Effects of hS100A2 and hS100A6 on Human Colorectal Cancer Cell ...

[Abstract]

BACKGROUNDS AND OBJECTIVES:The pathogenesis and development of tumor is a complex biologic process that involves various factors and numerous genes, including many signal molecules. Therefore, it is very important to study the effects and the mechanisms of tumor-related signal molecules, and the reciprocal relationships among these signal molecules for elucidating the mechanisms of the pathogenesis and development of tumor.The S100 proteins are small acidic signaling proteins (9?13 kDa) that are found exclusively in vertebrates.With at least 25 members found to date in humans, the S100 proteins constitute the largest subfamily of the EF-hand proteins. S100 proteins are proposed to have intracellular and extracellular roles in the regulation of many diverse processes such as protein phosphorylation, cell growth and motility, cell-cycle regulation, transcription, differentiation and cell survival. 21 members (S100A1?S100A18, trichohylin, filaggrin and repetin) of these proteins have genes clustered at chromosome locus 1q21, which has low stability. A potential role for S100 proteins in neoplasia stems from these activities and from the observation that several S100 proteins (including S100A2 and S100A6) have altered levels of expression in different stages and types of cancers. But the precise roles and the importance of S100 proteins in the development and promotion of cancer are poorly understood.The S100A2 is often downregulated in cancers, including melanoma, breast cancer, prostate cancer, lung tumors, so on. Furthermore, it has been indicated as a potential tumor suppressor by differential hybridization. But there are still opposite reports.The S100A6 protein was first purified from Ehrlich ascites tumor by Kuznicki and Filipek. The overexpression of S100A6 has been reported in several types of cancers, such as epithelial carcinomas (colon cancer, hepatocellular cancer, cholangiocarcinoma, pancreatic cancer and malignant melanoma), osteosarcoma, ovarian cancer, ras tansformed NIH3T3 cells and SV40 transformed mouse fibroblast. But the exact effects of S100A6 in cancers remain unknown.In order to elucidate the effects and the mechanisms of S100A2 and S100A6 on human tumors and to find some new markers for tumor diagnosis and prognosis, and some new targets for tumor therapy, the effects of S100A2 and S100A6 on cell proliferation, cell cycle, apoptosis and migration of human colorectal cancer cell line HCT116 and human osteosarcoma cell line MG63 were investigated; Furthermore, the effects of S100A2 and S100A6 on the level and distribution of?-catenin in MG63 cells were studied.METHODS1. The preparation and identification of recombinant hS100A2 and hS100A6 protein: pGST-Moluc-hS100A2 and pGST-Moluc-hS100A6 were digested by restriction endonucleases, Xho?and EcoR, and then analyzed by agarose gel electrophoresis. E Coli. (BL21) was transformed with the two plasmids respectively and the expressions of GST-hS100A2, and GST-hS100A6 were induced by IPTG. Recombinant hS100A2 and hS100A6 proteins were purified by Glutathione Sepharose 4B beads and identified by SDS-PAGE/ Western Blot. Their concentrations were determined by Bradford.2. The recombinant adenoviruse of S100A6 (AdS100A6) was amplified in human embryonic kidney cell 293(HEK293 cell); hS100A6 protein in the cell lysate of HCT116 infected by AdS100A6 was detected by SDS-PAGE/ Western Blot.3. HCT116 cells and MG63 cells were treated with GST, GST-hS100A2, GST-hS100A6, AdGFP, and AdS100A6, respectively. The final concentration of all proteins was 40?g/ml. Cell proliferation was measured by MTT method and trypan blue stain/cell counting; And cell cycle(PI stain) and cell apoptosis(annexin V-PE and 7AAD stain) were detected by flow cytometry; the cell migration ability was determined by transwell chambers; the changes of?-catenin expression were observed under laser scanning confocal microscope (LSCM) after immunofluorescence stain.RESULTS1. The two plasmids were digested into two fragments, about 300bp and 9kb, respectively. The results were consistent with their maps of restriction endonucleases. hS100A2 and hS100A6 were found in the two purified recombinant proteins ( SDS-PAGE/ Western blot), respectively. The yields of recombinant hS100A2 and hS100A6 were 5mg/L and 3mg/L, respectively (Bradford).2. hS100A6 protein was found in the cell lysate of HCT116 infected by AdS100A6( SDS-PAGE/ Western blot).3. The effects of exogenous S100A2 and S100A6 on cell proliferation of HCT116 cells and MG63 cells.The proliferation of HCT116 cell was inhibited from the 4th day and 5th day after respectively treated with recombinant hS100A2 and hS100A6 proteins (P0.05) of those in AdGFP group, respectively. The effects of hS100A2 and hS100A6 on MG63 cell cycle were similar to theirs on HCT116 cell cycle.These results indicated that both S100A2 and S100A6 might block cell cycles of HCT116 cells and MG63 cells by increasing G0~G1 stage cells and decreasing S stage cells. Recombinant hS100A6 protein and AdS100A6 had similar activities on cell cycles of the two cell lines.5 The effects of exogenous S100A2 and S100A6 on cell apoptosis of HCT116 cells and MG63 cellsAfter HCT116 cells were cultured with recombinant hS100A2 and hS100A6 protein for 48 hours, the apoptosis cell numbers were 2.3 times and 2.0 times as those of GST control groups, respectively (P0.05) and showed a tendency of increase.These results indicated that both S100A2 and S100A6 could promote the apoptosis of HCT116 cells and MG63 cells.6. The effects of exogenous S100A2 and S100A6 on cell migration of HCT 116 cells and MG63 cellsCompared with their own controls, the cell numbers of trans-membrane of HCT116 cells and MG63 cells after treated with recombinant hS100A2 protein for 60 hours decreased by 75 % and 83%, respectively(P

