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By Hank The Mechanic
By Hank The Mechanic CREATED Jan 30, 2013
KISS, this week, is celebrating 40 YEARS! MAN! I've always wondered, why or how, did KISS come to wearing makeup and crazy costumes? Well, in a recent interview, Gene Simmons explains all. Click HERE to read the interview, and see some cool videos. There's also some other cool KISS links below.
By Hank The Mechanic CREATED Jan 29, 2013
Our STATE, not the band. HA! Yeah, today, Kansas is 152 years old! The 34th state in the union, Kansas has a lot of fun trivia! Our great state has a bunch of things to do that you may have not known about! Click the links below to discover things about... KANSAS!
By Hank The Mechanic CREATED Jan 28, 2013
I came across this today, and am keeping my fingers crossed! Every year, American's DON'T claim money they have out there! It's not because they don't want it, it's because it's either not known about, address changed, or a ton of different reasons. Well, check out this story today, and see a bunch of links you can go to that will tell you if you have any unclaimed money. If you find any, SHARE! I'm the one who let ya in on this! HA! Click HERE.
By Hank The Mechanic CREATED Jan 25, 2013
So how many times has your girlfriend sent you naked pictures of herself? C'mon, we all have recieved some of these pix, and most of the time they come to us after a whole lot of begging and coaxing. It's true. Hell, I've done it. HA!! There is absolutely nothing wrong with that, because two people in love are having fun!
By Hank The Mechanic CREATED Jan 24, 2013
The wacky French people at Peugeot Citroen have developed a car that runs on freakin AIR! At high speeds, the car runs on gasoline with the small engine. Then when the car dips down to 43mph or less, it runs on AIR! They say the average driver of this car would get 141 miles A GALLON! WTF?!
By Hank The Mechanic CREATED Jan 23, 2013
I have found myself as a one-use guy so far with my iPad. I had thought my days would be filled with a wide assortment of helpful, entertaining, productive apps. Um, well, it seems to be primarialy used as a video game iPad. HA! I have asked you to send me apps that you really love, and the vast majority seems to be GAMES. I'm not complaining! I love to play, but sometimes a couple hours can go by before I get my ass up from the couch!
By Hank The Mechanic CREATED Jan 22, 2013
As you may have gathered, I am a electronics gadget junkie. I LOVE the newest, latest electronic stuff, whether it's for my mobile dj service, my personal use, or whatever. Well, now, some of these gadgets have made their way into my home. I have integrated my ipad and my iphone to my home entertainment. It's awesome. NOW, the Consumer Electronics Show has shown us new items for your home and more. From digital screen washing machines to wireless electric blenders, these new items are sweet! Click HERE for the list of the latest greatest.
By Hank The Mechanic CREATED Jan 21, 2013
Today, President Obama is inaugurated into his second term, and it's MLK day. Whether you like the President or not, we continue to grow as Americans and live in a wonderful country. We are FREE. We are becoming more integrated as a people. And the steps Martin Luther King took to spark equality shows more and more every day. Below are several links dedicated to todays events and history.
By Hank The Mechanic CREATED Jan 18, 2013
I'm not talking pocket pool either. I'm talkin' good ole' American,slate-top, green felt, manly pool! When I was in my late teens in Hutch, I thought I was a bad-ass pool player. My friends and I had our own sticks, bags, accessories, and we kicked ass. Or at least we thought. I even joined a weekly pool league. Granted, our team won, but not due to any fancy stick work from Hank.
By Hank The Mechanic CREATED Jan 17, 2013
This Friday thru Sunday, Sunday, Sunday, it's the 56th annual Starbird-devlin Car Show at Century 2! It's is always a great show that my Dad took me to when I was a kid, and now I'm taking my daughter. It is a great show that you can make a tradition out of with your family! Ticket prices are lower this year, kids are FREE, and more cars than EVER! Find out more at their website, www.starbird-devlin.com
By Hank The Mechanic CREATED Jan 16, 2013
So we sit here today sticking our collective tounges out at the Myans for the failed "end of the world" prediction. Now, just when we thought it was safe to go back into the world without fear of a catastrophe, here's another. Yeah. The "doomsday clock" lets the world know of imminent danger due to nuclear or biological weapons, climate change and other human-caused disasters. The hands of the clock have been placed at 11:55. THAT'S CLOSE! So what in the hell are scientists predicting?! Click the links for the story and other doomsday clock facts.
By Hank The Mechanic CREATED Jan 15, 2013
Ok, I'm bitchin' again. I am sick and tired of people NOT going the speed limit! It's weird! Usually, people speed, but for the past 6 months or so, people are going under. UGH! The best road/highway in Wichita? 235. I never encounter bad or slow drivers on 235. Now, 35? AHHHH! Speed up, stay left, something! And my drive home takes me down Seneca every day. From Kellogg to Pawnee, it's slow goin on a daily basis. I don't drive aggressive, but yesterday I was almost forced to. I was on my way to a live broadcast, going down Seneca, and this older lady was doing 25mph. Yeah. SENECA IS 40MPH! I was a car legnth behind her, people behind ME were honking thinking I was slow, so I simply went around her once I was able to. SHE STARTED YELLING AT ME AND SWINGING HER ARMS LIKE I HAD A PROBLEM!!!! This is CRAP! Slow drivers can not only be irritating, but can cause accidents! Please Wichita, drive the speed limit when you can.
By Hank The Mechanic CREATED Jan 10, 2013
Through tomorrow, the Consumer Electronics Show is happening in Las Vegas. What is the CES? It's the annual "unveiling" of the new gadgets for the year. Things are presented to the world at this time, and manufacturers hope that what they unveil will be the "next big thing". Missing from the show this year is Apple. They feel products get lost in the shuffle at this show, so they're gonna stick with stand-alone, away from the CES, product launches.
By Hank The Mechanic CREATED Jan 9, 2013
So I've told you about my Christmas present that was a big hit with me. If you haven't heard, cuz it's big news, Andrea bought an iPad Mini for me! IT'S BADASS! She told me last night she thought it was money well spent, but now she says she shouldn't have got it because I love IT more than HER! Ok, I'll admit, I do love the iPad Mini, but I DO love Andrea MORE. Just to clarify. HA!
By Hank The Mechanic CREATED Jan 8, 2013
Have you ever wondered where certain foods have gone? How come you don't see them in the store anymore? Had cravings for certain things you just can't find?! Maybe they have been discontinued! I found some lists of items that have been cut from your shelves. Click the links to see what they are. When you see them, you'll start craving some of em'! I did! I also added a link of McDonalds food items they tried out, but failed horribly.
By Hank The Mechanic CREATED Jan 4, 2013
By Hank The Mechanic CREATED Jan 2, 2013
Get fit? Stop smoking? Eat less crappy foods? What is your 2013 resolution? After having a stroke in late 2012, my goal is to get healthier. It's hard to do, but I have to do it. For me. For my family.
By Hank The Mechanic CREATED Dec 31, 2012
There were many highs and lows for me this year. My relationship with Andrea and my daughter, Gabby, continues to grow each day, and for that I'm thankful. 2012 also brought the bad with me having a stroke in August. I spent a week in the hospital. That was horrible. But the good news is that with every day that passes, I seem to get better. I need to lose weight, etc, etc, and 2013 will definately be better. I am also thankful for you. You are the people who make it possible for me to continue working at T95, in a job that is what I love and love doing each day. Let's have a good 2013 together!
By Hank The Mechanic CREATED Dec 21, 2012
So it appears we're still here! I haven't noticed anything unusual other than the normal stupidity around here! HA! For the latest end of the world news, click HERE.
By Hank The Mechanic CREATED Dec 20, 2012
Well, today is the eve to the end of the world! Really??? So what do you believe? Are you making preperations? Are you saying your goodbye's? Today, I'm keeping it simple with a few links from both sides of the fence. Check em' out, and see ya tomorrow!
Copyright © 2015 T95 The Rock Station
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text: Born in Detroit, attended college in Orlando, frequented Chicago, worked several years in New York and presently live in Los Angeles; I can proudly say that my portfolio is as diverse as my life – and with it is a passion is as big as my heart. I pride myself in being aware of my sensitive side and how it can be a tool that helps create a better experience for everyone around me. And it's those experiences as an contributor and a leader that have led me to win the awards I've won in collaboration with the talents of those around me.

subset: commoncrawl


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Jhene Aiko (@JheneAiko) In GQ magazine
Jhené Aiko will make an appearance in the May issue of GQ magazine.
Top 10 Dunks of the 2013-2014 NBA Season
A ook at the best slam dunks, alley-oops and high-flying action from the 2013-2014 NBA season.
Brian Williams Rapping Gin and Juice
NBC Nightly News managing editor and anchor Brian Williams raps Snoop Dogg's classic Gin and Juice.
