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|By Jeremy Geelan
Article Rating:
|May 27, 2012 11:00 AM EDT
With Cloud Expo 2012 New York (10th Cloud Expo) now under three weeks away, what better time to introduce you in greater detail to the distinguished individuals in our incredible Speaker Faculty for the technical and strategy sessions at the conference...
We have technical and strategy sessions for you every day from June 11 through June 14 dealing with every nook and cranny of Cloud Computing and Big Data, but what of those who are presenting? Who are they, where do they work, what else have they written and/or said about the Cloud that is transforming the world of Enterprise IT, side by side with the exploding use of enterprise Big Data – processed in the Cloud – to drive value for businesses...?
CLOUD EXPO SPEAKER NAME: Henry Fastert
TWITTER: @SHICloud
COMPANY: SHI Enterprise Solutions Services Group
10TH CLOUD EXPO OPENING KEYNOTE TITLE: Mission-Critical Applications in the Cloud – Myth or Reality?
KEYNOTE DESCRIPTION: http://cloudcomputing.sys-con.com/node/2209206
BRIEF BIOGRAPHY: Henry Fastert is Chief Technologist and Managing Partner at SHI, where he has overall responsibility for the SHI Enterprise Solution Services (ESS) Group. Leveraging over 25 years of IT experience, his responsibilities include both strategic and operational direction for ESS, as well as providing expertise on infrastructure design, data center consolidation, and technology consolidation to SHI's largest customers.
Fastert comes to SHI from Unisys, where he was responsible for their North American professional services organization. His responsibilities included business development and service delivery, as well as providing deep expertise on ERP, Business Intelligence, infrastructure design, and data center consolidation. Prior to joining Unisys, he spent five years as VP and Chief Technologist for Melillo Consulting..
BLOG: blog.shicloud.com/p/bloggers.html
"The ability to flexibly and quickly adjust infrastructure allows IT to respond to line-of-business requirements in a way that was impossible prior to the advent of cloud computing. SHI has seen a very rapid growth in cloud demand in the last two quarters."
[from Exclusive Q&A with Henry Fastert, Cloud Computing Journal]
Unless your company can spend a lot of money on new technology, re-engineering your environment and hiring a comprehensive cybersecurity team, you will most likely move to the cloud or seek external service partnerships. In his session at 18th Cloud Expo, Darren Guccione, CEO of Keeper Security, revealed what you need to know when it comes to encryption in the cloud.
Jul. 24, 2016 05:00 PM EDT Reads: 2,340
We're entering the post-smartphone era, where wearable gadgets from watches and fitness bands to glasses and health aids will power the next technological revolution. With mass adoption of wearable devices comes a new data ecosystem that must be protected. Wearables open new pathways that facilitate the tracking, sharing and storing of consumers' personal health, location and daily activity data. Consumers have some idea of the data these devices capture, but most don't realize how revealing and...
Jul. 24, 2016 05:00 PM EDT Reads: 2,015
What are the successful IoT innovations from emerging markets? What are the unique challenges and opportunities from these markets? How did the constraints in connectivity among others lead to groundbreaking insights? In her session at @ThingsExpo, Carmen Feliciano, a Principal at AMDG, will answer all these questions and share how you can apply IoT best practices and frameworks from the emerging markets to your own business.
Jul. 24, 2016 04:15 PM EDT Reads: 1,538
Ask someone to architect an Internet of Things (IoT) solution and you are guaranteed to see a reference to the cloud. This would lead you to believe that IoT requires the cloud to exist. However, there are many IoT use cases where the cloud is not feasible or desirable. In his session at @ThingsExpo, Dave McCarthy, Director of Products at Bsquare Corporation, will discuss the strategies that exist to extend intelligence directly to IoT devices and sensors, freeing them from the constraints of ...
