet's are painted in what appears to be expresso / black...there is NO hint of green at all on this piece. Also...I love that the inside of the drawer and cabinet is a complimentary neutral beige color and it is easy to find things inside. You just can't beat it!! It arrived via Fed Ex. The weight on the manufacturer's box states 63 lbs. My husband and I were able to take it up 13 steps. If you need to carry it up steps...Just a tip..empty the inside of the cabinet...a box with the legs are stored inside...it will be lighter to carry. It you look closely you can adjust the shelf or take it out to carry. The drawer does not come out.
- How did the color on site compare with the actual product? Much darker than the picture on Overstock's website.
- Please tell us about the quality of the product. Excellent quality at this price point.
- How accurate was the on site description of the product? Not exact...there is a shelf that is adjustable!


text:
Phoenix, AZUSA
October 18, 2013
i typically prefer wheat beers but i recently acquired a taste for IPA's at a beer tasting i went to a few months ago.
I like my wines sweet, but not so much as that i feel like i need to chase it down with water. i also enjoy a nice dry red wine.
Anywhere with a good margarita and street tacos
Ohioan lived out in AZ since 2001. Studying biology at ASU.
Already a member? Log in.
Already a member? Log in.
Meetup members, Log in
By clicking "Sign up" or "Sign up using Facebook", you confirm that you accept our Terms of Service & Privacy Policy

subset: commoncrawl


text:
Originally posted on MyTeamRivals.com | Last updated 1/8/12
Drew Brees and his New Orleans Saints have officially secured a match-up between them and the 49ers on January 14th. The win for the Saints comes after a beat down on Matthew Stafford and the Detroit Lions on saturday. Final score? 45-28. The Saints were down by four points at halftime, but came back very strong in the second half, scoring 35 points and holding the Lions to one touchdown per quarter.The NFC Wild-Card game between these two teams was no small, unnoticeable matchup. In fact it was one for the record books, if that what you want to call it. Only four quarterbacks in NFL history have ever thrown for over 5000 yards in a single NFL season, and ironically, two of them were playing in this game.Brees, who set a new single-season passing record earlier this season and passed Miami Dolphin great Dan Marino, completed 33 of 43 passes for 466 yards and had three second-half touchdowns. Those three touchdowns were 56-yarder to Robert Meachem, 41-yarder to Devery Henderson, and ...
NFL News
Delivered to your inbox
You'll also receive Yardbarker's daily Top 10, featuring the best sports stories from around the web. Customize your newsletter to get articles on your favorite sports and teams. And the best part? It's free!
Follow us on Twitter
|Best of Yardbarker
|NFL News
|MLB News
|NBA News
|NHL News
Today's Best Stuff
For Publishers
Company Info
Follow Yardbarker
|© 2017 Yardbarker, Inc. All rights reserved. Use of this website (including any and all parts and components) constitutes your acceptance of these Terms of Use and Privacy Policy. (What's New?)


text:
Give us a call to get our secret deals 1-800-468-3578
Property Location
With a stay at Holiday Inn Express Hotel & Suites Syracuse North - Cicero in Cicero, you'll be convenient to Paul deLima Roasting Plant & Factory Store and Big Don's Wild River Mini Golf. This hotel is within close proximity of The Greens at Beaumont and Cicero Historical Society.
Make yourself at home in one of the 74 air-conditioned rooms featuring refrigerators and microwaves. Complimentary wired and wireless Internet access keeps you connected, and cable programming provides entertainment. Conveniences include desks and complimentary newspapers, as well as phones with free local calls.
Be sure to enjoy recreational amenities including an indoor pool and a fitness center.
A complimentary buffet breakfast is included.
Business, Other Amenities
Featured amenities include a business center, dry cleaning/laundry services, and multilingual staff. A roundtrip airport shuttle is complimentary (available on request).


text:
Popular Now
Popular Now
Popular Now
Popular Now
01803 865913
+44 1803 865913
Edited By: Vangie Fradera de Hernandez
37 pages, B/w line illus
Language: Spanish
Spanish identification guide to the most common fish species of Puerto Rico and the Caribbean.
There are currently no reviews for this product. Be the first to review this product!
View all titles in Puerto Rico combined with Fishes: General
Your orders support book donation projects
my first port of call for ordering books
Search and browse over 110,000 wildlife and science products
Multi-currency. Secure worldwide shipping
Wildlife, science and conservation since 1985


text:
Vladimir Vladimirovich Nabokov, 1899 - 1977
TIME magazine cover (23 May 1969)
Born: 22 April 1899 (O.S. 10 April), St. Petersburg, Russian Empire
Died: 2 July 1977, Montreux, Switzerland
Biography from Wikipedia and Authors' Calendar
Additional quotes from Wikiquote. Wikiquote entries are "sourced" and may include items longer than those included here, particularly for poets, lyricists, and dramatists.
Vladimir Nabokov quotes:
Quotes found: 59 — (15 per page, this is page 1 of 4) 1 2 3 4 Next
Click here to find books by Vladimir Nabokov at Amazon.com
- A novelist is, like all mortals, more fully at home on the surface of the present than in the ooze of the past. permalink
Strong Opinions (1973)
- A sense of security, of well-being, of summer warmth pervades my memory. That robust reality makes a ghost of the present. The mirror brims with brightness; a bumblebee has entered the room and bumps against the ceiling. Everything is as it should be, nothing will ever change, nobody will ever die. permalink
Speak, Memory (1951)
- A wise reader reads the book of genius not with his heart, not so much with his brain, but with his spine. It is there that occurs the telltale tingle. permalink
- A writer should have the precision of a poet and the imagination of a scientist. permalink
- After Olympia Press, in Paris, published the book, an American critic suggested that Lolita was the record of my love affair with the romantic novel. The substitution "English language" for "romantic novel" would make this elegant formula more correct. permalink
afterward to Lolita (1956)
- And presently I was driving through the drizzle of the dying day, with the windshield wipers in full action but unable to cope with my tears. permalink
Lolita (1955)
- And the rest is rust and stardust. permalink
Lolita (1955)
- As far as I can recall, the initial shiver of inspiration was somehow prompted by a newspaper story about an ape in the Jardin des Plantes who, after months of coaxing by a scientist, produced the first drawing ever charcoaled by an animal: this sketch showed the bars of the poor creature's cage. permalink
afterward to Lolita (1956)
- Beauty plus pity — this is the closest we can get to a definition of art. permalink
discussing Kafka's Metamorphosis in Lectures on Literature (1980)
- Coincidence is a pimp and a cardsharper in ordinary fiction but a marvelous artist in the patterns of facts recollected by a non-ordinary memorist. permalink
- Curiosity is insubordination in its purest form. permalink
- Curiously enough, one cannot read a book; one can only reread it. A good reader, a major reader, and active and creative reader is a rereader. permalink
Lectures on Literature (1980)
- Direct interference in a person's life does not enter our scope of activity, nor, on the other, tralatitiously speaking, hand, is his destiny a chain of predeterminate links: some 'future' events may be linked to others, O.K., but all are chimeric, and every cause-and-effect sequence is always a hit-and-miss affair, even if the lunette has actually closed around your neck, and the cretinous crowd holds its breath. permalink
Transparent Things (1972)
- Don't cry, I'm sorry to have deceived you so much, but that's how life is. permalink
Lolita (1955)
- For me a work of fiction exists only insofar as it affords me what I shall bluntly call aesthetic bliss. permalink
afterward to Lolita (1956)
Quotes found : 59 — (15 per page, this is page 1 of 4) 1 2 3 4 Next
Please report any problems on this page! If you see any typos, incorrect attributions, deformed characters, or any other problem with this page, we want to fix it as soon
as possible. Please click here to report errors.
Note: Do not use titles in author searches, we don't use them, including president, senator, prime minister, king, queen, saint, or doctor, or abbreviations thereof. See explanation here.


text:
Wish residence of any person commonly is exactly a property which includes a charming pattern, just as Wood Desks Home Office photo collection indicates for you. You can use the Wood Desks Home Office snapshot collection for an inspiration to obtain your Dream property. By having a pattern that is owned, Wood Desks Home Office picture stock is going to be your specific wonderful inspiration. You may look Wood Desks Home Office photograph stock diligently, and so a lot of useful ideas could easily become procured. Several uncomplicated details including accents can be viewed plainly inside Wood Desks Home Office pic stock. Furthermore, various highlights such as the election of motifs and fascinating wall colour choices can even be observed in Wood Desks Home Office photo gallery.
To build your specific perfect dwelling, certain factors of Wood Desks Home Office image collection can be utilized as a guide. A single important thing you have to have could be the idea, and pssibly one of many illustrations or photos which were exactly in Wood Desks Home Office pic collection will probably be your solution. Next, something you may take from this Wood Desks Home Office snapshot stock is exactly the selection of walls designs, since appropriate wall structure colors would provide a comfortable atmosphere to your residence. Those factors must be applied perfectly so it could create a great design and style just like Wood Desks Home Office pic gallery indicates.
We wish you will be able to embrace the particular items coming from Wood Desks Home Office graphic collection really perfectly, so it s possible to construct your own rream property. Along with a lot of info which Wood Desks Home Office photograph collection provides, subsequently you will have a so much more model choices for your dwelling. Besides Wood Desks Home Office photograph gallery, this personal site at the same time gives lots of image gallery that will impress you personally, which means that always browsing this personal blog. Please enjoy visiting the following Wood Desks Home Office snapshot stock.