Title: Effects of hS100A2 and hS100A6 on Human Colorectal Cancer Cell Line HCT116 and Human Osteosarcoma Cell Line MG63

Category: Pituitary Tumor

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Source: http://www.tumorres.com/pituitary-tumor/34457.htm

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NJ gov strongly defends lowering flags for Houston (AP)

NEWARK, N.J. ? Gov. Chris Christie has strongly defended his decision to have flags flown at half-staff Saturday for Whitney Houston despite receiving emails and other messages disparaging the singer and criticizing him.

The Republican governor said Wednesday that he rejects complaints that Houston "forfeited the good things that she did" because of her struggles with substance abuse.

"What I would say to everybody is there but for the grace of God go I," he said.

Christie ordered flags flown at half-staff at state government buildings Saturday, the day funeral services are held for Houston at the Newark church she sang at as a child.

Twitter was abuzz Wednesday with reaction to the decision by Christie, whose home is in Mendham, a wealthy town where Houston also had lived.

In online postings, there were two main arguments against the honor for the Grammy Award winner who died over the weekend in California at age 48: One was that it should be reserved for members of the military, first responders and elected officials. The other was that it's wrong to honor a drug addict.

Heather Clause, a Richmond, Va.-based blogger who writes about teen moms and was tweeting critical comments, said in a telephone interview that she was appalled by the planned flag-lowering.

"It's just such a bad example for people," said 23-year-old Clause. She said the decision was like saying if someone sings well, drug use doesn't matter and "you can still be an idol."

In upstate New York, Rebecca Eppelmann, a newspaper copy editor, also tweeted her disgust at the Houston honor, then discussed her views.

"It should be for major events," she said. "It's horrible that she passed away. It's not something that should warrant this."

Christie said he was not saying that Houston, who was born in Newark and was raised in nearby East Orange, is a role model. Instead, he said, Houston deserves the honor because of her huge cultural impact and as "a daughter of New Jersey."

"I am disturbed by people who believe that because her ultimate demise ? and we don't know what is the cause of her death yet ? but because of her history of substance abuse that somehow she's forfeited the good things that she did in her life," said the governor during a briefing in northern New Jersey. "I just reject that on a human level."

The cause of Houston's death at a Beverly Hills hotel has not been determined, and the results of toxicology tests are pending. Investigators found several bottles of prescription medication in the hotel room where she died Saturday, but authorities said they weren't an unusually large number.

The governor noted he has ordered flags flown at half-staff for all 31 fallen New Jersey soldiers and every fallen police officer during his time in office.

He also ordered flags lowered last year for Clarence Clemons, the saxophonist for Bruce Springsteen's E Street Band.

In Newark, residents were irritated to hear of criticism of the governor's decision to honor Houston, who acknowledged her drug abuse in interviews.

Resident Anna Simpson said she admires Christie for honoring Houston because "if it were Bruce Springsteen or Jon Bon Jovi, nobody would bat an eye."