Iamsu! (@IAMSU) Feat Wiz Khalifa (@wizkhalifa) & Berner (@berner415) – What You Bout
Iamsu! connects with Taylor Gang members Wiz Khalifa and Berner for his latest offering called What You Bout'. This will appear on Iamsu! upcoming album titled Sincerely Yours, hitting stores on May 13th.
Iggy Azalea (@IGGYAZALEA) Live On Good Morning America
Iggy Azalea made an appearance on Good Morning America to perform her single Fancy which features Charli XCX. This track is featured on Iggy debut album titled The New Classic which is in stores and also Itunes now.


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Ra 1980
Man, 45 years old from Amman, Jordan
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xTip: Sending a positive vote will save this user to your account, like a personal bookmark. Additionally, they will see this vote and may like you back, which will allow the exchange of your private contact information.
Signed up: September 20th, 2012 (1114 days ago)
Last activity: 73 days ago
Profile hits: 2,751
Subscribers: 11
Age:45 years old
Region:Al Balqa'


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NPR Newscast
4min 45sec
The latest guidelines for cholesterol-lowering drugs emphasize their use to manage patients' risk for cardiovascular disease rather than simply hitting numerical cholesterol targets.
Roy Scott/Ikon Images/Corbis
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July 15, 2015 The latest cholesterol guideline identifies more people as candidates for treatment with statins. But the decision about taking a daily pill to reduce heart disease risk is more personal than ever.
This fungus among us — baker's yeast, aka Saccharomyces cerevisiae — is useful for more than just making bread.
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May 21, 2015 Genetically, at least, not that much has changed in the billion years since you two last shared a relative. Roughly half the 500 genes yeast need for life are interchangeable with the human versions.
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Lipitor and other statin drugs are commonly prescribed to lower cholesterol.
Paul J. Richards/AFP/Getty Images
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March 10, 2015 People with a hereditary form of very high cholesterol are much less likely to get diabetes, a study finds. And that offers clues as to why cholesterol-lowering drugs sometimes raise diabetes risk.
A daily cup of joe (or two) may help protect against Type 2 diabetes and cardiovascular disease. And an egg a day will not raise the risk of heart disease in healthy people, according to a panel of nutrition experts.
Premshee Pillai/Flickr
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February 19, 2015 A panel of top nutrition experts is recommending that Americans adopt a more plant-based diet and eat less meat and sugar. It also found that most people are not consuming too much cholesterol.
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Researchers say they think there's something in the avocado — other than just the healthy fat — that may lower bad cholesterol.
Tastyart Ltd Rob White/Getty Images
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January 7, 2015 Researchers have long known that avocados have "good" fats. But they didn't know if daily avocado consumption might affect cardiovascular disease risk factors. So they designed a study to find out.
Fresh data show that Vytorin, a combination pill for cholesterol, cuts the risk of heart attack and stroke.
Getty Images/Getty Images
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November 17, 2014 Merck's Vytorin reduced the risk of heart attack and stroke more than a cholesterol-lowering statin alone. The findings come from an international study of more than 18,000 people in 39 countries.
Film CT scans show these people have suffered strokes.
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July 15, 2014 Deaths from stroke are dropping too, a study suggests. But don't celebrate just yet. Diabetes, a big risk factor for these "brain attacks," is still on the rise.
May 10, 2014 No single concept has permeated American medical culture to the extent that our anxiety about cholesterol has. Old or young, man or woman, rich or poor — everyone wants a cholesterol test.
Getting kids to exercise regularly and eat right can reduce their risk of heart disease down the road.
Robert Brown/iStockphoto
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March 28, 2014 High cholesterol isn't just a problem for adults; roughly 1 in 3 kids may have abnormal levels, too, a large study suggests. Kids as young as 9 or 11 should get a baseline check, pediatricians say.
Statins are widely prescribed to reduce the risk of cardiovascular disease, but we may not be getting a clear picture of side effect risks.
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March 13, 2014 An analysis of research on the cholesterol-lowering drugs shows that most side effects are no more common with them than with placebo drugs. But statins can raise the risk of diabetes.
Not covered by Obamacare, but still sweet.
Cristian Baitg/iStockphoto
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December 18, 2013 The Victorian proverb touting the health benefits of daily apple consumption has data to support it, British researchers say. And cholesterol-lowering statin drugs do, too. People who eat plenty of fruits and vegetables and take statins when directed would be healthier still.
November 18, 2013 Updated just last week by the American Heart Association and the American College of Cardiology, the new guidelines are based on old data, some heart doctors say, and may overestimate the real risk of heart attack and stroke. That could result in overtreatment with cholesterol-lowering drugs.
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Statin drugs to lower cholesterol have become among the most widely prescribed prescription medications in the United States.
Bill Gallery/ASSOCIATED PRESS
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November 12, 2013 For decades, knowing your cholesterol number was the first step in preventing heart disease and stroke. Heart specialists would shift away from specific cholesterol targets under new guidelines. A risk-based approach tailored to each patient would become the new norm.
Lipitor and other statin drugs are among the most commonly prescribed drugs in the United States.
Mel Evans/AP
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April 3, 2013 A study of statin use in the real world found that 17 percent of patients taking the pills reported side effects, including muscle pain, nausea, and problems with their liver or nervous system. Many of those people quit taking the pills, at least temporarily.
Before filling one of these tubes with blood for a cholesterol test, you're supposed to keep your stomach empty. But that may not be necessary.
Nancy Louie/iStockphoto.com
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November 13, 2012 For most people, a nonfasting cholesterol test will do just fine, a Canadian study suggests. A meal beforehand is unlikely to change key ratios of fats in the blood that doctors use to assess a person's risk for heart disease and stroke.
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Chris Gethard
Few comedians are having as good a 2014 as Chris Gethard is so far. Late last year, Comedy Central ordered a pilot of his more-than-a-little-zany Chris Gethard Show. The live cable-access program has aired in New York City since 2011, developing a cult following thanks to its lovable underdog ringleader, unusual themes (one episode, titled "The Beef Off," settled once and for all which regular cast member was the beefiest), and special guest appearances from the likes of Amy Poehler, Andrew W.K., and Zach Galifianakis. The pilot was shot a few weeks ago, and its fate is now in the hands of Comedy Central. On top of that, Gethard's first album, My Comedy Album, drops April 22. But that's not all: He was also recently selected to do one of Comedy Central's half-hour specials, which will be taped at the end of March. Before that, he'll be at Acme Comedy Co. for five nights and, due to the timing of the gig, this week will be a testing ground for material. "A lot of what I do in my Comedy Central half-hour special will be decided based on these performances," Gethard says. 18+. (Photo by Zac X Wolf)
March 4-8, 8 p.m.; March 7-8, 10:30 p.m., 2014
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I just arrived in Punta del Este and will stay here until the 18th January. My spanish is close to 0 so its not easy to communicate with the people here.
If anybody from overseas speaking english likes to catch up for a beer, drop me an email!
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Results 116 to 120 of 159
November 30th, 2013 09:03 PM #116Senior Member
- Join Date
- Apr 2011
- Posts
- 367
December 23rd, 2013 12:58 PM #118BoysWilliam, James, Thomas, Isaac, Elias, Oisin, Henry, Nolan, Ronan, Orson, Ephraim, Arthur, Robert, Finn, Jude, Kieran, Maxwell, Kellan, Leander, Josiah, Edmund, Gideon, Samuel, Oscar, Vincent
GirlsAlice, Orla, Eleanor, Celia, Audrey, Genevieve, Honora, Cordelia, Ivy, Marigold, Sybil, Anna, Moira, Susanna, Veronica, Harriet, Aria, Vivienne, Edith, Helena, Charlotte, Isla, Magdalena, Rosalind, Violet
December 25th, 2013 09:57 PM #120Senior Member
- Join Date
- Dec 2013
- Posts
- 208
Names are all your choice. When deciding birth orders, roll the dice for how many months/years the children are apart from each other in age. Feel free to add pictures, descriptions, etc.
First roll the dice...
If you rolled an EVEN number, roll it again here:
5. You have five sons.
1. Hudson Leo Forbes is born first
2. Noah Duke Forbes is born 4 years later
3. Dashiell Theo Forbes is born 4 years later
4. Finn Sebastian Forbes is born 4 years later
5. Emmett Tobias Forbes is born 10 months later
What do your kids look like? (your choice)
Hudson: http://www.shutterstock.com/pic-2157...sz0aXgpSw-1-16
Noah: http://www.shutterstock.com/pic-9696...lZySeO1NA-1-13
Dash: http://www.shutterstock.com/pic-1048...60LdaznMw-1-11
Finn: http://www.shutterstock.com/pic-7538...Zar8jIYCA-1-18
Emmett: http://www.shutterstock.com/pic-9141...YeYp_T8-w-1-10
January 4th, 2014 04:33 PM #122Senior Member
- Join Date
- Nov 2013
- Posts
- 2,730
Lily Claire Blackwell
-birthday: July 14th
My name is Lily Claire Blackwell and I'm from Cambridge, Massachusetts, where I was raised by my grandparents in a teal, cobblestone house. I have a younger brother and younger sister, James and Charlotte. After high school, I went to the University of Hartford in Hartford, Connecticut. I majored in journalism.