text:
The Perfect Patties: 8 Great Burger Joints around Virginia
by Patricia Keppel | Posted: Mar 28, 2016 | Updated: May 31, 2016
Smoked, char-grilled, griddled…there really isn't one right way to make a burger. But by using fresh, locally sourced ingredients, crafting the patties by hand, and . . .Read Article


text:
Stuck in Love (2012)
Directed by
Starring (View all)
Greg Kinnear
Bill Borgens
Bill Borgens
Jennifer Connelly
Lily Collins
Samantha Borgens
Samantha Borgens
Nat Wolff
Rusty Borgens
Rusty Borgens
Kristen Bell
Logan Lerman
Liana Liberato
Michael Goodwin
Professor Abbott
Professor Abbott
Written by
Josh Boone
(written by)
(written by)
89 votesMovies without villains.. (66 items)
list by monclivie
Published 7 years, 8 months ago 12 comments
45 votes2014 (668 items)
list by popguns
Published 3 years, 3 months ago 3 comments
36 votesFavorite films of 2013 (15 items)
list by lotr23
Published 3 years, 3 months ago 8 comments
34 votesWhen Critics and Audiences Disagree (235 items)
list by l8t3r
Published 4 years, 3 months ago 5 comments
33 voteswatched in 2013 (293 items)
list by rona sweetlove
Published 3 years, 11 months ago 1 comment
View all Stuck in Love lists (43 more)
Stuck in Love Videos
Stuck in Love Trailer 2013 Movie - Official [HD]
24 Views
Added 3 years ago
View all Stuck in Love videos
Cover art, photos and screenshots
View all Stuck in Love pictures (34 more)
Three layers of the love stories of a family.
2 years, 12 months ago at May 3 21:10read more
An acclaimed writer, his ex-wife, and their teenaged children come to terms with the complexities of love in all its forms over the course of one tumultuous year.
Release date: 2 May 2013
Tags:My tags: Add tags
Add Review
Update feed
AJJ added this to a list 8 months, 2 weeks ago
✦ 2012✦ movies✦ (70 movies items)
Sasa added this to a list 9 months, 2 weeks ago
Watched in 2016 (56 movies items)
DarthTrino added this to a list 10 months ago
Películas que he visto y aumentando. (3617 movies items)
johanlefourbe added this to a list 1 year, 2 months ago
My Queue 2 - Movie backlog (934 movies items)


text:
NZXT Manta mini-ITX Overview
The NZXT Manta mini-ITX case is currently available online for $139.99 (Amazon | Newegg). Inside of the accessory box, we find plenty of mounting screws for the motherboard, storage drives, and the power supply unit. A few zip ties along with a brochure are also included.
The exterior of the Manta mini-ITX looks astounding with its curved and simple design. There is nothing much to see at the front of the case except for a very faint NZXT branding logo towards the bottom of the case; trust me, it's there.
On the main side panel, there is a large window to show off basically everything inside of the case. NZXT claims this is the largest side panel window for a mini-ITX case at the time of this review. It is non-tinted and both side panels include their own captive thumbscrews.
The rear of the Manta mini-ITX maintains a standard layout with a 120mm exhaust fan pre-installed. Two expansion slots are included and the power supply unit mounts directly at the bottom. The power supply unit is protected from dust by a removable dust filter at the bottom of the case.
At the top of the case, we can see the front panel ports along with the power LED and power button. The top maintains a very simple look just like the rest of the exterior.
The Manta mini-ITX stands on four rubberized feet that are tall enough to allow air to pass under the case and to the power supply unit. Here we can see the removable dust filter for the power supply unit.
Now that we have seen a quick look, let's go ahead to show more about the features the Manta mini-ITX has to offer.


text:
Is this attraction free to the public?
Is gratuity suggested at this attraction?
Is this attraction a "must-see" location?
Is this attraction good for couples?
Is this activity accessible without advanced planning or reservations?
Is this attraction good for small groups (less than four)?
Is this attraction suitable for all ages?
Would this be a good sunny day activity?
Would this be a good hot day activity?
Would this be a good cold day activity?
Thanks for helping!
Lovely church with beautiful stained glass windows. The Lucy chapel is of particular interest in that it accommodates the 17th century tombs of three successive Sir Thomas Lucy...
read more
Lovely church with beautiful stained glass windows. The Lucy chapel is of particular interest in that it accommodates the 17th century tombs of three successive Sir Thomas Lucy. Dame Judy Dench and her husband lived in Charlcote and Michael is buried in the churchyard.
We popped into this Church as it was near the Hotel and it was raining. It has lovely architecture and amazing stained glass windows. Judi Dench's husband Michael is buried in the churchyard and judging by the lovely red roses on the grave I think she must visit regularly. The church is in a lovely location next to the National...
Staying in Charlecote Pheasant Hotel this church is just across the road alongside the National Trust property. The entrance to the church is at the back away from the road which we presume dates back to olden days when the people from the big house would have come across their land to the church. As well as being the buriel...
A great example and well preserved
The Lucy family tomb and the history that goes with it is excellent and interesting
Very interesting and picturesque church located just by the from entrance to the NT property. Is the burial place for members of the Lucy family and their tombs are certainly impressive. Well worth checking out.
Church is far more interesting than we expected.
Lovely setting.
Parking very close to church in little lay-by.
Lots ot see in the church and much of interest.
Information leaflets inside so you get to know the history.
Highly recommend.
Nice visit. Very quiet. Lights come on as you move around the Church. Small congregation I imagine who do their best to welcome visitors.
for a tiny village, this church is a must see. Its on the edge of the large charlecote estate, quaint church yard surrounding it.
lovley village church, with some very nice features. But as this is the 25 th January ,why are the Christmas decorations still up?
small church,enjoyed the peacefulness.same name as church in my small town,where my daughter got married.lovely setting.
Own or manage this property? Claim your listing for free to respond to reviews, update your profile and much more.


text:
Really. This happened to me. Thank goodness for some menfolk who worked really hard to beat it out. But...there's gotta be a better way!
sort by best latest
I'd keep a badger on hand for the occasion.
In all seriousness, I have no clue how I'd do it. I'm in Vermont so I never thought about snakes, scorpions, etc. That must have been an experience.
You can help the HubPages community highlight top quality content by ranking this answer up or down.
Close the windows and park the car in direct sunlight - if the temperature outside is above 80 degrees the car will heat up very quickly and the snake being cold blooded will heat up and become very active and aggravated - he will look for a way out of the heat and when you see him moving around open the door nearest him, he'll crawl out! But don't leave the car unattended because a couple hours in that heat inside the car could kill the snake.
You can help the HubPages community highlight top quality content by ranking this answer up or down.
I must advise against tsadjatko's advice on this subject. Many snakes are not able to escape the maze of a car's insides fast enough when in direct sunlight, nor are they wise enough to realize that any dropping temperatures means they must find a warm blanket and a nice fireplace. Many snakes will die in a car because they simply cannot find a way out. And then you have a huge smell on your hands... You can only expect so much from an animal.
I have heard two success stories of owners with lost snakes in cars who recovered their lost pets by placing a small cardboard box (big enough to house the snake, but small enough so that it would be a cozy fit) on the floor of the car right before you go to bed. The box would have a hole cut into the side large enough for the snake to enter, and a warm, thawed mouse would be placed either nearby or sometimes inside the box. This setup is intended to cater to a snake's night-time activities and their superior sense of smell.
The idea is that during the darkness and quietness of the night, the snake may be coaxed into exploring its surrounding, and may become enticed by the smell of food. After consumption, snakes seek a small, dark space (like a box) to curl up in and hide until digestion is finished. Again, I have known this to work twice in the past.
You can help the HubPages community highlight top quality content by ranking this answer up or down.


text:
The Tudor Heritage Black Bay 41 watch will enter stores this month after launching at this year's Baselworld.
VIDEO OF THE DAY: Hublot shows how Ferrari learned how to design watches
Hublot Techframe Ferrari Toubillon Chronograph takes design inspiration from the architecture of supercars.
VIDEO OF THE DAY: AVI-8 presents its 2017 Flyboy Automatic Camouflage watch
Aviator watch brand repaints dial of its 2017 model.
VIDEO OF THE DAY: Breitling presents its 60th anniversary Superocean Héritage II
Swiss watchmaker dedicates the 2017 models to modern-day explorers.
VIDEO OF THE DAY: Sci-fi film shows precise construction of Zenith Defy El Primero
Defy El Primero 21 described by Zenith as the master of extreme precision.
VIDEO OF THE DAY: Omega unveils trilogy of diamond anniversary collections
The Seamaster 300, the Railmaster and the Speedmaster were all introduced in 1957.
VIDEO OF THE DAY: Rolex presents its 50th anniversary Sea-Dweller
43mm Sea-Dweller includes a cyclops lens above the date and its name depicted in red for the first time.
VIDEO OF THE DAY: WatchPro's favourite watch of Baselworld, the Porsche Design Actuator
The precision of the Actuator can only be appreciated with it in your hand, but a video is second-best.
VIDEO OF THE DAY: Watches of Switzerland grills Tudor head of design at Basel
Ander Ugarte, head of design at Tudor, was interviewed by the team about his favourite timepieces.
VIDEO OF THE DAY: Patrick Dempsey shows his love for Jack Heuer creation
Hollywood actor describes his passion for Autavia timepieces in video poem for its creator.


text:
Already a member ? Please Log In.
Not a member yet? Go to the quick Sign Up !
Save time and effort by comparing these leading Web Hosting software.
Quickly access the latest reviews to read actual user opinions and ratings. Get amazing coupons and discounts, access Free Demos or Trials immediately.
DNSimple provides hosted DNS and domain management via a simple web interface, REST API or iPhone application. DNSimple supports a wide variety of top... Learn More
SingleHop is a leading global provider of hosted IT infrastructure and Cloud computing. We are pioneering the global IaaS movement by enabling SMBs an... Learn More
500 domains
Easy to use web interface for managing domain records.
Free iPhone App for adding and managing domains and records from your iPhone.
Support for A, CNAME, MX, SPF, URL (forwarding), TXT, NS, PTR, SRV, NAPTR, SSHFP and AAAA records.
Unlimited number of records for each domain.
Wildcard DNS records
One-click DNS setup for services like Google Apps, Heroku and many others
Native support for DNS lookups over IPv6.
REST API for creating and managing domains and records.
Free iPhone App for adding and managing domains and records from your iPhone.
Optional domain registration or transfers starting at $14/year (for com, net and org). Prices vary for other TLDs.
WHOIS Privacy Protection for $8/year per domain.
Single site SSL certificates for $20.00 for year and wildcard certificates for $100.00 per year, with the easiest certificate interface ever.
Quick-turnaround email support from DNS and domain experts who are here to help.
The satisfaction of not having to use GoDaddy!
Tuesday 27 November 2012
DNSimple is, without question, my favorite domain name registrar and DNS provider. They've been able to live up to their name and really make DNS--simple. If you're shopping around for a new registrar, I'd highly recommend you give DNSimple a shot, I guarantee you'll like them.The only gripe I have about DNSimple is only that they don't (yet) have an Android app! (They do have an Iphone app, if you're an IOS guy.)More about DNSimple
Friday 20 June 2014
Single Hop has created a reputation in the web hosting industry as the fastest growing hosting company that offers great hosting services. On top of that, the company uses powerful state of the art technology equipment to power their hosting services. Great up time statistics and outstanding customer care are some of its strong points.More about SingleHop
Enter your email and press continue to start downloading.


