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Meteorological Monitoring Companies in Wisconsin
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Telkonet Inc.
based in Waukesha, WISCONSIN (USA)
Telkonet, Inc. offers EcoSmart energy management technology as well as EthoStream high-speed Internet access solutions. Both product lines satisfy Telkonet`s commitment to product innovation and technological expertise, and have been installed in several hundred thousand rooms worldwide. Telkonet`s EcoSmart products, along with the EcoCentral management platform, reduce ...
In-Place Machining Company
based in Milwaukee, WISCONSIN (USA)
In-Place Machining Company designs and develops specialized machine tool systems, and provides precision field machining services as well as optical and laser alignment and measurement to a wide range of industries including Nuclear, Fossil, Hydro and Wind Power Generation; Steel & Aluminum Production; Marine; Pulp & Paper Manufacturing; Mining & Cement Manufacturing; ...
Ben Meadows
based in Janesville, WISCONSIN (USA)
Ben Meadows Company provides quality environmental products and supplies for the outdoor professional. From soil augers to dissolved oxygen meters... everything you need for soil, water and atmospheric testing, sampling and monitoring is included in our environmental section. Ben Meadows Company also offers a wide selection of products for drum and tank sampling plus a ...
Rice Lake Weighing Systems
based in Rice Lake, WISCONSIN (USA)
Rice Lake Weighing Systems is an international leader in the manufacture and distribution of weight-related products and process-control equipment. Within this ISO 9001 registered company, progressive new technologies merge with the wisdom of industry experience and create the future of weighing through cutting-edge research and design. Our roots are planted firmly in the ...
Engineered Fiberglass Composites Inc
based in New Lisbon, WISCONSIN (USA)
High quality fiberglass composite buildings designed for all-weather, year-round protection of your equipment and/or instrumentation. The modular buildings offer corrosion resistant materials of construction for an extra long life. Modular-type construction enables flexibility and variation in sizing and design. These light weight, easy to install buildings are insulated ...
DAF Corporation
based in Kaukauna, WISCONSIN (USA)
DAF Corporation has years of experience and expertise to recommend, manufacture, rebuild and install wastewater treatment equipment. We can provide services such as process consultation and conduct wastewater characterization and pilot equipment studies for our customers.
ChemIndustrial Systems, Inc.
based in Cedarburg, WISCONSIN (USA)
ChemIndustrial Systems, Inc. was founded in 1992 by a group of engineers and other professionals with decades of experience in process design and liquid handling industries. The ChemIndustrial group rapidly expanded coverage of the process, irrigation, chemical and liquid waste treatment industries, meeting customer requirements through: Development and application of ...
based in Cedarburg, WISCONSIN (USA)
Energenecs provides application engineering, equipment, and field services for water and wastewatertreatment and control systems. We serve municipal and industrial customers in Midwestern USA. Energenecs, with numerous registered Professional Engineers on its staff, is certified by the State of Wisconsin Department of Regulations & Licensing to practice engineering in ...
RTP Environmental Associates, Inc.
Office in WISCONSIN (USA)
RTP Environmental Associates, Inc. provides professional environmental consulting services to meet client needs for today and for the future. We develop timely and cost effective approaches for permit application preparation, training, modeling analyses, ambient monitoring needs, as well as testing and analytical services to satisfy all regulatory requirements.
WEM Automation LLC
based in New Berlin, WISCONSIN (USA)
WEM is recognized around the world as the premier provider of innovative control and data integration solutions. WEM Automation LLC is recognized around the world as the leader in designing, installing and servicing process automation systems. Our experience enables us to provide cost effective solutions for the smallest of operations up to the largest full plant control ...
Extech Instruments
Distributor in WISCONSIN (USA)
Extech Instruments is based in Nashua, New Hampshire USA, just north-west of Boston. Recognized for the breadth and depth of its Test & Measurement offering and strong distribution network, Extech was acquired by FLIR Systems in November 2007. Extech Instruments' high-value, handheld test instruments and FLIR-branded infrared thermal imager are sold through a network of ...
Strand Associates, Inc
based in Madison, WISCONSIN (USA)
Specialized services in the areas of civil, environmental engineering, and science. As a firm that specializes, we concentrate all our resources – time, personnel, and money – on select areas. This has enabled us to build a reputation for quality engineering and science that extends throughout the country. Our firm, established in 1946, has now grown to become a thriving ...
Onset Computer Corporation
Distributor in Janesville, WISCONSIN (USA)
Onset's mission is to be a global leader in data logger and monitoring solutions used to measure, record, and manage data for improving the environment and preserving the quality of temperature-sensitive products. We are committed to a culture of continuous improvement, innovation, and world-class customer support. Our award-winning HOBO data logger and weather station ...
Badger Meter, Inc.
based in Milwaukee, WISCONSIN (USA)
Badger Meter helps you measure what matters today, protecting precious resources for tomorrow's generation. You can depend on our industry-leading flow measurement solutions to help you optimize your operations, leading to a better bottom line and a better world. We help you leave a mark to be proud of. Badger Meter offers an end-to-end solution that helps water utilities ...


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Hourly - Intermediate ($$) - Est. Time: Less than 1 month, 10-30 hrs/week - PostedSkills: Building Estimation Data Entry Xactimate
You will be given a scope of work and need to translate that into Xactimate. You must be able to send an .esx file after the job is complete. Most of the work will consist of taking estimate 1 and inserting into estimate 2.

subset: commoncrawl


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The jacket and pants didn't match in color or fabric type. I liked the "OLD" Track/Training suits from the 1970-80's. They zipped in the legs so one could get the pants on or off without having to remove your shoes. Also the fabric was like a satinine outside and had a really comfortable and soft inner fabric. They were easy to keep clean and maintain. I was the Athletic Trainer ATC for 7 high schools in the Des Moines Unified High School District in Des Moines, IA; and I also worked at Drake University in Des Moines, IA; Iowa State in Ames, IA; and University of Iowa in Iowa City, IA. A Anyway, I wish you would manufacture the old suits again as they were fantastic. I had a couple of suits for each school in their colors, about 20 in all. ... read more
I wanted something that was fitting so I can wear it by itself or on top of something. This is really nice! It's very thin, so it's not exactly a good "warm" jacket, but more of like a track jacket, which is what it is. The zipper pockets are nice as well- they hold a lot of things! I like this jacket overall, and would recommend it! ... read more
These shorts have a big white band at the bottom, one that didn't look as prominent online. I wanted shorts that were more plain, and the online photo didn't make it seem they were decorated so elaborately. I returned them. ... read more
I really like this shirt. It's perfect! It's if the highest quality and is actually light! It's not a heav y shirt, especially around the neckline. There is nothing negative I can write about this product. Absolutely nothing! Nothing, ... read more
my problem with the product is how bad the product was I threw them in the washer and the threads started to come out. I thought it was just that prayer that I got so I ordered another pair and they were the exact same way. They look nice they fit good but I'm a professional fighter and I trained in them and they fell apart. ... read more
This jacket is really nice, and I loved it when I first received it, but it runs a bit long and slightly large. I am about 6'2 medium frame (broad shoulders slim waist) and a small fits me well. Excellent style and fit - liked it so much ordered another one. ... read more
Perfect jacket for all purposes. It's super comfortable and looks awesome. The collar isn't too high either, so you can easily zip it up all the way and not have it bugging your chin. It's very light weight and has cooling ventilation in high wear areas so do not purchase if you are looking for warmth. It is a light weight cooling warm up jacket. I wear it to the gym and to the field and even out on the town because it looks great. Like I said, it's my new favorite jacket. It does run a tad bit big but not big enough to order a size up - it's a little loose not tight fitting. ... read more
They're snug, but keep the bits from bouncing too much. No butt-crack-oriented sweat marks (come off it; it happens to everyone and you know it); and for those of us who refuse to wear a silly looking iPod armband, the pockets zip well; so you needn't worry about your keys/phone/ipod/whatever bouncing out. Also, they're not bad on a day where it's cold out. Above all, they look good and fit comfortably. I'd recommend them. ... read more
Great pants. Bad sizing. I have the Condivo 14 and tiro 15 and they're great. I'm 6ft 0in and I wear a medium, which is great fitting, but the waist for the Condivo 16 is tighter and feels straining. It doesn't stretch like the previous editions. I tried going up one size but the legs are too big and the waist is good which isn't good. ... read more
I got 2 pairs for my son. He wears them to soccer practice and around the house all the time. They look great, the color is vivid, the material moves and wear well. All around awesome shorts. Add the Drydye environmental friendly dye process, makes you feel as good about buying them as you do wearing them. ... read more
I purchased this jacket for my teenage daughter, to match the pants I got for her over Christmas. She absolutely loves it. Has worn it many times already and she tells me she gets many compliments. The feel is great and the fit it's perfect. She's about 5'3" and 130 lbs, so I purchased the medium size, which fits her perfectly. ... read more
The quality on this jersey is great. It fits true to size. However, it does run just a little long. If you're a medium, I suggest to get a small. (Go a size under your general size). I love the overall design. I could not ask for more at all. I'm completely satisfied. ... read more
This jersey gets a lot of attention. People love the sleek look of the black and white colors, and are drawn to the dragon graphics. From the chest to waist of the jersey the fit tapers, providing a tailored look. The sharp shoulder seams are square when your arms at your sides, but puckers when you raise them. Some disappointments: the RM crest is plastic and lacks in quality to other jersey I own, I really pretty upset that this jersey didn't come with Champions League patch. ... read more
Nice and light. Love the fit so much that I am tempted to also buy the black, as well. The logo on the front is worth every penny! It's one size fits all, so women will need to adjust the velcro on the back. Very well made. ... read more


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Today, I was at a party with my crush. The collar on his shirt was sticking up so I fixed it for him. He gave me a hug and said, "Aww you're so good to me. You're like my mother. You can be my college mother." I got mother-zoned. FML
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Young Adult Epic Fantasy - Book 3Dragon Ore is book three of The Dawning of Power Young Adult Epic Fantasy trilogy. Clinging to life, Catrin Volker struggles to regain her strength as her foes go in search of even greater power. Ancient enemies threaten and forgotten alliances emerge in the exciting conclusion of The Dawning of Power trilogy.