Simpson was at New Hope Baptist Church Wednesday, where Houston's funeral will be held, to pay her respects on her way home from her airport job.

"She never forgot where she came from," Simpson said. "She was real. We would see her and be like, `She's one of ours,' and she always made us proud, no matter what happened."

After Houston became famous she continued to make regular trips to the public school she attended in East Orange and to which her family has directed donations be sent in lieu of flowers.

___

Associated Press writer Geoff Mulvihill in Haddonfield, N.J., contributed to this report.

Source: http://us.rd.yahoo.com/dailynews/rss/celebrity/*http%3A//news.yahoo.com/s/ap/20120216/ap_en_mu/us_whitney_houston_flags

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Janice Voss, veteran of 5 space shuttle flights, dies at 55

Astronaut Janice Voss, a veteran of five spaceflights and a former science director for a NASA exoplanet-hunting spacecraft, has died after a battle with cancer. She was 55.

"Just got the very sad news that U.S. astronaut Janice Voss passed away last night," the Association of Space Explorers, an international organization representing more than 350 individuals who have flown in space, wrote on Facebook. "Our thoughts go out to her family and friends."

NASA confirmed Voss' death in a statement issued on Tuesday, saying she had passed away overnight.

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Chosen by NASA for the astronaut corps in January 1990, Voss served as mission specialist on five space shuttle missions, including the only repeat flight in the shuttle program's 30-year history. She flew with the first commercial laboratory, rendezvoused with Russia's Mir space station and helped create the most complete digital topographic map of the Earth.

Five-time shuttle flier
Voss launched on her first and final missions aboard the shuttle Endeavour. As a member of the STS-57 crew in June 1993, she helped conduct biomedical and material science experiments in the first commercially-developed Spacehab module, a pressurized laboratory mounted in the orbiter's payload bay that more than doubled the work area for astronaut-tended activities.

In February 2000 Voss again launched on Endeavour, this time for NASA's Shuttle Radar Topography Mission. After deploying a nearly 200-foot mast, Voss and her crewmates worked around the clock in two shifts to map more than 47 million square-miles of the Earth's land surface. [ Shuttle's Best Science Missions ]

Voss' second flight to space marked the first time a space shuttle came within the vicinity of Russia's space station Mir. Flying on the shuttle Discovery, Voss and her STS-63 crewmates ? including Eileen Collins, the first woman to pilot a U.S. spacecraft ? rendezvoused with the Russian outpost to verify flight techniques, communications, and navigation and sensor aids. The February 1995 "Near-Mir" mission set the stage for the first shuttle-Mir docking later that year.

Voss' two other spaceflights, STS-83 and STS-94, were the only time in the shuttle program's history that an entire crew was launched twice to achieve the same mission. The crew's first attempt began with a liftoff on Columbia on April 4, 1997. Three days into the mission however, a problem with one of the orbiter's three power-generating fuel cells resulted in the flight being cut short and the crew members returning to Earth.

Three months later with Columbia back in working order, Voss and her six STS-83 crewmates launched again, this time as the STS-94 crew. During the successful 15-day flight, Voss and her fellow fliers worked inside a European Spacelab module, conducting experiments as part of the Microgravity Science Laboratory (MSL) mission.

In total, Voss logged over 49 days in space, traveling 18.8 million miles while circling the Earth 779 times. Her five missions tied her with the record for the most spaceflights by a woman.

Four years after returning to Earth for a final time, Voss transferred from Johnson Space Center in Houston, Texas, to NASA's Ames Research Center at Moffett Field, Calif., where she headed the science program for the agency's Kepler space observatory. Designed to search for Earth size planets orbiting distant stars, Kepler was launched in March 2009 and to date has confirmed 61 exoplanets and identified more than 2,000 planetary candidates.

Voss left Ames in 2007 and most recently served as the payload lead in the astronaut office's space station branch at the Johnson Space Center.

"As the payload commander of two space shuttle missions, Janice was responsible for paving the way for experiments that we now perform on a daily basis on the International Space Station," chief astronaut Peggy Whitson said in the NASA statement.

"By improving the way scientists are able to analyze their data, and establishing the experimental methods and hardware necessary to perform these unique experiments, Janice and her crew ensured that our space station would be the site of discoveries that we haven't even imagined."