text:
Fashion editorial "Insomnia" by Minsk (Belarus) based photographer Denis Golovey.
"Things happen or they don't happen, that's all. Nothing is accomplished by sweat and struggle. Nearly everything which we call life is just insomnia, an agony because we've lost the habit of falling asleep. We don't know how to let go. We're like a Jack-in-the-box perched on top of a spring and the more we struggle the harder it is to get back in the box." - Henry Miller, Tropic of Capricorn
Photography: Denis Golovey
Hair & makeup: Malango (Yulia Ivanova)
Models: Marie


text:
Streak's men were heading for humiliation when South Africa reduced them to 74 for six with Makhaya Ntini striking four times.
But Streak worked his way to 50, from 90 balls, to lift Zimbabwe to 173 for eight. That total looked well within South Africa's reach. England's opponents in Saturday's NatWest Series final at Lord's were rattled, however, as Douglas Hondo removed Herschelle Gibbs (2) and Martin van Jaarsveld (5).
- More about:
- NatWest
- South Africa
- Southern Africa
- Zimbabwe


text:
Adesiju Food Catering Enterprises is located in , Ojo, Lagos. Our company is mainly into Beverages,Catering Institutes,Continental Cuisine and offering Foreign Dishes, Small Chops, Catering Services, Indoor And Outdoor Catering Services. If you have any questions, do feel free to contact us on the details in our profile or view our website for more details.
Are you the owner of this business?
Claim your business today and interact better with your customers.
Claim business Template Design
What's wrong?
Help us improve the business information


text:
Acer Aspire ATC-280-UR11 Desktop Black Acer Aspire ATC-280-UR11 is a late 2016 model, mid-range Windows 10 desktop PC with mid-range to high-end specs, including very large storage and memory, a capable quad-core AMD A10 series processor with a benchmark score … Continue reading
Category Archives: Acer
14 inch Acer Swift 3 SF314-51-52W2 Full HD Laptop Acer Swift 3 SF314-51-52W2 is a brand new release, lightweight and slim computer in a stylish aluminum chassis, with a 14 inch Full HD display that moves on a 180 degree … Continue reading
Acer Aspire ES 15 ES1-572-31KW 15.6″ HD Laptop Acer Aspire ES 15 ES1-572-31KW is a practical and powerful, 15.6-inch mid-range laptop with a sixth generation Intel i3 processor, a generous amount of storage, decent amount of battery life, the latest 802.11ac … Continue reading
Acer Aspire E 15 E5-575-33BM 15.6 inch FHD Laptop Acer Aspire E 15 E5-575-33BM (NX.GG5AA.005) is a new release 15.6 inch mid-range laptop with the latest 7th generation Intel i3 processor, Full HD screen, great modern connectivity and up to … Continue reading
Acer Aspire AXC-704G-UW61 Compact Desktop Acer Aspire AXC-704G-UW61 is a hot new release compact desktop PC with just one-third the size of a regular desktop, modern design and entry level specs and features for the efficient performance of daily tasks. Technically … Continue reading
Acer Aspire ATC-780-UR61 Desktop Computer Acer Aspire ATC-780-UR61 is a modern, sleek and functional mid-range desktop computer as part of the TC series for general daily use and heavy multitasking of multimedia and other tasks. Technical specs include the sixth generation … Continue reading


text:
Here is our review of the Fabian Strategy, one of the most polarizing Titan Exotics. Some people love it, and.
Telesto is an exotic Fusion Rifle that deals Void damage. Halo fans will find this gun somewhat nostalgic, as its exotic perk is akin to Halo's.
Rise of Iron: Year 3 Thorn Exotic Review: Best Perks and Build Guide. How to get the best possible stats and perks out of the Year 3 Thorn Exotic Hand Cannon!. Destiny Taken King: Sleeper Simulant Exotic Review It means that if you get a exotic review guide kill, you get a little help with the next one. Low RoF Hand Cannons are often given greater base Ranges, so this is a disappointment. Charging before rounding a corner, free sothink dvd off an explosive volley, and then shifting to a new area consistently worked for me in terms of clearing out people defending rooms or hallways. They are often overlooked and replaced by snipers and shotguns in both the Crucible and in PvE activities. Iron Banner: Finnala's Peril and Deidris's Retort God Rolls.
Free: Exotic review guide
|How to Get Touch of Malice - Exotic Raid Quest Guide. All Calcified Fragment Locations. Stick to using it in the Crucible if you want to make the most of the Unrepentant perk. Locations on The Reef. Everything you need to know about Sealed Ahamkara Grasps and their stats, perks, and playstyles! Iron Banner: Finnala's Peril and Deidris's Retort God Rolls. View Gallery Every so exotic review guide an exotic comes along that redefines what it means to be an exotic.
|Is bovada safe and legal
|Exotic review guide
|Once I got comfortable with it, however, the transition to pop-in-and-out was easy. I set out to take a look at whether this gun is rightfully overlooked, or a well kept secret. The Erotic Reviewalso known as ntclibyaus.orgis a review site that allows customers to rate their experiences with sex workers referred to as "providers" on the website. The difference here is borderline negligible. He prides himself on his ability to fail at almost any exotic review guide, regardless of difficulty.


text:
74103Re: [TexasCzechs] Searching for Employer of Maid-my ancestor
Expand Messages
- Apr 26 3:24 AMSorry to be so late in responding, due to unforeseen circumstances I'm juggling to many roles lately.First and far most I want to respond to Paul, it was never my intentions offending anyone, by using the term "that" in my writing. I find all of my blood lines interesting even if I wasn't aware of any particular ethnicity, I myself refer to me as a Heinz 57, because I have a little of everything. I embrace and I am proud of every ounce of it.Rosemary, I didn't have a family tree made until afterwards, because I wondered, but it did connect me to people later showing me moderately related, when nothing in their tree nor mine link but 1 last name being same???? so I wondered how they were determining relationships. I did the DNA primarily to match to a 2nd family that my 3rd grt grandfather deserted my grandmother in 1870's never to be heard of again. The 2nd family and I know it's him, but we thought it would hit to confirm us from him, and it did not....but we hit on 2 same cousins so we know we have a connection. I plan on doing a grandparent lineage DNA I found on line and they specifically gene marker to prove he is the same man.Peggy Harmon, CPC"Texas is My Heart"New Braunfels "is my home"On Sunday, April 20, 2014 2:18 PM, Leroy Janda <ljanda@...> wrote:
I agree 1000%, wife had the same results.I think they try and go and find all family trees where she was posted, and used that to calculate the %. Probably did not even do a dna.
- << Previous post in topic


text:
Welcome to The Advanced Materials Research Laboratory (AMRL)
The Advanced Materials Research Laboratory is a collaborative initiative between Departments within the Faculty of Engineering to develop new materials characterisation and testing research facilities. The facility comprises 2 designated Advanced Materials Research Laboratories within the Department of Mechanical and Aerospace Engineering;
Materials Characterisation Laboratory (JW224) comprising forensic analytical techniques including SEM/ FESEM with EDS, WDS, EBSD, STEM, coolstage and in-situ Experimental Mechanics (Deben microtest 2kN tensile testing).
XRD powder diffraction, residual stress, texture and 20000 C heat stage. with multi sample holder
Laser Flash thermal diffusivity plus TGA / DSC and Dilatometry/ TMA
Intrusion Porosimetery 900 micron to 3.5 nm pore diameter.
Nikon XT H 320 XRay CT scanner .
Experimental Mechanics Laboratory (JW124) houses a complete suite of mechanical testing equipment for tensile, compression and fatigue testing from tens of Newtons to 250kN
Additional AMRL equipment is housed within the Department of Bioengineering and a third designated laboratory hosted within the Department of Civil Engineering (JW109) including AFM (with nano indenter)


text:
Expert Service 1-661-257-9993 [Hours]
12V Compact Smart De-Sulfating Battery Charger for Lead Acid Batteries
Get FAST answers from customers ASK a Question See questions answers
General Features:
- State of the art compact car battery charger for smaller 12V lead-acid batteries
- Perfect for charging smaller batteries found in motorcycles, jet skis, ATVs and lawnmowers
- Suitable for maintenance charging of 'normal-sized' batteries like car batteries
- Provides fully automatic charging and maintenance in 6 steps to maximize battery life and performance
- Unique display allows you to follow the entire charging process
- Water and dust resistant design allows for use in nearly any environment
- Charger is non-sparking, reverse polarity protected and short circuit proof
- Charging Voltage: 14.4V
- Charging Current: 0.8A Max
- Battery Capacity: 1.2-32 Ah
- Up to 100 Ah for maintenance
- Authorized Internet Dealer
- 5 Year Manufacturer's Warranty
Also found as: US 0.8, US 08, 56865
Battery Charger Specifications
Warranty and Return Policy
Summary of Customer Ratings & Reviews
Product Summary
Model Number
Internal SKU
Internal Product ID
Wiring Diagram Legend
Warning: The image depicted shows the resistance change when wiring multiple subwoofer terminals. Please refer to your subwoofer's owner's manual for the proper wiring of its terminals. Sonic Electronix, Inc. is not responsible for damage caused to your audio system or vehicle due to improper installation. Please call tech support at 1-877-289-7664 if you require additional assistance.
Product Name: CTEK US 0.8 (56-865)
Returns Made Easy
We want your online shopping experience to be easy and convenient. In case you need to return an item, our customer friendly return policy makes it easy.
60-Day Money-Back Guarantee
Have a problem with your order and need to return an item? Our generous 60-day return policy allows you to return items for your money back. Should your product have a defect, we will exchange, issue store credit, or repair the product for the entire length of the warranty. Learn More
No Restocking Fees
That's right, 100% refund on complete, like new returns. Learn More
365-Day Coverage
Let's face it, electronics break. We've got you covered for a full year. Learn More
Easy Return Shipping
Request a return, use our shipping label, and ship it back. Learn More
In-House Experts
Each of our employees are true experts with real life experience and passion for the products. Our team is always committed to providing you with electronics you'll enjoy.
Always available to answer your questions
Our trained install professionals can tackle any install challenge
Research & How To
Browse our online library of educational resources to learn more about the items you are looking for. Our experts have developed many tools, written many articles, and numerous videos to help you find the best item for your needs.