Henry Connor Montgomery
-birthday: May 2nd
His name is Henry Connor Montgomery and he's from New York City, New York, where he was raised by both parents in a cobblestoned house in the country. He has one older brother and two younger sisters, Charlie, Grace and Alice. After high school, he went to Lynchburg College in Lynchburg, Virginia. He majored in architecture.
We met through a mutual friend, Ruby, and moved to Boston few years later. Our first date was ice skating at the local rink and dinner after. We dated for three years before getting engaged. He proposed over dinner in our favourite restaurant and we married a year later in the park. We honeymooned in Athens, Greece, and then moved to Hartford, Connecticut to a big, old fashioned house. We start having kids 7 months after we get married.
DD/DD: Rose Katharine "Rory"/Kiera Meredith (6)
DS: Hugo Benjamin (3)
DD: Hazel Bridget (1)
DD: Thea Olive (nb)
We have four daughters and one son. First came Rose Katharine and Kiera Meredith, twins. They both have curly, brown hair, but Rory has brown eyes and Kiera green. Then, three years later, Hugo Benjamin was born. He has straight, brown hair and green eyes. Two years later, we had Hazel Bridget, who has curly, blonde hair and brown eyes. A year after her birth we had Thea Olive, who has straight, brown hair and brown eyes.
January 4th, 2014 07:25 PM #124Auntie to Connor Douglas (June 2012) & Parker Isabella (Oct 2013)
~*Ariadne*Celeste*Esmé*Deirdre*Linnéa*~I believe that maturity has more to do with what types of experiences you've had and what you've learned from them and less to do with how many birthdays you've celebrated.
My full list here. Please comment and I'll comment on yours
(Not currently expecting or TTC)


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The ATA's mandate requires it to work on two levels:
- At the national level – ATA is expected to work with all stakeholders to identify solutions to address systemic bottlenecks that can have a catalytic effect to transform the lives of smallholder farmers across the country; and
- At the geographic level – ATA is expected to work with implementation partners to ensure that a focused set of catalytic interventions are integrated around a prioritized set of commodities around the country.
I. Support to Address Systemic Bottlenecks for National Impact
Over the past decade, the Ethiopian agriculture sector has primarily focused on ensuring food security and identifying new technologies that will increase farmers' production and productivity. While this approach has brought about major growth of the sector, it will be necessary to do more in order to create the structural transformation of the agricultural sector necessary to support Ethiopia's industrial and overall economic growth ambitions. As such, the ATA has been asked to focus on the systemic bottlenecks that constrain Ethiopia's smallholder farmers from moving from subsistence toward commercial agriculture.
Identifying and Resolving Systemic Bottlenecks
By consulting international case studies, and leveraging best practices from other problem solving organizations, the ATA has developed an Ethiopia-specific approach to identify and resolve systemic bottlenecks. This includes:
Senior policy makers prioritize a narrow set of specific issues that require immediate attention into the Agricultural Transformation Agenda;
Identify solutions for prioritized bottlenecks through deep engagement with technical experts in Ethiopia and across the world;
Support local organizations that are implementing solutions to ensure that analyses and strategies lead to results on the ground; and
Robust monitoring and performance management to ensure that interventions are actually executed on schedule and within budget.
II. Coordinate Implementation of an Integrated Set of Solutions around Prioritized Commodities and Geographies
Ethiopia is an extremely diverse country with over 30 different agro-ecologies, as well as a variety of cropping and livestock systems. All systemic solutions prioritized in the Transformation Agenda will have national application.
However, the greatest impact on the ground will only come when these systemic solutions are integrated into a coordinated set of interventions applied to specific commodities and geographies. As such, a second aspect of ATA's mandate is to work with stakeholders at the Regional level to identify and support the implementation of a narrow set of systemic interventions in an integrated fashion. This requires ATA to work closely with Regional governments and implementation partners both from the public, private and civil society sectors. It also requires ATA to be deeply involved in Regional planning and resource allocation processes.
Geographic integration has been done on a limited basis during the initial stage of ATA's efforts. However, rapid expansion of ATA's work in this area is expected in the coming months due to the development of the Agricultural Commercialization Initiative (provide link to that highlighted deliverable) and the focus of GTP2 on increased smallholder farmer commercialization and the market orientation of key commodity value chains.


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Dr. Jeffrey Weddington graduated from the University of California, Los Angeles David Geffen School of Medicine in 1996. He works in Fort Washington, MD and 1 other location and specializes in Dermatology. Dr. Weddington is affiliated with Fort Washington Medical Center.
Medical School: University of California, Los Angeles David Geffen School of Medicine; Graduated 1996
*This physician has multiple office locations. See Office Information for details.
703 Giddings Ave Ste M
Annapolis, MD 21401
11701 Livingston Rd Ste 302
Fort Washington, MD 20744
6192 Oxon Hill Rd Ste 404
Oxon Hill, MD 20745
6130 Oxon Hill Rd Ste 201
Oxon Hill, MD 20745
19735 Germantown Rd Ste 210
Germantown, MD 20874


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Morning, I ended up just having the bread with a dippy egg!" And have lost another Pound!" Woohoo!"
Morning, I ended up just having the bread with a dippy egg!" And have lost another Pound!" Woohoo!"
thats great news lou, Dukan is a fab diet xxx
well done, going great x
Thank you!" Quick question should I be loosing as much as I am in the first week of cruise? I don't want to be getting me hopes up & then all of a sudden stop...
wow your weightloss is great
dont knock it, loosing them lbs is a buzz. Enjoy
it all seems to go fast to start with, at least it did for me, it will soon even itself out and settle down to a steady 1-2lb a week. Just let it happen hun, well done x
Hello Everyone!" I fell off the wagon at weekend again!" and gained 3/4lbs!" I can't seem to help myself at work it's so much easier to just get on with it but at weekends I seem to just go wild!" It doesn't help that my boyfriend is eating normally.....
I had a PP day yesterday and todays PV.. Am I doing it right or should I do a week of Attack to get back on track??
No definately don't do attack again. Its only designed to be a starter. You could possibly do a couple of days of PP, but honestly just get back to normal. some people do have 'eating' weekends but stay good the whole week though.
its hard when other people are eating 'normal' but what you have to tell yourself is that it isn't forever, its only till youreach TW and can then start reintroducing.
Thanks for the advice Dukannotsofatty!" I've Lost 7lbs again so hopefully I wont go mad at weekend and eat really sensibly.... Hopefully!"
I would find it hard to do two days of eating with 5 days of not. eating loads and norty stuff I mean LOL
Argh!" I have done it again with only myself to blame after this weekend coming all weekends will be Dukan!" Gained 3lbs!" Grrr!"
I've just changed my goal weight to 11stone for now as at this moment in time 10st feels abit unobtainable and I'm not feeling the diet today...obv cravings from weekend!" Boooo!"
Weight is just a number it's how you feel within yourself that's important! X


text: Mashup: Music Anchor Comments Share via FacebookShare via TwitterShare via LinkedinShare via Google+Share via RedditShare via Mail Music discovery on your PC using Last.fm, MusicBrainz, Amazon, eBay. Create your own playlists and stream them directly from YouTube. Track this Mashup Submit a mashup Summary Comments Followers Summary Comments Followers Comments (0) Comment * Leave this field blank Post Comment


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Volition Photos
Volition ReviewsVolition 13
"Best AAA Culture in the industry "
RecommendsPositive OutlookApproves of CEO
- Work/Life Balance
- Culture & Values
- Career Opportunities
- Comp & Benefits
- Senior Management
I have been working at Volition full-time
Culture, events, quality of life (rarely do we crunch, and not for long), great leadership, cheaper area to live.
Midwest isn't for everyone. Weather is rough in the winter. There's honestly no major con about the studio itself - it's a dream company.
Advice to Management
High-level vision holders dip into the specifics of the project's details in a way that sometimes disrupts the low-level work, but nothing majorly wrong here.
Volition Interviews
Interview Experience
Interview Experience
Getting an Interview
Getting an Interview
Interview Difficulty
- Accepted OfferPositive ExperienceAverage Interview
I applied online. The process took 2 weeks. I interviewed at Volition (Champaign, IL).
Job posting on their company website, hiring for temp gig for 1-2 years. Submitted resume and cover letter, got a response asking to write answers to a questionnaire- things like briefly review a game and talk about its strengths and weaknesses, how do you approach challenges, how much do you game, how would you troubleshoot a technical issue. Called back to set up interview, met with two QA Supervisors; pretty straight-forward conversation about previous experience and interests. Played a buggy build of an old project while they watched, and was expected to call-out bugs as they came up.