Jul. 24, 2016 03:45 PM EDT Reads: 1,718
You think you know what's in your data. But do you? Most organizations are now aware of the business intelligence represented by their data. Data science stands to take this to a level you never thought of – literally. The techniques of data science, when used with the capabilities of Big Data technologies, can make connections you had not yet imagined, helping you discover new insights and ask new questions of your data. In his session at @ThingsExpo, Sarbjit Sarkaria, data science team lead ...
Jul. 24, 2016 03:30 PM EDT Reads: 886
Extracting business value from Internet of Things (IoT) data doesn't happen overnight. There are several requirements that must be satisfied, including IoT device enablement, data analysis, real-time detection of complex events and automated orchestration of actions. Unfortunately, too many companies fall short in achieving their business goals by implementing incomplete solutions or not focusing on tangible use cases. In his general session at @ThingsExpo, Dave McCarthy, Director of Products...
Jul. 24, 2016 03:15 PM EDT Reads: 1,666
Traditional IT, great for stable systems of record, is struggling to cope with newer, agile systems of engagement requirements coming straight from the business. In his session at 18th Cloud Expo, William Morrish, General Manager of Product Sales at Interoute, outlined ways of exploiting new architectures to enable both systems and building them to support your existing platforms, with an eye for the future. Technologies such as Docker and the hyper-convergence of computing, networking and sto...
Jul. 24, 2016 03:00 PM EDT Reads: 1,002
WebRTC is bringing significant change to the communications landscape that will bridge the worlds of web and telephony, making the Internet the new standard for communications. Cloud9 took the road less traveled and used WebRTC to create a downloadable enterprise-grade communications platform that is changing the communication dynamic in the financial sector. In his session at @ThingsExpo, Leo Papadopoulos, CTO of Cloud9, discussed the importance of WebRTC and how it enables companies to focus...
Jul. 24, 2016 02:30 PM EDT Reads: 754
With an estimated 50 billion devices connected to the Internet by 2020, several industries will begin to expand their capabilities for retaining end point data at the edge to better utilize the range of data types and sheer volume of M2M data generated by the Internet of Things. In his session at @ThingsExpo, Don DeLoach, CEO and President of Infobright, discussed the infrastructures businesses will need to implement to handle this explosion of data by providing specific use cases for filterin...
Jul. 24, 2016 02:00 PM EDT Reads: 1,186
IoT generates lots of temporal data. But how do you unlock its value? You need to discover patterns that are repeatable in vast quantities of data, understand their meaning, and implement scalable monitoring across multiple data streams in order to monetize the discoveries and insights. Motif discovery and deep learning platforms are emerging to visualize sensor data, to search for patterns and to build application that can monitor real time streams efficiently. In his session at @ThingsExpo, ...
Jul. 24, 2016 01:30 PM EDT Reads: 804
Early adopters of IoT viewed it mainly as a different term for machine-to-machine connectivity or M2M. This is understandable since a prerequisite for any IoT solution is the ability to collect and aggregate device data, which is most often presented in a dashboard. The problem is that viewing data in a dashboard requires a human to interpret the results and take manual action, which doesn't scale to the needs of IoT.
Jul. 24, 2016 12:00 PM EDT Reads: 1,873
Internet of @ThingsExpo has announced today that Chris Matthieu has been named tech chair of Internet of @ThingsExpo 2016 Silicon Valley. The 6thInternet of @ThingsExpo will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA.
Jul. 24, 2016 12:00 PM EDT Reads: 1,886
What does it look like when you have access to cloud infrastructure and platform under the same roof? Let's talk about the different layers of Technology as a Service: who cares, what runs where, and how does it all fit together. In his session at 18th Cloud Expo, Phil Jackson, Lead Technology Evangelist at SoftLayer, an IBM company, spoke about the picture being painted by IBM Cloud and how the tools being crafted can help fill the gaps in your IT infrastructure.
Jul. 24, 2016 11:00 AM EDT Reads: 2,024
SYS-CON Events announced today the Enterprise IoT Bootcamp, being held November 1-2, 2016, in conjunction with 19th Cloud Expo | @ThingsExpo at the Santa Clara Convention Center in Santa Clara, CA. Combined with real-world scenarios and use cases, the Enterprise IoT Bootcamp is not just based on presentations but with hands-on demos and detailed walkthroughs. We will introduce you to a variety of real world use cases prototyped using Arduino, Raspberry Pi, BeagleBone, Spark, and Intel Edison. Y...