text:
|The convenient store which gathers anything from Tokyo
Please enjoy high quality shopping in Japan !
|Product name
|Phytomer oligopeulpointjel * 30 ml
|Product description
|Essence of the skin due to excess sebum secretion and acne;
|Water PG polyquaternium--37 capriloilglycine dimethyl Isosorbide di (Caprylic / capric acid) PGTEA panthenol methacrylic acid alirclosporimmer PPG-1 trideceth-6 phenoxyethanol fragrances salicylic acid acriratescopolymer oleic acid sorbitan methylparaben asparagopsisalmata extract citric acid maternally ethylparaben, isobutylparaben propylparaben
|How to use
|Use after conditioning skin with lotion. Trouble area with the tip of the tube, gently apply a small amount. One day many times as available from the make-over. * Hurt the skin symptoms such as skin peels easily, you may not.
|Area / country
|And foreign-made cosmetics
|"Thalassotherapy sea marine cosmetic brands Spa and beauty salon in Megumi's favorite saltwater and seaweed sea mudflats and efficacy PHYTOMER Phytomer.
Start Date:03/04/2017 00:00 JST
End Date:30/06/2017 23:59 JST
This product cannot be shipped to United States from the store.
Please contact the store for further information.


text:
Project for KatelynJB -- 4
This project was awarded to KatelynJB for ₹9000 INR.Get free quotes for a project like this
Project Budget₹9000 INR
Total Bids1
Project Description
Hi, As discussed earlier, I would need a Coming Soon Landing Page and a 404 page, I do need it HTML5 since I can do it on Wordpress myself. Deliverables would be full HTML5 page + CSS and Photoshop designs used if any, thank you
Awarded to:
Skills Required
Looking to make some money?
- Set your budget and the timeframe
- Outline your proposal
- Get paid for your work
Hire Freelancers who also bid on this project
Looking for work?
Work on projects like this and make money from home!Sign Up Now
- Forbes
- The New York Times
- Time
- Wall Street Journal
- Times Online


text:
You have Adblock enabled. Adblock has been known to cause issues with site functionality. If you experience any difficulties, please try disabling Adblock. Close
Add to Favorite List:
Create a new folder:
Cindy Vega in the Bathroom
Recognize a pornstar in this video?
Help make pornstars easier to find on Tube8 by telling us who is in this video.
Thanks! The pornstar names will be added to this video when enough users submit the same names.
You must enter a pornstar name that already exists in our database.
- Category: Teen
- Views: 72,839
- Added: 2 years ago
- Description: Super skinny and tall Cindy Vega fucks her boyfriend in the shower.
- Tags: skinny, tight
- Pornstars: Sindy Vega Add Pornstar
- Favorite by: 111
- Featured: 2 years ago
- Report This Video
Video Comments
All Video Comments (1)


text:
|Half Baths
|3,300 sq. ft.
|Property type
|Minimum Stay
|7 nights
Our home is a large (3300 sq. feet), fully renovated Victorian single family house located in Brookline, MA - just steps from the Boston border. The master bedroom has a queen size tempurpedic bed and an en-suite master bathroom with a jetted tub. There are two additional bedrooms - each with a queen size bed and a dedicated full bathroom. Additionally we have a queen size air-mattress that can be set up in the 2nd floor office. This is our primary (and only) residence so the house comes complete with everything you will need for a comfortable stay including a fully equipped renovated kitchen with stainless steel appliances and granite countertops, 3 flat screen HD TVs with premium channels (HBO, Showtime, etc.), high speed WIFI, large capacity washer and dryer, linens and towels, outdoor deck and gas grill. 7 night minimum rental. Discounted rates available for stays of 30 nights or more.
Our family of five had a wonderful stay here during the 6 weeks we were in Boston for medical treatments. It was a perfect place to relax and ideally located for easy access to anywhere in Boston. The house had everything we could want -- from printer and toiletries to books, games and laundry detergent. Literally everything was provided (even winter ice melt!). And as the photos show, it's a very beautiful and well cared for home. What we appreciated most, however, was the owners' flexibility, excellent communication, and obvious concern with our comfort. Nancy and Jeff checked in periodically to see if we needed anything, and were always immediately responsive when we had questions. They clearly care a great deal about their home and renters. We can't recommend this house highly enough.
Nancy and Jeff were very available if we needed anything. Their home was beautiful and we truly enjoyed our stay there. We appreciated their attention to detail. They made sure that we were very comfortable. Thank you! It could not have been better.
Nancy and Jeff are awesome! They help you in everything they can to make your vacation more easy.
The house is very clean, new, big!
Short wallking distance to restaurants.
Nice playgrounds two blocks away.
My family had a great time during our stay! The home was lovely and had plenty of space for sleeping, cooking and working whilst vacationing to Boston! Nancy was very helpful when we needed extra serving dishes, and her responses were quick. The home was clean, had all of the staples, including coffee, in the kitchen. We would stay here again!!
The pictures do not do this home justice! The house is simply breathtaking. It was the perfect Boston home base for our vacation. Jeff and Nancy went out of their way to make us feel welcome and provided everything we needed for the 7 of us to enjoy our week. Staples like coffee, tea, sugar, cream, paper plates, napkins, laundry soap, dryer sheets and such were all provided. The kitchen is well stocked with dishes, flatware, pots, pans, cutting boards, toaster oven, etc. The cabinets contain games and plenty of movies and books. There was not a single thing that we needed that was not provided. This is their personal home and the attention to detail is obvious. The beds were very comfortable. No a/c, but we didn't need it. Hot water was plentiful and VERY hot! Water pressure was good in all the bathrooms. The house is very conveniently located to the T-station and many shops/restaraunts in Brookline Village. We enjoyed walking to Fenway Park, the Museum of Fine Arts, and the Isabella Stewart Gardner museum from the house. We were even able to use membership passes for the museums that were left for our use. Their is a Stop-n-Shop grocery store within easy walking distance and a very nice Wegmans just a short drive away, discount cards were also available for our use. A nice binder of local information is available and a ton of carry-out/delivery menus as well. No complaints whatsoever and we would definitely stay there again if our travels take us back to Boston.
We spent two great weeks in this lovely home. The house has been meticulously cared for and is furnished with everything you could possibly need and more.
It is perfectly located for getting to all areas of Boston and also suburbs. One of the unexpected pleasures was that the house is close to the "Emerald Necklace" park and Jamaica Pond, so we were able to take beautiful long walks every day.
Nancy was great to work with-- very helpful and responsive about our questions.
Brookline is a small town with a very urban feel, surrounded on 3 sides by the city of Boston. Our neighborhood is made up of single and multi-family homes located close to one-another. There are sidewalks and several parks/playgrounds nearby. Our home offers easy access to all Boston attractions as we are conveniently located within walking distance (0.3 miles) to the Brookline Village 'T' stop (Boston's Mass Transit system). We are also walking distance to many local shops and restaurants.
|Rate Period
|Weekend Night
My Standard Rate
7 night minimum stay
|Cleaning Fee
|Property Damage Protection
Guests can choose to pay a $5,000 refundable security deposit in lieu of the $99 insurance protection. Rates are subject to change until reservation is confirmed. No refunds for cancellations within 60 days of check-in.