Interview Questions
- If you were sealed in this room, with no form of communication with the outside, how would you attempt to get out? Answer Question
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Our implementation of CombatCounter is incorrect. It should be triggered for every possible-combat actions and count down from a value (IESDP says 150, real value is perhaps nearer 100) until it reaches 0.
See devSin's commentary at http://forums.gibberlings3.net/lofiversion/index.php?t5489.html
I tested this in bg2 (patched ToB). It resets itself to somewhere between 140 and 150 (the IESDP's 150 would seem the right answer, given this) whenever melee combat is started (eg, when you click on an enemy) or after a few(?) seconds of combat (it doesn't seem related to round time, proximity to the actor, hitting or attacks). It stops resetting itself when the combat is stopped. It also increments itself to a low value (I think below 30) when dialog is attempted, whether successful or not - this is when the actual dialog attempt happens, not when the click happens. I can't get it to change at all for spellcasting, whether hostile or not.


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All Items : Dinnerware : Metlox : Pre 1960 item #761993 (stock #safn-201)
Matching cup with saucer in the antiqua pattern made by Metlox, poppytrail, Vernonware of California. Soft creamy oatmeal color with antiqued effect showing off the relief of fruit and berries bordering. I actually have 9 sets available and all are perfect. Price is for 1 cup with it's matching saucer.
All Items : Cookie Jars : Pre 2000 item #761980 (stock #safn-199)
This is a cute little jar for the Halloween holiday. THe little pumpkin is covered with fall foliage, and has a jack-o-lantern face. Made of hand painted glazed ceramic. Small in size, measuring just over 6" tall. Would be great for candy and treats. Perfect condition.
All Items : Cookie Jars : Pre 1980 item #761967 (stock #safn-197)
This is a great cookie jar for the owl lover. Made of hand painted glazed ceramiic. Measures 11" tall. Cookie jar is a tree stump, and has one owl peering from inside a hollow in the stump, and a small owl sits on top of the stump and is the knob for the cookie jar. The jar does have a chip repair under the rim of the jar. (They done a nice job, as I almost didnt see it). Otherwise, great condition. No marks on this cookie jar. It almost looks Otagiri made.
All Items : Cookie Jars : Pre 1990 item #761960 (stock #safn-196)
Adorable hand painted and glazed cookie jar is the shape of an old fashioned knit Christmas stocking. Stitches, patches , buttons, and frills. Filled with toys candy and gifts. Measures 10 1/2" tall and is in perfect condition. Made by GKAO.
All Items : Kitchen : Salt and Pepper Shakers : Pre 1960 item #761955 (stock #safn-195)
Cute salt and pepper elephants. One is pink and the other is blue. Measures 3" tall. Perfect condition other than one has a missing stopper. Made of heavy ceramic.
All Items : Cookie Jars : Pre 1990 item #761949 (stock #safn-194)
Nice big cookie jar measures 10" tall, and is made of hand painted ceramic.Made by Sherwood Brands. Nice condition. There seems to be a few tiny rough glaze chips that looks like they have been touched up, upon close inspection. The cookie jar has two sides, one side a Sweet shop and the other side looks like a regular victorian building. Email if you need to see the other side photo.
All Items : Pottery and Stoneware : Pre 1960 item #761916 (stock #safn-190)
Bright and colorful, this little vase measures just 3 3/4" tall. Made of red clay, and hand painted with a heavy glaze. My guess is that it is Mexican. No chips, cracks or repairs.
Cute little vintage doll measures 7 1/2" tall. She has blonde rooted hair and rooted lashes. Eyes open and close. original dress and panties but no shoes. Marked made in Hong Kong with an odd logo mark on her back. She is in good condition with no marks or damages.
All Items : Dinnerware : Franciscan : Pre 1990 item #761569 (stock #ba-28-10)
Here is the Franciscan ivy gravy boat with its seperate underplate. Made in England. Both pieces are in pristine condition. Sold together for one price.
All Items : Dinnerware : Franciscan : Pre 1990 item #761563 (stock #ba-27-10)
Beautiful Franciscan tulip dinner plates measure 10 5/8" in diameter. Microwave, freezer, oven and dishwasher safe. I have 8 available and all are in pristine condition. Price is for each plate.
All Items : Dinnerware : Franciscan : Pre 1990 item #761557 (stock #ba-26-9)
Awesome lasagne bake dish in the ivy pattern. Dishwasher,freezer, microwave, & ovenproof. Measures 10 3/8" x 13" and is 2 7/8" deep. This is very heavy, and a wonderful practical piece to add to your dinnerware set. made in Portugal and is in pristine condition.
All Items : Dinnerware : Franciscan : Pre 1940 item #761535 (stock #ba-20-6)
Perfect cover for the 1/4 lb butter dish if you need a replacemet top. THis is in the franciscan ivy pattern. California USA. I have 2 perfect ones available. Price is for each.
All Items : Dinnerware : Franciscan : Pre 1940 item #761531 (stock #ba-19)
Here we have a nice replacement lid for your 1/4 lb Franciscan poppy butter dish. Yellow and lavender poppy decoration. Perfect condition. USA.
All Items : Dinnerware : Franciscan : Pre 1950 item #761527 (stock #ba-18-6)
This cup is pristine! Franciscan ivy pattern made in the USA.
All Items : Dinnerware : Franciscan : Pre 1950 item #761526 (stock #ba-17)
Franciscan ivy pattern cup, green trim. Older USA. Perfect other than a fleabite on the inner rim you can only feel, but not see.
All Items : Dinnerware : Franciscan : Pre 1950 item #761523 (stock #ba-16-6)
Single saucer in the Franciscan ivy pattern by Gladding McBean. 6 1/8". Perfect condition.
All Items : Dinnerware : Franciscan : Pre 1970 item #761520 (stock #ba-15-5)
Franciscan ivy bread and butter plate measures 6 3/8" in diameter. There are 6 available. All six are in mint condition. Marked with the Franciscan F on the back, USA. Price is for each plate. Please let me know quanity when ordering.
All Items : Dinnerware : Franciscan : Pre 1940 item #761517 (stock #ba-14-5)
Hard to find pair of salt and pepper shakers for the Franciscan Ivy dinnerware set. Each is in the shape of an ivy leaf. One is light the other dark green, and each measure 3" tall. Original rubber stoppers. Pristine condition. I have a picture of the bottom if you need to see it, email me. Old stock made in USA.


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Anderson no longer at law firm, council votes to put open seat on ballot
Chuck Anderson no longer works at the law firm that bears his name and Anderson city leaders have moved forward with efforts to fill his vacant council seat.
He stood before a judge this week and admitted lying to federal agents who were investigating a large-scale cocaine case. Anderson had been representing a client, Lonnie Maddox, in the drug case in 2013. Federal agents asked Anderson the whereabouts of two cars belonging to Maddox. Anderson lied to agents about having a GMC Denali and a classic Chevrolet Chevelle that belonged to Maddox, prosecutors said.
The Denali was ultimately recovered from the home of one of Anderson's law partners.
Anderson has not returned repeated messages from the Independent Mail. He has changed his personal voice mail so that it no longer includes his name.
Lia Bantavani, a spokeswoman for the U.S. Attorney's Office in the Western District of North Carolina, said Tuesday that no one else has been charged in connection to Anderson's case.
Federal prosecutors in North Carolina are handling Anderson's case because the U.S. Attorney's Office in South Carolina is overseeing the Maddox case and has recused itself from Anderson's.
According to court documents, Maddox agreed last week to admit being part of a conspiracy to distribute cocaine, and to acknowledge that he helped move around money that was gotten by illegal means.
Anderson, 44, has not yet been sentenced for lying to investigators. A woman who answered the phone at his practice Tuesday said Anderson no longer works at the firm.
His wife, Floy Kenyon Anderson, is also a partner in the firm, along with attorney Robert Lusk.
Kenyon Anderson was not available Tuesday and Lusk did not return a message seeking comment.
Chuck Anderson could face sanctions from the South Carolina Supreme Court, said Lesley Coggiola with the state's Office of Disciplinary Counsel. An attorney facing criminal charges has 15 days, from the day that the charges are issued, to report the matter to state officials, Coggiola said.
She said she would not say whether Anderson had followed that requirement.
Once a lawyer reports being charged, Coggiola's office will make a recommendation that the attorney be placed on interim suspension.
"The interim suspension is usually done quickly," Coggiola said. "While they are on suspension, we begin our case. We try to resolve the case from a disciplinary standpoint. He could volunteer to relinquish his law license. If a lawyer is in trouble, he or she might say, 'I just want to turn my license in.'
"But that does not make the investigation go away, from our standpoint," she said. "That is still pursued."
When the Office of Disciplinary Counsel finishes its investigation, it makes a recommendation that the state's highest court can accept or reject.
