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The Department of Defense announced today the identities of two soldiers who were killed while supporting Operation Iraqi Freedom.
Cpl. Tomas Sotelo Jr., 20, of Houston, Texas, was killed on June 27 in Baghdad, Iraq. Sotelo was traveling in a convoy when a rocket propelled grenade struck his vehicle.
Sotelo was assigned to Headquarters Troop, 2nd Armored Cavalry Regiment, Fort Polk, La.
Sgt. Timothy M. Conneway, 22, of Enterprise, Ala., was injured on June 26 in Baghdad, Iraq and he later died of wounds on June 28. Conneway was traveling in a government vehicle when an explosive device detonated and struck the vehicle. Conneway was assigned to the 3rd Battalion, 75th Ranger Regiment, Fort Benning, Ga.
Both incidents are under investigation.


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Basketball: VC men fall out of first place with 76-71 loss to Hancock
Pirates get pushed around by first-place Hancock
The Ventura College men's basketball team didn't so much lose control of the Western State Conference on Wednesday night as much as it was taken away.
Forward Demetrius Thomas scored 19 points, guard Khufu Najee had 18 points and Seth Kena had 15 as No. 9-ranked Hancock took control of the WSC North by holding off No. 5 Ventura, 76-71, at the VC Athletic Event Center.
"It was (Kena's) heart that won the game," Ventura coach Joey Ramirez said. "He came in and pushed us around. They bullied us."
Ivan Dooley scored 19 points, Zalmico Harmon had 10 points and Jaylin Airington had nine points and eight rebounds as the No. 5-ranked Pirates (22-6, 6-2) fell into a second-place tie with Los Angeles Pierce ahead of Saturday's rivalry game with Moorpark.
"We've got to move on and focus on Saturday," Ramirez said. "Everyone in our conference is looking at one game at a time."
Najee Salaam returned after missing last Saturday's trip to Santa Barbara City with a sprained ankle and had 15 points and 10 rebounds.
Thomas, Hancock's star forward, didn't start the game. Yet the Bulldogs scored the first six points.
Sophomore Eric Mitchell spurred the Pirates' immediate 6-0 response.
When Thomas finally entered the fray, the 6-foot-8 forward keyed an 18-5 run as Hancock took control, 24-11.
Ventura dug out of a double-digit deficit to hammer Hancock, 78-58, in Santa Maria on Jan. 19. But, this time around, the comeback was much tougher because the Pirates went 35 minutes without a 3-pointer.
"You can't live or die by the 3," Ramirez said.
Down 33-26 at the half, Ventura started the second half well. Dooley's 3-point play and Salaam's finish closed the gap to 37-35, but Thomas buried a 3 and the Pirates never pulled even.
VC women cruise to 75-33 victory
Cynthia Hernandez had 17 points, Hanna Burge had 10 points and seven rebounds and Lisa Maria Sanchez had eight assists as the No. 2-ranked Ventura College women's basketball team rolled to a 75-33 win over Hancock at the VC Athletic Event Center.
Freshman forward Adrianne Sloboh, a Royal High product, became the 10th player in school history to pull down 16 or more rebounds in a game.
Freshman Carly Maciel had eight points, including back-to-back 3s early in the second half.
Playing without two injured starters, guard Imani Dixon and forward Ashanti Sweezer, first-place Ventura (25-3) won for the 18th time in 19 games, improving to 7-1 in WSC North play
"It makes a difference," Ventura coach Ned Mircetic said. "It's a tough thing to change lineups mid-stride."
Meanwhile, in San Luis Obispo, sophomore Aria Johnson made two free throws in the final minute as Cuesta (18-9, 6-2) toppled Santa Barbara City, 54-52, to move into sole possession of second place.
Ventura will travel to San Luis Obispo on the final day of the season, Feb. 23.
Moorpark women beat Pierce
Riley Abraham had 27 points and 11 rebounds, Allison had 15 points and eight rebounds and Camilla Chun had seven points and seven rebounds as the Moorpark College women's basketball team beat Los Angeles Pierce, 61-50, at Raider Pavilion.
Avenging a loss in Woodland Hills on Jan. 19, the Raiders (16-11, 6-2) won for the third time in four games to move within a game of third-place Santa Barbara City.
Moorpark visits cross-county rival Ventura on Saturday.
Moorpark men fall to 1-7 in WSC
Despite 19 points from point guard Shayan Boloor, the Moorpark College men's basketball team fell to Los Angeles Pierce at Raider Pavilion.
Spencer Levy had 14 points, D.J. Saul had 12 and Sahdi Odu added 10 for the Raiders (6-16, 1-7).
Donte Williams scored 29 points and Victor Evans had 24 for Pierce (17-8, 6-2), which turned a 28-28 game into a 57-46 halftime lead.
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Tourist/Visitors Information
Hours of Operation/Ticket Information
Historic Pensacola
We are open Tuesday-Saturday, 10:00 a.m. to 4:00 p.m. Tickets for the complex may be purchased at the Tivoli High House Shop, the T. T. Wentworth, Jr. Florida State Museum, the Pensacola Children's Museum, and the Voices of Pensacola Multicultural Center in Downtown Pensacola. For more information about the Historic Pensacola museum complex and UWF Historic Trust, call 850-595-5985.
Admission Fees:
- Adult Ticket - $6.00
- Child (ages 3-14) - $3.00
- Discount Ticket (Senior Citizens 65+, Active Military and Dependents, AAA Member) - $5.00
- Members FREE
Historic Pensacola Village
Tour Times in Historic Pensacola Village: 11:00, 1:00, and 2:30.
Guided Tours
We offer guided tours of our historic homes and buildings three times daily at 11:00 a.m., 1:00 p.m., and 2:30 p.m. Guided tours are part of admission ticket. Tours will last approximately 1 to 1.5 hours.
Visitors will receive an in-depth tour of the 1805 French-Creole Lavalle House, the 1871 Dorr House, the 1832 Old Christ Church, and the 1890 Lear-Rocheblave House.
Guided Tour of the Barkley House
Ticketed visitors to Historic Pensacola Village may now enjoy a 45-minute walking tour of a portion of the Pensacola Historic District and the Barkley House. Tour guides will lead guests past historic structures along Zaragoza Street, highlighting significant architecture and local history. Located on Pensacola Bay, the circa 1825 Barkley House is the oldest surviving example of a high-house in the city. Visitors will have the opportunity to learn about the Barkley family, catch a glimpse of Pensacola during the American Territorial Period and enjoy the vista of Pensacola Bay from this unique home. Tours leave from the Tivoli High House at 2 pm.
Tours are subject to change due to renovation and repair, please contact us for the most current and updated information. You may contact the Tivoli High House Store at 850-595-5993 or our administrative office at 850-595-5985.
T. T. Wentworth, Jr. Florida State Museum
Our flagship museum in the UWF Historic Trust complex houses three floors of exhibit space. It is open Tuesday through Saturday, 10 a.m. until 4 p.m. Located at 330 South Jefferson Street. For questions, contact the T. T. Wentworth, Jr. Florida State Museum at 850-595-5990.
The Wentworth Museum is open for Half-Price Sundays from 12:00 until 4:00. Admission is half-price and is good for seven days. For more information contact 850-595-5990.
Pensacola Children's Museum
The Pensacola Children's Museum is located in the Arbona Building at 115 East Zaragoza Street. The Museum is on Zaragoza Street between Jefferson and Tarragona Streets, directly behind the Pensacola Cultural Center. The museum is open Tuesday through Saturday from 10 until 4. It is a hands-on exploration of Pensacola history for ages 2 to 13. For more information, contact the Museum at 850-595-1559.
Voices of Pensacola presented by Gulf Power
The Voices of Pensacola Multicultural Center is now open at 117 East Government Street. This multicultural center highlights Pensacola's diverse history and rich cultural heritage. It is open Tuesday through Saturday 10 until 4. For more information, contact the Multicultural Center at 850-595-5840.
How to Find Us
Get driving directions, view nearby restaurants and hotels at Google Maps.
From 1-10, Take Exit 12 to I-110 South. Exit I-110 at 1C (Garden Street). Turn left at the first light (onto Tarragona Street). Follow Tarragona Street four blocks south to Church Street. The parking lot is on the right. Historic Pensacola Village will be to your left and the T. T. Wentworth, Jr. Florida State Museum will be to your right.
From Hwy 98 East of Pensacola, Cross the Pensacola Bay Bridge exiting left on Bayfront Pkwy. Follow Bayfront to Tarragona Street; turn right, north two blocks to Church Street. The parking lot is on the left. Historic Pensacola Village will be to your right and the T. T. Wentworth, Jr. Florida State Museum will be to your left.
- T. T. Wentworth, Jr. Florida State Museum
- Museum of Commerce
- Tivoli High House - Ticket and Information Center
- Manuel Barrios Cottage
- Museum of Industry
- Julee Cottage
- Lavalle House
- Lear-Rocheblave House
- Old Christ Church
- Dorr House
- J. Earle Bowden Building - Administrative Offices
Tell us what you think!
Please let us know how much you enjoyed your trip to Historic Pensacola with Tripadvisor.
- Write a review of Historic Pensacola Village


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Thursday, January 17, 2013
Robin Scott Shannon and Margaret Morgan Shannon of Summerville, South Carolina are pleased to announce the engagement of their daughter Jessica Michele Shannon to Justin Michael Sims of Prescott, Arizona. Mr. Sims is the son of Michael Lee Sims and Brandie Lee Sims of Prescott.
The bride-elect is a 2010 graduate of Summerville High School and attends Trident Technical College. She is employed with Target in Summerville.
The groom-elect is a 2008 graduate of Prescott High School in Arizona and attended Yavapai Community College in Arizona. He is a member of the United States Navy.
The bride-elect is the granddaughter of Mary Lucille Niesslein and the late Kenneth Ray Morgan, Patricia Rose Morgan, Mickey Deloris Morgan, Leroy Eugene Shannon and Virgil Alvin Niesslein all of Summerville.
The groom-elect is the grandson of Cole Luther Kite and Tawny Lynn Kite of Black Canyon City, Arizona; and Peggy Lee (Sims) Kemp of Tucson, Arizona.
The wedding is planned for May 18, 2013 at Boone Hill United Methodist in Summerville.
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Grandparents Day
(Pittsburgh) (September 2011)— A day spent with your grandkids is a memory that you keep forever, so why not it spend at the Pittsburgh Zoo & PPG Aquarium, Sunday, September 11, for UPMC For Life. Grandparents Day. Grandparents are admitted free to the Zoo when accompanied by their grandchildren. Grandparents also receive one free-ride-all-day tram bracelet, and a free coffee or tea. The day is sponsored by UPMC For Life
The Pittsburgh Zoo & PPG Aquarium is open year round. For directions, hours, tickets and group sales information, call 412 665-3640. Visit the Zoo's website at www.pittsburghzoo.org The Pittsburgh Zoo & PPG Aquarium is an accredited member of the Association of Zoos and Aquariums Zoo. Look for the AZA logo whenever you visit a zoo or aquarium as your assurance that you are supporting a facility dedicated to providing excellent care for animals, a great experience for you and a better future for all living things. For more information visit www.aza.org.