$5.99Feral is book two of The Balance of Power Young Adult Epic Fantasy trilogy. Reeling from the loss of the Regent dragons and her son to the black armies, Catrin Volker can take no more. Armed with power never before seen on Godsland, she goes in search of her son and her enemies. May the gods have mercy on those who stand in her way.
$5.99Regal is book six of The World of Godsland Young Adult Epic Fantasy series and is the much anticipated conclusion of The Balance of Power trilogy. Going in search of solace and something to give his life meaning, Sinjin Volker will find wonders he would never have imagined and dangers from his deepest fears. Armed with only artifacts he's powerless to use, Sinjin must change the world by first
$5.99Dragonhold's secrets come at a steep price. No one knew just how many mysteries the ancient mountain keep held. None could have guessed what they would find so far underground, locked away from the world. Many would wish they had never found it. For others, long awaited retribution is at hand. Book Eight of the Godsland Series The World of Godsland fantasy series in order: The Dawning of Power
$0.99The stars have gone... Long dormant relics bask in the light of a hundred comets, restoring magics of a previous age. Objects inert for eons become dangers and tools no one can ignore. The use of these artifacts will put everything at risk while at the same time providing the greatest opportunity for human advancement in history. No matter what happens to the people of Godsland, the dragons have
$0.99Regent is book one of The Balance of Power trilogy and book four of the Godsland Young Adult Epic Fantasy Series. In the years since the end of The Herald War, the world has become complacent despite Catrin Volker's warnings. When the darkness of her visions comes to pass, no one is prepared, and the fate of humanity will rest in her hands.
Young Adult Epic Fantasy - Book 2Inherited Danger is book two of The Dawning of Power Young Adult Epic Fantasy trilogy. Catrin leaves her homeland behind as she goes in search of knowledge and peace, unaware that she will face the greatest evil her world has ever known.
Young Adult Epic Fantasy - Book 1Call of the Herald is book one of The Dawning of Power young adult epic fantasy trilogy. Echoes of the ancients' power are distant memories, tattered and faded by the passage of eons, but that is about to change. A new dawn has arrived. Latent abilities, harbored in mankind's deepest fibers, wait to be unleashed. Ancient evils awaken, and old fears ignite the fires of war.
$4.99The world of Arkaria is a dangerous place, filled with dragons, titans, goblins and other dangers. Those who live in this world are faced with two choices: live an ordinary life or become an adventurer and seek the extraordinary. When their allied settlement in Emerald Fields is attacked by the mighty titans, the guild of Sanctuary is drawn into a centuries-old conflict between the races of
Rise of the Blood Masters: Book Five of the Dragon Stone SagaAudience: Young Adult Fantasy Series Bestsellers! Over 100,000 copies sold! The brave dragon rider, Tallin Arai, continues his fight against the Balborites, a cult of religious fanatics and deadly mageborn assassins that have been wreaking havoc across the continent. In the east, the dwarf kingdom crumbles into chaos. The dwarf clans go deeper into civil war, while an even greater danger rises
$3.99The second volume of the epic fantasy saga that began with The Hawk and His Boy takes us back to the story of the thief Jute. The emissaries of the Darkness have infiltrated the city of Hearne in search of him. Desperate to escape, the boy flees the city and heads into the wilderness of the north. But the ghosts of the past have other plans for him and, soon, Jute and his friends must choose


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TearawayShared publicly -
Dear MediaMolecule! I need your help. My wife is pregnant and my main target right now is to help her be happy and have a good way to spend her free time. I bought a bluray disk with your amazing game "Tearaway Unfolded". But it is an US market version, I suppose. Because it has only English language there. We are living in Ukraine and my PS4 account has the same region. The main problem is that the English level of my wife is poor and it's making hard to play. Could you give us Russian language, please? She is loving your game very much. My e-mail is "email@example.com" and PSN account is "vanya lugovskiy" (Ukraine, Sumy). Please help us. I'm waiting for your answer soon Thank you!
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The Talbots are evacuating their home amidst a zombie apocalypse. Mankind is on the edge of extinction as a new dominant, mindless opponent scours the landscape in search of food, which just so happens to be noninfected humans. This book follows the journey of Michael Talbot; his wife Tracy; and their three kids, Nicole, Justin, and Travis. Accompanying them are Brendon, Nicole's fiancé and former Wal-Mart door greeter, and Tommy, who may be more than he seems. Together they struggle against a ruthless, relentless enemy that has singled them out above all others.
The Talbots have escaped Little Turtle, but to what end? For, on the run, they find themselves encountering a far vaster evil than the one that has already beset them. As they travel across the war-torn countryside they soon learn that there are more than just zombies to be fearful of: With law and order a long-distant memory, some humans have decided to take any and all matters into their own hands. Can the Talbots come through unscathed, or will they suffer the fate of so many countless millions before them?
It's not just brains versus brain-eaters anymore. And the stakes may be higher than merely life and death, with eternal souls on the line.
©2010 Mark Tufo (P)2012 Tantor
"Mark Tufo has written the first zombie series that kept me up reading all night." (John Ramsey Miller, author of The Last Family)
Accountant addicted to Audiobooks. I was never a reader until I started my Audible Subscription.
"Zombie Story that now has Vampires?"
Never read the print version, but Sean Runnette does a nice job with Mike's sarcastic humor!
Mountain Man by Keith C. Blackmore is really good. It is narrated by R.C. Bray
He does a great job with the sarcasm on Mike and I really like Tommy's voice he does.
Little of each! Some Humor and some sadness as some key characters have died.
I will be buying book 3 soon.
"Tufo does it again!!"
Tufo continues to bring on the raw survival story of the Talbots and company..It is hard to rate this one book against all of the others once I have gone through all of the Fallout books.. This story helps set the hook in for this zombie series.
Runnette's performance is spectacular..he has an uncanny way of portraying Mike and bringing out Marine Corps memes. Talbot reminds me of a few of my buddies from the Corps.
indy music guy
"Favorite Apocalypse Series of All-Time"
Perfection. PERFECT narrator for this series!
I cannot say enough great things about this series! All of Tufo's Zombie Fallout series are equal parts of excellence...add them all together and get my favorite series - BAR NONE - of all-time! If you, even remotely, enjoy Apocalypse stories and/or like Zombies or Supernatural stories...even a little, you should REALLY enjoy this! I've recommended this series to more than 20 readers and listeners so far...the overwhelming majority of them react with a statement like, "Dude, why didn't you tell me to start this on a Friday night! I can't stop reading/listening...I haven't slept in days...but man, I love the part where..."
The ones who don't respond that way to me are the ones who "didn't have time to start it yet!"
"I tried... And I failed."
I like some humor in the novels that I most enjoy but it's not always a necessary element. this series just takes it a bit too far. It's not slapstick (as has been said in other reviews) but more of a constant commentary on day to day life by a smartass. That's fine but in this book it takes it to the point of hindering the story. The tale itself i find interesting but you have to dig thru countless comments, asides and silly stories to get to the meat.Some reviewers have fawned over the "McDonald's" tale which baffles me. I understand that this is a work of fiction but to have the main character go on and on about an obviously over-the-top unbelievable experience at Mickey D's was the last straw for me. I couldn't sit thru it and I just couldn't find it funny... Mainly because it just doesn't ring true as anything other than a play on the stereotypes of the Indian, the teenage girl, the bad mother, the roach infested fast food restaurant, the defeated manager, blah, blah, blah... I came to read an apocalypse novel but at this point in the book i finally came to realize I was getting about 25% zombie novel and 75% rant. Pass.
Stay on the subject and not so much on the internal ramblings of the main character. But, hey, I'm obviously in the minority here so who am I to judge the writer's technique?
The reader was great and really makes the main character come alive. He IS Michael Talbot. It's a very good reading of the tale.
The McDonald's scene would hit the cutting room floor faster than you could blink your eye.
Meh. If you want a comedy along the lines of Archie bunker, this is for you! If you want a Zombie Apocalypse novel, there are much better books available.
Love me some creepy, scarey, suspenseful, thrilling reading - when discovered I could hear some of my favorites read TO me...Audible had me!
"Another great book in the "Fallout" series"
While this is my first "audiobook" review, I have read the entire Zombie Fallout series, and listened to the first of this series....well, first.
It is so difficult to listen to one of your favorites, after "hearing" the characters in your head while reading them all originally - but Sean Runnette did a great job, portraying the personality of the main characters, and really bringing me back to the enjoyment of when I was reading them.
Mark Tufo is a fav writer of mine, but I do think you have to have a particular sense of humor to always "get" full effect of his stories (at least the ones I have read). Another reason why I appreciated this narrator so much - I felt like he "got it:.