text:
|Home | About | Journals | Submit | Contact Us | Français
Missing phenotype data can be a major hurdle to mapping quantitative trait loci (QTL). Though in many cases experiments may be designed to minimize the occurrence of missing data, it is often unavoidable in practice; thus, statistical methods to account for missing data are needed. In this paper we describe an approach for conjoining multiple imputation and QTL mapping. Methods are applied to map genes associated with increased breathing effort in mice after lung inflammation due to allergen challenge in developing lines of the Collaborative Cross, a new mouse genetics resource. Missing data poses a particular challenge in this study because the desired phenotype summary to be mapped is a function of incompletely observed dose-response curves. Comparison of the multiple imputation approach to two naive approaches for handling missing data suggest that these simpler methods may yield poor results: ignoring missing data through a complete case analysis may lead to incorrect conclusions, while using a last observation carried forward procedure, which does not account for uncertainty in the imputed values, may lead to anti-conservative inference. The proposed approach is widely applicable to other studies with missing phenotype data.
Interest in identifying the genetic basis of variation in quantitative traits continues to grow. In humans, family-based linkage studies were once the prominent study design to identify quantitative trait loci (QTLs), but the advent of genome-wide association studies (GWAS) using unrelated subjects has made QTL identification more feasible than ever, and numerous medically relevant traits have been mapped using this approach in the past few years. The use of model organisms to map QTLs is complementary to studies in humans and also offers some notable advantages. The vast genetic diversity of different natural populations can be exploited to uncover different regions of genome that harbor QTLs that would otherwise be obscure, and at the same time experimental populations can be designed to avoid potential biases due to population structure. Additionally, environmental variables can be tightly controlled to reduce undesired variance. Finally, model organisms provide the opportunity for experimentation that could not be performed in humans.
The mouse is often used for such purposes. Divergent parental strains are crossed to form F1 heterozygotes, and these F1 mice are then intercrossed (or backcrossed to a parental line) to form F2s with recombinant chromosomes. The trait, or phenotype, of interest is quantified, and statistical methods are employed to assess the association between the genotype and phenotype (Broman, 2001). Since the mice are studied in a common environment, differences in phenotype may be attributed to genetic discordance. This approach has been used for numerous traits of biomedical relevance, and through further experimentation, investigators can identify the specific genes underlying the phenotype of interest. Because a large effort is expended to breed mice for these types of studies, the mouse genetics community has developed resources to facilitate and expedite QTL mapping. In particular, recombinant inbred lines (RILs), generated by repeated (e.g. > 20 generations) brother-sister matings of mice produced from intercrosses, have been developed and are commercially available. These offer the advantage of being a renewable resource; that is, mice of the same genotype can be phenotyped for multiple traits across space and time. The Collaborative Cross (CC) is the newest and most powerful resource of this type. The CC was designed to overcome many limitations of previous QTL mapping approaches by capturing the maximal genetic diversity of inbred strains while creating a balanced population structure (Chesler et al., 2008; Churchill et al., 2004). Specifically, the CC is a panel of recombinant inbred lines derived from eight-way crosses of classical (A/J, C57BL/6J, 129S1/SvImJ, NOD/LtJ, and NZO/H1LtJ) and wild-derived inbred strains (CAST/EiJ, PWK/PhJ, and WSB/EiJ). The strengths of the CC compared to other approaches (Roberts et al., 2007) and initial applications with the developing lines of the CC have recently been described (Aylor et al., 2011).
Here we use unrestrained whole body plethysmography (WBP) to phenotype a respiratory system trait in developing lines of the CC. WBP measures the amplitude and frequency of breathing and, from the features of the breathing waveform, an empirical parameter known as enhanced pause (Penh) is calculated. Measurements of Penh in the presence of aerosolized saline and then increasing concentrations of methacholine, a bronchoconstrictor, are used as measures of breathing effort, and this changes in the context of lung inflammation due to allergen challenge (Hamelmann et al., 1997). Penh is therefore often used in mouse studies of allergic airway disease, a model of human asthma. Results from previous QTL mapping experiments have been nicely summarized in two recent papers (Camateros et al., 2010; Leme et al., 2010). Zhang et al. (1999) used an F2 intercross, applied an ovalbumin-induced allergic model, and identified QTLs on chromosomes 9, 10, 11 and 17. Ackerman et al. (2005) identified interacting loci on chromosomes 2 and 6, and most recently Camateros et al. (2010) identified loci on several different chromosomes. We sought to exploit the diversity of the CC to identify new QTLs for Penh. This diversity was immediately apparent during the phenotyping process, as some mice had Penh values that were very high even at low doses of the bronchoconstrictor methacholine, and hence phenotyping was stopped in order to protect the well-being of the mouse. This led to a situation in which there was missing data for some mice, a result that has not been described in previous studies with the same goal.
Without missing data, the standard QTL mapping strategy would be to (i) fit a genetic model at each marker (for a large number of markers), (ii) obtain a test statistic that quantifies the discrepancy in the phenotype value across different genotype strains at each marker, and (iii) apply a permutation test to determine if the largest observed test statistic is statistically significant. If statistical significance is achieved, then this is considered evidence of a QTL at the marker having the maximal test statistic. In the situation where data are missing for some of the mice, this analysis must be adapted to appropriately account for the missing data.
If data necessary for quantifying the phenotype of interest is missing for a subset of mice, a significant loss of power and bias may be introduced unless the missing data are appropriately handled. Frequently the phenotype value is a numerical summary of several observed characteristics of the mice, e.g., area under the curve (AUC) values obtained from dose-response data in drug metabolism studies. In this case, when one or more of these observed characteristics are missing for a subset of mice, the desired phenotype measure (AUC) cannot be computed. An analysis that only uses data from those mice with complete measurements may yield biased results, depending on the missingness mechanism. Rather than perform the desired QTL mapping analysis on just the subset of mice for which the phenotype quantity is available, a model that describes the characteristics used to compute the phenotype summary may be developed and used to impute the values for these characteristics. Then, the complete dataset obtained from the imputation model may be used to impute the missing phenotype summary. The approach of multiple imputation (MI) is preferred to imputing a single dataset, because it provides a way to incorporate uncertainty in the imputed values into the analysis (Little and Rubin, 2002; Schafer, 1997).
While genotype imputation has become a widespread tool in genetic association studies (Li et al., 2009), imputation of missing phenotype data has been less commonly applied by the QTL mapping community. In particular, previous studies of Penh did not observe the same level of phenotypic diversity as seen in the CC mice, and so missing data did not pose a major challenge for these studies (Ackerman et al., 2005; Camateros et al., 2010; Leme et al., 2010; Zhang et al., 1999). Some prior work beyond the literature studying breathing effort in mice has explored methods for handling missing phenotype data in linkage analysis, family-based studies, and pedigree analysis (Ding and Laird, 2009; Fridley and Andrade, 2008; Fridley et al., 2003; Xing et al., 2003). In these studies the phenotype investigated was a univariate, continuous parameter rather than a function of incompletely observed dose-response curves, and so the development of a suitable phenotype imputation model was not a primary focus. Additionally, one feature not emphasized by these studies is how inferential methods that account for the imputation of missing phenotype data should control for multiple comparisons, which is an important goal in mapping studies seeking to identify significant QTLs among a large number of potential markers.
In this paper we describe the methodological challenges involved in incorporating MI in QTL mapping when there is substantial missingness of a complex phenotype. We focus on a particular experiment looking at the genetic factors that contribute to breathing effort, as quantified by Penh, in mice. Our work provides two main contributions. First, we develop a novel imputation model that captures the unique features of the data. Second, we develop a methodology for combining MI with a commonly used statistical analysis for QTL mapping. One of the features of this approach is that it is a generalization of the analysis that would have been conducted had there been no missing data. While our imputation model is specific to the experimental protocol for measuring Penh over increasing doses of the bronchoconstrictor, the conjoining of MI and QTL mapping may be generally applied to other experiments with missing phenotype data.
The outline of the paper is as follows. In section 2 we introduce the phenotype and genotype data and in section 3 we define the phenotype summary for QTL mapping. We describe the phenotype model used for MI in section 4 and the methodology for combining imputed datasets with the QTL analysis in section 5. The application of the methodology to QTL mapping of the Penh data is detailed and the results presented in section 6. We conclude with a discussion of the methods and results in section 7.
One hundred and sixty-two male mice that were part of a collaboration, described in Aylor et al. (2011), were obtained and housed singly at the National Human Genome Research Institute. To distinguish between the fully inbred lines of the CC and the mice used for this study (which are not yet fully imbred), we refer to these mice as "preCC" mice. Given that the mice were not yet fully inbred, each mouse's genome was unique, and biological replicates were precluded.
Genotyping and haplotype assigment methods have been described by Aylor et al. (2011). Briefly, mice were genotyped using the Affymetrix Mouse Diversity Array (Yang et al., 2009a). SNP data were then used to infer haplotype probabilities using a hidden Markov model (Mott et al., 2000). That is, for each region of the genome, the probabilities that the region descended from each of the eight parental strains of the CC were estimated, and these founder haplotype probabilities were used (not genotypes) in our QTL model described in Section 5.1. For this study we consider just chromosome 8 in order to demonstrate the application of the proposed methodology. Because the marker density exceeded the total density of recombinations in the cross, it was possible to reduce the chromosome 8 genotype data to approximately 1,380 intervals within which estimated genotype probabilities were essentially constant, indicating a single haplotype for that region. Interval boundaries were defined at transitions in highest probability genotype, based on Baum-Welch output from a Hidden Markov model. In most intervals, haplotype probabilities were near 1 for the inferred states. The haplotype probabilities for each mouse sum to two (because each mouse inherited two CC alleles), and since the mice are the products of inbreeding but not yet fully inbred, most regions are homozygous for one CC strain.
A longitudinal study design (Lofgren et al., 2006) was conducted in which each mouse was phenotyped for Penh using WBP at three time points, the first two being baseline measurements, and the third subsequent to an allergen challenge, as shown in Figure 1. The two baseline measurements were conducted within one week, and hence we assume that the baseline phenotypes are exchangeable. After acclimatization to the WBP chamber over 15–20 minutes, mice were phenotyped for Penh as follows. The mice were administered increasing doses (0, 3.1, 6.2, 12.5, and 25 mg/ml) of methacholine by nebulization over a 2.5 minute period. Following the nebulization period at each dose, mice were followed for five minutes, and average Penh over the five minute period was calculated.
The protocol we employed is a slight modification of that described by Kelada et al. (2011). Low endotoxin Der p 1, from the Dermatophagoides pteronyssinus species of house dust mite, was purchased from Indoor Biotechnology. Mice were sensitized with 10 g Der p 1 by intra-peritoneal injection on days 22 and 30 of the study. On day 37, mice were challenged by oro-tracheal aspiration with 50 μg of Der p 1 diluted in 40 μl of saline. Peak inflammatory responses occurred 72 hours after airway challenge, and so mice were phenotyped at this time point.
The phenotyping protocol just described was designed to yield three methacholine-Penh dose-response curves for each mouse, including two replications at baseline (before the allergen challenge) and a single curve measured after the allergen challenge. The phenotype summaries, which are functions of the three dose-response curves, are described in section 3.
We next introduce some notation. Let Yijkl denote the Penh response for mouse i at dose dj of methacholine, for replication k, of condition l, where l {pre, post} is the pre- or post-allergen challenge condition. Denote the methacholine-Penh dose-response curve for mouse i of replication k and condition l by Yikl(d), where d is the dose, and note that Yijkl = Yikl(dj) for j = 1 to 5.
Table 1 summarizes the distribution of missingness of the dose-response curves. Given that a Penh value is missing for a particular dose, it is missing for all subsequent doses. Thus there are six possible missingness patterns for each methacholine-Penh curve: Penh missing at all 6 doses of methacholine; Penh observed at the first dose, but missing at doses 2 through 5; Penh observed at the first two doses, but missing at doses 3 through 5, and so on. Values at higher doses are missing in order to protect the well-being of the mouse. High concentrations of methacholine can lead to overstimulation of the airways, causing excessive narrowing and hence extreme difficulty breathing, and/or systemic cholinergic crisis. Additionally, we note that a subset of 40 mice were not phenotyped for the first repetition of the pre-condition (i.e. were only phenotyped at a single timepoint) due to a change in the WBP protocol.
To understand the potential impact this missing data may have on QTL mapping, we first provide a brief overview of commonly used terminology from the statistical literature on missing data. Historically, three classes of missing data mechanisms have been defined, which serve as a useful tool for selecting appropriate statistical analyses for handling missing data (Rubin, 1976; Little and Rubin, 2002). The first mechanism, missing completely at random (MCAR), occurs when the missing data come from the same underlying probability distribution as the observed data. The second mechanism, referred to as missing at random (MAR), arises when the probability that a data point is missing depends only on observed variables, which can include covariates (e.g. mouse body weight) and outcomes (e.g. observed Penh values Yijkl). Missing data are classified by the third mechanism, missing not at random (MNAR), when the probability that a data point is missing depends on unobserved variables. If the missingness mechanism is MCAR, then statistical analyses based only on individuals with complete data will yield unbiased results, though substantial loss of power may result under high levels of missingness. When the less stringent assumption of MAR holds, it is sufficient to posit a model for the outcome process and draw inference using multiple imputation or maximum likelihood methods. However if the mechanism is MNAR, then one must model the missingness mechanism as well.
For the 40 mice that are completely missing dose-response curves from the first repetition of the pre-allergen challenge condition, it is plausible that these curves are MCAR, as the decision to change the WBP protocol was independent from the data-generating process. For these mice, the missing curves were completely imputed based on the imputation model described in section 4, which assumes the less restrictive MAR mechanism. We next investigate the potential missingness mechanism for the partially observed curves. Figure 2 displays the observed data for all mice in both the pre- and post-allergen challenge conditions. We observe that curves that have missing Penh values at the higher doses of methacholine tend to be increasing faster than those that are not missing Penh values. If the data were MCAR, then the rate of increase of curves with missing data would be the same as the rate of increase of the completely observed curves. It follows that the missingness mechanism is not MCAR for these partially observed dose-response curves, and so a complete case analysis would be inappropriate. However, the observed data cannot similarly be used to distinguish between MAR and MNAR. The approach for integrating multiple imputation with QTL mapping described in section 5 is valid under the MAR assumption.