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The 2012 International Green Construction Code (IgCC) is expected to help make newly constructed or remodeled buildings more efficient, reduce waste, and have a positive impact on health, safety, and community. Developed with input from code and construction industry professionals, environmental organizations, policy-makers, and the public, IgCC is for communities pursuing sustainable construction above and beyond the traditional level of International Code Council codes. www.iccsafe.org
Sponsored CEU
This course is registered for 1 HSW hour with AIA and will focus on gates and gate...
Building Codes
William "Jay" Hall returns to the Portland Cement Association (PCA) as manager of codes...
Codes and Standards
The revision (ANSI/AWWA C651-14) is the result of multiple years of review, research,...
International Code Council
If you know an exemplary individual or group whose contributions to building safety,...
PUBLIC WORKS Staff
Please read our Content Guidelines before posting.
Copyright ©
2015 Hanley Wood Media, Inc. All rights reserved.


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The American Austin Automobile Company was founded in 1929 in Butler Pennsylvania. As the name would imply they built mildly Americanized versions of the... More
Bantam 1939 roadster, finished in red, white coves, red interior, frame-off, every nut & bolt by marque restorer, AACA Sr, 1st, 3-spd, 4-cyl, 20... More
BANTAMS: 1930-41 Austins, American vehicles, parts, largest active source; MC, Visa, Discover, all services. 2439 S Bradford St, Allentown, PA 18103, PH/FAX: 610-791-4497; www.austinbantam.com;... More


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C&N Sayre Location Expanding HoursPosted Wednesday, 29 July 2015 19:05 | Filed under C&N News
WELLSBORO, PA - The Citizens & Northern Bank office in Athens remains closed due to flooding. To accommodate customers impacted by the recent flooding, the bank's Sayre office at 1827 Elmira Street is temporarily offering extended hours. The Sayre branch office is now open 8:00 a.m. – 6:00 p.m. Monday-Friday until further notice. The Sayre branch will maintain regular Saturday hours of 8:30 a.m.-noon.
"Many of our customers are struggling to clean up their homes and businesses and can't always get to the bank during normal business hours," said Thomas Rudy, Jr., C&N EVP and Director of Branch Delivery. "We want to make sure we're accessible when those customers need us. Sayre is our closest location to our Athens branch and we're hoping the extended hours will prove more convenient for those affected by this devastating flood."


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Jason Purvis
A dad's journey of discovery and (many) diapers
Monthly Archive for November, 2010
Talkin' P-U on the TU
Whoever said that babies that are breastfed don't have stinky diapers is just plain out of their mind. Yesterday morning, I went in to get Abbi out of her crib. I walked into the room and was blown away by the horrific stench that was emanating from my daughter. I had to rub my eyes […] [Read More]
Looking back
Before I started blogging with the TU, I did some writing on my own where very few followed along. Em reminded me of a post that I had once written. It was a letter to Abbi (we didn't know she was going to be a she at that time). It was written in the 34th […] [Read More]
Sunday Snapshots 11-28-10
We did quite a bit of spitting up on Thanksgiving and Black Friday. The turkey bib only lasted a few minutes. [Read More]
Em on Breastfeeding
Today, Em is offering insight to the whole breastfeeding thing. From my point of view, breastfeeding is miraculous, appreciated, at times difficult, and something I will never fully experience. I'm okay with that. My role is to be supportive, feed Abbi from a bottle after Em pumps, and stay clear of shooting streams of milk. That stuff sure […] [Read More]
2 short months
I am so happy to say that Abbi is 2 months old today! Well, depending on how you measure time, she is either 2 months or 8 weeks and a few days. I'm still confused about that. It's so hard to believe how much she has grown. Remember this little baby? And now she's transformed […] [Read More]
The shot(s) heard around the Capital Region
If you were anywhere in the Capital Region earlier today and heard a blood curdling scream, you can rest assured that everything is now okay. Abbi had her two month wellness visit which included her first round of vaccinations. The wellness part of the exam went fine. Abbi weighs exactly 12 pounds and is in […] [Read More]
Freshman 15
Freshman 15…probably a title you'd expect to see over at This is College. It's no secret that many young men an women can, and do, gain 15 pounds (or more) that first year. They go from high school where they may have been active in sports, had home-cooked meals and were at the top of their […] [Read More]
Show More


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Surviving Holiday Beer Season: A Cold-One for the Rest of UsPosted 3 years ago
Rihanna: "Obsession of the Year" YES! we agreePosted 3 years ago
Star Trek Into Darkness Movie ClipPosted 3 years ago
Call of Duty: Black Ops 2 zombie trailerPosted 3 years ago
First Image of Hugh Jackman as 'The Wolverine'Posted 3 years ago
Superman Villain Black Zero to Appear in "Man of Steel"?Posted 3 years ago
'The Hobbit: An Unexpected Journey' The 2nd Full Trailer ReleasedPosted 3 years ago
Robocop Set photos, Joel Kinnaman and Abbie Cornish on the setPosted 3 years ago
'Dark Knight Rises' 13-Minute FeaturettePosted 3 years ago
Judge Dredd Movie Update: Photos, Poster and Plot InfoPosted 3 years ago
'Dark Knight Rises' 13-Minute Featurette
Got your google alerts set for anything new to come out of Camp Nolan? And you cant wait to see Dark Knight rises on the 20th of July? Well you dont have to wait that much longer but to tease...
- Posted 3 years ago
NEw Dark Knight Posters + Bonus Cat women poster.
New Dark knight Cat Women poster. Apart of a set of Main character posters, Check out the rest Below. Here's the official synopsis for The Dark Knight Rises: Warner Bros. Pictures' and Legendary Pictures' "The Dark Knight Rises" is...
- Posted 3 years ago
Funny avengers photos
Incoming search terms:funny avengersfunny avengers picturesfunny avengers picsavengers funnyavengers funny picturesfunny avengers tumblrfunny avenger picturesfunny avengers comicsfunny avengers pictures tumblrfunny avengers photosavengers funny comicsavengers funny pics
- Posted 3 years ago
Dark Knight Rises Trailer #2 and The Prologue Video!
OK UPDATE NEW LINK AND VIDEO OLD ONE DELETED KEEP CHECKING BACK! So this weekend the official Trailer for the Dark knight rises has hit Theaters across the country. Although the full Quality release has yet...
- Posted 4 years ago
Dark Knight Rises Bane Poster
HOLY YOU GOT YOUR FACE CRACKED BATMAN! Hey Guys check out this poster to just hit the net of Bane and what seems to be Left of the Dark knights Helmet. Its not the tradition "Breaking of...
- Posted 4 years ago
Dark Knight Rises in New york – Occupy Trump Tower
Batman Occupies Trump Tower with a Hot Sports Car that looks like our dream car of the week, The "Lamborghini Reventón". Not sure exactly whats going on in the scene hopefully the Dark knights car isn't being towed … that would suck lol....
- Posted 4 years ago
Sneak Peek- Dark Knight Rises – Bat Wing!
Its nice to see all the Leaked Videos out there on the web for this upcoming film. But ether C. Nolan dosent care about fans getting a peak and plans to change most of what we think...
- Posted 4 years ago


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I'm developing a site with a horizontal menu that has two sub-levels. The second level appears below the primary item. Then when you hover over that, the third level flies out to the right of that `
I'm used to seeing the third level align next to its parent. But is there a way to get the third level to align to the top of the parent's `
Yes. Instead of giving the parent li a 'position:relative', give the parent ul a 'position:relative'. That would align the child ul (I'm assuming you're using absolute positions for the sub-levels') with the parent ul, instead of the parent li. An absolute position element is positioned relative to an element that has a relative position in which it is placed.
Edit: Wrong answer sorry, since I realize that you're talking about the third level submenu.
I'm not sure if there's a better way to do this, but you could use a div as the drop down element(thus absolutely positioned). Then the ul placed inside the div could have a relative position. The third level position would then be absolutely positioned in relation to this ul.
Let me know if you need a codepen example.
You must be logged in to reply to this topic.


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2:51 pm
Mon November 11, 2013
'Honor Flights' Race To Bring WWII Vets To D.C. Memorial
Originally published on Mon November 11, 2013 3:52 pm
More than 16 million American's fought in World War II. There's only about a million of them who are still alive and they're all older than 80. Hundreds are dying each day. A non-profit group has made it their mission to honor these remaining veterans by flying them to Washington, D.C., to visit the World War II memorial. The trip isn't something many veterans at this age can do — or afford — on their own. Since the first "Honor Flight" in 2005, groups in almost every state have followed suit and more than 100,000 vets have taken the journey.