There isn't a book, or series of books that compares to the Zombie Fallout series, specifically A Plague Upon Your Family. I have purchased so many "zombie" books trying to capture this style again, and though there are many other great zombie books - this is certainly unique.
Another writer I enjoy for his quirky humor in his zombie books, is Chris Philbrook, and his "Adrians Undead Diary" series is fabulous - but very different from the "Fallout" series.
Sean Runnette did a wonderful job narrating this book. When I heard him reading the book, it took me no longer then a few paragraphs to imagine his voice as the one I could hear when I was reading them myself. He did an excellent job with the Michael Talbot character, and that can't be easy - Michael Talbot is an endearing, quirky, weird, funny, neurotic guy - I have no idea how Mr. Runnette pulled him off!
If I didn't have a 6 year old, and a 3 year old around listing for swear words over the speaker, or tugging at my sleeves for attention when I had my ear buds on - I most definitely would have listened to it in one seating. No doubt!
Also enjoyed by Mark Tufo:
Lycan Fallout: Rise Of The Werewolf
The Book Of Riley A Zombie Tale ebook set 1-4
Zombie Fallout Pt 1 - Pt 7
"good story line, but...."
I don't like his reading but I guess maybe he is growing on me. I thought he did pretty a good job. He just doesn't seem to fit the sardonic character he is reading for.
Ok, I did like the book and the story line. Even though the story line is picking up into a more fantasy style of zombie book where there are Higher forms of ancient undead, that was kind of the lead-in from book one anyway so you kindo of knew where they were going with it. It should not have been a bad surprise if you liked the first book. But what was my issue with the book well...
****BIGTIME RANT*****How can zombie's not freeze in -10 degree weather? I mean serious?!?!! Warm blooded, out door, animals FREEZE in that temperature if they do not find shelter. Going north should be the safest place on earth for them during the winter time. The author totatally messed up the time frame. Maybe have them cuaght off guard in the spring or something, but being attacked in one of the coldest part of the country at the harshest part of the winter season? C'mon dude you really need research at least a tiny bit.
For instance, I don't know anything about guns so when i read the books i take it for granted that the author has researched a little to get his facts straight about how the guns work, type, and range, etc...But not having a zombie freeze in -10 degree weather makes me feel the author didn't research crap and maybe he just made up the guns and they don't exist and such. Well Thats my rant but i did like the story line. But dude, DO SOME RESEARCH!!!
gluten-free and happy
"Second book even better"
This book is fun, scary, exciting and well performed.
Warm bodies, but scarier!
I listened to the first book in the series. I really enjoy his performance. My favorite reader of all the books I've listened to.
I have recommended this audible book to several others. Very enjoyable if you like this genre.
looking forward to the next book!
Came on here for Star Wars books, now its time to get to history, politics, and religion.
"Going a bit further to the left"
The narrator for the writing was perfect. The book's natural speech and the narrator's nonchalant way of reading them meet well in this book that has some dumb, crazy, and attempted frightening points. I would place this narrator for this book up with Mark Thompson narrating a Star Wars book. The book isn't great, especially since it's going into vampire territory now, but the narrator puts it up there.
The son being used to speak for the vampire.
Mike...everyone is Mike, I guess.
"More Gross-Out, Less Humor"
I loved the first book in this series - it has a great gross-out factor tempered with a lot of humor, and a dead-pan delivery (ha ha) that reminds me of Ray Romano (love him).
The second book is much darker, dramatically ups the gross-out factor, and has way less humor. Plus, it introduces vampires, which feel very out of place in this story, and I had a hard time reconciling it. I don't think I'll continue with the series; I wish I'd stopped at book 1. (Still read book 1 - it's an awesome ride!)
I enjoy reading fantasy, science fiction, and horror the most. To improve, I read about language, psychology, spirituality, and art. I read about computer science and business for professional reasons.
"Great book, obnoxious name"
You may wonder why I'd give a book with such a horrific name five stars. Such a nightmarish idea in the second person is sure to rile anyone up. But I don't judge books by their cover, and this was was an incredible horror book.
The main character and his family continue to develop in the midst of a horrific setting, with a plot line based on family group survival. The plague creates mindless herds of violent masses seeking to eat the flesh of the family. Society appears to have lost the battle, and these large groups of subhumans are able to roam the country in groups attacking who they please.
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text: My wife and I reserved a room at this location, strictly due to price. What a mistake!! The rooms are in poor condition - the "new" flooring is already cracking, with seam separation. Our toilet back-up and overflowed - the mgr's response was to give me a broken plunger!! I had to push for a new room, which they grudgingly... 更多


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Mickelson on course for maiden US Open triumph but Donald and Rose remain in the hunt, while Woods and Rory fall by the wayside
Phil Mickelson has the chance to write a true American fairytale on Sunday after taking the lead going into the final round of the 113th United States Open at storied Merion.
Five times a runner-up in his national championship but never a winner, the four-time major champion begins with a one stroke advantage and will try to finally claim victory on his 43rd birthday, and on Father's Day to boot.
The doting dad, of course, made a now celebrated 6,000 mile trip in his private jet to California earlier in the week to attend his 13 year old daughter Amanda's graduation from middle school. He only arrived back hours before his opening round.
In charge: Phil Mickelson goesi nto the final round at Merion with a one-shot lead over the chasing pack
Fist pump: Luke Donald celebrates on a great day for him at the US Open and collects his ball (below)
Sitting pretty: Justin Rose smiles after making a birdie on the 10th hole and hits his tee shot on the 13th (below)
It won't be easy of course, and particularly given the calibre of players all queuing up to play the role of villain. They include the English pair, Justin Rose and Luke Donald.
If truth be told, it looked as if
they would have the lead for themselves, only to become two more victims
of the brutal pair of closing holes. Neither did much wrong, but Rose
walked off with a bogey-bogey finish and poor Donald ended up feeling
still worse after a bogey and double bogey.
For both it was cruel reward after 16 holes of skilful graft and grind.
Still, the outcome is that both are only two off the lead after rounds of 71. But it was Mickelson who showed the class of a four-time major champion. He finished birdie, bogey to make up three shots on his playing partner Donald and two on Rose to finish as the only man under par after 54 holes.
Mickelson shot a level par 70 to stand on one under par. Then come Hunter Mahan, Charl Schwartzel and Steve Stricker who, at 46, is attempting to become the oldest winner of this event. One stroke further back are Rose and Donald.
'I feel a bit hard done by after 17
and 18 but that's the US Open and I will have my shot at winning in the
final round,' said Rose.
As for Mickelson, he said: 'It was a lot of fun out there and I just can't wait to get back out there and play the final round.'
Ian Poulter was the third Englishman who began the day firmly in the mix but his chances faded over the brutal closing stretch. What part did having to set the alarm at 4.45am play in him losing concentration and making mistakes at the end of a marathon day? It was a surely a factor given how well Poulter was playing for his first 14 holes. He was one over par for the tournament and, at that point, just three off the pace.
Shocked: Mickelson reacts after his ball hit the metal pin and did not fall, on the 14th green
Run into trouble: Tiger Woods did not have much fun during the third round of the US Open
Frustrating: Rory McIlroy never got going during the third round and endured a difficult day
Plotting: Charl Schwartzel lines up his putt on the 14th green and hits from a bunker (below)
But a wildly hooked drive at the 15th landed on the road known as Golf Close and that might have closed his chances as well. He ended up with an ugly double bogey six and followed it with another bogey at the 16th. A final bogey at the 18th meant a bitterly disappointing 73 and, at five over, Poulter might be too far back. What an unlucky break it was not being able to finish his second round on Friday night owing to all the rain delays. Never underestimate the part good fortune plays in winning majors.
'It's a shame because I feel like I played lovely out there,' said the 37-year-old.
What of the two best players in the
world? For the third day in a row Tiger Woods and Rory McIlroy were
paired together and for the third day in a row they failed to inspire
one another.
Neither has a shot at winning on Sunday,
and their reaction at the finish, as they embraced and consoled each
other on their futile efforts, said it all.
Woods shot a dismal 76 to finish on nine over par while McIlroy was only one better. 'I just couldn't get the right pace on the greens all day,' Woods lamented. His five year wait to win a major goes on.
One hundred years on from a 20-year-old American amateur shocking the world and winning the US Open, 19-year-old Michael Kim was threatening to follow in Francis Ouimet's footsteps when he moved to within one of the lead with four holes to play. He looked like he was having the time of his life.
Mixed fortunes: Garcia ran into trouble but later hit a string of birdies to help himself out
Strong showing: Rickie Fowler barely put a foot wrong on a stunning day at the US Open
Actually, three dropped shots in the last four holes as reality set in didn' t reduce the smile by much. The American amateur, playing in his first US Open, finished with a 71 and will start the final round in the top 10.
Meanwhile, good rounds from forgotten man Paul Casey (71) and Lee Westwood (69) means there are no fewer than five Englishmen in the top 16.
For two, of course, there is the chance of glory today and to emulate the glorious achievement of Tony Jacklin, the last English winner of this event way back in 1970. Not to mention the chance of becoming the first Englishman to win a major of any kind since Sir Nick Faldo won his third Masters in 1996.
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Tab Books!
Tab Book Announcement! Hello everyone, we have some news regarding the Tab Books. Unfortunately, our projected manufacturing costs were unrealistic and the manufacturing of the physical books is far too... Read more
Ghost Ship Octavius sailing through the East Coast in JUNE!
World Entertainment Inc. and Ghost Ship Octavius are proud to announce our "Sick and Frozen Tour" East Coast Dates! Supporting us on this trek will be our brothers in metal,... Read more
Ghost Ship Octavius sailing through the East Coast in JUNE!