"During the last few years, Janice continued to lead our office's efforts to provide the best possible procedures to crews operating experiments on the station today," she said. "Even more than Janice's professional contributions, we will miss her positive outlook on the world and her determination to make all things better."

Path to space
A native of Rockford, Ill., Voss received her bachelor of science in engineering science from Purdue University in 1975, a master of science degree in electrical engineering and a doctorate in aeronautics and astronautics from the Massachusetts Institute of Technology in 1977 and 1987, respectively.

Voss' first work with NASA was during her undergraduate studies at Purdue. A member of Johnson Space Center's co-op program, she worked on computer simulations in the engineering and development directorate in the years leading up to the start of the shuttle-era. Voss returned to the Johnson in 1977 for a year, working as a crew trainer teaching entry guidance and navigation.

Before becoming an astronaut, Voss worked at the Orbital Sciences Corporation, supporting mission integration and flight operations for the Transfer Orbit Stage that launched the Advanced Communications Technology Satellite from the space shuttle in September 1993, and NASA's ill-fated Mars Observer from a Titan rocket in 1992.

A multiple recipient of NASA's Space Flight Medal, Voss donated her personal papers documenting her spaceflight career to Purdue Libraries' division of archives and special collections in 2009.

"Knowing that someone else got from here to there brightened many of my days at Purdue," Voss said at the time, referring to the university's earlier astronaut alums. "Maybe my papers will help someone else feel that they aren't that different from me."

"If I can do it, then so can they," Voss said.

Follow collectSPACE on Facebook and Twitter @ collectSPACE and editor Robert Pearlman @ robertpearlman. Copyright 2011 collectSPACE.com. All rights reserved.

? 2012 Space.com. All rights reserved. More from Space.com.

Source: http://www.msnbc.msn.com/id/46302650/ns/technology_and_science-space/

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Sports Recreation: Minimize Stress and Have Fun | Where To ...

Therefore, everyone needs some time to unwind and have some entertainment. Our modern way of
living also tends to be inactive, and the meals most people eat are often not very healthy. Exercise is a
very important component in restarting us. For these reasons, sports recreation pursuits are some
of the best fun we can choose. There are wide ranging options for hobbies to offer sports
recreation. You can even want to learn some new leisurely sports skills.

Training systems to learn new sports recreation skills can be found online and in much bigger cities. You can understand
and participate in workforce sports through the community Y and other businesses. Team sports recreation
functions you might like to participate in comprise softball, baseball, hockey, and sometimes
soccer designed for younger people. Another squad sport you might appreciate is bowling. Regardless of the
skill level you can find your bowling team to have fun with.

Particular person sports recreation pursuits you can learn and participate in include golf, tennis games, running,
aerobic party, gymnastics, and horseback riding. Don?t overlook bicycling and also weight training too.
Many of these sports can be figured out at the local Gym as well. For some of these, you need to have
a friend to relax and play against or share the exerience with. Countless runners, for instance, prefer to run with a
colleague. And games like football and handball really need cooperation for best results.

If you would rather, you can learn concerning sports recreation expertise online. You can find training to teach
almost any outdoor activity or activity you can think of including underwater handbags, fly-fishing, and
para-gliding in the Alps! Needless to say, unless you can get some hands-on knowledge you?ll only have
any theories of the outdoor activity. Still, online courses are a way to get a taste of sports
and recreational activities you might someday would like to try out.

A really enjoyment sports recreation activity you might get a chance to participate in is ping pong. Ping pong, or
table tennis as it is sometimes called, is played on a table with rubber coated paddles and
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You ought to hit the ball best honeymoon destinations with your paddle so it strikes the meal table and clears the online before your
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Source: http://www.wheretosubmitarticles.in/sports-recreation-minimize-stress-and-have-fun/

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Judge allows 'Sister Wives' suit to proceed

SALT LAKE CITY (AP) ? A federal judge has ruled there's sufficient evidence to allow a polygamous family made famous by a reality TV show to pursue a lawsuit challenging the constitutionality of Utah's bigamy law.

U.S. District Judge Clark Waddoups on Friday dismissed Utah's governor and attorney general from the case, but allowed the suit to proceed against Utah County Attorney Jeffrey Buhman, the Deseret News and Salt Lake Tribune report.

Buhman threatened to prosecute Kody Brown and his four wives ? Meri, Janelle, Christine and Robyn ? after the TLC show "Sister Wives" debuted in September 2010, but his office has not filed charges.