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Front elevation
Plan detailsPlan type: Craftsman
Bedrooms: 3
Bathrooms: 2.5
1st Floor sq ft: 2,489
2nd Floor sq ft: N/A
Total heated sq ft: 2,489
Bonus room sq ft: 327 (in addition to total heated sq ft)
Width: 66'-6"
Depth: 69'-0"
Garage: 2 car
Ceiling hts: 1st floor - 9'-0" 2nd floor - 8'-0"
Foundations: Basement
Modify this house plan
Brick, stone, cedar shakes, and board-n-batten siding blend with great harmony in this modern craftsman home plan. Big cub appeal but, cozy on the insde. A split bedroom house design allow for parents to keep distance from noisy teenagers. In addition to the three bedrooms, there is a bonus room which can be converted to a fourth bedroom.
Similar designs: DE012 | DE014 | DE017B | DE072 | DE082 | DE099
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Peter Steele 1962 - 2010
For the few Type O negative fans here ..... It's a sad day.
Re: Peter Steele 1962 - 2010
Rock In Peace, Mr. Steele.
Wolfmoon.............Thanks, for letting us know.
Re: Peter Steele 1962 - 2010
I'm a huge Type O Negative fan.
I've been pretty crushed by the news.
Re: Peter Steele 1962 - 2010
I was shocked and bummed out by the news of peters passing.
Re: Peter Steele 1962 - 2010
Yes . . same here. There will never be another like him. Statements from band members below.
Josh Silver: "Peter, My endless source of frustration, (as I'm yours) you have really done it this time. You have changed and touched countless lives through music, comedy and often brutal honesty. You've made life both interesting and irritating and I could not imagine not having known you for 37 years. It still isn't true in my mind but in time I will miss you and the creating that we all endured together. We certainly disagreed constantly and I believe (and hope) we all learned from each other. Should I call you my brother, friend or neighbor? I can only call you Peter (and usually after 2 PM). We laughed at ourselves more times then I can count. Knowing humans are preposterous creatures and I know we reveled in that fact. I will miss you in time, but at this moment your premature departure seems surreal and has pissed me off to no end. Though I never told you that I harbor a deep respect for you, I do. Goodbye my friend."
Kenny Hickey: "Peter Steele was one of the most brilliant and funny personalities in music and it was all for real. Half the time people thought he was joking, but he was actually telling the truth. Part of me died with him."
Johnny Kelly: "It's impossible for me to put into a few sentences what I am feeling at the moment Peter. I'm not sure if I should eulogize or roast you. Both good and bad, we went on one hell of a ride together and sadly, the ride has come to an end. You truly were a unique person. Your music touched many people. Myself included. Whether it was talking about The Beatles, power tools, how Pluto was no longer considered a planet or calling me at 3am asking me to drive to your house to have a fistfight with you, you always kept it interesting. It was a privilege to have been your bandmate. It's something that I will always cherish."
Re: Peter Steele 1962 - 2010
From Bill Ward, Black Sabbath.
Peter Steele / Type O Negative
Posted on Apr.16, 2010, under Misc Items
The flamboyant ambassador of Metal's substratum is dead. I am diverted from my next indicated thing to do. I am compelled to pause and digress and re-listen to one of my all-time favorite bands, Type O Negative. I fall into memorable moments, to enhance my first discoveries of Peter Steele.
I don't want to believe he's died. I've felt in shock since the early morning sad news of today arrived. Peter Steele – a brilliant musician, songwriter, singer has died. His death is a tragedy and heartbreaking. His passing is an enormous loss to the Metal community and to music period.
In music, he challenged life itself in a poetic, prophetic, charismatic and extraordinarily insightful way. A world observer, a teller of tales, a renaissance man, a man from the future, a powerful force in today's maladies and life challenges.
This man will not be forgotten. I raise my goblet of water and praise him. I thank him, and I burn candles and desire incense aromas to fill our house, now illuminated in his honour. Candles burning outside flicker, and the sea crashes onto the shore. We are still. We are in memory.
Peter, you will be celebrated forever.
–Bill Ward
PS: A profound statement.
Re: Peter Steele 1962 - 2010
Re: Peter Steele 1962 - 2010
He was unique, those kind of people that are so messed up but at the same time so brilliant are very few and far between. He had a weird way of looking at things that was so brutally honest and refreshing you couldn't help but agree, no matter how depreciating it was. Life was messed up, so messed up all you could do about it was make fun of it to take the sting off.
Peter has been more than a musical inspiration over the years for me. Type O were one of my first favourite bands when I was younger and I still listen to their music and enjoy it to this day.
I have more respect for him then almost anyone else I have never met and many that I have. His music will always continue to be a huge influence on my life.
Re: Peter Steele 1962 - 2010
An outstanding commitment from a true hardcore fan.
Re: Peter Steele 1962 - 2010
Saw Type O back on October 27th in Atlanta. Peter tossed a water bottle in the audience and it hit me on the head.
Type O is one of my favorite bands..I was looking foward to a new album
RIP Peter.
Re: Peter Steele 1962 - 2010
I had a similar thing happen at 1 of there shows but instead of getting clocked on the head with a water bottle . . . . . it was a roll of toilet paper. It was a Halloween show and they ended the show by throwing out roll after roll of Toilet paper. The place was a mess ;) All in good fun. I'll sure miss those times.
Re: Peter Steele 1962 - 2010
another one bites the dust....... I'm sure he's already jammin in a band on the other side!
Re: Peter Steele 1962 - 2010
Peter Steele 1962 2010
Hehe, ill say its more like JLo & BAffleck, or right now, Brad and Angelina, as both arent/werent married just like Peter & Camilla BradP&JenniferA were married and ended up split..
But yes, in terms of popularity, looking good together etc., yah PGade&CMartin were similar to BradP&JenniferA..
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Narrow your search
The redesigned luxury SUV features a new twin-turbocharged V-6 option and four designer Black Label themes.
Pricing not available
The all-new Lincoln MKC shows interior and exterior styling competitive in the luxury set, along with a stellar range of driver assistance features, but it should go further in insulating passengers from the rigors of the road.
The 2014 Lincoln MKZ Hybrid offers a good set of high tech features that ease driving and increase comfort, but the car falls short of the luxury feel of competitors from the likes of Mercedes-Benz and Lexus.
With excellent fuel economy, a comfortable interior, and really excellent cabin tech, the 2011 Lincoln MKZ Hybrid makes for a near perfect commuter car and general transportation.
The 2011 Lincoln MKX is a vehicle that shows a good deal of potential as a tech car, but its MyLincoln Touch cabin tech suite feels unfinished. Wait for news of a software update.
The 2010 Lincoln MKS is quite a tech cruiser, with features that will thrill any gadget lover while traveling in comfort.
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With three rows of seating, the 2010 Lincoln MKT is a big, powerful cruiser capable of carrying many people in high-tech comfort.
Pricing not available
While we love the 2010 Lincoln MKZ's cabin tech, the next model update should take advantage of more efficient engines currently under development at Ford.
Pricing not available
Lincoln Continental Concept makes it's debut at NY Auto Show.
Lincoln's new Continental Concept is looking lovely on the show floor of the New York International Auto Show. Let's take a closer look.


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This is my collection of things that catch my eye, from interior design and pretty places to good food and generally quirky cool stuff. Come take a look around, leave me a comment and say hello - I'd love to hear from you! Read more about me
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Category Archives: Travel
…Could there be a nicer time to visit? Cherry blossom, sunshine and plenty of pancakes and ice cream. We've just got back from spending a week in the Big Apple. Ever since I first tried AirBnB, I've become hooked – … Continue reading
It looks like all I've been doing is taking one trip after another. That's not untrue, but there are a few disclaimers, namely that a) I was working in between, and b) I've not given myself much time off for … Continue reading
We took a week away to celebrate our first anniversary. Ages ago, we thought we'd go to New York (I even found a little Brooklyn bolthole for us to stay) but boy-oh-boy am I glad we dropped that idea (Sandy. … Continue reading
I'm pretty lucky that I get to review some really nice places for this blog and the latest place I got invited to stay at this weekend, The Crown in Amersham, wins hands-down as my favourite place so far. Hopefully, … Continue reading
This weekend, R & I escaped to the Cotswolds. I was invited to stay at The Old Swan & Minster Mill, in the tiny village of Minster Lovell about half an hour or so from Oxford. And when I say tiny, … Continue reading