All hands on deck Ghost Ship Crew, GSO is coming through the East Coast in June and we want to meet you! We hope to see all of our east... Read more
TAB BOOK Completion
Hello GSO Crew!
For those of you that have ordered the TAB BOOK, it is getting proofread by me right now and will put together immediately! I will be sending... Read more
Ghost Ship Octavius Debut Live Shows
Hello Everybody!
The time has come and we're finally going to start bringing GSO on the road! We have a list of shows that we've booked with infinity concerts, and... Read more
Vinyl Coming!
Hello everybody!
The Vinyl has been made and we are waiting for the shipment to arrive so that we can send them out to you!!(if you ordered vinyl). So... Read more
So today I sent out the remaining Kickstarter backer packages! Keep an eye out for your package in the next few days! Europe, you know the drill, customs likes to... Read more
Monday & Tues March 24th! Check the list!
Hey guys, some packages went out yesterday and some today, to get more orders packaged faster here is a list of went out. If you don't see your name yet... Read more
More GSO ON THE WAY!
Hello peeps! So today a bunch more packs are going into the mail today so I will send you an email at some point today/tonight if your package was on... Read more
Mailing and Downloads
Hello and happy Saturday! Just a quick update on mailing and digi-downloads. I'm working as fast as I can to get everything out and into your hands. Packages are going... Read more
Digi coming!!
Hey guys so we're working hard to get everyones packages out the door so in the meantime, the packages that have not left the building will be getting a download... Read more
My Fellow Ghost...Shipians!..
Hello ladies and Germs! Well, our first backer pledged on April 28, 2014 and here we are almost a year later bringing you our first labor of love, the Ghost... Read more
Manufacturing Time!!!!
Update!!! We are very pleased to announce that the completed debut Ghost Ship Octavius album and it's final Artwork will be sent off for manufacturing this coming Monday morning!!! It... Read more
Mixes nearly complete! Off to manufacturing next!
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I realized that it has been a few weeks since the last update so I wanted to take a minute to keep everybody up to date! This... Read more
Instructions on how to change your address for shipping if needed!
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I have been getting several messages asking what to do if you have moved and you need to update your shipping address with us. Here is a link... Read more
The T-shirts are in!
Hey GSO Crew!
Update time! I picked up the T-shirts and they are AWESOME! :). Album is being mixed and everything else is getting sorted. This is so cool...can't wait... Read more
Album Tracking Complete! Entering Final Production Phase!
Hey Ghost Ship Crew, a little update for you all! Guitar solos are done! All of the orchestral sections are being added today and then it's re-amping time with the... Read more
GSO Rhythm Guitars Studio Update!
Hello GSO Crew Members and Supporters!
We are currently in the Recording Studio finishing up all of the rhythm guitars with Producer Aaron Smith in Seattle, WA. Everything is sounding... Read more
Recording Begins July 6th!
In just under one week, on July 6th, Van enters the studio in New Jersey with producer Aaron Smith to officially begin the recording of the debut Ghost Ship Octavius... Read more
'Get Drawn Into The Album Art' - Last Week Price Reduction
We have reduced the price of the 'Get Drawn Into The Album Art' perk and 'Become a Ghostly Character On The Album' perk for our last week to help us... Read more
A LA CARTE MENU ADDED for ADD-ON to Existing Rewards!
Greetings GSO Backers! We have decided to add an "A La Carte" Menu where you can add on individual items to your order! We have taken the time to speak... Read more
Paypal Option Added for Backers without Credit Cards!
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NEW BACKER REWARD ADDED- Caparison C2 Horus Guitar Played on the Album!!!
Get the Caparison Horus C2 Guitar that Matt will use to play solos on the upcoming GSO debut album! This guitar is not yet available to the public and will... Read more
FULL GUEST ARTIST LIST ANNOUNCED for Debut Album!!!
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We have an eclectic bunch of extremely talented lead guitarists... Read more
New Song THE MILLS OF THE GODS on Soundcloud!
We set up a SoundCloud Page where you can rock out to 'Alive', 'Saturn & Skies' and a BRAND NEW SONG called 'The Mills of the Gods'!!! Check it out,... Read more
Refer 2 Friends and get THE FROZEN TICKET!!!
IMPORTANT ANNOUNCEMENT to all CURRENT and FUTURE BACKERS!!!
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JEFF LOOMIS & PER NILSSON to Play Guest Guitar Solos on GSO Debut Album!!!
BREAKING NEWS!!! We are very pleased to announce that our very good friends Jeff Loomis and Per Nilsson will be making GUEST SOLO appearances on our debut album! This is... Read more
GSO Interview Podacst on KISW Metalshop with NEW SONG!!!
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Here it is folks, as promised, a NEW ANIMATED LYRIC VIDEO for 'ALIVE'! We passed the $5K mark so here is your gift! Video animations and layout by Adōn Fanion... Read more
Adōn Explains Creation of Limited Edition GSO T-shirt Art!
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26 Spring Crafts for Kids
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Use pink crepe paper and pearls to create a clever spring bouquet, displayed on an egg ornament from Plaid Enterprises. This spring ornament makes a fun host(ess) or thank you gift.
- 4019 - FolkArt ® Enamels™ - Fresh Foliage, 2 oz.
- FolkArt Enamels - Azure Blue (item #4037)
- 50mm clear glass egg ornament Sunshine #MS-DP-2614-27
- Script Liner
- Pink Crepe Paper
- 5-White Pearls
- White Satin Ribbon
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Squirt Azure Blue inside the egg. Gently shake the egg until the inside is coated in paint. Let dry.
Fold the crepe paper in half and cut small flower shapes out. Put the end of the paintbrush handle into the center of the doubled flowers and wrap around to give some shape.
Hot glue the two flowers together and glue a pearl in the center.
Hot glue 3 of the flowers onto the egg in a bouquet shape. Using the script liner and Fresh Foliage paint the stems. Let dry.
- Glue the last two flowers onto the egg and finish painting the stems. Let dry. Glue the white bow to the ornament as shown.
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text: Randgold Resources (GOLD) Is Rebounding In Early Trade 4/9/2012 9:49 AM ET Randgold Resources (GOLD) has gapped open sharply higher Monday morning and has continued to gain ground in early trade. Shares are now up 7.74 at $89.18 on above average volume.Randgold Resources has rebounded off of support this morning, following over a one-month decline. by RTT Staff WriterFor comments and feedback: email@example.com News Biotech Stocks Facing FDA Decision In October Top Ten Highest Paid CEOs World's Top Ten Oil Producers, Ranked


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"The education that I received at Lake Superior State University has opened many doors for me. I have worked with Firefighters and Paramedics for over thirty years as a co-worker, educator, and student. I have been very blessed in my life to have these opportunities. I truly believe that education is the cornerstone of everyone's career and LSSU provides the best.
My future goals are to become the Director of a Fire Science program and follow in the footsteps of the great educators that I have had the privilege of learning from. This will not be easy, but I believe that these Professors have prepared me to be successful. My achievements will be a direct result of the knowledge that they have bestowed upon me. It will then be my obligation to share this knowledge with others. "
Keith M. Campbell '07
Fire and hazardous materials response coordinator for the U.S. Navy based in Bahrain
Lake Superior State University staff members Kathy Danielkiewicz and Heather Ferguson, and faculty member Sherilyn Duesing have been chosen by their co-workers and students as recipients of employees of the year and distinguished teaching awards.
The awards are presented annually to a member of each of the university's employee groups: faculty, Steelworkers union (food service, physical plant and clerical workers) and administrative/professional staff.
The award for Duesing, an assistant professor in the School of Mathematics and Computer Science, was announced during commencement ceremonies today on campus. The honors for Danielkiewicz, secretary for the School of Business, Economics and Legal Assistant Studies, and Ferguson, director of Advising and Retention, were announced during the annual LSSU Retirement and Service Awards program earlier in the semester.
The Distinguished Teaching Award has been presented to LSSU faculty since 1971. Students, faculty and staff submit nominations that are considered by a committee of students at the top of their classes and past recipients of the award. Bruce Harger Ph.D., LSSU vice president of academic affairs and provost, quoted from an essay and nomination for Duesing written by an education student.
"As a senior, I have had the pleasure of continuing to be taught by this professor, although I'm not sure if the word 'taught' captures the essence of my experience. I felt that I was being mentored. It was as if I was in a room with my colleagues and we were being guided by excellence; epitomized by the values and attributes of the professor…Henry Adams tells us that 'A teacher affects eternity' and I for one can assure you that Prof. Duesing has affected eternity through my life and the lives that I will touch as a teacher."
Duesing has taught math at LSSU since 1984, when she was hired as an adjunct professor. She became a full time instructor in 1993 and was promoted to assistant professor in 1998.
She has helped develop course modules for pre-algebra and introductory algebra courses at LSSU and has assisted with the teacher education program as a mathematics liaison at Brimley Elementary School. She has served on several university committees, including the developmental education committee, in which she was chair, and the advising committee. She was also vice president of the LSSU Faculty Association.
Duesing received an associate's degree from North Central Michigan College and a bachelor's degree in education, with a mathematics major, from Central Michigan University. In 1998, she earned a master's degree in mathematics education at Northern Michigan University, where she was also selected as the outstanding graduating graduate student in NMU's Dept. of Mathematics and Science Education.