It is important to point out that there may be a philosophical objection to considering Penh values at the higher doses to be missing. In some cases, it is possible that the mouse may not have survived exposure to the bronchoconstrictor (methacholine) beyond a particular dose, but this dose of maximal tolerance is not observed by the study design. Thus the concept of "missing" may not be well-defined at the doses where Penh values have not been measured, if those values could never exist. Nonetheless, it may be reasonable to assume based on discussions with our collaborators that a Penh value at least one dose beyond the largest observed dose does exist. We will address this concern by considering alternative specifications of the phenotype summary (section 3) which do not depend on the highest doses, so as to mitigate reliance upon imputed Penh values at doses where their existence is questionable.
There are two primary phenotype summaries of interest for each mouse. The first phenotype , defined as the area under the curve for the pre-allergen challenge condition, quantifies breathing effort under normal physiologic circumstances. The second is the area under the curve for the pre-allergen condition subtracted from the area under the curve for the post-allergen condition. This latter phenotype quantifies the increased breathing effort when the lung is affected by airway inflammation. The two summaries and are correlated but are both unique biological entities of interest to the lung research community. Since mice have two replications of the pre-condition, we use the average area under the curve over the two replications. Specifically, we define the phenotype summaries of interest by
where is the average of the two pre-condition Penh measurements at dose d. For the purposes of calculating AUC for (1) and (2), we assume a linear interpolation of each dose-response curve between consecutive doses. The phenotypes of greatest scientific interest are and , with the integration through the fifth dose.
To demonstrate the potential gain achievable by an imputation approach versus a completers only analysis, we calculate the number of mice for whom and could be calculated using the observed data. Note that there are two ways to define a complete case for the phenotype summaries (1) and (2), since there are at most two repetitions of the pre-allergen challenge condition for each mouse. First, one could consider an exclusive criterion, where it is required that both precondition curves be observed at each dose up through d in order for (and hence since it is a function of ) to be computed. Alternatively, one could consider an inclusive criterion where is computed if either of the two pre-condition curves is observed through dose d. For this latter case, if there is only one measurement at a particular dose d, the value i(d) is set to be that measurement, and if there are are two measurements at that dose, the value i(d) is set to be the average of the two measurements. Using the inclusive criterion, out of the 162 mice, one could calculate for 127 mice and for 79 mice. Requiring the exclusive criterion, and could be calculated for only 71 mice and 53 mice, respectively.
If one were to conduct an analysis with only the complete cases, results may be biased since mice with complete data tend to have lower Penh values. To increase power and reduce the potential for bias in our analysis, we build a model for the methacholine-Penh curves that incorporates scientific knowledge of the shape of the relationship and use multiple imputation within the QTL mapping framework.
We considered sensitivity to the particular phenotype specifications and , which were calculated by integrating up to the fifth dose. There are a few reasons why alternative specifications to the original phenotypes might be desired. First, since the doses were not equally spaced, calculating area under the curve is more heavily influenced by observed Penh values at higher doses than at lower doses. Second, since there is more missing data at higher doses (Table 1), phenotype specifications based on fewer doses would be less sensitive to the imputation scheme and more subjects would be included in a complete case analysis leading to reduced bias. Finally, there is the potential philosophical issue regarding the imputation of Penh values that may not exist (described above). To address these concerns, we considered alternative phenotype specifications that are functions of Penh values at only the first three or four doses, namely and where we consider both j = 3 and j = 4.
We developed a flexible parametric model to capture the nonlinear shape of the methacholine-Penh exposure-response relationship over the five dosage levels, before and after the allergen challenge. The model also incorporates mouse-specific covariate data, namely body weight, measured three days prior to the first Penh phenotyping (Figure 1), and the year phenotyping was conducted, as both were significantly associated with phenotype values. Since Penh must be strictly greater than zero, and since scientific knowledge of the range for Penh necessitates a finite upper bound, we modeled the transformed Penh as
where Zi1 is an indicator of year (2008 or 2009), Zi2 is body weight, and K is the upper bound. The maximum Penh value that we observed in the dataset was 28.2, and so we set the upper bound to be K = 30.
In the model (3), each mouse and each pre- or post-challenge condition was allowed to have its own knot til, which may be located at any of the last four doses. Note that a knot at the last dose corresponds to the case where a knot is not needed. The slope after the knot β4(til) was allowed to depend on the location of the knot. To account for the correlation of repeated Penh measurements from the same mouse, while at the same time accounting for the heterogeneity of the mouse-specific curves before versus after the allergen challenge, we assumed a random-effect distribution on the model coefficients. Specifically, we set β3il = β3l + bi and assumed (β0i, bi)′ ~ N((β0, 0)′, Σ). We assumed the residuals to be independent with eijkl ~ N(0, ) to capture the heterogeneity of the Penh response at different dose levels before and after the allergen challenge. Since some of the observed curves were non-monotone, the term eijkl can be thought of as measurement error from the true underlying curve, which is assumed to be monotonically increasing.
We assigned prior distributions for locations of the knots til; the model coefficients β1, β2, and β4(til); the parameters for the random-effect distribution β0, β3l, and Σ; and the heterogeneity parameters (j = 1, . . . , 5, l = pre, post). Specifically, we assumed that the knot locations come from a multinomial distribution, where the prior probability that the knot is at each of the four potential dose levels is 1/4. The parameters β0, β1, and β2 were each assigned independent N(0, 106) prior distributions. For the variance parameters, we assumed σjl ~ unif(0, 100) and for Σ = var((β0i, bi)′), we assumed that σ(β0i) ~ unif(0, 100), σ(bi) ~ unif(0, 100), and cor(β0i, bi) ~ unif(0, 1), where σ(·) denotes the standard deviation.
In order to incorporate the requirement that each mouse's curve be monotone, we would require that β3il ≥ 0 and β3il + β4(til) ≥ 0. Rather than introducing this strong restriction on each curve, we elected to just have the population average curve be monotone by incorporating the restrictions β3l ≥ 0 and β3l + β4(t) ≥ 0, (j = 1, . . . , 5, and l = pre, post) on the prior distributions for these parameters. Thus we specified the prior distributions as
where I(·) is an indicator function.
The model was used to impute M = 10 complete datasets from the posterior predictive distribution of Yijkl. Justification for the choice of M is provided in section 6.1. In particular, the mth complete dataset was obtained as follows. First randomly select a posterior sample s. Then, for each value of Yijkl that was missing in the original dataset, simulate a sample . Finally, set .
We first review a common approach for QTL mapping when there is complete data, and then we outline our methodology to integrate multiple sets of imputed data into the QTL mapping algorithm. We also consider some more routine methods for handling missing data that may be compared with the proposed multiple imputation approach.
Consider the case of no missing data. Let Yi be the phenotype summary for the ith mouse, i = 1, . . . , I. Let Zi be a vector of covariate data for mouse i and let Z be the matrix of covariate data having rows Z′i. Further, let Xp be the matrix of data for marker p (p = 1, . . . , P), where each row represents a mouse and each column (g = 1, . . . ,G) is the corresponding haplotype probability. In particular, a row Xip of the matrix Xp represents the vector of haplotype probabilities of mouse i having each of the eight CC parental haplotypes (see Section 2.1 and Aylor et al. (2011) for a description of haplotype inference methods). The goal of QTL mapping is to ascertain for which markers there is a difference in the phenotype value across the different genotypes. To do this, one conducts a hypothesis test for each marker and then computes a p-value that accounts for multiple testing of the P different markers.
To test the null hypothesis that there is no difference in the mean phenotype value across strains for marker p, we used an additive genetic model (described in Aylor et al. (2011)),
Note that G = 2 corresponds to the common scenario in which only two genotype classes are under consideration. The null hypothesis H0 : βgp = 0 for g = 1, . . . , G − 1 may be tested using an appropriate test statistic (e.g. likelihood ratio test, score test, or Wald test). For the remainder of this section, we will consider the multivariate Wald statistic, as this is the statistic that will be generalized in the analysis that takes into account multiple imputation. Denote the desired test statistic for the pth marker by . Thus, for the Wald test of the null hypothesis, we compute
where βp = (β1p, . . . , βG−1,p) and Σp is the estimated variance of the estimated parameter p. A p-value may be computed from the appropriate asymptotic distribution (in this case ) or from a permutation distribution. Since a permutation test may be readily adapted to account for the testing of multiple markers, it is a practical choice.
A permutation test may be implemented as follows. For iterations r = 1, . . . , R, first permute the phenotype summaries for the I mice to obtain , keeping the design matrix Xp as well as the matrix of covariate data Z fixed. Then calculate the test statistic for the permuted summaries for each marker p. Identify the largest of the test statistics across markers at the rth iteration and denote this by . The p-value for the largest observed test statistic is then calculated as the proportion of iterations for which . P-values may similarly be obtained for the second largest observed test statistic, and so on.
For each marker p and each imputed dataset m, the genetic model (4) is fit, yielding estimates of and the variance-covariance matrix . The generalization of the Wald test statistic for the pth marker that accounts for the variability in the imputed datasets is defined by
This test statistic is the multivariate analogue to the test statistic used for hypothesis tests of a one-dimensional parameter for multiply imputed datasets. However, unlike the univariate case, finding a suitable reference distribution for the multivariate analogue (6) is not straightforward. Approximate reference distributions have been proposed that are based on making additional assumptions about the between- and within-imputation covariance matrices p and Bp, and which use an alternative estimate of the total variance Vp (for details, see Little and Rubin (2002) or Schafer (1997)). In addition to requiring additional assumptions, as with any asymptotic approximation it is also assumed that the sample size is large enough so that the approximation is reasonable.
An alternative to invoking large sample properties and applying the approximations necessary to derive an appropriate reference distribution is to use a permutation test. In addition to requiring fewer assumptions about the data, permutation testing may be conveniently employed to account for the testing of multiple genetic markers, as outlined above. The QTL mapping algorithm for complete data may be seamlessly integrated with multiple imputation by replacing the test statistic (5) used in the permutation testing procedure for complete data with from (6). In particular, we run the algorithm for R iterations as follows. For r = 1, . . . , R,
The p-value for the largest observed test statistic is then calculated as the proportion of iterations r for which , where is the maximal test statistic of the observed data. Note that the imputation model is fit just once prior to permutation testing, and that the same M imputed datasets are used throughout. In addition, as in the complete data setup, the order of the covariate data Z and marker data Xp remains unchanged throughout the procedure.
Rather than developing an application-specific imputation model and incorporating the variability of multiple imputed datasets into QTL mapping, there are several alternative approaches to handle missing phenotype data.
We first consider a complete case analysis (CCA) where we perform the QTL mapping procedure separately for the subset of mice for which could be computed and for the subset for which could be calculated. With regards to the two repetitions of the pre-allergen challenge condition phenotyping, we used the inclusive criterion for defining a complete case (section 3). Of the 162 mice that were both genotyped and phenotyped, the statistic could be calculated for 127 mice and could be calculated for 79 mice. Additionally, we considered the last observation carried forward (LOCF) method (Heyting et al., 1992). While each of the 162 mice had at least one measurement in the pre-allergen condition, there were 6 mice without any post-allergen measurements. Thus, while the phenotype summary could be computed for each mouse under the LOCF imputation scheme, could be calculated for 156 of the 162 mice. For both CCA and LOCF, QTL mapping was performed using the permutation testing method for complete data described in section 5.1.
The phenotype model from section 4 was fit using WinBUGS version 1.4.3 (Lunn et al., 2000). We ran a single chain for 50000 iterations, with a burn-in of 10000, and we kept every fifth sample, for a total of S = 8000 posterior samples. The fitted methacholine-Penh curves are shown in Figure 3. Specifically, the fitted value of Yijkl (on the non-transformed scale) was estimated as K exp(μ^ijl)/{1 + exp(ijl)} where μ^ijl is the posterior mean of μijl = β0i + β1Zi1 + β2Zi2 + β3ildj + β4(til)(dj – til)+ from model (3). Note that since μijl is the median of the transformed Penh value log{Yijkl/(K – Yijkl)}, by taking a monotone transformation, it follows that K exp(μijl)={1 + exp(μijl)} is the median of Yijkl. We see that the Penh values in the post-allergen challenge condition are generally larger than in the pre-condition across the five doses of methacholine, as expected based on the change in lung function due to allergen challenge. However, this trend is not true for all mice; as shown in Figure 3, for example, values of the dose-response curve in the pre-condition exceed values in the post-condition for mouse 138. Additionally, these plots show that the variability in the methacholine-Penh curves increases at larger doses of methacholine.
To assess goodness-of-fit of the imputation model, we examined the posterior predictive intervals (PPI) of the methacholine-Penh dose-response curves. In particular, for each mouse i, each dose j, and each challenge condition l, we generated 8000 samples from the posterior predictive distribution of Yijkl using the retained MCMC samples, from which we calculated the 2.5th and 97.5th percentiles. We found that for the pre-condition, out of the 284 observed Penh values at the first dose, 97.2% were contained within the respective posterior-predictive interval; of the 284 observed Penh values at the second dose, 97.2% were within the PPI; at the third dose 97.9% of the 281 Penh values were within the PPI; of the 269 values at the fourth dose, 98.1% fell within the PPI; and of the 198 values at the fifth dose, 98.0% were contained in the PPI. For the post-condition, out of the 156 observed Penh values at the first dose, 98.7% were contained within the respective posterior-predictive interval; of the 156 observed Penh values at the second dose, 94.9% were within the PPI; 98.0% of the 150 Penh values at the third dose were within the PPI; of the 125 values at the fourth dose, all fell within the PPI; and of the 81 values at the last dose, 98.7% were within the PPI.
As described in section 4.2, M = 10 complete datasets were imputed. Figure 4 shows, for a sample of three mice, the fitted curves from the model and the corresponding imputed curves obtained from the model. We then applied the phenotype summary map from equations (1) and (2) to obtain for each mouse i and each complete dataset m (where Yi denotes either or ). As justification for the adequacy of M = 10 imputations, we applied the theory developed by Rubin (1987) to estimate the relative efficiency of an estimate based on M imputations to one based on an infinite number of imputations. We found that for the analysis with , the relative efficiency of multiple imputation with M = 10 datasets for estimating βp was ≥ 99% for each of the P markers; for the relative efficiency was ≥ 97% for each of the P markers.