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Hundreds opposed to airport development scheme
Posted: December 21, 2012|
HUNDREDS of people have told the city council what they think of plans to allow development on Plymouth airport land.
And the overwhelming majority are strongly opposed to moves by Sutton Harbour Holdings to build a massive new community with shops, offices and homes on the site.
Their comments will be sent to a government planning inspector today as part of Plymouth City Council's area action plan for Derriford and Seaton.
It comes almost a year after Sutton Harbour closed the airport, on December 23 last year.
In October Plymouth City Council was told by the planning inspectorate to reopen its consultation on the draft Derriford and Seaton area action plan after Sutton Harbour submitted a masterplan to build on the airport.
The airport had been protected in the draft area action plan, but was removed by the new Labour city administration, which said its fate should be decided as part of the wider Plymouth Plan.
A council spokeswoman said: "We received a total of 542 representations on the Sutton Harbour plan, of which 521 expressed concerns, six supported and 12 made other comments."
There were also 10 late submissions and another 92 incomplete representations which had incomplete details and were inadmissible but will be forwarded to the planning inspectorate.
The area action plan and supporting information will be submitted to the planning inspectorate today.
A planning inspector will hold a public examination, possibly in March or April next year, and publish a report in the summer.
CONSULTATION: What you said
EDITED extracts from a few of the 542 comments on the council's consultation website about the Sutton Harbour Holdings masterplan to build on the airport site:The site must stand ready to be re-opened as an airport when the economic conditions improve.The city will never forgive SHH or our elected officers if they collude in the destruction of our Airport. – RICHARD GRIFFITHSI am involved with a small business in the … wave energy sector …. My colleagues from overseas view travel access to Plymouth, which should be a future hub in marine renewables, as less than satisfactory. – NICHOLAS STRACHAN I am employed in oil-related work in the Shetland islands and travel by air on a regular basis. I used Plymouth airport in the past 14 years to get to my place of employment.We need an airport for the long term future. – MR V CAIN Although I understand that it has been difficult to operate the airport successfully as a going concern, I am concerned as a small business owner that removing it permanently will cause considerable downturn to the local economy. – MARK FUGEI live at Southway and the last thing I want is this monstrosity on my doorstep. Congestion is already a problem in Tavistock Road and this will only add to it. – EDNA COLES The SHH masterplan … is in the wrong location … Derriford and Seaton are best served by PCC's favoured location [Seaton Barracks].Transportation infrastructure is pathetic, particularly in the case of the PCC option [Seaton Barracks]. Both plans maintain the fiction that they will be adequately served by public transport, walking and cycling or modal change. This is a ridiculous assumption. – RICHARD SROKA Plymouth Civic SocietyI used the airport a lot in the past … to fly … to Scotland and other airports as part of my job in the Royal Navy. Plymouth Airport plays a vital role for the Royal Navy and ships at sea on the South Coast both British and foreign. – ALAN ROWE Since the property developers closed the airport at the end of 2011 Plymouth has been withering on the vine. This Sutton Harbour plan should be rejected out of hand for what it is - a ... lunge for profits without any consideration of what is actually beneficial to Plymouth. – JASON HARGREAVES Please retain the land at Plymouth Airport as an Airport – once it's built on its gone for ever. Don't let Sutton Harbour Holdings [profit] from a poorly conceived lease! – PHIL BROWN University College Plymouth St Mark & St JohnHow many times do you want the communities and people to say give us our airport back? – CHRIS BURTI do not support the application by Sutton Harbour Holdings to develop the airport site. The road infrastructure is not suitable for the envisaged increase in cars and lorries. The A386 is already gridlocked as is Novorossisk Road at peak times. – JULIE POOLEAny development of any size in the north of the city would be hugely detrimental to the existing city centre. There are about 15% of shops vacant in the city, most of which will never re-open. – NICHOLAS RINGThe college "firmly believes that the airport is essential to the continued development of Plymouth students' employment relationships and the global economy". – BARBARA JONES, Deputy principal Plymouth College of ArtIt is sad to see the lack of future thinking in the old council and new council to sell off the airport to a company such as Sutton Harbour. They have consistently put hurdles up against anyone wishing to operate an air service from Plymouth. – SHAUN PARNELL, Totem Timber Plymouth Sutton Harbour Holdings should be able to proceed with their plan for the airport … they should be forced by Plymouth council to include at least 20% affordable housing available to locals. The only people that could afford to use the airport all year round were the people that did not have to pay for their air ticket, ie. they could claim the fare back on company expenses. 'Viable' collected 37,000 signatures to save the airport, but when Tudor Evans leader of the council asked people to write to the Prime Minister through him he only got 400 letters and e-mails. The airport land should not stand empty for any length of time just waiting for someone to come along and operate the airport, which is not going to happen. SHH should be allowed to implement their housing plan for the Airport site. – KEITH BODKIN I am writing to you on behalf of the older and less able people of Plymouth. It is difficult to manage to get on and off a train with luggage when you walk with sticks or crutches. Having used the airport yearly in the past I know how much easier it is to go to Plymouth Airport, hand in your luggage and have a quick, safe journey to Gatwick where you can be met by a wheelchair or buggy and be taken to collect your bags, ready for your onward journey. – LINDA SHIPPINGWe are utterly opposed to any plans to reinstate the airport.1) Air travel is not the future. It is very polluting, inefficient and expensive. To support such development contradicts Plymouth's declared policies on sustainability, low carbon communities. 2) Air travel is likely to be taxed via international agreement in the near future. 3) There is already a good airport just 40 miles away in Exeter. Good public transport links could be made to this airport if necessary. 4) Sustainable businesses that provide long term jobs will not be reliant on an airport in the city. 5) The previous airport did not make money … and was only ever used by a small minority. 6) We should instead campaign for excellent, affordable and sustainable public transport links via greatly improved bus and rail services. 7) The city should actually make a feature of the fact that it is a green progressive city that does not need an airport. – ANDREW PRATT Plymouth Green Party
Politicians: Speak up
THE HERALD is calling on city politicians to nail their colours to the mast over the future of the airport site.This newspaper has made its position perfectly clear – the airport site must be protected from development to preserve it for a time when we will once again need an air link.Now we are asking our leaders to speak as clearly on the issue and say whether they believe the Roborough site should be protected from the bulldozers for the foreseeable future.We are asking the straightforward question: Should the land at Roborough be retained as an airport for the foreseeable future, or should development be allowed on the land?Online at www.thisisplymouth.co.uk/pcalist we have published the names of those who have answered with a clear yes. It is far from being a comprehensive list and over the coming weeks we are calling on all councillors to add their name to the list and state their position. Ultimately we will publish the full list of responsesCouncillors wishing to state their position to be added to the list should contact Keith Rossiter on 01752 765529.


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Limited edition pre-order of 12″ vinyl with free digital download. Black vinyl with 4 color center label and plain jacket with center cut hole. **Please note vinyl is for pre-order until August 31st, 2015, ships September, 2015. If a minimum of 40 pre-orders are not met, all orders will be canceled and money refunded. Please include your email in your order, thanks!**
Includes unlimited streaming of Root Down via the free Bandcamp app, plus high-quality download in MP3, FLAC and more.


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Originally published on Wed December 19, 2012 11:03 am
Many pregnant women and potential adoptive parents are bypassing traditional adoption agencies and connecting online. This trend raises a number of ethical concerns about the regulation of the online process and interactions between birth mothers and hopeful adopters.
View the discussion thread.


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Campus Crusade for Christ, the parachurch ministry of Bill and Vonnette Bright, has decided to drop the name of Christ from their organization. You can find the news coverage here. According to the article, 9 percent of Christians and 20 percent of non-Christians were alienated by the name, "Campus Crusade for Christ." A representative for Campus Crusade
for Christ, now called Cru, said that the name brought the Middle Ages to mind, a time when Christianity was forced upon people.
Why not drop "crusade" from the name? Militarily, the crusades might have been necessary, but theologically, they were a disaster. Why invoke the crusades in an effort to promote the gospel? At the same time, if a motivating factor behind the name change is not wanting to alienate people by the name of Christ, then there are bigger problems involved. Surely we herald the forgiveness of sins and the life-giving work of Christ that comes by grace alone through faith alone in Christ alone as we proclaim the gospel. But there is no amount of sugar that can be placed on the lip of the bitter cup of salvation that can somehow sweeten the drink. There is no way to blunt the call to die to yourself, take up your cross, and follow Christ. Anything less than cruciform discipleship is cheap grace.


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New TV on DVD releases this week
- Here are the new TV DVDs, in stores tomorrow. The Adventures of Ozzie & Harriet - Best of Ricky and Dave The Batman - Season 5 Batman: The Animated Series - Goth...
- Read more
TV continues uninterrupted in comic books
- What are you going to do? The Office, How I Met Your Mother, and even new shows like Pushing Daisies and Chuck are either out of fresh episodes or quickly running out. W...
- Read more
Tony Jay dead
- Yes, the headline would usually include the "at (age)," but I can't find anything that says what his age was. Nothing on the IMDB, and nothing on his site either. You wo...
- Read more
Leading the Teen Titans to protect Earth is Robin, formerly Batman's sidekick. He has no special powers, just a utility belt and his mind to help solve problems. His team members Starfire, Raven, Cyborg, and Beast Boy live together in Teen Tower, and each of them has supernatural abilities to fight a new generation of villains.