She has long been interested in mathematics education and has taught at the middle school and high school level before coming to teach at LSSU. Outside of teaching, she enjoys quilting, sewing, skiing and hiking. She and her husband Paul, an engineering professor at LSSU and also a recipient of the Distinguished Teaching Award, have three children, Alice, Sarah and Peter.
Majoring in Nursing...
Katie Farnsworth
Big Rapid, MI
Small campus and small class sizes. [ more ]


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- The NCIC follows the Alternative Dispute Resolution provision as per the constitution to address hate speech offences, as was in the case against Chirau Ali Mwakwere, where he apologized to those he had offended, and the case subsequently dismissed/dropped. Is such a move counter-effective? Does it embolden hate speech propagation, once it is known to be an option to 'resolve' a hate speech offence case?
- What lessons have Kenyan media learnt from the outcome of the 2007 election, the ensuing violence, and the role they have been perceived to have played?What is the media's responsibility in triggering or curbing hate speech, beyond the news (and money) value that some of the triggers eg politicians' names and ethnic group mentions on headlines hold? As Aisha Ali said, media and the public have a symbiotic relationship, because they (media) need the public goodwill to do their job. Case-in-point: media needed the people's support when the'government came after them', and therefore they have a responsibility to citizens over and above making money.
- Does referring to Members of Parliament as 'MaVultures' (vultures were defended vehemently) and more recently as 'MPigs' constitute hate speech? Kenyan law does not recognize politicians as a group that can be targeted through such speech, but neither does it recognize gender, sexual preference, religion nor any other group category; it only mentions 'ethnic hatred': hatred againsta group of persons defined by reference to colour,race, nationality (including citizenship) or ethnic ornational origins. (NCI Act, s.13(3) )
- We asked Boniface Mwangi whether he considers such terms to constitute hate speech; he referred to cultural folklore, Animal Farm in particular, justifying that it's about the traits that MPs showcase that relate to the animal analogies. "You can't hide behind culture or hate speech when someone describes you for who you are," he said. Boniface, however, clarified that his has not been a campaign preaching hate; instead it's about advocating for behavioural change by asking our leaders-elect to 'stop behaving like greedy pigs'. But what is the difference when such analogies are used to refer to people based on other group belongings? What makes it 'acceptable' to compare people who constitute a political group, to animals, but not ethnic groups or any other groupings to which people belong? ? Is it more acceptable to target and criticize those in power, vs a minority who may not be able to fight back?
- There's a normalization of discrimination (especially along gender lines) that has erstwhile existed offline and now manifests online. There also exists a culture of intolerance in Kenya complicating integration and cohesiveness, that needs to be addressed.
- Is there a thin line between hate speech and cyberbullying, and where/how do we draw it? (The example of the online attacks that renowned radio presenter, Caroline Mutoko, has come under were referenced. Has she come under constant attack because she's prominent, or because she is a woman who happens to be prominent? Here, it was explained that women are the largest 'minority group' , and that attacks against Caroline easily cross from cyberbullying to hate speech as a result).
- We also got a first hand account from Dennis Itumbi who has previously been summoned by NCIC over 'alleged hate speech on his Facebook account'. He passionately narrated his version of events, highlighting some of the shortcomings in the NCIC's operating mechanisms. "You cannot conduct cybercrime investigations using analog methods," he said, while referring to NCIC's flaws in digital evidence recovery.
- How do we achieve cohesion in Kenya, over and above curbing hate speech, which is a symptom of a bigger issue? "We cannot keep coexisting when we are litigating," an NCIC official reminded us.
- Way forward?Online community policing, the responsibility of online communities towards a cohesive society was discussed. This has already been observed in the Kenyan online space, and we have highlighted some examples in our Umati reports.


text: Pregnancy Pregnancy Tips Don't Miss This Why Some Women Can't Get Pregnant How to Tell if Your Baby's Vision Is Normal Sign Up for Our Women's Health Newsletter Thanks for signing up! Sign up for more FREE Everyday Health newsletters. Oops! Please enter a valid email address Submit We respect your privacy. Pregnancy Basics Make a Belly Cast First Trimester Face Facts Workout Don'ts What to Wear? Make a Belly Book Second Trimester A Better Back Makeup 101 Make It Well-Done Expectant Mom Mantra Third Trimester Neck Know-How How Dry I Am Stubble Trouble Getting Pregnant Know Your Family History Childbirth and Beyond Five Beauty Boosters Decoding Your Baby's Cries top


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|Dharambir K. Sanghera* and Piers R. Blackett
|University of Oklahoma Health Sciences Center, Oklahoma City, USA
|Corresponding Author :
|Dharambir K. Sanghera, Ph.D
FSB, FAHA, Associate Professor of Pediatrics
Department of Pediatrics, Section of Genetics
University of Oklahoma Health Sciences Center
Oklahoma City, OK 73104, USA
Tel: 405- 271-6026
Fax: 405-271-6027
E-mail: firstname.lastname@example.com
|Received May 17, 2012; Accepted June 19, 2012; Published June 23, 2012
|Citation: Sanghera DK, Blackett PR (2012) Type 2 Diabetes Genetics: Beyond GWAS. J Diabetes Metab 3:198 doi: 10.4172/2155-6156.1000198
|Copyright: © 2012 Sanghera DK. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
|Related article at
Pubmed Scholar Google
Visit for more related articles at Journal of Diabetes & Metabolism
|The global epidemic of type 2 diabetes (T2D) is a major public health problem of 21st century and the fifth leading cause of death worldwide [1]. The disease is also a leading cause of morbidity and contributes to development of premature coronary heart disease (CHD), stroke, peripheral vascular disease, renal failure, and amputation. According to latest statistics released by the International Diabetes Federation, the number of people living with diabetes is expected to rise from 366 million in 2011 to 552 million by 2030; 80% of these people with diabetes will live in developing countries (http://www.idf.org/diabetesatlas/5e/the-global-burden). According to these predictions, in three leading countries with diabetes populations USA, India, and China the approximate estimate of 23.7, 61.3 and 90 million people with diabetes in US, India, and China in 2011 will increase to 29.6, 101.2, 129.7 million by 2030 (Figure 1). The global health expenditure on diabetes is expected to increase from 376 billion dollars in 2010 to 490 billion in 2030 [2].
|Substantial evidence suggests that T2D is a multifactorial disease with a strong genetic component. High concordance rate obtained in monozygotic twins (96%) supports a substantial contribution of genetic factors to T2D [3-7]. Furthermore, 40% of first-degree relatives of T2D patients develop diabetes as compared to 6% in the general population [8]. Segregation analysis also points to the polygenic nature of T2D, in addition to the existence of a major genetic component [9,10]. The general estimates of heritability (h2) of T2D is 0.49 and the relative recurrence risk for a sib of an affected person (λs) to develop T2D is 3.5 [11,12]. However, the findings and the results of recent genome-wide association studies (GWAS) have significantly underestimated these heritability estimates and presents a challenge for ongoing and future investigation.
|Clinical Phenotype of T2D
|Unlike simple characterization of type 1 diabetes (T1D) which is primarily due to autoimmune mediated destruction of pancreatic beta cells resulting in insulin deficiency, the pathogenesis of T2D is more complex and remains a matter of debate. Hyperglycemia in T2D is a consequence of complex interplay between insulin resistance (sensitivity) and abnormal insulin secretion [13]. It is initially characterized by compensatory insulin secretion associated with insulin resistance. However, the β-cell's response is inadequate for the increased demand during progressive resistance to insulin-mediated glucose disposal, eventually resulting in β-cell failure and overt diabetes. There is also accumulating evidence that the β-cell is adversely influenced by influx of fatty acids [14] and cholesterol, which accumulates and exerts toxic effects when efflux by HDL is limited [15].
|The progression of insulin resistance often associated with obesity and leading to mild glucose intolerance preceding an increase in glu cose levels above defined thresholds for diabetes, has made it difficult to define the true phenotype, possibly because of early involvement of multiple tissues such as muscle, fat and pancreatic β-cells. Furthermore, the insulin resistant state, usually preceding diabetes by several years, is associated with a cluster of co-morbidities including obesity, dyslipidemias, and elevated blood pressure [16,17], and the presence of three or more traits is currently recognized as the metabolic syndrome which is also predictive of diabetes [18].
|More than 80% of diabetic subjects are obese and these individuals typically have an android body type (upper body obesity) manifesting as an increased waist circumference [19]. Obesity, the metabolic syndrome, and T2D are becoming increasingly prevalent even in children and adolescents living in rapidly developing countries in different parts of world [20-24]. The prevalence of T2D in the US is higher among minorities and ethnic populations like African Americans, Native Americans, Hispanic Americans, Asian Americans, and Pacific Islanders than in the general population [25-28]. High prevalence is even seen in children belonging to ethnic minorities [29-32]. Although environmental factors play an important role in determining the risk of disease, overwhelming data support that genetic factors influence the disease susceptibility [33]. Lifestyle modifications (weight reduction and physical activity) delay or even prevent the development of T2D [34,35], such changes are exceedingly difficult to sustain outside the research setting and seem to have contributed little to control the diabetes epidemic [36].