Comparing the distribution of the simulated from the posterior predictive distribution of Yi based on model (3) to the phenotype summaries calculated from mice with complete dose-response data shows that these distributions differ (Figure 5). In particular, the distribution of the imputed phenotype summaries has higher variability and wider tails than the observed phenotype summary values for both and . This can be explained by the fact that mice with missing Penh values tend to be those with larger Penh values at the higher doses, since phenotype summaries will only be large in absolute value if at least one of the pre- or post-allergen challenge curves has high Penh values.
We implemented QTL mapping using the methods described in section 5 for the phenotype summaries and , j = 3, 4, 5. For the QTL mapping model (4), the covariates were Zi = (Zi1, Zi2) where Zi1, Zi2 were the covariates used in the imputation model (section 4).
We first describe results from our primary analysis of incorporating multiple imputation within the QTL mapping framework through permutation testing (section 5.2) for the phenotype summaries and , and then we compare these results to results based on using CCA and LOCF as well as results based on the alternative phenotype specifications. Figure 6 shows the − log10(p-values) at each marker along the genome, for each of the different approaches and for each phenotype specification, that do not account for multiple testing. The p-values of the maximal observed test statistic for each approach, adjusted for multiple comparison, are included in the legend in the top right of each plot (Figure 6).
Using equation (6)x, we calculated the observed test statistics (p = 1, . . . , 1380) that account for the multiple imputation for each marker p. Permutation testing was based on R = 10000 permutations. We first examine the p-values that do not adjust for the fact that 1380 hypothesis tests (corresponding to the 1380 markers) were performed. For , there are two regions for which the p-value is less than 0.05, at chromosomal positions 95.7–109.0 megabases (Mb) and at 109.3–112.3 Mb, which consists of 124 and 24 consecutive markers, respectively; there is also a single marker having an unadjusted p-value below 0.01. The smallest p-value of 0.009 occurs for the marker with the maximal test statistic at 98.4 Mb. When we accounted for multiple comparison in the permutation test, the p-value for the largest observed test statistic was 0.17.
For , four regions of the genome were identified as having consecutive markers with unadjusted p-values less than 0.05, at 9.5–9.8 Mb, 76.2–79.8 Mb, 82.0–88.9 Mb, and 91.2–92.0 Mb, which consisted of 6, 36, 56, and 9 markers, respectively. The largest test statistic was (p unadjusted = 0.018) at 87.4 Mb. When we accounted for multiple comparison in the permutation test, the p-value for this largest observed test statistic was 0.29.
For the alternative approaches of complete case analysis (CCA), last observation carried forward (LOCF) imputation, and sensitivity analyses based on calculating the phenotypes using the first three or first four doses, we repeated the QTL mapping procedure.
We first observe that, consistent with the fact that there is much less missing data for calculating and than for calculating and , the discrepancy in results across the CCA, LOCF, and MI approaches is reduced for phenotype specifications based on fewer doses (Figure 6). The p-values adjusted for multiple comparison suggest that there may be a QTL for between chromosomal positions 95 Mb and 113 Mb. For a particular specification of (e.g. calculated by integrating up to the third, fourth, or fifth dose), the location of the QTL does not vary much across the approach used for handling missing data. However, using one of the more naive approaches (CCA or LOCF) may lead to overstating the statistical significance, since p-values are lower under both CCA and LOCF than under MI for and . For the phenotype , given the high adjusted p-values for the largest test statistic, and the inconsistency in the location of the largest effect across the three phenotype specifications, there is little evidence of a QTL. However, the CCA approach does identify a QTL at 34.3 Mb for , demonstrating that restricting to complete cases introduces bias that may yield false positives.
In this paper we propose a framework for handling missing phenotype data in QTL mapping experiments through multiple imputation (MI), which we then apply to a study of genetic markers for allergic airway disease in mice. To conduct MI we develop a novel phenotype model describing the methacholine-Penh dose-response relation. We also compare the permutation method for QTL mapping with multiply-imputed datasets to two less sophisticated, but more commonly used, approaches for dealing with missing data. Moreover, we perform sensitivity analysis based on the phenotype summary specifications to assess the robustness of our conclusions to the degree of missingness present and to address a philosophical objection to the imputation of Penh values at doses where those values may not exist. This study focused on chromosome 8 as a "proof of concept," in order to illustrate the proposed methodology for integrating MI with QTL mapping. In the future this approach will be applied to conduct QTL mapping for each of the remaining chromosomes.
The expectation-maximization algorithm (Dempster et al., 1977) and hidden Markov models have been used for dealing with missing genotype data in QTL mapping experiments (Broman and Sen, 2009). However, though some previous genetic studies have investigated approaches for handling missing phenotype data (Ding and Laird, 2009; Fridley and Andrade, 2008; Fridley et al., 2003; Xing et al., 2003), these types of approaches have not been widely adopted by the QTL mapping community. Generally, the experiment has either been conducted so that missing data are considered to be minimal and analyses ignore the problem, or ad hoc methods such as LOCF have been used without justification. However, these approaches may yield questionable results. Restricting to the subset of subjects (e.g. mice) for which there is complete data has the potential to lead to biased results if the missingness mechanism is not missing completely at random. On the other hand, though a single imputation approach precludes bias induced by a complete case analysis, it may introduce other biases if the imputation at the larger doses underestimates the values of the true curve. Further, by treating imputed values as observed data, single imputation may understate the variability underlying the imputed values which may lead to incorrect inference (Rubin, 1987).
In this study the range of Penh values observed (~1–28) greatly exceeds those published in the literature (maximum values ~10). This suggests that the genotypic diversity of the Collaborative Cross (CC) produces remarkable phenotypic diversity. In our case, the diversity of breathing effort phenotypes (Penh) was so high that the protocol had to be curtailed to prevent undue stress to the mice. This resulted in missing data, necessitating the phenotype imputation approach we have developed. Our results indicate that on chromosome 8 there is a potential QTL for baseline Penh when integrating up to 12 mg/ml methacholine (dose 4), between chromosomal positions 95 Mb and 113 Mb. The identified region contains 203 genes, including 195 that are protein-coding genes and several that are microRNA genes. This QTL is near a QTL previously reported by Camateros et al. (2010), lending support that it may be real. Validating this QTL and identifying the causal variants within this region will require additional experimentation as well as bioinformatic analyses incorporating complementary data (e.g. gene expression data, SNP data, as described in Aylor et al. (2011)).
It is worth noting that the experiment we conducted was limited by the availability of a relatively small set of CC lines (n = 162). When the CC lines are fully developed, more than 300 will be available, thereby providing a considerable increase in sample size as compared to the current study. At that time, biological replicates will also be available. Experimental noise or measurement variation can be reduced using multiple replicates, and therefore power will also increase.
The imputation model we developed flexibly captures the nonlinear shape of the methacholine-Penh dose-response curves, and achieves a good fit to the data. We additionally assessed sensitivity to the specification of the phenotype imputation model (3) and prior distributions, finding that the fitted curves were not highly affected and that the location of the potential QTL remained consistent across specifications. In comparing the proposed methods to two simpler approaches for handling missing phenotype data, our results demonstrate the potential pitfalls of using complete case analysis (CCA) in the presence of substantial missing data when the data are not missing completely at random. We found that while the CCA method implied the presence of a QTL for the phenotype , imputation approaches did not yield consistent results. Similarly, failing to account for the uncertainty in imputing a single dataset, as in LOCF, may lead to anti-conservative inference as suggested by the results for and .
The approach we described for conjoining MI with QTL mapping is easily generalizable to different study designs and QTL mapping analyses. For example, QTLs for diabetes-related traits are an area of considerable interest (Clee and Attie, 2007). Glucose tolerance tests are routinely performed in these studies, yet these measurements are inherently limited by an upper bound of the assay of ~400–600 mg/dL (Jarvis et al., 2005; Pawlak et al., 2004). When such out of range values are observed, investigators typically invoke simple approaches to deal with the missing data, potentially leading to false positive or false negative results. In this situation, an imputation model suitable to the application may be developed, and then multiple imputation may be incorporated into QTL mapping using the methodology we have described. Additionally, while this paper focused a particular QTL mapping analysis, the approach for integrating multiple imputation with QTL mapping is not limited to the chosen analysis. For example, rather than testing the statistical significance of the largest observed test statistic max( ), one could easily adapt the permutation testing procedure to assess the significance of the largest series of consecutive markers having test statistic above a particular threshold.
We considered a two-stage analysis where, in the first stage we developed a model for the methacholine-Penh dose-response curves and performed multiple imputation, and in the second stage we performed QTL mapping to assess the association between a collection of genetic markers and the mouse-specific phenotype summaries. To incorporate multiple imputation within the QTL mapping analysis, we applied the combining rules developed by Rubin (1987) to obtain a test statistic that accounts for the variability of the imputed datasets. An alternative method is the "fuzzy p-values" approach of Thompson and Geyer (2007). Adapting this approach to our application would be an important direction of future work. A possible limitation of applying a two-stage analysis as we have done is that, given the hypothesis that genotype and phenotype are associated, it could be argued that the phenotype model for multiple imputation should include available genotype information. However, including the totality of genotype data would yield an ill-posed problem, with more predictors than responses. One solution would be to develop an overarching model that jointly describes both phenotype data (observed and unobserved) and genotype data, which would be used for QTL mapping.
Another direction of future work is to explore models for nonignorable missingness, as it is likely the assumption that the unobserved phenotype data are missing at random (MAR) is too simplistic. Since the decision to proceed to a higher dose was based on whether the mouse was observed to be struggling at the beginning of the phenotyping of that dose, the assumption that the data are MAR may not be plausible. This could be addressed using mixture models (Little, 1994) with models similar to (3) specified for each missing data pattern as given in Table 1. To implement this, one would need to assume that many of the parameters are the same across patterns. Sensitivity analysis could be conducted by varying the slope after the last observed dose (which is not identifiable from the observed data within a pattern).
There are several possible extensions of the methods developed here. Rather than consider a summary statistic (such as area under the curve) for the methacholine-Penh curves, one might conduct QTL mapping directly on the dose-response curves. This could be done either through a longitudinal analogue of (4) where the multiple Penh measurements Yij for mouse i at dose dj (j = 1, . . . , J) are modeled, or by adopting a functional data analysis framework (Ramsay and Silverman, 2005) to consider the entire mouse-specific curve Yi(d) as a functional outcome in the QTL mapping model. Functional mapping methods have been developed for other applications (Ma et al., 2002; Yang et al., 2009b), and approaches for incorporating multiple imputation within functional mapping could be examined in future work. To our knowledge the degree to which missing data differentially impacts QTL mapping when the target is a scalar summary (e.g. or ) versus repeated measurements (Yij) versus a continuous function (Yi(d)) has not been explored. Additionally, while our comparison of inference under the proposed multiple imputation approach to inference under CCA or LOCF is illustrative of the potential problems that may arise by using these more naive approaches, future work might conduct simulation studies to quantify the loss of power and bias that may result.
In previous QTL mapping experiments, we have seen little mention of approaches for dealing with missing phenotype data when the phenotype summary of interest is a complex biological parameter that is a function of missing data. While clearly it is preferable to design and conduct experiments in order to minimize the frequency of missing data, its occurrence is often unavoidable in practice. Consequently, developing statistical analyses that use as much information as possible, that limit potential biases, and that have high power to detect a QTL when one exists is an important task. The methods described in this paper, though developed to study airway disease in mice, are broadly applicable to QTL mapping experiments in the presence of missing phenotype data.
Jennifer F. Bobb, Johns Hopkins Bloomberg School of Public Health.
Daniel O. Scharfstein, Johns Hopkins Bloomberg School of Public Health.
Michael J. Daniels, University of Florida.
Francis S. Collins, National Human Genome Research Institute, National Institutes of Health.
Samir Kelada, National Human Genome Research Institute, National Institutes of Health.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
… CN Sharma child molesting saga
In an interesting twist to the saga involving television station owner, Chandra Narine Sharma, relatives of the girls involved are reporting that they are being offered a whopping $10M to discontinue proceedings that could lead to charges.
Sharma is being accused of sexually molesting a 13-year-old girl and impregnating her older sister when she was 14 years old.
But according to the aunt of who brought the story to light, someone who she claims is a relative of Sharma, has been approaching her family with the huge cash offers.
The first offer was for five million dollars but this was doubled the following day.
The woman told media operatives in an interview that she was at work on Saturday when she received a telephone call from home, informing her that an offer of $5M was proposed.
"Yesterday (Sunday) it raise to $10M," the woman said.
She said that she did not meet the person making the offer but from the description given, she is certain that the man is a relative of Chandra Narine Sharma.
"The person say that we could take the money and finish the story and we must take the girl to Berbice," the woman added.
Sharma in an interview with Kaieteur News had claimed that the woman was no relative of the girl and that she was only out to extort money from him.
But the woman dismissed the claim pointing out that her father had introduced the teenager as his daughter, but because of the age difference, the girl usually refers to her as 'Aunty'.
"I own this child as my sister," the woman related. According to the woman, the teenager approached her on Easter Sunday and begged her to take her kite flying.
The woman said that she agreed and took the teenager kite flying on Easter Monday.
It was then that the child revealed the shocking details of her alleged abuse at the hands of Sharma.
The woman said that the child was reluctant to return home after the Easter outing.
Following the teenager's revelations, the woman reported the matter to the Ministry of Human Services, and an immediate investigation was launched.
"I don't need the money. The child is in the custody of the Ministry of Human Services. I don't know who the person is offering the money because nobody came to me," the woman stated.