Jul. 24, 2016 11:00 AM EDT Reads: 1,308
Much of IT terminology is often misused and misapplied. Modernization and transformation are two such terms. They are often used interchangeably even though they mean different things and have very different connotations. Indeed, it is somewhat safe to assume that in IT any transformative effort is likely to also have a modernizing effect, and thus, we can see these as levels of improvement efforts. However, many businesses are being led to believe if they don't transform now they risk becoming ...
Jul. 24, 2016 11:00 AM EDT Reads: 1,108
CenturyLink has announced that application server solutions from GENBAND are now available as part of CenturyLink's Networx contracts. The General Services Administration (GSA)'s Networx program includes the largest telecommunications contract vehicles ever awarded by the federal government. CenturyLink recently secured an extension through spring 2020 of its offerings available to federal government agencies via GSA's Networx Universal and Enterprise contracts. GENBAND's EXPERiUS™ Application...
Jul. 24, 2016 11:00 AM EDT Reads: 1,793
SYS-CON Events announced today that LeaseWeb USA, a cloud Infrastructure-as-a-Service (IaaS) provider, will exhibit at the 19th International Cloud Expo, which will take place on November 1–3, 2016, at the Santa Clara Convention Center in Santa Clara, CA. LeaseWeb is one of the world's largest hosting brands. The company helps customers define, develop and deploy IT infrastructure tailored to their exact business needs, by combining various kinds cloud solutions.
Jul. 24, 2016 09:30 AM EDT Reads: 1,081
The best-practices for building IoT applications with Go Code that attendees can use to build their own IoT applications. In his session at @ThingsExpo, Indraneel Mitra, Senior Solutions Architect & Technology Evangelist at Cognizant, provided valuable information and resources for both novice and experienced developers on how to get started with IoT and Golang in a day. He also provided information on how to use Intel Arduino Kit, Go Robotics API and AWS IoT stack to build an application tha...
Jul. 24, 2016 08:45 AM EDT Reads: 911
Whether your IoT service is connecting cars, homes, appliances, wearable, cameras or other devices, one question hangs in the balance – how do you actually make money from this service? The ability to turn your IoT service into profit requires the ability to create a monetization strategy that is flexible, scalable and working for you in real-time. It must be a transparent, smoothly implemented strategy that all stakeholders – from customers to the board – will be able to understand and comprehe...
Jul. 24, 2016 08:30 AM EDT Reads: 2,067
It's 2016: buildings are smart, connected and the IoT is fundamentally altering how control and operating systems work and speak to each other. Platforms across the enterprise are networked via inexpensive sensors to collect massive amounts of data for analytics, information management, and insights that can be used to continuously improve operations. In his session at @ThingsExpo, Brian Chemel, Co-Founder and CTO of Digital Lumens, will explore: The benefits sensor-networked systems bring to ...
Jul. 24, 2016 08:30 AM EDT Reads: 1,473

subset: commoncrawl


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Safe Haven Hardcover – September 14, 2010
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Intrusion: A Novel
A loving couple, grieving the loss of their son, finds their marriage in free fall when a beautiful, long-lost acquaintance inserts herself into their lives. Learn More
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Safe Haven may be a bit predictable, but what makes it stand out for me was delving into the mind of a control freak. Katie Feldman has a secret she's running from and it's a big one. She's found a safe haven in a small South Carolina town where she can just live life on her own terms, safe from fear. Of course it doesn't last because stories need endings, and too much happiness can ruin a good story like nothing else.
The hero in this story is a man who does the right thing and loves his children. With only this guy, the story would be pure vanilla. Throw in a 'bad guy' who's good at his job and loves his wife, and you've got some swirls of chocolate thrown in that are delicious.