|T2D remains undiagnosed for many years because hyperglycemia is usually not severe enough to provoke noticeable symptoms, unlike T1D which often presents with keto-acidosis requiring admission to hospital for correction and initiation of treatment. But hyperglycemia can cause significant pathological and functional changes, which can cause organ damage before the diagnosis of T2D is made. The long term effects of diabetes include micro-vascular and macro-vascular complications and those who have genetic susceptibility are at greater risk [37]. Micro-vascular complications are progressive development of disease in fine capillaries supplying blood to the kidneys and retina of the eye that results in blindness. Macro-vascular complications include hypertension, coronary artery disease, peripheral vascular disease, cerebral vascular disease, and hyperlipidemia. Diabetic patients also have neuropathy, which may lead to foot ulcers, amputations, sexual dysfunction, and non-healing skin wounds. Certain infections such as staphylococcal sepsis is more common in diabetics and infections of the ear, nose, throat as well as reactivation of tuberculosis associated with high rate of mortality and morbidity are more likely with poor blood glucose control. Consequently, diabetes is a disproportionably expensive disease. The economic impact of diabetes in the US is enormous, and is expected to increase from $113 billion to $336 billion [38].
|Linkage and Candidate Gene Association Studies
|Progress in identifying the genetic basis of simple Mendelian (monogenic) diseases during the past decade has been substantial. More than 3000 monogenic disorders have been successfully mapped by linkage and family based studies (http://www.ncbi.nlm.nih.gov/omim). On the other hand, complex diseases like T2D do not segregate in simple Mendelian fashion but rather are affected by multiple genetic and environmental factors. Linkage and candidate-gene focused studies were successful in identifying some rare familial forms of T2D presenting at young ages called maturity onset diabetes of young (MODY), mitochondrial diabetes and neonatal diabetes. However, linkage and association studies on the common form of T2D provided inconsistent results and failed replication in multiple populations [39]. Only PPARG, KCNJ11 and TCF7L2 were identified as established genes associated with common forms of T2D [40]. Apparently, inconsistencies across populations were due to the heterogeneity of the disease itself or its pathogenesis, incorrect candidate selection because of incomplete knowledge of molecular mechanisms, variation in study design, sample size, population-specific linkage disequilibrium, choice, analytical methods, or over-interpretation of results [40-43].
|Genome-Wide Association Studies (GWAS)
|Completion of the Human Genome Project in 2003 [44] led to subsequent advances in biomedical research. Since 2007, a new technology in the form of 'genome-wide chips' has facilitated remarkable progress in T2D genetic research with the first publication of five large GWA scans within the span of four months, showing that more than 500,000 SNP markers distributed across the genome [45-49]. This approach has been successful in locating genes for other diseases besides T2D and obesity [40] namely, type 1 diabetes [50], prostate cancer [51], rheumatoid arthritis [52], Crohns disease [53,54], and cardiovascular disease [55] and is being applied to other complex disorders. Use of this 'hypothesis-free' approach involved in GWAS has opened new areas of biology to explore as discoveries of more than seventy entirely new T2D loci clearly suggest that associations are not limited to candidate genes and by applying GWAS and re-sequencing approaches, new genes involved in disease pathogenesis can be identified [56] (Table 1).
|The number of risk alleles for complex diseases identified by GWAS since 2007 exceeded those identified in the entire preceding decade, and these studies can effectively detect multiple common variants with small effects with odds ratios (ORs) <1.2 [36,39,57] but offers limited power to detect rare variants with stronger effects [58]. However, GWAS have the best chance of detecting and identifying additional genes for T2D and are being pursued in different population groups, to provide a more realistic 'genetic risk landscape' of the disease [59], and account for variation in population-specific environmental interactions. Notably, most immediate advances from GWA studies are not disease prediction or prevention, rather these studies are providing a better understanding of the disease pathophysiology since further studies can lead to defining the role of the newly identified genes in key metabolic pathways involved in pathogenesis. Also GWAS, together with targeted resequencing of the identified loci allows recovery of a more complete inventory of the sequence variation leading to identification of novel metabolic pathways and proteins as potential targets for interventions [57]. Since substantial numbers, maybe thousands, of genes with small effects are operative in human disease and can be identified by this approach, their combined effect may be substantial [56]. Furthermore, gene-gene interaction may not only be additive but effects of a mutation can be more than doubled when compounding mutations are present. The phenomenon known as epistasis was first shown to occur in a rodent knockout model in which the insulin receptor when combined with IRS-1 knockout resulted in 40% increase in diabetes onset compared to isolated knock-outs [60]. The phenomenon could contribute to effect size and in part account for the low heritability accounted for by classic GWAS. It follows that multi locus association testing could be effective in the presence of epistasis. With this in mind the Welcome Trust Case Control Consortium used twolocus tests of association and detected epistatic signaling of association at 79 SNP pairs [61]. However the large number of possible associations in GWAS requires filtration strategies based on knowledge of the biochemistry and known interacting pathways [62]. However, several models for analysis have been proposed and consensus on the optimal methodology has not been reached [63].
|The common strategy used for elucidating the inherited components of complex disease is based on common disease-common variant hypothesis [64,65]. The hypothesis has held true in the case of putative causal variants in APOE, including APOE ε4, association with Alzheimer's disease, interleukin 23 receptor association with Crohns disease (the at-risk allele has a frequency 93% in the general population), and PPARG2 association with T2D (at-risk (pro) allele frequency ~85% in general population). The assumption with the use of common variants is that either they will identify the association with the common disease, or a quantitative trait (QT) directly, or indirectly being in linkage disequilibrium (LD) with a functional variant. Most of the T2D loci identified using GWAS approach are common variants with small effects. The first GWAS in T2D was published by a French group of investigators who identified a zinc transporter and member of solute carrier family SLC30A8 and HHEX along with confirming the association of TCF7L2 and KCNJ11 with T2D [49]. Three additional GWAS in their jointly published findings confirmed TCF7L2, KCNJ11, PPARG, SLC30A8, and HHEX. They also discovered novel T2D loci called CDKAL1 and a variant near CDKN2A-B [47,48,66]. These findings were also simultaneously confirmed by GWAS performed on an Icelandic sample by the deCODE researchers [67]. Association of FTO with T2D was also first discovered by GWAS for obesity [OR for T2D being 1.27, p < 10-8] [68] and was later confirmed in replication studies [46].
|Additionally, a joint meta-analysis of three previously conducted T2D GWA scans by the Diabetes Genetics Replication And Meta-analysis (DIAGRAM) consortium [69] has detected an additional six novel loci with strong evidence of association; JAZF1, CDC123, TSPAN8, THADA, ADAMTS9, and NOTCH2 [69] in 10,128 individuals using 2.2 million SNPs. Also, another European study reported a strong association of an intronic SNP (rs560887) in G6PC2 with fasting plasma glucose levels by screening 392,935 SNPs in 654 non-T2D participants using GWAS [70]. Association of G6PC2 with fasting plasma glucose levels was replicated in the FUSION study [71]. Further, a variant near MTNR1B has been identified to increase FBG levels in two separate GWA studies [72,73]. Association of MTNR1B variants (rs10830963 and rs1387153) with FBG or T2D in our Punjabi cohort from India has not been confirmed. Instead a less common variant on 5' UTR (rs1374645) of MTNR1B revealed a strong association with low FBG levels in normoglycemic individuals with low BMI (<25 kg/m2). Furthermore, our data showed strong interaction of this variant with BMI with respect to FBG levels [74]. Most recently, a meta-analysis was performed on 21 GWAS comprising 46,186 participants to identify loci influencing glycemic traits. They performed follow-up studies on 25 loci in up to 76,558 additional subjects and identified nine loci associated with fasting glucose and HOMA-B (ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (IGF1) in European populations [75]. Another study analyzed combined GWAS data from 8,130 individuals with T2D and 38,987 controls of European descent, and in follow-up meta-analysis found signals in a further 34,412 cases and 59,925 controls and identified 12 new T2D associations including new signals at KCNQ1, KLF14, DUSP9, CENTD2, HMGA2, HNF1A and PRC1 [76].
|Genetic Diversity among Populations
|The majority of GWAS so far (96%) have been predominantly performed in Caucasians [77,78]. Given the existence of a strong genetic diversity among the world communities due to biological traits, cultural histories, languages, caste system, physical appearance, food habits etc., the information from GWA findings on Caucasians should not be transferred and used to predict risk in other populations [79,80]. Even the largest and best executed GWAS is unlikely to provide a complete assessment of "the genetic risk landscape" of the disease [59]. These loci neither account for all cases of T2D, nor do they sufficiently account for the variation in quantitative sub-phenotypes of T2D. Extending the use of new methodologies such as GWA to study non-European ancestries with different mutational spectra and demographic and cultural histories is important for identifying the population-specific patterns for allele frequency and LD relationship of the susceptibility loci, and population-specific environmental factors for disease risk or protection [57]. Several GWAS in other ethnic groups are currently ongoing and will be capable of revealing novel susceptibility loci. GWAS have proven more successful in small homogenous populations like Finland, Iceland and Costa Rica, and have potential to detect regionally common variants which may be missed in outbred populations [78]. It is of interest to note that many of the common loci originally associated with diabetes in European populations have not been replicated in other non-European populations [49,67,77,81]. For example, loci associated with Crohns disease in Caucasian GWAS were not associated in Japanese [81], also the association of CDKAL1 locus with T2D was not seen in African Americans [67]. As discussed earlier, the common intron 1 FTO variant (rs9939609) was initially associated with diabetes in Europeans, and phenotypic interactions appear to be diabetogenic, but the associations were due to obesity [68,69]. However, the question has been explored in independent association and meta-analyses of South Asian populations in whom BMI and waist association with FTO is similar to that seen in Europeans, but a strong association with diabetes is only partly accounted for by BMI [82,83]. A recent large scale meta-analysis conducted on 96,551 individuals from East and South Asia confirmed that the association of rs9939609 with T2D was independent of obesity [84].