The family sued Buhman, Gov. Gary Herbert and Attorney General Mark Shurtleff in July 2011, claiming Utah's bigamy statute violates its constitutional rights to due process, equal protection, free exercise of religion, free speech and freedom of association.

Waddoups, in his 21-page ruling, wrote that he dismissed Herbert and Shurtleff from the case because Shurtleff assured the Browns that they wouldn't be prosecuted. Shurtleff has a policy of not prosecuting consenting adult polygamists as long as they're not committing other crimes.

But the judge wrote that Buhman conducted interviews with the news media that made it clear he intended to investigate and prosecute the Browns. The fact that no charges have been filed does not matter, he added.

"The entirety of actions by the Utah County prosecutors tend to show either an ill-conceived public-relations campaign to showboat their own authority and/or harass the Browns and the polygamist community at large, or to assure the public that they intended to carry out their public obligations and prosecute violations of the law," Waddoups wrote.

There's reason for the Browns to believe they could face prosecution in Utah County, the judge continued, and that could have a "chilling effect" on their ability to practice their First Amendment rights in the state.

But the Browns must show that there's a real and viable threat to their constitutional rights for the lawsuit to hold up in court, Waddoups wrote.

Brown moved his wives and 16 children from Lehi to the Las Vegas area in January 2001.

Buhman said he had not yet reviewed the ruling and was not prepared to comment.

Associated Press

Source: http://hosted2.ap.org/APDEFAULT/4e67281c3f754d0696fbfdee0f3f1469/Article_2012-02-04-Sister%20Wives-Lawsuit/id-11858ff52143488990ae1b19ef31981e

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Mad Catz Cyborg M.M.O. 7 gaming mouse hands-on

Mad Catz' Cyborg RAT series of adjustable computer mice has seen its fair share of minor revisions, and while wireless upgrades and whitewashed DPI tweaks have given consumers a hair of choice in transforming desktop pests, the differences between these devices has been modest, at best. Until now, at least. Enter the Cyborg M.M.O. 7, the outfit's latest addition to its aggressively styled line of gaming mice, minus the RAT moniker. Despite shedding the name of its predecessors, Mad Catz' latest point-and-clicker is as much of a RAT as the mice that came before it, but distinguishes itself with more buttons, more features and more color. Read on to see what's new, what's different and what should have stayed the same.

Gallery: Mad Catz Cyborg M.M.O. 7 gaming mouse hands-on

Continue reading Mad Catz Cyborg M.M.O. 7 gaming mouse hands-on

Mad Catz Cyborg M.M.O. 7 gaming mouse hands-on originally appeared on Engadget on Mon, 06 Feb 2012 10:00:00 EDT. Please see our terms for use of feeds.

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Networking tips offered to get next job ? Latin Business Today

By | Feb 5, 2012

UPLAND, Calif., Feb. 5 (UPI) -- It's purposeful networking, not simply schmoozing, that lands a person their next job, career coach Ford R. Myers says.

Myers' advice is timely, since Monday through Friday is International Networking Week, an initiative of BNI, an Upland, Calif., business networking and referral organization.

Myers, president of Career Potential LLC and author of "Get The Job You Want, Even When No One's Hiring," says job seekers can get hired through skillful networking, regardless of economic conditions.

"Networking is not about 'schmoozing,' or selling someone on the idea of hiring you," Myers advises in a release. "Rather, it's about purposefully and gracefully asking peers for help, advice, input and contacts -- and offering real value in return. Networking boosts job seekers' self-esteem and helps them connect people with opportunities -- especially when those opportunities are hard to come by."

Here are some tips from Myers to position yourself to get a job offer:

-- Schedule face-to-face meetings with your top tier contact list of people in the best position to help you.

-- Schedule phone meetings with other people who might lead you to key industry influencers.

-- Get at least 3-5 names from each person with whom you speak.

-- Be confident and purposeful.

-- Bring a list of questions and ask if it's OK to take notes.

-- Establish good rapport by getting the other person to talk about himself/herself.

-- Don't use the word "fired," be positive and focus on the future.

-- Don't make excuses or sound apologetic for your current career situation.

-- Look for opportunities to offer something of value in return for the other person's help and support.

-- Always send a "thank you" note immediately after your networking meeting.

Source: http://latinbusinesstoday.com/2012/02/networking-tips-offered-to-get-next-job/

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