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A capillary electrophoresis system incorporates an application specific cassette having run buffers and capillary column selected for the specific separation procedure, with buffer reservoirs sealed for storage and shipment. A serving apparatus is provided to accept the cassette and perform the specific electrophoresis procedure until buffer is depleted or the performance of the separation column deteriorates. The serving apparatus comprises liquid transfer apparatus, removable covers for the reservoirs of the cassette, electrodes for establishing voltage potential, and a control system.
1. An application-specific cassette for capillary electrophoresis comprising:
a base for supporting and positioning elements of the cassette;
an inlet buffer reservoir fastened to the base, said inlet buffer reservoir having a first upper opening and containing a first run buffer;
an outlet buffer reservoir spaced apart from said inlet buffer reservoir and fastened to the base, said outlet buffer reservoir having a second upper opening and containing a second run buffer;
a capillary electrophoresis column sealed through a sidewall of and opening at one end into said inlet buffer reservoir below the surface of said first run buffer, and sealing through a sidewall of and opening at the other end into said outlet buffer reservoir below the surface of said second run buffer, said capillary electrophoresis column having a separation matrix therein; and
removable seals for sealing said first and second upper openings after placing buffer therein.
2. An application-specific cassette as in claim 1 wherein said inlet and outlet buffer reservoirs and said base are formed into a single injection molding.
3. An application-specific cassette as in claim 1 wherein said removable seals comprise one or more liquid-impervious films mounted to said upper openings by adhesive.
4. An application-specific cassette as in claim 1 comprising one or more auxiliary reservoirs each containing a chemical solution for use in an electrophoresis procedure, and sealable by a liquid-impervious film with an adhesive.
5. An application-specific cassette as in claim 1 wherein said inlet and said outlet buffer reservoirs are configured to engage cover assemblies at said upper openings, said assemblies for providing an electrode and a waste tube in the covered reservoir, the engagement of one of the cover assemblies at an upper opening being capable of sustaining a positive pressure above atmospheric pressure across the cover assembly.
6. A serving apparatus for receiving and using an application-specific cassette having an inlet and an outlet buffer reservoir, run buffers therein, and a capillary column, said serving apparatus comprising:
a probe apparatus including a probe tip and a pump for aspirating and dispensing liquids to and from the buffer reservoirs and containers, said probe apparatus comprising a first electrode placed to contact liquid within said probe apparatus;
translation apparatus supporting said probe apparatus for translating said probe apparatus in three dimensions;
a first removable cover assembly configured for covering said inlet buffer reservoir and immersing a first waste tube and a second electrode in buffer solution therein, and providing an ability to support a positive pressure across said first cover assembly;
a second removable cover assembly for covering said outlet buffer reservoir and immersing a second waste tube and a third electrode in buffer solution therein, and providing an ability to support a positive pressure across said second cover assembly; and
a high voltage power supply connected to said first, second, and third electrodes for supplying electrical potential selectively and independently to each;
said serving apparatus configured to accept and hold said application-specific cassette with said inlet and outlet buffer reservoirs within the translation range of said translation means.
7. A serving apparatus as in claim 6 wherein said pump comprises a syringe cylinder driven by a linear actuator.
8. A serving apparatus as in claim 6 wherein said first and second removable cover assemblies comprise pass-through seals for passing said probe tip within said inlet and outlet buffer reservoirs while maintaining ability of said cover assemblies to support a positive pressure differential across said removable cover assemblies.
9. A serving apparatus as in claim 6 further comprising auxiliary vials and containers of fluids within the range of said translation means.
10. A serving apparatus as in claim 6 further comprising a temperature controllable enclosure for enclosing said application-specific cassette when mounted in said serving apparatus to control the temperature of said capillary column.
11. A serving apparatus as in claim 6 further comprising a microprocessor-based control system for managing operations of said serving apparatus, said control system comprising an operator interface for entering commands and data.
12. A system for performing capillary electrophoresis comprising:
an application-specific cassette having spaced-apart reservoirs fastened to and spaced apart on a base and containing run buffers, and a capillary column opening below the surface of said run buffers; and
a serving apparatus configured to mount said application-specific cassette, said serving apparatus having a liquid transfer apparatus for presenting and withdrawing same materials and chemical solutions to and from said cassette, a first electrode placed to contact liquid within said liquid transfer means, a second electrode immersible in one of said buffer reservoirs and a third electrode immersible in the other, and a high voltage power supply connected to said first, second, and third electrodes for supplying electrical potential selectively and independently to each.
13. A system for capillary electrophoresis as in claim 12 wherein said application-specific cassette before mounting comprises removable seals for closing an upper opening of each of said buffer reservoirs, sealing said run buffers therein until ready for mounting to said serving apparatus.
14. A system for capillary electrophoresis as in claim 13 wherein said removable seals comprise liquid impervious film mounted to said upper openings by an adhesive.
15. A system for capillary electrophoresis as in claim 12 wherein said spaced-apart reservoirs each have an upper opening configured to engage a cover assembly in a manner to sustain a positive pressure differential across the cover assembly.
16. A system for capillary electrophoresis as in claim 12 comprising auxiliary vials and reservoirs, wherein said liquid transfer apparatus comprises a probe with a probe tip connected by fluid conduit to a syringe pump activated by a linear actuator, and translation apparatus for translating said probe tip in three dimensions to reach into said buffer reservoirs and said auxiliary vials and reservoirs.
17. A system for capillary electrophoresis as in claim 16 wherein said serving apparatus comprises cover assemblies each with a cover for closing upper openings in said buffer reservoirs, said cover assemblies each comprising an electrode sealed through the cover, a waste tube sealed through the cover, and a pass-through for passing a probe of said liquid transfer apparatus into the covered buffer reservoir, while maintaining an ability to sustain a positive pressure differential across the cover.
18. A system for capillary electrophoresis as in claim 12 further comprising a temperature controllable enclosure for enclosing said application-specific cassette.
19. An application-specific cassette for a specific capillary electrophoresis procedure involving post-separation detection comprising:
an inlet buffer reservoir with a first upper opening and a first run buffer therein;
an intermediate buffer reservoir spaced apart from said inlet buffer reservoir with a second upper opening and a post separation buffer therein;
an outlet buffer reservoir spaced apart from said intermediate buffer reservoir, with a third upper opening and having a second run buffer therein;
a first capillary electrophoresis column opening at one end into said inlet buffer reservoir below the surface of said first run buffer and opening at the other end into said intermediate buffer reservoir, said capillary electrophoresis column having a separation matrix therein;
a second capillary electrophoresis column opening at one end into said outlet buffer reservoir below the surface of said second run buffer and opening at the other end into said intermediate buffer reservoir, said capillary electrophoresis column having a separation matrix therein; and
removable seal means for sealing said first, second and intermediate upper openings after placing buffer therein.
This invention is in the field of apparatus and methods for separating macromolecules by capillary electrophoresis, and pertains in particular to high-performance automated apparatus and methods.
There has been rapid growth in recent years in apparatus and methodology for biochemical enterprise, particularly for cleavage, separation and analysis of macromolecules such as DNA, including human DNA. There are a number of uses, such as determining the presence of genetically induced conditions such as Sickle-Cell Anemia, Huntington's Chorea, and others. Fluorescent gel scanning has also become important as an identification tool. Genetic code from a human hair or a flake of skin can be matched to a single human being, supplementing older procedures, such as fingerprint analysis in criminal law.
Macromolecules are typically separated and identified by electrophoresis. In electrophoretic separation electrically charged macromolecules of different size and structure move at different velocities in a specific medium under the influence of an electrical field. Typical mediums and geometries in use include agarose slabs, polyacrylamide gel films, matrix-filled columns, and micro columns (capillary tubings) either with or without a gel matrix.
The use of micro-column, (capillary) electrophoresis for macromolecule separation and analysis is a relatively more recent development than slab and film techniques. A principle advantage of micro-column techniques is their suitability for analysis of very small sample volumes, e.g. microliter or sub-microliter amounts of sample. Being able to analyze such small amount of sample has become very important because, oftentimes, the available volume of sample for analysis is extremely small. For example, in forensic and criminal investigations, the only material that may be available for analysis may comprise a fingernail scraping or a stain on an article of clothing.
Another advantage of microcolumn techniques is that the relatively smaller and simpler apparatus lends itself to techniques of automation more readily than do slab and film systems. For example, a detector to monitor bands passing a point in a micro-column is a relatively simple device compared to a scanning detector for monitoring a plurality of channels in an agarose slab. The column itself is small and easier to manipulate. The reservoirs for storing buffers and other solutions are smaller as well and more easily managed. There are many other examples of the advantages of micro-column electrophoresis.
With all of the advantages and advances in electrophoresis by the use of micro-columns, there remain problems that have not been solved before the present invention. Many of these are simply variations of difficulties encountered in the effort to automate all types of electrophoresis apparatus. For example, considering only capillary electrophoresis, not all starting samples respond in the same way to the same set of electrophoretic circumstances. By circumstances is meant the value set of all of the variables that effect electrophoresis of sample material.
The variable set includes the length and bore of the capillary, the separation matrix in the column, the electrical potential maintained across the ends of the column, the types of buffers used, the geometry of the physical apparatus, the arrangement and techniques for sample injection, ability to control temperature and to dissipate heat generated in the electrophoresis operations, arrangements for fraction collection and more.
Variable sets for specific samples and purposes have been developed empirically over time and tested. New circumstances for new sample compositions continue to be developed.
Some circumstances are relatively easy to manage. For example, the electrical potential across the ends of a column. Others are quite difficult. The only way to change the capillary bore, for example, is to change the capillary, and to do so is a relatively clumsy operation in many existing systems.
What is clearly needed is an apparatus wherein the circumstances difficult to change are incorporated into a module that can be placed in a serving apparatus to provide the circumstances more generic and relatively easier to manage. In this invention, the module would provide circumstances specific to a pretested specification, for separations of a certain type and class of sample. The module, which is called an application-specific cassette by the inventors, includes a support block, or framework, designed to interface physically to the serving apparatus; a capillary of the correct length, bore, material, and wall thickness, and incorporating a particular gel or liquid matrix; buffers solutions at the ends of the capillary; and one or more reservoirs for liquids needed in the specific procedure. In a specific and preferred embodiment of the present invention, the application-specific cassette is disposable. When the buffers are depleted, the matrix ages and deteriorates, or any other change renders the cassette unusable, the cassette is simply removed, and a new one substituted to the serving apparatus.