Before resigning from his elected office this week, Anderson had been on the Anderson City Council for a year and a half. He took office in July 2012 and was supposed to serve a four-year term. Now, the city council must amend an ordinance so Anderson's vacant seat can be included in an already-scheduled election to be held in April. The mayor's seat and the seats in odd-numbered city wards were already scheduled to be part of that municipal election.
City council members took less than two minutes Tuesday to amend the ordinance so that Anderson's empty seat will also be on the spring ballot. That amendment has to be voted on two more times before the ballot can be changed.
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Governor Snyder calls for adding LGBT protections in Elliot-Larsen Act
June 2, 2014 by Lynne Bowman, Regional Field Director
On Thursday Michigan's Governor Rick Snyder joined a growing coalition of voices calling for the inclusion of LGBT people in the state's civil rights protections. Joining Snyder in his announcement were the Detroit Regional Chamber of Commerce and the Grand Rapids Area Chamber.
For decades, pro-equality forces in Michigan have worked to amend the state's existing civil rights law, the Elliot-Larsen Act, to include sexual orientation and gender identity. The efforts have gained momentum recently with the launch of the Michigan Competitive Workforce Coalition under the leadership of Jim Murray, President of AT&T Michigan. Others in the coalition include Chrysler Group LLC, Kellogg, Herman Miller, and Steelcase.
HRC has a long-standing partnership with activists on the ground in Michigan who are working tirelessly to see success before the end of the 2015 legislative session. The ACLU of Michigan, Equality Michigan and Unity Michigan are all deeply involved in the work and we are pleased to join them in this effort.
HRC is making and investment in the effort by placing a statewide Field Coordinator in the state to work with the organizations helping to raise the voices of Michiganders to share why nondiscrimination protections are important to their lives. Candidates much have no less than one year of issue or candidate political organizing experience and have the ability to travel across the state. This is a temporary position. If you or someone you know is interested, please contact me at firstname.lastname@example.com for further information.
Top Posts
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June 1, 2015
Frequently Asked Questions about the Upcoming Marriage Equality Case at the U.S. Supreme Court
April 24, 2015
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June 15, 2015
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June 5, 2015
Watch: HRC Answers Your SCOTUS Questions
June 19, 2015


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Truworths Boosts Chief Executive's Pay 24% as Profit Increases
Sept. 27 (Bloomberg) -- Truworths International Ltd., South Africa's largest clothing retailer by market value, raised Chief Executive Officer Michael Mark's pay by 24 percent in its last fiscal year as profit increased.
Mark earned 19.3 million rand ($2.45 million) in the 12 months through June 26, including base salary of 4.87 million rand and a 9.88 million-rand performance bonus, Cape Town-based Truworths said in its annual report, published on its website. He was paid an additional 10.8 million rand as part of a performance share-based plan, up from 10.5 million rand in the prior period.
Annual net income climbed 21 percent to 1.94 billion rand as the retailer opened more stores, Truworths said on Aug. 18. Sales rose 13 percent to 8.68 billion rand. The company's stock has gained 12 percent in 12 months.
To contact the reporter on this story: Janice Kew in Johannesburg at firstname.lastname@example.com
To contact the editor responsible for this story: Antony Sguazzin at email@example.com


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#655 of 1480
Frisco, TX
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Expert Printing and Assembly
Shipping Within 2 - 3 Business Days
Millions of Satisfied Customers
Fairy Tale Forest
Inge Johnsson
Photograph - Digital Capture Watermark Not On Actual Prints
Forest at Sol Duc Falls in Olympic National Park, Washington
Olympic National Park is a United States national park located in the state of Washington, in the Olympic Peninsula. The park has four basic regions: the Pacific coastline, alpine areas, the west side temperate rainforest and the forests of the drier east side.
U.S. President Theodore Roosevelt originally created Mount Olympus National Monument in 1909. It was designated a national park by President Franklin Roosevelt on June 29, 1938. In 1976, Olympic National Park became an International Biosphere Reserve, and in 1981 it was designated a World Heritage Site. In 1988, Congress designated 95 percent of the park as the Olympic Wilderness. The western side of the park is mantled by a temperate rain forest, including the Hoh Rain Forest and Quinault Rain Forest, which receive annual precipitation of about 150 inches (380 cm), making this perhaps the wettest area in the continental United States (the island of Kauai in the state of Hawaii gets more rain).
As opposed to tropical rainforests and most other temperate rainforest regions, the rainforests of the Pacific Northwest are dominated by coniferous trees, including Sitka Spruce, Western Hemlock, Coast Douglas-fir and Western redcedar. Mosses coat the bark of these trees and even drip down from their branches in green, moist tendrils.
Valleys on the eastern side of the park also have notable old-growth forest, but the climate is notably drier. Sitka Spruce is absent, trees on average are somewhat smaller, and undergrowth is generally less dense and different in character. Immediately northeast of the park is a rather small rainshadow area where annual precipitation averages about 16 inches.
June 10th, 2012
Viewed 3,491 Times - Last Visitor from Mountain View, CA on 10/05/2015 at 12:02 PM
Sales Sheet
More from Inge Johnsson
Cuiaba, Ma - Brazil
Nice work Inge.
Nagpur, Ma - India
Wonderful and congratulation on sale announcement.
Colorado Springs, CO - United States
Congrats on your sale!!!
Surrey, BC - Canada
Beautiful mood and light. Congratulations on your sale.
Tower City, PA - United States
So beautiful! Love the light! Congrats on your sale!
Nearest Town Is Gillette, WY - United States
Oh my I used to live in NW Oregon and had friends up in Aberdeen ... Love the Peninsula with all it's old trees ... Paradise (smile) ... Wonderful capture ... That's for bringing back the memories (smile)
Farmington, NM - United States
Lovely and the way the light comes through is fantastic...L/fav
Overpelt, Li - Belgium
Amazing capture, really a fairy tale image!
Burnaby, BC - Canada
Stunning light. Congratulations on your feature in Photographic Excellence. v
Loveland, CO - United States
Awesome images Inge! Congratulations on being selected Artist of the Month!! f/v
Camberley, Su - United Kingdom
Beautiful atmospheric image v
New Iberia, LA - United States
Beautiful capture! I would love to see that in person one day, so dreamlike. F/V
Seattle, WA - United States
Oh my goodness this is breathtaking! All of us locals know this isn't made up. The park really is that beautiful all the time. Thank you for bringing this to our eyes. Voted
Central Point, OR - United States
Etna, NH - United States
Inge you have an amazing portfolio full of fantastic landscapes! Hur s�ger man "incredible" p� svenska?
Thanks Edward for the kind comment. By the way, it's "otrolig" in Swedish. Next you're gonna ask me how to pronounce that... ;-)
LICHFIELD - United Kingdom
I Love this Inge...so atmospheric...!!!!
Thanks so much, Jean. I was quite happy to find this wonderful fog that morning in the forest.
Elgin, IL - United States
Very magical!! I love this!! :^D---jrr
Many thanks :-)
Simi Valley, CA - United States
I love this one, too! Pure magic!! f/v
Thanks Lynn. I was lucky to have this nice foggy morning there :-)
Bhopal, Ma - India
Beautiful fav
Many thanks!
Lambertville, NJ - United States
Beautiful! fav
Thanks so much, Val :-)
Please Wait...
Beverly Hills, CA (United States)
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Opinion: Why the Tea Party's Squeaky Wheel is a Good Thing
For people who pay a lot of attention to politics, it is hard to understand how some people still don't know who Tea Party Movement types are, and what they stand for. Over the last couple years, they've gone from an unknown entity, to a major driver in American politics, taking over the driver's seat in the Republican base and eating up a huge share of the political media's attention.
But that hasn't done good things for its popularity.
Neither party has been willing to touch the "third rails" of politics, entitlement programs for the retirement aged, for generations. In fact, with a handful of notable exceptions, both parties have been more than willing to pander to this older demographic, that votes in higher numbers than any other, by adding more benefits to programs like Medicare and social security.
But with the threat of Tea Party backlash in primaries across the country a constant worry for Republican officeholders, their extreme budget-cutting views have helped push nearly the entire GOP caucus in the House and Senate to vote for Paul Ryan's Medicare slashing budget proposal. Only the three remaining moderate New England Republicans (Olympia Snowe, Susan Collins and Scott Brown) are standing against Ryan's budget.
This would have not been possible without the pressure of the Tea Party, and it could lead to massive backlash in next year's election.
Polling data shows that, after peaking near 40 percent last year, just a third of the American people support the Tea Party. To put this in perspective, these numbers are even lower than approval ratings of the two major parties, both who are hovering in the mid to lower 40's. Dig into the data a bit deeper though, and you find even more interesting tidbits.
The media, who has always been obsessed with squeaky wheel groups that make a lot of noise, is convinced the Tea Party will harm the Republicans' chances of gaining seats next year. This may certainly be true if they have to make far right campaign promises and votes to win their primary support, but most of the public doesn't seem to care about them. When given the option, around half of people polled say that a candidate saying they are a member doesn't change their view of them. Just 26 percent say it makes them less likely to support them, versus only 14 percent that says it would make them more likely to.