|There is now a strong consensus that a single population is not sufficient to uncover all the variants underlying complex diseases [77]. Recent GWAS studies in non-European populations have yielded intriguing new variants, for instance, the KCNQ1 signal (first noticed in Japanese) provided insight into novel etiological pathways leading to insulin resistance and T2D. Joint meta-analysis of Caucasian GWA studies later discovered a second independent signal at the KCNQ1 locus (rs231362) and an overlap between loci implicated in monogenic and multifactorial forms of diabetes at HNF1A [76]. The new Caucasian signal at KCNQ1 was later confirmed in our study on Asian Indians [85]. These findings provide new insights into the pathophysiology of this extremely complex disease by discovering previously unknown variants and often replicating the findings but not always. Association of UBE2E2 with T2D was first described in East Asians [86]. Recent larger meta-analysis using East Asian GWAS cohorts yielded eight new loci for T2D in (GLIS3, PEPD, FITM2-R3HDML-HNF4A, KCNK16, MAEA, GCC1-PAX4, PSMD6 and ZFAND3) [87]. Six novel loci for T2D (GRB14, ST6GAL1, VPS26A, HMG20A, AP3S2 and HNF4A) were first discovered in our South Asian population GWAS by Kooner et al. [88]. To date, GWAS and meta-analyses in T2D and related quatitative traits have together identified approximately 75 susceptibility variants which together explain about 10% of the observed familial aggregation (Table 1). Although these effect sizes detected so far do not reflect biological or clinical significance of these variants, they highlight a particular genomic region likely to be associated with the trait. Of all the complex traits studied by GWAS , Crohns disease, has so far had the strongest association with genetic components of risk; up to 20% being explained by variants detected by GWAS [89]. Furthermore, GWAS provided important biological insight with direct clinical relevance adding to significant understanding of the disease compared to what was known about Crohns disease five years back.
|Hidden Heritability and Rare Variant Hypothesis
|Despite these successes, results of multiple GWAS and meta-analysis studies (based on the common disease-common variant hypothesis [64,65]) show that common variants discovered in these studies explain only a small fraction with odds ratios (ORs) ranging from 1.1- 1.2 (p values <10-8) of the heritable component of T2D risk, and these studies have not pinpointed with certainty the causal variants at the associated loci [90]. Importantly, commercial arrays used in GWAS have captured much less than 60% of common SNP information in the majority of the GWAS conducted to date [91]. One approach could be to expand the sample sizes to detect more variants of moderate effects, however, the GWAS approach is usually underpowered to locate those variants with a frequency of <5% [57], and is considered unlikely to yield more information on the heritable control of disease than what has already been revealed by GWAS [92]. Moreover, association studies, based on the common disease-common variant hypothesis would have less relevance if the disease is caused by less common mutations at multiple sites and each arising on a different ancestral haplotype [93].
|Many studies have suggested the large role of rare variants in complex diseases [93,94], and that the genes carrying common variants with small effects also are likely to carry rare variants with large effects [69,90] which can only be detected by targeted resequencing [57,95]. Also, several independently and dominantly acting rare variants from different genes, together can confer a detectable relative risk for a given "common disease-rare variant" hypothesis [96]. These low frequency variants with stronger penetrance would be expected if trait alleles have undergone purifying selection [94]. Resequencing of certain candidate genes have shown that certain rare variants can be identified in individuals with extreme phenotypes [97]. These rare variants when clustered together can produce a significant genetic effect in a complex disease [56]. For instance, rare functional recessive mutations contribute to HDL and triglyceride levels [98,99], suggesting the possibility of discovering such variants in a common disease such as T2D [100]. Fearhead and colleagues [101] detected several rare functional variants in five genes among 124 subjects with multiple adenomatous polyps showing combined ORs of 2.2; p = 0.0001. Furthermore, 25% of these individuals had a single rare variant and none of them had two or more. Their findings suggested that mainly missense and possibly also nonsense, promoter, and splicing variants can collectively explain a significant portion of inherited susceptibility to colorectal adenomas. It is realistic to expect similar findings in other chronic diseases [94,96] including diabetes. It is also important to note that rare variants are often founder variants caused by genetic drift and can be population-specific and less cosmopolitan. For instance, the rare variants discovered in the APC gene for colorectal adenomas in Ashkenazi Jews were not present in Koreans [102]. These findings therefore suggest that the search for genetic variation in diabetes-susceptible populations should be carried out in multiple ethnicities in carefully chosen candidate genes, and in clearly phenotyped disease groups. Further use of whole genome rare variants from the 1000 Genomes Project (www.1000genomes.org) and T2D-GENES Consortium with data from multiple ethnic groups will substantially improve power of these studies and capture heritability associated with rare variants from specific regions or even from the other regions of the genome, especially those (deletions/insertions, duplications, inversions) that have escaped detection in GWAS [103]. Additionally, imputation from 1000 Genomes Project will allow discovery of new association signals from the missing "dark matter". It will also assist in post-GWAS fine mapping and functional characterization of known associations and enhance our ability to fine map signals of local adaptation as has been successfully achieved for refining the locus for smoking quantity at 15q25 (P = 9.4 × 10−19) using imputed data from 1000 Genomes which increased marker density to five-fold compared to HapMap2 [104].
|In addition to major gene mutations and SNPs, changes in the normal genome sequence can result in significant miscoding with disease association [105]. Submicroscopic deletions, duplications, insertions, inversions and translocations can occur in a small DNA segment and are known as copy number variations (CNV) [106]. Since methodology for CNV detection is relatively new [107], few studies have looked for CNV associations with T2D. Since copy number variation in the leptin receptor gene (LEPR) is associated with T2D [108], it is possible that genome-wide detection of CNV could improve association with T2D as shown for insulin resistance in the HyperGEN study, in which joint tests of association for SNPs and CNVs were performed resulting in detection of association in the T-cell receptor gene (TCRVB) in African Americans [109]. Furthermore, generation of CNV maps [110] supports potential for studying CNV for missing variation in T2D and possibly accounting for the inter-ethnic variation found among diabetes- prone populations [111]; however, common CNVs may be tagged by SNPs and have been indirectly explored in previous SNP-based studies [112]. Further, the event resulting in a CNV may also disrupt regional genes and their regulatory elements accounting for disease association [106].
|Role of Epigenetics in Missing Heritability
|The assumption of SNP-based GWAS has been that genomic sequence variations account for disease phenotypes, however recent developments in the field of epigenetics has shown altered expression of genes that could lead to enhanced prediction of obesity and diabetes [113]. The modifications include DNA methylation and histone modifications such as ubiquitination, adenosine diphosphate ribosylation, phosphorylation, acetylation and methylation. Since the modifications are often observed during cellular growth and differentiation, it follows that the changes may influence embryological development of cells involved in carbohydrate homeostasis such as the β-cells, adipocytes and myocytes. For example, DNA methylation occurs in the leptin gene promotor and in GLUT4 in 3T3-L1 cells [114]. The epigenetic mechanism mediate the influence of the environment on gene expression [115]. For instance, leptin plays an important role in programming of obesity-related traits [116,117], change in expression could be involved in the pathogenesis of T2D, in part supporting epidemiological findings that fetal growth and early nutrition in childhood influence adult disease incl , HNF1B (rs 4430796 uding T2D [118]. For example, increased methylation of PDX-1 results in decreased expression in pancreatic islets from patients with T2D suggesting that modification of PDX-1 plays a role in T2D pathogenesis [119]. Besides, epigenetic modifications are sensitive to change by nutrients, hormones and toxins that can operate during developmental windows according to tissue- specific and sex-specific mechanisms [120], which potentially can be reversed with treatment and prevention [121,122].
|Gene-Environment Interaction
|Evidence from the epidemiology of T2D overwhelmingly supports a strong environmental influence interacting with genetic predisposition in a synergistic fashion as has been recently reviewed [123], however current state-of-the-art methods for measuring environmental effects lack precision and can result in changes in statistical power to detect interaction [123,124]. Since lifestyle factors are important in preventing diabetes [125,126], interaction of gene variants with measures of dietary intake and exercise have been selected for studies on gene-environment interaction. For example, HNF1B (rs 4430796) was shown to interact with exercise; low levels of activity enhanced the risk of T2D in association with absence of the risk allele, but there was no protective effect of exercise when the allele was present. It follows that subgrouping by genotype may serve to enhance risk prediction while considering gene-environment interaction as has been done for exercise [127]. Also lifestyle including exercise modified the effect of a CDKN2A/B variant on 2-hour glucose levels in the Diabetes Prevention Program [128] but was not confirmed in the HERITAGE study using different measurements and phenotypes involving insulin sensitivity and β-cell function [129]. The pro12ala PPARG variant also interacts with physical activity for effect on 2-hour glucose levels [130], which was confirmed in the smaller HERITAGE study [129]. In addition, a relationship of dietary fat intake with plasma insulin and BMI differs by the pro12ala PPARG genotype [131].