The serving apparatus includes a robotic system with a liquid-handling apparatus having a probe translatable in three dimensions, and an electrode in contact with fluid in the liquid handling apparatus.
In an embodiment of the invention an application-specific cassette is provided for performing a particular electrophoresis procedure. The cassette comprises an inlet buffer reservoir with an upper opening and a first run buffer selected for the specific procedure, an outlet buffer reservoir spaced apart from the inlet buffer reservoir and with an upper opening and a second run buffer selected for the specific procedure, and a capillary electrophoresis column opening at one end into the inlet buffer reservoir below the surface of the first run buffer and opening at the other end into the outlet buffer reservoir below the surface of the second run buffer. Removable seal means are provided to seal the upper openings of the reservoirs for storage and shipment before use.
In a preferred embodiment, the buffer reservoirs are formed into an injection molded cassette with the column mounted horizontally and sealed through a sidewall of each of the buffer reservoirs. A preferred means of sealing the reservoirs before use is by a liquid-impervious film placed over the reservoir openings with an adhesive. There may also be auxiliary reservoirs formed into the cassette for reagents and buffers specific to the specific application.
A serving apparatus is provided to accept the cassette and perform the specific procedure until buffers are depleted or the column performance deteriorates. The serving apparatus has a probe with a tip and pumping means for withdrawing and dispensing liquids to and from reservoirs, a translation sub-system for moving the tip into and out of liquids in reservoirs, cover assemblies for engaging openings in the reservoirs and inserting an electrode and a waste tube in the reservoirs, and a high voltage power supply for providing electrical potential selectively and independently to each of the buffers, There is in addition an electrode in the liquid transfer apparatus providing an ability to sample injection and fraction collection by electromigration, and an detector interface to detect separated bands in the capillary column may be provided in some embodiments.
A system is provided for electrophoresis comprising a serving apparatus as described and an application specific cassette, and in an alternative embodiment, the application-specific cassette is configured with an intermediate buffer reservoir and two capillary columns, to perform post-separation detection.
FIG. 1A is an isometric view of an application-specific cassette 11 according to an embodiment the present invention.
FIG. 1B is a cross-section of cassette 11 along section line 1B--1B of FIG. 1A.
FIG. 1C is an isometric view of one buffer reservoir of a cassette showing a seal film.
FIG. 2A is an isometric view of a serving apparatus according to the invention with an application-specific cassette mounted for operation.
FIG. 2B is a cross section of a reservoir of a cassette along section line 2B--2B of FIG. 2A.
FIG. 2C is a cross section of a buffer reservoir of a cassette along section line 2C--2C of FIG. 2A.
FIG. 2D is a mostly schematic illustration of a liquid-handling apparatus of a serving system according to the invention.
FIG. 3A is a schematic showing specific operations of the apparatus of the invention.
FIG. 3B is another schematic illustrating specific operations.
FIG. 3C is another schematic showing specific operations of the apparatus of the invention.
FIG. 4 is a cross section showing a cassette according to the invention having an intermediate reservoir for post separation detection.
FIG. 1A is an isometric view of an application-specific cassette 11 according to an embodiment of the present invention. In this embodiment a base block 13 establishes the overall length D1 of the cassette, and end blocks 15 and 17 are securely fastened, such as by adhesive material, to the base block. Inlet and outlet buffer reservoirs 19 and 21 are machined into end blocks 15 and 17 respectively, and auxiliary solution reservoirs 23 and 25 are provided in end block 17.
It will be apparent to one with skill in the art that the choice of inlet and outlet for the buffer reservoirs is arbitrary and interchangeable, and the placement of the auxiliary reservoirs is also arbitrary. They could as well be placed in the block at the opposite end of the cassette, or divided between the blocks. Also, the use of auxiliary reservoirs in a cassette is optional, rather than necessary to the invention. The use of one or more auxiliary reservoirs provides a preferable feature of extending the useful life of the cassette.
There are a number of different suitable materials for the blocks. They may be phenolic, polycarbonate, or one of several other polymeric materials available that are machinable and relatively impervious to the chemicals and sample material used in micro-column electrophoresis, and non-contaminating to the electrophoresis chemistry. Moreover, the assembly need not be from separate blocks as shown. The cassette might be formed in a single piece by processes such as injection molding, or machined from a single piece. In the present embodiment, one of the main criteria is cost, and in a preferred embodiment the cassette is meant to be a disposable entity after use.
In addition to the provision of the several reservoirs, the block is prepared in this embodiment to mount a capillary tubing 27 between inlet and outlet buffer reservoirs 19 and 21. FIG. 1B is a section view of block 13 of FIG. 1A taken along line 1B--1B. Capillary tubing 27 is shown mounted through a bore 29 into buffer reservoir 21 and through a bore 31 into buffer reservoir 19. The capillary in this embodiment is permanently mounted by means of a thermosetting adhesive, or other suitable adhesive material compatible with the material of the blocks.
The position of the ends of the capillary relative to the position of the buffer reservoirs, as may be determined by the capillary length D3 and dimension D2, is carefully maintained to a predetermined position, for reasons of transfer of materials, which will be apparent in following descriptions.
Two capillary supports 33 and 34 (FIG. 1A and 1B), comprising blocks each having a groove to support the capillary, are provided adhered to base block 13. Capillary 27 is adhered in this embodiment to supports 33 and 34 by a suitable adhesive, not shown in the figure. If the capillary is produced with an opaque coating on the outer diameter, as is the case with many capillary tubings in the art, this coating is removed between supports 33 and 34, to provide a clear area for a detector to be described below.
As was described in the background section above, workable sets of circumstances for different applications for electrophoresis have been empirically developed. An application-specific cassette according to the present invention is provided for a specific known solution, and comprises therefore a gel or liquid matrix in the capillary, buffer solutions in the inlet and outlet buffer reservoirs, and optionally auxiliary chemicals in the auxiliary reservoirs. Additional buffer may also be provided in one or both of the auxiliary reservoirs, so fresh buffer solution may be transferred at some time in operation from an auxiliary reservoir to one or both of the buffer reservoirs.
In the present embodiment, after a cassette is assembled, the capillary is sealed into place, and buffers and other chemicals are placed in the various reservoirs, the top of each reservoir is sealed with a foil treated paper seal having an adhesive. This seal is similar to seals of this sort used for pharmecuticals, over-the-counter medicines and many other consumer items. The result is a cassette complete with capillary and run buffers that may be manufactured economically in quantity and stored for a considerable time before use. Cassettes may be thusly provided as a product to users having serving apparatus. New variations may also be provided as new specific chemistries are developed.
It will be apparent to those with skill in the art that the various dimensions of a cassette within the scope of the present invention may vary widely. For example, capillary bore may vary according to specific application, ranging from perhaps 10 to as much as 100 micrometers. Also, the length of a capillary can vary, depending on the length of a cassette, which may be designed to fit one or another of various serving apparatus, and on other factors as well. The length of a capillary can vary from as short as 1 cm. to as long as 50 cm., or even longer in some applications.
Although it is not readily apparent in the drawing figures, which are not meant to present dimensions to scale, there is an advantage in keeping the buffer reservoirs rather small, and the auxiliary reservoirs relatively large. Typical buffer reservoirs range from 0.1 to 10 milliliters. A relatively small buffer reservoir saves time in procedures for purging and replacing buffer solution in operation. A relatively large auxiliary reservoir provides relatively more material for use in operation, extending the period between necessary service procedures. Accordingly, in the present embodiment, the buffer reservoirs are much smaller than the auxiliary reservoirs.
FIG. 1C is an isometric view, enlarged, of block 15 from FIG. 1A, showing an exemplary seal 35 covering reservoir 19. A tab extension 37 from the foil seal in this embodiment allows a user to grasp and peel the seal from the top of the reservoirs when mounting the cassette to a serving apparatus for use, as described below.
FIG. 2A is an isometric view of an application-specific cassette 11 according to the invention mounted in a serving apparatus 41. Cassette 11 is registered to a surface 39 of serving apparatus 11 by virtue of engaging pins (not shown) extending from surface 39 by bores (not shown) provided for the registering purpose in the cassette. In this arrangement the cassette is held in place by screw clamps 43 and 45.
There are many alternative means of registering and holding the cassette in serving apparatus 41, and the means described is convenient, rather than required in the invention.
In the embodiment illustrated by FIG. 2A the serving apparatus comprises a frame 47 comprising end towers 49 and 51 rigidly connected to base 39 and supporting a carrier beam 53 positioned above the cassette and parallel to the long axis of the cassette. The carrier beam is for supporting and guiding a horizontal carriage 55 along the length of and above the cassette registered to surface 39.
In the embodiment shown in FIG. 2A carriage 55 is supported by linear bearings (not shown) on two precision shafts 57 and 59, and is translated by turning a worm shaft 61 that engages a precision worm nut (not shown) fixed in the carriage. The worm shaft is driven by a drive train powered by a stepper motor in an enclosure 63 at the end of the serving apparatus.
The linear drive is a precision drive capable of accurate positioning of the carriage along the length of carrier beam 53, and includes position sensors (not shown) for determining exact position for purposes of positioning the carriage and calibrating the translation apparatus.
Horizontal carriage 55 supports a vertical carriage 65 having drive means for limited vertical translation. Moreover, vertical carriage 65 carries a second horizontal carriage 67 that may be positioned in three horizontal positions along a horizontal axis at a right angle to the long axis of beam 53. Horizontal carriage 67 carries a demountable probe 69, which is in the nature of a hypodermic needle, and connects to a tubing leading to a syringe pump, described in more detail below.
When an application-specific cassette is mounted to the serving apparatus the sealing covers are removed and durable run covers are mounted to the openings of the buffer and storage reservoirs. For example, covers 71 and 73 are mounted to reservoirs 23 and 25, respectively.
FIG. 2B is a partial cross-section through reservoir 23 and cover 71 along section line 2B--2B of FIG. 2A. In the present embodiment, cover 71 incorporates an o-ring 73 in a groove, and the cover engages in a counterbore 79 provided at the open end of the reservoir. An o-ring groove 81 is provided in the counterbore to receive the o-ring of the cover, so the two snap securely together in engagement.
A flexible pass-through 77 engages in a bore 75 at the center of the cover, providing a passage whereby probe 69 may enter and exit the reservoir without exposing the contents in the reservoir, and sustaining a low pressure seal, allowing procedures to be employed requiring positive pressure above atmospheric pressure in the reservoir. Typical procedures are described below, and require pressure differential generally on the order of 100 psi and lower. Pass-through 77 in the present embodiment is a radially symmetrical, conically shaped gasket, molded from one of several available flexible polymers, such as PVC.