More than any other group, the Tea Party illustrates how a small portion of the population, in a democratic republic with low levels of political engagement, can get much more power than they should, through being loud, active and unreasonable. Groups like this don't need to convince the American people at large to get what they want, they only need to convince some elected members of one of the two major parties.
But this is the Tea Party's right. As much as it shows a fundamental flaw in our two-party system, if the rest of us were as engaged politically, the extremes wouldn't have such disproportionate power.


text:
gelatin powder used in making soft candy
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Gulaman, Gelatin, Seaweed
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Noted at http://liveunplugged.wordpress.com/2010/12/06/windows-live-mail-version-2011-options-menu/
what do i have to do to see option tabs or mail submenu in windows live mail
CNET's Forum on browsers, e-mail, and other Web applications is the best source for finding help, troubleshooting, or tips from a community of experts. Discussions cover types of Web browsers (ranging from Mozilla Firefox, Chrome, to Windows Internet Explorer), plug-ins, Latest e-mail providers ranging from G-mail to Yahoo mail, VoIP software, chat clients, fixing security issues, avoiding spam, toolbars, tabs, and anything related to Web applications.
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Centromeres, and the kinetochores that assemble on them, are essential for accurate chromosome segregation. Diverse centromere organization patterns and kinetochore structures have evolved in eukaryotes ranging from yeast to humans. In addition, centromere DNA and kinetochore position can vary even within individual cells. This flexibility manifests in several ways: centromere DNA sequences evolve rapidly, kinetochore positions shift in response to altered chromosome structure, and kinetochore complex numbers change in response to fluctuations in kinetochore protein levels. Despite their differences, all of these diverse structures promote efficient chromosome segregation. This robustness is inherent to chromosome segregation mechanisms and balances genome stability with adaptability. In this review, we explore the mechanisms and consequences of centromere and kinetochore flexibility as well as the benefits and limitations of different experimental model systems for studying them.
During growth and cell division, cells must balance the requirement for faithful chromosome replication and segregation with the need to adapt to changing conditions. Failure of centromere/kinetochore function can result in aneuploidy, a change in chromosome number [1]. Aneuploidy is often detrimental [2, 3], but it has the potential to be adaptive under some conditions [4, 5]. For example, defects in genomic stability are implicated in cancer and can cause miscarriages and birth defects, while moderate aneuploidy may contribute to increased proliferation of cancer cells and to resistance to antifungals in yeast [4, 6, 7]. Chromosome segregation errors can lead to DNA damage and chromosome rearrangements [8, 9].
The kinetochore, a complex of ~100 proteins, must assemble on centromere DNA to allow spindle microtubule attachment to the chromosome during mitosis. Genome stability requires efficient and accurate kinetochore assembly, ensuring that each chromatid attaches to a single pole and that sister chromatids attach to microtubules from opposite poles. Thus, centromeres, and the kinetochores assembled on them, must participate in an exquisitely regulated process to ensure proper chromosome segregation. However, kinetochore assembly must be robust enough to handle physiological changes, such as overexpression of kinetochore proteins in cancer cells [10], that could perturb the balance of kinetochore components available for assembly.
Recent work suggests that centromeres/kinetochores tolerate significant deviations from the canonical structural features. These deviations include: variations in centromere DNA length, frequent alteration of the underlying DNA sequences, shifts in kinetochore position on the DNA and changes in kinetochore complex number (Box 1). This review discusses mechanisms cells use to maintain accurate chromosome segregation despite alterations to centromere/kinetochore structure. We suggest that centromere/kinetochore flexibility provides a buffering mechanism that facilitates successful evolution by maintaining chromosome segregation fidelity.
Centromere DNA length—The length of DNA associated with CENP-A and kinetochore protein complexes varies significantly from yeast to humans.
Centromere DNA sequence – The DNA sequences where centromeres are found vary greatly between organisms and also within a single species. Additionally, studies of closely related species indicated that the DNA associated with kinetochores might undergo accelerated mutation, further enhancing centromere sequence diversity.
Kinetochore position – The position of the kinetochore on the chromosome is flexible in a wide variety of organisms ranging from the yeast C. albicans to humans. One of the characteristics of primate speciation is the appearance of evolutionary new centromeres: kinetochores that associate with chromosomal regions that differ from the position of the ancestral centromere. Additionally, acentric chromosome fragments can be rescued by neocentromeres, kinetochores that form at new chromosomal positions.
Kinetochore complex number – The kinetochore complex number is defined as the number of kinetochore protein complexes assembled per centromere DNA region. The kinetochore complex number varies with changes in centromere DNA length and kinetochore protein concentrations in the cell.
Eukaryotes exhibit diverse centromere organization patterns and kinetochore structures [11, 12] (Table 1). Many proteins that make up the kinetochore are conserved from yeast to humans. Centromeres are defined as chromosomal regions that bind CENP-A (CenH3), a histone H3 variant that replaces canonical H3 at centromeres, and assemble a functional kinetochore. Centromeres vary in length as well as inheritance mechanisms. Saccharomyces cerevisiae has point centromeres in which a DNA binding sequence is necessary and sufficient to drive kinetochore assembly [12]; in contrast, other fungi, plants and mammals form regional centromeres that usually occupy repetitive DNA and are inherited epigenetically. Many different models have been proposed to explain the mechanism(s) of epigenetic regional centromere inheritance [13]. Centromere regions vary in length from 125 bp in Saccharomyces spp. [12], to megabases of repetitive DNA found in mammalian and plant centromeres [11], and to entire chromosomes found in organisms that have holocentric centromeres, such as Caenorhabditis elegans [14]. As centromere structures have diverged, centromeric DNA sequences have also evolved rapidly.
Not only does centromere DNA differ dramatically between organisms [12], it differs between centromeres within a given organism [15, 16]. For example, different chicken centromeres vary in length from 30 kb to 500 kb, and contain either non-repetitive or highly repetitive DNA [15]. In contrast, human centromeres are all associated with alpha-satellite repeat DNA, although its length can vary from several hundred kb to 10 Mb [17, 18]. Diversity in the degree of repetitiveness indicates centromere "age" in evolutionary terms, as centromeres accumulate repeats that are maintained over time [19, 20]. The dog and horse genomes include high sequence divergence in centromeric satellite repeats on different chromosomes [21]. In the fungal pathogen Candida albicans, which has regional centromeres of ~ 3 kb, each centromere contains a unique, non-repetitive core sequence [22].
Consistent with the sequence diversity found between centromeres, centromere DNA evolves at rates much higher than rates at other non-coding regions. For example, C. albicans and Candida dubliniensis diverged more than 20 million years ago, and while the centromeric synteny in these two organisms has been maintained, the centromere DNA sequences diverged faster than other orthologous intergenic regions [23]. Similarly, maize centromere sequences evolve rapidly via high rates of gene conversion [24], and mouse Y centromere repeat DNA sequences have diverged from repeat sequences found at other chromosomes and in closely related mouse species [25]. The mechanisms directing rapid evolution of centromere DNA sequences are a topic of active research. Meiotic drive, an evolutionary force in which paired chromosomes compete for access to the egg during reproduction, has been proposed to promote centromere sequence divergence [26]. In S. cerevisiae, where meiotic drive would not be expected to be a substantial evolutionary force, as meiosis results in four equal spores rather than a single egg, rapid evolution of centromere DNA is consistent with an elevated mutation rate at centromere DNA [27], perhaps due to features of the centromeric chromatin that enhance mutational frequencies or inhibit DNA repair mechanisms.
In addition to having diverse primary DNA sequences, centromeres have evolved by moving to new positions in the genome. Different, evolutionary new centromere positions are a defining characteristic of primate species. Ectopic kinetochore assembly is a key step in their formation. For example, human centromere 6 was repositioned from an ancestral location to its current location in a common ancestor of hominids [28]. Indeed, approximately half of the human chromosomes have evolutionary new centromeres, based upon comparison with macaques [20].
Evolutionary new centromeres often arise in gene deserts [29] and accumulate repetitive sequences [20]. Similar events have been seen in other mammals [30] as well as in plants (e.g., rice [19] and cucurbits [31]). These evolutionary events, detected via altered synteny of centromere-associated satellite DNA as well as through karyotype analysis, require that kinetochore proteins assemble at a new locus and no longer assemble at the old locus. A similar process, termed neocentromere formation, occurs in modern humans as well as in model organisms.