|It is possible that there are genes that because of their known metabolic involvement are likely to interact with specific nutrients. For example, SLC30A8 which encodes a zinc transporter localized in secretory granules, interacted with dietary zinc to effect fasting insulin levels [132]. However, the majority of GWAS variants have not shown interaction with environmental factors for effect on diabetes or related traits. Therefore, it is likely that prospective future studies will utilize improved assessment methods to increase power and avoid false interpretation [133,134]. This could be enhanced by prioritizing variants that are most likely to have effects [135] or selective sampling according to extremes of the environmental factor could reduce the requirement for sample size [136]. These and other strategies such as meta-analysis, nested case control and genotype-based studies have been recently reviewed [123,133] and the difficulties in measuring environmental exposures have been emphasized, including the application of analyses based on logistic regression [124] and problems with instruments such as physical activity questionnaires [137]. Validated food frequency questionnaires are popular instruments for evaluation diabetes risk and are often used in conjunction with food analysis software [138,139]. Similar methodology has been adapted to assess two predominant food consumption patterns by Prudent and Western [140], and demonstrated synergistic interaction with genotype and a less healthy Western dietary pattern in determining male risk for T2D by showing that the gene-diet interaction was higher in men with a high genetic risk score determined by a gene counting method [141]. Also the effects of diet may predominate at specific developmental periods [142] suggesting that age and associated physiological changes are important as well as differences between genders. It has also been observed that homogeneity of an environmental factor such as physical activity in an Asian Indian study, may reduce ability to detect interac tion, but could be solved by subgrouping by the level of activity [143], but increased recruitment would be needed to maintain power.
|It is also speculated that several rapidly developing communities lived through centuries of famine and starvation and over the years have adapted to survival under conditions of unreliable food supply because they have "thrifty genes" which enable efficient storage of energy during periods of food deprivation [144]. The genes that once represented a survival advantage, now, in the modern environment with continuous low physical activity and dietary excess, result in obesity and T2D [145,146]. In addition, early protein deprivation, as indicated by low birth weight is considered as an important contributor to the high prevalence of T2D especially in people living in the under-developed world [147-150] possibly when followed by rapid catch-up growth in infancy and early childhood [151]. The "thrifty phenotype" hypothesis was based on data showing a positive association of low birth weight with cardiovascular disease and diabetes in adulthood [152], but the metabolic basis of gestational programming of the fetus is poorly understood. In Europeans, the risk of T2D increases with increase in birth weight whereas in Pima Indians and in South Asians T2D rates increase with both low and high birth weight [153] suggesting that the effective birth weight distribution is bimodal. Poor intra-uterine nutrition also may be associated with abnormal β-cell development and differentiation [154-156]. Therefore, it is imperative to develop a deeper understanding of genetic, epigenetic, environmental, cultural, and social factors that may interact to cause progression to T2D, and are responsible for the declining health of the fastest developing nations of the third world.
|Prospects of Translation of Genetic Findings into Clinical Practice
|As discussed in more detail in previous sections, investigations on genetic determinants of T2D have progressed substantially following increased characterization of the genome and use of GWAS, however a large portion of the heritability is unaccounted for, and many of the genes commonly found in GWAS have small effects [157-160]. Thus, state-of-the-art clinical use of genotyping is currently restricted to investigation. Also the use SNPs to predict disease onset or response to treatment is limited but can be done with some monogenic forms of diabetes [160]. Furthermore, metabolic pathways that have been responsive to treatment with pharmaceutical agents or exercise, or have been associated with disease have rarely contained genes that have been identified by large scale studies such as GWAS.
|However, pharmacogenetic studies on association with the effects of anti-diabetes drugs have provided a significant clinical link with epidemiological findings [161], and frequently used agents in the treatment of T2D such as metformin, sulfonylureas, and thiozolidinediones have been shown to have gene associations. The principal metforminassociated pathway involves adenosine-monophosphate-activated kinase (AMPK), a regulator of glucose levels [162], and therefore has potential as a drug target [163] for the control of T2D by metformin and possibly other agents. It serves as a sensor of cellular energy status via its phosphorylation and activation. Liver AMPK controls hepatic glucose production by inhibiting expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) and this action is down-regulated by the drug, metformin. Candidate genes encoding enzymes, cofactors and transcription factors involved in AMPK-associated pathways have potential for studies on therapeutic responses. Since gene variants coding for steps in metabolic pathways involving metformin's action may lead to defining treatment responses, investigators in the United Kingdom conducted a meta-analysis using the glycemic response to metformin as the phenotype. A variant at a locus containing ATM, the ataxia telangiectasia mutated gene (rs11212617) was associated with the response [164]. The importance of the variant was emphasized by showing that specific inhibition of ATM attenuated the phosphorylation and activation of AMPK in response to metformin in a rat hepatoma cell line [164]. Although the effect attributed to the variant was small, the study identified an important upstream AMPK regulator adding information on how metformin works, and in addition identified a possible mechanistic link with DNA repair and cancer prevention. Shu et al. have investigated polymorphisms in SLC22A1 which encodes organic cation transporter 1(OCT-1) which functions to facilitate absorption of metformin into hepatocytes and identified association with reduced responsiveness to metformin's effect on glucose levels during an oral glucose tolerance test [165]. Also association with HbA1c was shown with a variant in SLC47A1 encoding the multidrug and toxin extrusion protein 1 involved in the excretion of metformin into bile and urine [166]. Furthermore, the two genes both acting to increase intracellular metformin, have an interactive effect [167]. The roles of both genes were supported by the Diabetes Prevention Program investigators [168] who confirmed association of the glucose-lowering response to metformin and variants in SLC47A1, and interactions with SLC22A1 and the AMPK genes STK11, PRKAA1, and PRKAA2.
|Genes encoding the receptor and pathways for PPARγ agonists constituting the thiozolidinediones, are often required for glucose control because of their role in promoting insulin sensitivity. Of 133 PPARG variants tested in the TRIPOD study, eight showed evidence for association with response to troglitazone therapy defined as change in tolerance to intravenous glucose (IVGTT) [169] but no association with fasting glucose suggesting that the response shown in the IVGTT represents insulin resistance whereas fasting glucose is more likely to be a measure of β-cell failure.
|In studies where overt T2D has been the phenotype the majority of associated polymorphisms have encoded proteins known to be involved in β-cell metabolism; for example TCF7L2, KCNJ11 and HHEX have shown robust association [170,171]. This suggests that these genes could prove useful in predicting β-cell preservation during the course of T2D. The glucokinase gene (GCK) coding for the initial glucosesensing step in the β-cell can have activating mutations causing hypoglycemia that might provide structural and functional models leading to drug targets for treating T2D [172]. In the GoDARTs study, investigators examined the medication response of metformin and sulphonylurea based on the TCF7L2 variants mainly affecting the β-cell. The carriers of the at risk 'T' allele responded less well to sulphonylurea therapy than metformin [173]. Also it is of significant public health interest that in the Diabetes Prevention Program, lifestyle modifications were shown to reduce the risk of diabetes conferred by risk variants of TCF7L2 at rs7093146, and in placebo participants who carried the homozygous risk genotype (TT), there was 80% higher risk for developing diabetes compared to the lifestyle intervention group carrying the same risk genotypes [35]. These findings could herald significant future progress in the field of T2D pharmacogenomics, possibly leading to the development and use of agents tailored on the basis of genotype.
|Predictive Power of Genetics beyond Traditional T2D Risk Factors
|Traditional clinical assessment includes a complete history of the presenting traits including family history of the traits, and followed by laboratory studies that present either as continuous variables or dichotomous values according to a cut-point. From this basic clinical information it has been possible to derive risk scores that predict T2D such as the Finnish Diabetes Risk Score and the Diabetes Risk Calculator [174,175], which have been used to assess predictive ability using receiver operating characteristic curves or the area under the curve (AUC) as an index of discriminative accuracy. Using these scores prediction has only been considered to be modest [176], leading to the question whether genetic risk prediction might be superior. Studies have been based on genetic variants alone and in combination with environmental factors, but studies have differed in the number and choice of genes and environmental factors. However, disappointingly the genetic risk models had lower prediction capacity than the clinical environment-based models [176]. Differences in populations, BMI, age and duration of follow-up may offer some explanation [176,177], but improving genetic methodologies may be the main issue needing to be addressed.
|The escalating current and predicted world-wide prevalence of T2D underscores the need to develop effective strategy for early detection, prevention and targeted treatments. In this regard the field of genetic epidemiology has potential to progress to effective translation to patient care possibly within the next decade [178]. Current stateof- the-art approaches that may lead to this goal include defining the pathogenesis of T2D and the phasic nature of the pre-diabetes phenotype beginning in utero and progressing to overt T2D associated with β-cell failure. Thus far, genes identified by GWAS using diagnosed T2D as the phenotype have mainly been associated with β-cell failure but have only accounted for less than 5% of the heritability. Although the greater than 60% heritability appears clinically obvious, environmental interactions, biochemical DNA modifications (epigenetic), and the combined effect of rare variants and other genes present potential challenges in defining risk association. However, the contribution of GWAS over the past decade should not be minimized because of low contribution to heritability, since the genes showing association have been pointers to key metabolic pathways involved in pathogenesis and in the action of drugs used for T2D treatment. Future functional studies on model organisms or cell lines will enable functional characterization of identified risk loci and will provide better understanding of the biological mechanisms of disease development. Improved SNP- and expression-array technology, sequencing, and targeted re-sequencing are likely to enhance current understanding and progress. In addition, inclusion of "at risk" populations in the developing world is likely to improve understanding of gene-environment interaction and is more likely to lead to detection of rare variants and novel variants that associate with unique phenotypes and disease associations.
|This work was partly supported by NIH grants -R01DK082766 funded by the National Institute of Diabetes and Digestive and Kidney Diseases and NOTHG- 11-009 funded by National Genome Research Institute, USA. Superb technical assistance provided by Latonya Been in manuscript preparation is duly acknowledged.


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