FIG. 2C is a section taken along line 2C--2C through a cover assembly 83 assembled to buffer reservoir 19 in operation. Cover assembly 83 has an o-ring 85 mounted in a groove in the cover just as described above for cover 71, and the counterbore in the reservoir opening has a receiving groove for engaging the o-ring and providing a secure seal and maintaining the cover in place during use.
A platinum electrode 87 is mounted through a seal 89 in cover assembly 83 and provides a means of applying electrical potential to the run buffer in the buffer reservoir, as required for electrophoresis. The electrode is attached on the outside of the cover assembly to a high voltage line 91 leading away from the reservoir and eventually to a high-voltage power supply in enclosure 63 (FIG. 2A).
A waste tubing 93 is similarly sealed through the cover and attached to a flexible tubing 95 leading to a waste container or drain (not shown) away from the cassette. A pass-through 97 like pass-through 77 in FIG. 2B is engaged in a bore 99 through the cover assembly, and provides access for probe 69 (FIG. 2A) to the interior of the buffer reservoir.
For the buffer reservoir in the particular embodiment illustrated, the pass-through for the probe is off-center, and a locating pin 101 fixed in the cover assembly and engaging a bore 103 in the cassette determines the location of the pass-through in assembly. The cover and pin locations are configured to place the center of the pass-through above, in line with, and just beyond the end of capillary 27 in the buffer reservoir. Dotted outline 105 indicates the position of probe 69 inserted through pass-through 97 to the depth of the capillary beneath the surface 107 of run buffer in the buffer reservoir. It will be apparent to those with skill in the art that there are alternative means of locating the cover to the proper position.
The positioning of the probe relative to the capillary end is the reason for careful control of length D3 of the capillary and placement D2 of the capillary relative to the cassette (see FIG. 1B). As will be detailed below, transfer of sample material to be electrophoresed and fraction collection are dependent on this placement.
Waste tubing 95 and high voltage lead 91 in the present embodiment extend away from the reservoir cover within a common outer covering 109, which leads to enclosure 63.
A cover assembly 111 similar to cover assembly 83 assembles to the buffer reservoir 21 on the opposite end of cassette 11 from buffer reservoir 19. Cover assembly 111 is generally opposite hand from assembly 83, comprises a waste tube, a platinum electrode, and a pass-through for the probe, and places the pass through so probe 69 may be positioned at the end of the capillary in buffer reservoir 21 as well as in buffer reservoir 19. A high voltage wire for the electrode of cover assembly 111 leads away from cover assembly 111 along with a waste tube in a common cover 113. The electrode is powered by the high voltage supply in enclosure 63.
A detector cell assembly 115 mounted to the serving apparatus provides a means for detecting separated bands of materials passing through the capillary and passing the information to monitoring circuitry of a connected control system described more fully below. The detector cell assembly is mounted with a space 116 allowing the cassette to be inserted from the front such that capillary supports 33 and 34 span the width of the detector cell assembly providing an aid in placing and positioning the cassette.
When the cassette is registered in the serving assembly in the present embodiment, the capillary between supports 33 and 34 is positioned precisely against a detector beam focusing element 118. A detector unit incorporating light sensitive diodes is mounted in a pivotal element 120 that is pivoted up (as shown) for positioning a cassette in the serving apparatus, and down and latched (not shown) for operation, which places the detector assembly diodes in position to intercept light passed through or emitted from the capillary tubing. In a preferred embodiment the detector apparatus is that used in the Model 270 Electrophoresis apparatus marketed by Applied Biosystems of Foster City Calif.
It will be apparent to those with skill in the art that there are several alternatives in detection. Among them are electrochemical detection, UV absorbance, and fluorescent detection. Detectors of the different sorts may be adapted in the present invention.
In the presently described embodiment, referring to FIG. 2A, the translating carriages are in extent all within the outer boundaries defined by end towers 49 and 51. Although not shown, there are removable outer covers for enclosing the cassette, carriages, and other elements. Temperature control devices within enclosure 63 provide temperature management within the enclosure in operation, typically in a range of from 10 to 50 degrees C.
FIG. 2D is a mostly schematic diagram of some elements serving probe 69 in serving apparatus 41. A syringe pump 122 driven by a remotely controllable actuator 117 is connected by conduit 119 to an electrode housing tee 121 wherein an electrode 123 connected to a high voltage power source is capable of providing an electrical potential in liquid in the tee. The tee is connected to tubing 125 leading to probe 69. The syringe pump and actuator are located in enclosure 63 (FIG. 2A), and tee 121 with electrode 123 is located in the carriage assembly. A high voltage lead for the electrode passes through common bundle 70 along with tubing 119 leading from enclosure 69 to carriage 55 with sufficient slack to allow the carriage to travel to the full reaches of beam 53. Power and control lines for actuating mechanisms for the various motions of carriage 55 also pass through bundle 70.
The arrangement of the syringe pump, electrode 123, and probe 69 allows for a number of useful functions to be described more fully below.
Also registered to surface 39 of serving apparatus 41 is a supply block 127 providing multiple container positions arranged in three rows parallel to the long axis of cassette 11, such that the centerline of one row passes through the centerline of reservoir 23, the centerline of the center row passes through the centerlines of the inlet and outlet buffer reservoirs 19 and 21, and the centerline of the remaining row passes through the centerline of reservoir 25.
Supply block 127 provides a means of presenting samples to the system for separation, presenting fresh buffers without removing the cassette, and providing containers for fraction collection. Although not seen in FIG. 2A, there is also a wiper and waste tube outlet for the probe, and in some embodiments, a wash station may be incorporated.
The three directions of motion of the translation mechanisms integrated into serving apparatus 41 are capable of moving the tip of probe 69 into either of the inlet and outlet buffer reservoirs, either of auxiliary reservoirs 23 and 25, and into a container at any one of the container positions in auxiliary block 127.
In following descriptions translations of the probe tip will be referred to as X in the horizontal direction along the axis of the cassette, Y horizontal and at right angle to X, and Z as vertical translation.
Because of the registration of the cassette and the auxiliary block onto surface 39, there are a specific number of repeatable X-Y positions for the present embodiment. For example, carriage 55 must be able to travel to the X coordinate for each of the positions of auxiliary block 127, to the common X coordinate for reservoirs 23 and 25 in the cassette, and to the X coordinate for each of the inlet and outlet buffer reservoirs. In the embodiment shown this is ten X positions, but there may be more or fewer in other embodiments.
In the Y direction, the arrangement of all the elements is such that there are just three repeatable Y coordinates. This arrangement for X and Y positions provides a simplifying situation that allows sensors to be used for flagging X positions, and Y carriage 100 is actuated to a back position, a center position, and a forward position, which are fixed positions for the actuator.
In the Z direction for the probe (vertical) there needs to be an uppermost position at a height sufficient for probe 69 to clear all elements of the cassette and the auxiliary block and any containers, and at least one lower position wherein the tip of the probe is at the level of the end of the capillary in the cassette, which is a common height for each of the buffer reservoirs. It is useful as well to have at least one other vertical position to allow the probe to reach very near the bottom of the buffer reservoirs, and there may also be one or more different Z coordinates needed in using the probe in conjunction with the sample and fraction collection reservoirs, and with wash, wipe and waste stations if used.
There are then as many as five vertical positions to be repeated, and these are set as a function of adjustable sensors provided with the actuator for vertical carriage 65. In other embodiments there may be more or fewer than 5 vertical coordinates to be repeated.
Control of the serving apparatus in operation may be accomplished in several ways. In the apparatus shown an I/O interface and a microprocessor controller are built into the serving apparatus in enclosure 69, and a communication cable 129 leads to an LCD display and input interface (not shown). Through the interface an operator may enter pre-programmed sequences or series of sequences, and also may control the apparatus manually. Alternatively, cable 129 may be a serial communication cable connected to a general-purpose computer having a dedicated program for operating the apparatus. A power cable (not shown) connects also to enclosure 63 to furnish the power requirements.
The serving apparatus illustrated principally by FIG. 2A is but a single example of a serving apparatus that may be used with an application specific cassette according to the present invention. In an alternative embodiment a cassette is provided without the counterbores at the openings of the reservoirs, and covers are provided mounted on vertically traversing carriages with the serving apparatus, including o-ring seals for sealing to the top of the cassette around the reservoir openings.
In yet another alternative embodiment an X-Y-Z apparatus for moving the probe is used comprising stepper motor drives and a capacitance sensing means associated with the probe. An apparatus of this sort is disclosed in copending application Ser. No. 07/927,254, incorporated herein by reference. There are other such systems in the art that might be used with an application specific cassette, and it will be apparent to one with skill in the art that there are many ways that the requirements of the serving apparatus might be met. There are, indeed, commercial robotic devices known to the inventor which might well be adapted to function as the serving apparatus in the present invention.
Once an application-specific cassette according to the invention is fixed in position to a serving apparatus, the system comprising the cassette will perform a finite number of separations until buffer is depleted or capillary performance deteriorates. At this point the user may remove the used cassette and replace it with another for the same or for a different application.
FIG. 3A is a partially schematic elevation view of cassette 11 and auxiliary block 127 with sample vials and fraction collection vials, such as vial 131, in auxiliary block 127 registered on surface 39, showing probe 69 in an idle position. Similar views are used below to illustrate various operations of the system. In FIG. 3A buffer reservoir 19 has waste tube 93, and an electrode 87. Buffer reservoir 21 has a similar waste tube 135 and electrode 133. Electrode 123 is resident in tee 121. It is important to the following operating descriptions that the three electrodes may be biased independently, and important too, to remember that, although only X and Z translations may be illustrated in the schematic of FIG. 3A and similar schematics, there is also a Y translation (into the plane of the paper in the figure) to reach more auxiliary and sample positions.
The operation of sample injection is illustrated in FIG. 3B. Probe 69 passes to a selected sample S in a vial 131 in block 127 (or in any other similar sample vial) shown as position A, by operation of the X-Y-Z translation subsystem. Probe 69 submerges in the sample, and by operation of syringe pump 122 withdrawing a finite amount, a volume of sample is withdrawn from the vial into the probe tip. The probe is then translated from the sample vial to the inlet buffer reservoir, which may be either of the buffer reservoirs. In this case, buffer reservoir 19 will be the inlet buffer reservoir.
The probe tip is lowered through pass-through 97 to a depth in the run buffer where the probe tip is immediately adjacent to the end of the capillary, as shown in position B. An electrical potential is now applied across electrodes 123 and 133, or 87 and 133, or both. The potential causes electromigration of charged analytes from the probe tip into the capillary. Selected operation of syringe pump 122 to expel sample at a predetermined rate, usually very slowly, may also be incorporated in some instances.