Neocentromeres are new kinetochores that assemble at DNA loci not previously associated with kinetochore proteins. Neocentromere formation can occur in the absence of chromosome rearrangements (repositioned neocentromeres) or it can rescue chromosome fragments, amplifications or rearrangements on either linear or ring chromosomes (rescue neocentromeres) [32]. Importantly, neocentromeres can be inherited from one generation to the next. In two documented cases of repositioned neocentromeres, the new centromere position has been maintained through at least three generations [28, 33]. While repositioned neocentromeres are rare [32], it is likely that their incidence is underreported, because they are only identified by cytogenetic screening and cause no obvious phenotypes, as the chromosome is still segregated during mitosis and the genome remains essentially intact. The majority of the ~100 reported neocentromeres are rescue neocentromeres [32] and the accompanying amplifications, deletions, or gene disruptions cause phenotypes including developmental disabilities as well as cancers such as non-Hodgkin's lymphoma [34], acute myeloid leukemia [35], and liposarcomas [36].
Neocentromeres map to at least 21 different human chromosomes, including X and Y, although they are particularly common on specific chromosome domains including 3q, 13q and 15q [32, 37]. Little is known about how neocentromeres form or why they form at particular positions. Human centromeres are always found in highly repetitive regions; the same is true for some neocentromeres [38], but not for others [39]. Chromosome regions that maintain neocentromere function show little similarity, and none of them are associated with alpha-satellite DNA, making it difficult to ascertain the sequence elements that might contribute to a good site of neocentromere formation. Interestingly, even among neocentromeres found in the same chromosomal band, no two neocentromeres have formed at exactly the same underlying DNA sequences [32, 38, 40]. LINE retrotransposon sequences appear to be important for establishing neocentric chromatin at some neocentromeres, as knockdown of transposon transcript levels impaired neocentromere function during mitosis [41]. ChIP-chip analyses failed to define common features of human neocentromeres [30], but higher resolution analysis in the future with ChIP-seq or ChIP-exo on CENP-A may provide additional insight. To date, the only sequence data available does not reveal any obvious patterns, supporting the idea that centromere position is remarkably flexible in humans.
Unlike evolutionary new centromeres that have persisted and that always formed in regions that were gene deserts [29], human neocentromeres form either in gene deserts or in regions that include actively transcribed genes [32, 39]. Of course, it is unknown which recently formed neocentromeres will or will not persist to become evolutionary new centromeres. It is tempting to speculate that those formed in gene deserts are more likely to persist, while those formed on actively transcribed regions may cause phenotypes that interfere with their selection and evolutionary persistence. Indeed, some neocentromeres are more prone to chromosome missegregation than native centromeres. Human neocentromeres are more defective in recruiting Aurora B kinase and in correcting chromosome segregation errors than native centromeres [42]. Additionally, some patients exhibit neocentromere mosaicism (presence of the neocentromere in a fraction of somatic cells), suggesting that the chromosome carrying the neocentromere was lost in a subpopulation of the cells and implying that, in vivo, some neocentromeres are less stable than native centromeres [32]. Determinants of the chromosome segregation efficiency of different neocentromere positions remain to be identified.
Kinetochore position flexibility occurs in diverse species (Table 2), with some neocentromeres near the native centromere [43], others at the boundaries of euchromatin and heterochromatin [44, 45] and yet others within subtelomeric regions in a manner dependent upon heterochromatin formation [46]. In contrast, human neocentromeres exhibit no apparent association with heterochromatin [39]. In plants there are two types of neocentromeres: neocentromere knobs' and rescue' neocentromeres that, like human and C. albicans neocentromeres, appear following centromeric deletions or rearrangements [16, 48]. Neocentromere knobs' drive selection of specific chromosomes during meiosis [49]. Thus, they are likely to involve different mechanisms of formation and/or maintenance [50].
Genetic model organisms can be used to test hypotheses based on the analysis of clinical neocentromeres. For example, neocentromere mosaicism suggests that some neocentromeres are more functional than others [32]. Neocentromeres in C. albicans resemble those in humans in that they are flexible in their position and not associated with heterochromatin [47]. Furthermore, as a unicellular eukaryote, C. albicans is highly amenable to selection of low frequency events such as chromosome loss. Thus, it can be used to compare the efficiency of chromosome segregation of different neocentromeres.
Features shared by neocentromeres with high chromosome segregation accuracy and native centromeres are candidates for properties necessary for kinetochore function. In C. albicans, centromeres co-localize with the earliest, most efficient replication origins, and DNA regions that acquire a neocentromere become the earliest, most efficient replication origins on the chromosome and associate with the origin recognition complex [51]. S. pombe CEN1 also replicates earliest of all loci on chromosome 1 [51], and in several other fungi the sequence features (DNA skew [52]) suggest that replication and centromere function are linked [51]. Additionally, in S. cerevisiae, cohesin binding is enriched at pericentromere regions, and cohesin binding to pericentric DNA requires a functional kinetochore [53]. Neocentromeres with different levels of stability may be useful for examining the roles of DNA replication, cohesin binding, and chromatin structures in promoting accurate chromosome segregation.
In vitro, telomere-telomere fusions produce dicentric chromosomes that subsequently delete alpha-satellite repeats to inactivate one of the centromeres [54]; this process has also been detected in clinical specimens [55]. This is particularly interesting because it may be analogous to events that initiate evolutionary new centromeres [56]. A similar mechanism has been documented in plants: during the evolution of maize and grass genomes, chromosomal rearrangements that produced dicentric chromosomes were resolved by inactivation of one centromere [57, 58]. Drosophila may be a particularly interesting model for examining repositioned neocentromeres that resemble the evolutionary new centromeres in primates [20, 28]. Drosophila cells form dicentric chromosomes following overexpression of CENP-A [45], and mechanisms that determine which centromere remains functional may provide insights into which human neocentromeres are destined to become evolutionary new centromeres.
The overall structure of each kinetochore-microtubule complex is remarkably conserved from yeast to humans, although the number of complexes per centromere varies substantially, and some kinetochore proteins have specialized roles. S. cerevisiae and other organisms with point centromeres canonically contain one kinetochore-microtubule complex per centromere [59, 60], and organisms with larger kinetochores, such as fission yeast, have multiple iterations of essentially the same kinetochore-microtubule complex [59]. There are some kinetochore proteins found specifically in organisms such as S. cerevisiae with point centromeres (e.g., Ndc10) and others found specifically in organisms such as S. pombe and humans with regional centromeres (e.g., CENP-H/Fta3) [12], but overall the sub-complexes that comprise the kinetochore are conserved in composition and architecture.
Recently, tethering experiments with kinetochore proteins have defined the minimal requirements for centromere function, elucidating multiple mechanisms that can result in kinetochore assembly and, by implication, neocentromere formation. Interestingly, these requirements appear to differ in different organisms. For example, in S. cerevisiae, tethering Ask1, a member of the fungal-specific Dam1 complex, forms a synthetic kinetochore and improves plasmid segregation [61, 62]. In the Xenopus egg extract system, CENP-A nucleosomes recruit other kinetochore proteins, resulting in kinetochore-microtubule formation [63]. In fact, the 6 C-terminal amino acids of CENP-A when fused to the rest of conventional H3, are sufficient to bind CENP-C and to recruit the other kinetochore components [63], suggesting that CENP-C may be sufficient to initiate Xenopus kinetochore assembly. Similarly, the Drosophila CENP-A protein, CID, is sufficient for kinetochore formation when CID is tethered to LacO operator sequences and thereby incorporated (together with other histone proteins) into previously non-centromeric chromatin [64].
In humans, overexpression of CENP-A alone is not sufficient to form complete kinetochores [65], although implications of this negative result are unclear. Interestingly, several different routes to kinetochore assembly in human cells have been demonstrated using elegant tethering experiments to induce synthetic kinetochores. First, alpha-satellite DNA, presumably via binding of CENP-B can recruit kinetochores to human artificial chromosomes (HACs) [66]. However, HAC formation is inefficient, occurring only in a small number of cell lines [67]. Second, tethering of HJURP, a CENP-A chaperone initially identified as a Holiday JUnction Recognition Protein, to DNA promotes CENP-A incorporation and the specification of centromeric chromatin in human cells [68]. Third, tethering both CENP-C and CENP-T to the same DNA nucleates kinetochore-microtubule attachments and partially rescues an acentric chromosome [65]. Such tethering experiments provide insights into the order of assembly events and the dependencies involved. For example, tethering of both CENP-C and CENP-T bypasses the requirement for CENP-A, supporting the idea that a normal function of CENP-A is to recruit CENP-C and CENP-T, which together direct kinetochore assembly. Accordingly, such experiments may reveal the proteins most important for neocentromere formation and tethering or engineering the levels of critical kinetochore components may facilitate the efficient production of human artificial chromosomes.
Centromere flexibility also manifests through variation in the amount of CENP-A bound per centromere and in the number of kinetochore-microtubule protein complexes assembled per centromere DNA region, defined here as the kinetochore complex number. Kinetochore complex number varies greatly on evolutionary time scales. Kinetochore complex number can also change rapidly as a physiological response to alterations in the centromere DNA or levels of kinetochore proteins.