text:
Frequently Asked Questions
What is it exactly that you do? **
Simply put, we help clients get rid of negative listings from Google, Yahoo & Bing. Negative listings can be a result of losing a case in court, questionable business ethics, marital problems, conviction etc.
Chances are, if you search up your (or your company's name) and the results you see are less than pleasing, we can help you fix it.
Our aim is to get the negative entries for your or your business name suppressed so they are hidden from public's eye.
What makes you so different? **
Well, we are totally different for a lot of reasons.
Firstly, this company is managed by a well-known reputation management consultant who has helped over 37 clients get rid of negative listings.
Reputation Enhancer is a small company with a virtual office so we have lower overhead, enabling us to provide cost-effective solutions to people in dire need of these services.
Also, all our services are extremely personalized and I can easily be reached over the phone or via email.
Finally, our reputation management methods simply work and when you contact us, we would be happy to provide you proof for this.
How soon can I see results, generally speaking? **
Each client is different & depending on where your negative listings are showing up, it might take less or more effort on our part to get rid of them.
Generally speaking though, you should start seeing results within 3-4 months. Some clients have been able to get rid of all bad entries on Page 1 of Google within 3 months.
Who are your most common clients? **
Well, it depends. Generally speaking, our clients comprise of celebrities, medical and health professionals, small business owners or people who are under litigation or servicing sentences.
Disclaimer: Results may vary **