After sample injection, the electrical potential is removed, and the probe tip returned to idle position, shown as position C. Run potential is then applied across electrodes 87 and 133 causing charged analytes to migrate through the capillary, forming separated bands, past detector assembly 115, and eventually into outlet buffer reservoir 21.
Fraction collection may be accomplished by moving the probe tip during an electrophoresis run to the outlet buffer reservoir, in this case reservoir 21, and down through pass-through 137 to a position immediately adjacent to the outlet end of the capillary, similar to the position taken for sample injection in the inlet buffer reservoir. This position is shown as position D in FIG. 3B. At an appropriate time during a run, electrical potential across electrodes 87 and 133 is removed, and a potential is established across electrodes 123 and 133, causing analytes eluted from the end of capillary 27 to migrate into probe tip 69. Again, the process may in some instances be aided by selected operation of syringe pump 122.
After a fraction is collected from the capillary into the probe tip, the tip is removed from the outlet buffer reservoir, electrical potential re-established across 87 and 133 to continue the run, and the probe tip is translated to one or another of fraction collection vials in block 127.
When buffer depletion occurs before capillary deterioration, buffers may be replenished in the buffer reservoirs, either with the same, or with different buffer. The replenishment sequence is made possible by the ability of the seals and pass-throughs to countenance a positive pressure above atmospheric pressure.
FIG. 3C shows the basics of buffer replenishment. Buffer reservoir 19 is used as illustrative. Probe 69 is first moved to a buffer storage reservoir, which in this example is reservoir 25 in the cassette. Buffer might also be stored at auxiliary positions in the serving apparatus reachable by the probe. The probe passes down through pass-through 139 below the surface of the buffer in the reservoir, and the syringe pump is operated to withdraw fresh buffer into the probe, lines 125 and 119, tee 121, and syringe pump 122, in sufficient quantity to replenish the buffer in reservoir 19. This position is shown in FIG. 3C as position E.
After taking up fresh buffer, the probe is moved to position F through pass-through 97 below the surface of buffer in the reservoir. Syringe pump 122 is then operated to expel the fresh buffer into the reservoir, forcing the depleted buffer out of the reservoir and to waste by means of waste tube 93 (connected to waste containers or drain).
For large-diameter open-tube capillaries, expulsion of fresh buffer from the probe into reservoir 19 might well result in buffer transfer through the capillary rather than to waste. For this reason valves are provided independently in waste lines 109 and 113 although the valves are not shown (see FIG. 2A). To avoid transfer through the capillary in the replenishment procedure described above, the valve in line 113 for waste tube 135 in reservoir 21 is closed, and the valve in line 109 for waste tube 93 in buffer reservoir 19 is open.
An open-tubed capillary may be flushed by a procedure similar to the buffer replenishment procedure. To flush capillary 27 from buffer reservoir 19 to buffer reservoir 21, the valve for waste tube 93 is closed and the valve for waste tube 135 is opened while buffer is injected into reservoir 19,
Probe 69 may be rinsed between sampling, buffer replenishment or replacement, or other reagent use, by moving the probe to an auxiliary vial containing a rinse solution, drawing up a finite volume, moving the probe to an optional wash station, and expelling the rinse solution in a bore providing back-flow of the solution around the outside of the probe. An optional wipe station may also be employed.
By virtue of the translation ability of the serving apparatus a variety of sample preparation, buffer manipulation, and other procedures may be performed during and between electrophoresis runs. By transferring samples, buffers, diluents, or reagents from one vial to another, the system may accomplish dilutions, addition of reactants, sampling reactions at timed intervals, desalting of samples, and other chemistries requiring liquid transfers.
It is also within the scope of the invention to automate cassette replacement to the serving apparatus, so that cassettes may be automatically inserted, interfaced, and removed, permitting unattended operation of multiple and different applications.
FIG. 4 shows an embodiment of the invention useful for post-separation detection. In this embodiment a cassette 141 is provided with an intermediate reservoir 143 and two capillaries 145 and 147, and outboard buffer reservoirs 149 and 151. All three buffer reservoirs are provided in the serving apparatus with waste tubes, electrodes connected to high voltage sources, and passthroughs for the probe to maintain pressure ability, as described above for the cassette with two buffer reservoirs.
In operation the probe would deliver a post-separation buffer or reactant to the reservoir in the manner described above for buffer replenishment. Separation is performed in a first capillary section, either capillary 145 or 147, separated analytes exit the first column (capillary), move across the gap between columns in the intermediate reservoir reacting with the buffer or reactant in the intermediate reservoir, enter the second capillary column, then move past detection cell 153 on the way to reservoir 151. All the other techniques described for the two reservoir cassette are available to be used with the three-reservoir cassette.
In the cassette aspect of the invention, there are specific combinations of separation matrices, buffer solutions, and the like that may be provided to accomplish specific types, kinds or classes of separations. Some examples are included here.
As one example, for the separation of SDS-denatured proteins the capillary of an application-specific cassette according to an embodiment of the invention contains a cross-linked, polymerized gel of acrylamide in a buffer. The gel is 5.1% total acrylamide and 2.6% bis-acrylamide crosslinker (i.e. 5.1% T and 2.6% C) in a buffer of 300mH Tris (pH 8.8), 3.2 mM sodium dodecylsulfate (SDS), and 2.3M ethylene glycol. The inlet and outlet buffer reservoirs contain the buffer only. The auxiliary reservoirs optionally contain the same buffer, allowing buffer to be replaced in operation until the separation matrix deteriorates, at which time the used cassette is removed and discarded and a new cassette containing the same materials is inserted in a serving apparatus.
As another example, for the separation of DNA sequencing products the capillary contains a cross-linked polymerized gel of acrylamide in a buffer. In this case the gel is 6% T and 5% C, 8% M urea, and a buffer of 89 mM Tris, 89 mM boric acid, and 2% mM EDTA at pH 8.3. The inlet and outlet buffer reservoirs contain the buffer solution but no gel. Again, the auxiliary reservoirs (if any) may contain additional buffer solution for replacing used solution as needed during operation, extending the useful life of a cassette.
There are many more examples known to the inventors and representing separation matrix and buffer solutions use for specific types of separations. The following list of references contains information presenting specific sets of variables for several more specific types of separations, but is not meant to be limiting, as there are many more such sets of circumstances, and new sets may be incorporated to application specific cassettes as they are discovered and developed.
For DNA restriction fragments and for polymerase chain reaction (POR) amplified products, see Heiger, D. N., Journal Chromatography, 516, 33 (1990). For DNA sequencing products see Swerdlow, H. et el., Journal Chromatography, 516, 61 (1990). For separation of synthetic single-stranded DNA, see D. Demorest et el., Journal Chromatography, 559, 43 (1991).
For SDS-proteins see Tsuji, K., Journal Chromatography, 550, 823 (1991). For native proteins see Cohen, A. S., et al., Chromatograhia, 24, 15 (1987). For oligosaccharides see Liu, J., et al., Journal Chromatography, 559, 223 (1991).
For materials for an application-specific cassette for separating DNA restriction fragments in an application having a liquid polymeric matrix (entangled polymer solution), where the matrix is the same in the column and in the reservoirs, see Chin, A.M., et al., Am. Biotech. Lab./News Ed 7 (10A), 16 (1989). For SDS proteins in a liquid polymeric matrix, see Widhalm, A., et al., Journal Chromatography, 549, 446 (1991).
It will be apparent to those with skill in the art that there are many changes that might be made without departing from the spirit and scope of the invention. Many variations have already been described above, such as material and construction variations for the cassette. There are even more variations that might be made in the serving apparatus. The level of automation is also a variable option, and a system according to the invention may be manually operated or fully automated.
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Detroit Getting RoboCop Sculpture; St. Louis Needs Snake Plissken Statue
|Snake Plissken: You know this guy's statue would be a major tourist draw.
No knock against RoboCop, but in our minds a more statue-worthy, '80s flick depicting a crime-ridden city of the future was the 1981 film Escape from New York -- a movie shot largely in St. Louis.
Kurt Russell played the main character of Snake Plissken, a criminal sent into the penal colony of New York City to rescue the president of the United States. St. Louis was chosen as a filming location because writer/director John Carpenter (Halloween) needed a decrepit urban core that could pass for a burned-out New York of the future.
As documented on IMDB and this fantastic website, several St. Louis landmarks made their way into the movie, including the Fabulous Fox Theatre (prior to its renovation), Union Station (ditto the Fox parenthesis) and the downtown streets of Locust and Washington avenues.
So, how about it, St. Louis? Wouldn't a big statue of Snake Plissken look great somewhere downtown? Perhaps at the corner of Washington and Tucker to -- you know -- scare the criminal element away from the nightclub and loft district?
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Tuesday, August 13, 2013
A brand new improv comedy troupe, The Charleston Improvables will perform their first full-length show at 8 pm on Aug. 24 at the historic James F. Dean Theatre in downtown Summerville. "Whose Line Is It Anyway?" style improv games will entertain, amuse and engage the audience.
As the Improvables are a troupe of eight local improvisers and comedians, most of whom are also stand-up comedians, an Improvables show is "stand-up comedians meet improv games." The show contains games that can be seen on the Emmy Award Winning U.S. TV show, "Whose Line Is It Anyway?" Every improv game is based on input from the audience. At least one game will include entire quotes and phrases that were written by the audience before the show begins. One of the most exciting games for the audience begins with the Improvables interviewing an audience member onstage and then a comedic parody of that person's daily life.
While some improv shows aim for dramatic accomplishment at the expense of comedy, the Improvables are entirely focused on creating laughter. All but one of the Improvables are local stand-up comedians, and several have extensive training and performance backgrounds with the iO Theatre in Chicago and Theatre 99 in Charleston. The Improvables troupe consists of: Humorous keynote speaker Brian Carter (Theatre 99 trained), Moey Conway (iO Theatre trained), 2nd Lieutenant Lain Healey, Deshawn Christopher Mason, Kari Hanson, Robert Thomas, Kat LeeHong and Steve LaRowe.
Audiences at Improvable shows will recognize many games from "Whose Line Is It Anyway?" and are certain to be engaged and amused. The Improvables performed a short preview set during a comedy show July 30th at Castaway's Grille, and audience response was overwhelmingly positive.
If an Improvables show were rated by the MPAA, it would be PG-13. The show is basically family-friendly, but, for the cautious parent, the Improvables recommend leaving children below high school age at home.
The Improvables will perform at 8pm on August 24th, September 19th and October 11th at the historic James F. Dean Theater. Tickets are $12 and may be purchased online at http://www.flowertownplayers.org/improvables, in person or over the phone Wednesday through Saturday from 12 pm to 6 pm by calling 843-875-9251. More info is available at http://charlestonimprovables.com/.
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Finally, the Rock has come back to video games, and for the first time ever, on the iPhone. Well, he did for a little while, anyway.
WWE Legends Of Wrestlemania was released on March 24 and being an avid wrestling fan, I immediately bought a copy. Meanwhile, on the March 30, TNA Wrestling was released, bringing the wrestling promotion wars to the iPhone and taking me to potential heaven. So here I am, ready to pit the two games against each other in a wrestling showdown of epic proportions, one not seen since the glorious WWF vs. WCW days of the late 90s.
Continue reading the smackdown at The Apple Blog