The amount of CENP-A bound to the centromere DNA defines the region with the potential to assemble kinetochore complexes and to bind microtubules. Centromere regions in most organisms contain a mix of CENP-A and histone H3 nucleosomes [69]. Even organims with small point centromeres, may contain a mix of CENP-A and H3 as recent evidence suggests that there may be 2–4 CENP-A nucleosomes per centromere in S. cerevisiae [70, 71], although this is controversial as there appears to be one major CENP-A nucleosome [72, 73], and other CENP-A nucleosomes may be randomly interspersed with H3 nucleosomes in the surrounding region [70].
In humans, CENP-A binds to alpha-satellite tracts, the length of which varies between chromosomes and between individuals. The amount of CENP-A bound to centromere DNA also changes in proportion to alpha-satellite tract length: longer repeats are associated with more CENP-A (Fig. 1a) [18]. Intriguingly, CENP-A and kinetochore protein levels are higher in cancer cells than in non-transformed somatic cells [10, 74, 75]. The amount of CENP-A bound to centromere DNA increases with increasing cellular CENP-A levels, generated either by transformation of a tissue culture cell line with oncogenes or by direct overproduction of CENP-A (Fig. 1b) [18]. Importantly, the CENP-A binding remains restricted to the alpha-satellite repeat tract.
In fungi, overproduction of CENP-A also alters kinetochore complex number, including increased numbers of kinetochore proteins and increased numbers of spindle microtubules at C. albicans centromeres (Fig. 1c) [76, 77]. Here, too, the larger kinetochore remains limited to within a core region of centromere DNA [76]. In C. albicans, as in fission yeast [59, 78], upon CENP-A overexpression [76], H3 binding to the centromere region decreases, suggesting that extra CENP-A nucleosomes can displace canonical nucleosomes. Because experiments with C. albicans CENP-A overexpression were performed on populations of cells [76], whereas experiments with human CENP-A overexpression analyzed individual cells [18], it is likely that the proportion of centromere DNA bound by CENP-A relative to H3-containing nucleosomes increases upon overproduction of CENP-A in humans as well. Flexibility in the precise positioning of CENP-A nucleosomes in individual cells would result in an increase in CENP-A binding per basepair in the population and a resulting increase in the number of kinetochore-microtubule attachments with potential accompanying changes to chromosome segregation.
Surprisingly, in humans and in C. albicans, cells engineered to express excess CENP-A recruit extra kinetochore complex proteins per centromere, yet in both cases chromosome segregation appears to be normal [18, 76]. Thus, the number of kinetochore complexes, and presumably microtubules bound, per centromere does not have an obvious effect on kinetochore function. However, the requirement for other kinetochore and cell cycle components may be altered by changes in kinetochore complex number. For example, increasing the kinetochore complex number from ~1 per centromere to >1 per centromere in C. albicans reduces the requirement for the Dam1 complex [76], consistent with the role of the Dam1 complex as a processivity factor for attachment of kinetochores to microtubules (Fig. 2a). Overexpression of CENP-A in transformed cancer cells may be a driving force for aneuploidy. Retinoblastoma protein (pRb)-depleted cells have elevated CENP-A, altered expression of many spindle checkpoint proteins, and high levels of aneuploidy. Interestingly, reduction of CENP-A to normal levels decreased the severity of mitotic defects in pRb-depleted cells [75]. Thus, altering CENP-A levels in combination with conditions permissive for cancer may have a profound effect on chromosome stability.
In organisms with multiple kinetochore-microtubule attachements per centromere, decreases in kinetochore complex number are also observed. Human kinetochores can function with just a small fraction of the normal CENP-A bound [79, 80]. Thus, human cells with multiple kinetochore-microtubule attachments per centromere have a buffering mechanism that permits chromosome segregation even when CENP-A levels are decreased. Interestingly, decreased levels of kinetochore proteins cause different responses in different cell types. For example, depletion of CENP-A protein in fibroblasts eliminates kinetochore function, while the same knockdown permits cell cycle progression in human pluripotent stem cells, presumably because they have stored CENP-A mRNA, further protecting them from CENP-A depletion [81].
An open question is whether increased kinetochore complex number alters the requirement for checkpoint pathways. In C. albicans, the reduced requirement for the Dam1 complex in cells with larger kinetochores suggests that multiple microtubule attachments per kinetochore increase the probability of a chromosome successfully attaching to a spindle pole. However, multiple microtubules per centromere also could increase the probability of merotelic attachments, where a single centromere attaches to both poles potentially causing chromosome missegregation. Accordingly, error-correction pathway components, such as Aurora-B-like kinases may have increased importance in cells with increased kinetochore complex number (Fig. 2b). Future studies combining altered levels of CENP-A and other kinetochore proteins should reveal the interactions between mechanisms regulating the number of kinetochore-microtubule attachments and checkpoint pathways.
We propose that centromere DNA, kinetochore position, and kinetochore complex number are more flexible that previously appreciated, because adaptation requires that chromosome segregation fidelity be maintained despite changes in cell physiology that alter kinetochore protein levels or disrupt native centromere DNA. The contribution of DNA sequence, if any, to regional kinetochore function is likely to become clearer as larger numbers of individuals are analyzed via high-throughput sequencing and as tools for the analysis of repetitive DNA emerge. In addition, model organisms with regional centromere sequences that are readily manipulated will reveal possible relationships between centromere sequence flexibility and kinetochore function [47, 76, 82].
DNA sequences or structures that support kinetochore assembly may also become evident as more neocentromeres are generated in model organisms. Identifying the underlying sequences of additional neocentromeres in clinical samples [38, 39], coupled with the development of techniques to manipulate human tissue culture cells to form neocentromeres, will provide a more complete picture of the mechanisms by which neocentromeres form. For example, overexpression of CENP-A alone is not sufficient to form ectopic centromeres in human cells when the native centromere is present [83]. However, whether extra CENP-A or other kinetochore proteins facilitate neocentromere formation in the absence of tethering remains to be determined.
Kinetochore complex number flexibility appears to be a common feature shared across diverse species. For example, when CENP-A is overproduced, kinetochore complex number increases in fungi and humans [18, 76]. Changes in kinetochore protein levels are associated with cancer cells [10, 74], which are particularly prone to alterations in kinetochore position [36] and to increased kinetochore complex number [18]. Whether chromosomes with altered kinetochore positions and/or complex numbers are more prone to segregation errors or are more dependent on specific checkpoint pathways also remains to be determined. Thus, larger kinetochores, combined with secondary stresses that increase the numbers of merotelic attachments, may contribute to chromosome instability in cancer cells.
The study of centromeres and kinetochores is advancing very rapidly, and models of structure and function are evolving much more quickly than the centromeres themselves. Initial mechanistic insights were based on studies of point centromeres, which have specific sequence requirements and were thought to adhere to a rigid structural model; yet recently, the composition, number and position of CENP-A-associated nucleosomes in S. cerevisiae has become controversial [70, 71, 73, 84, 85]. Flexibility in centromere sequence, kinetochore position and complex number is emerging as an inherent property of centromeres in virtually all eukaryotes. Thus, future models will need to account for this remarkable flexibility. The next few years will likely reveal many insights into mechanisms that segregate chromosomes under a range of conditions, including physiological and genetic stresses.
We apologize to authors whose work we did not have room to cite due to space limitations. We thank Berman lab members for helpful discussions and Simon Chan, Barbara Mellone, and Beth A. Sullivan for very useful comments on the manuscript. This work is supported by a Ruth L. Kirschstein NRSA Fellowship F32 AI800742 and a Postdoctoral Fellowship, Grant #PF-12-108-01-CCG from the American Cancer Society to L.S.B. and by NIH/NIAID AI075096 to J.B.
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Summer Music from Greensboro kicks off TuesdayJuly 31,2013
Staff Report
GREENSBORO — Summer Music from Greensboro will present violinist Jaime Laredo and cellist Sharon Robinson, Vermont's first family of music, joined by Montpelier flutist Karen Kevra, at 8 p.m. Tuesday, Aug. 6, at the downtown United Church of Christ.
The program includes Ravel's Sonata for violin and cello, the Handel-Halvorsen Passacaglia, Hindemith's Eight Pieces for solo flute, C.P.E. Bach's Duo in e minor for flute and violin, and Haydn's Divertimento III in C major for flute, violin, and cello.
The storybook romance of the Bolivian-born violinist Jaime Laredo (Vermont Symphony Orchestra's music director) and American cellist Sharon Robinson has, over the past 35 years, been expressed in every facet of their lives. Joyous and sophisticated music making, both separately and together, has earned them many devoted fans the world over. Combining international careers and a multifaceted home life both in Vermont and Cleveland, theirs is a bond of both close personal affection and strong dedication to music.
Tickets are $20 (free for 18 and younger); go online to to www.summermusicfromgreensboro.net.MORE IN This Just In
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