I like that the main character of the story realizes that the world if full of shadows where things aren't as simple as they might seem. I won't give away anything else in the story because I read one review and it spoiled the ending for me.
Safe Haven is just what you'd expect from Nicholas Sparks, but I think that's a good thing.
I haven't necessarily enjoyed every book he's ever written, but there have been a few that stand out. I enjoyed Message in a Bottle and Dear John. However, I'm beginning to wonder what's so great about his writing these days.
I found Safe Haven slow, plodding and, my goodness, I honestly found most of the book boring and repetitious, especially where Kevin, Erin's husband, comes in. I did appreciate the relationship between Katie and Alex, that they were taking time to get to know each other. I'm weary of the jumping to the next intimate level so quickly after introductions. Anyway, simply didn't warm up to this book, or the characters, at all. It was not a "I can't put it down" read. Matter of fact, I had to force myself to finish it. When I have to do that, I know I'm not involved. I found the ending contrived and just plain too fantastical. While I enjoy a nice twist,and I'm all for things of a Heavenly nature, it just felt twisted the way Sparks worked it.
Also, I guess I'm just tired of the beautiful woman/man who everyone can't take their eyes off of. Why can't a woman/man be OK but there's an essence about her/him that draws people. I'm tired of the beautiful people descriptions. Surely writers can use their imaginations to work believable characters, especially along the romance line, who are ordinary yet extraordinary?
If you're a fan of Nicholas Sparks regardless of what he puts out, you'll most definitely feel rewarded. If you've always been lukewarm to the monotony he can often write up,I'd suggest steering clear.
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True to life, touching, wanted the children to replace what they lost so early in their lives. Single Dads can make a difference.Published 6 days ago by rona craddock
i thoroughly enjoy the book. I really enjoy Nicholas Sparks booksPublished 8 days ago by Amazon Customer
I loved the movie so I wanted to read the book. The book was amazing!! It followed the movie and was extremely detailed!!Published 9 days ago by Tamara McKinney
The book was very interesting it had some different points covered compared to the movie. Not sure which I like better the movie or the book!Published 10 days ago by Sally Kiecker
This books was very good... But so much different from the movie.Published 12 days ago by Amazon Customer
A beautiful, loving story. Enjoyable reading, excellently written like always by Nicholas Sparks!Published 12 days ago by Florence Hatfield
Awesome book, loved the surprise at the end! Didn't see it coming.Published 13 days ago by Amazon Customer


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Linux Out of the Real World
The data produced by the payload is sent over a serial connection to the Rack Interface Computer. It bounces around on the orbiter for a little while, then is beamed to ground side via a satellite in NASA's Tracking and Date Relay Satellite System (TDRSS, everyone calls them Tetris satellites). The data goes through some other NASA systems (including communications-relay vessels in the Pacific Ocean) and, finally, makes it to MSFC. At MSFC, it enters a machine named (in good NASA style) the "Virtual Remote Users Gateway", VRUG for short. The VRUG is connected via a dedicated NASCOM line to our Remote Payload Operations Command Center (Remote POCC) in Boulder. The data then goes into a pile of ISDN-to-Ethernet routing hardware and into a network card in our ground side support computer (another Linux machine, used for development of the experiment's software and the analysis of returned data). On its screen, the ground side support computer displays squiggly lines (which the biologists like to look at) and pictures of plants. Another channel going up to orbit from ground side exists using the same hardware interfaces (RIC and VRUG).
Figure 9. The plants at the end of the mission
The data from the payload describes the conditions in the orbiter. From Boulder, it is sent over the Net to the ground control experiment in Florida. The ground control is similar to the payload in orbit, but it has an Ethernet card instead of the third serial port and a fragile (but cheap) and spacious magnetic hard disk of the garden variety. The ground control produces data of the same form as the orbiting experiment, but with (hopefully) different content. This data is sent back over the Net to the support machine in Boulder for analysis.
An unusual instance of a common problem affects communications with the orbiter. Each TDRSS satellite can see a small portion of the sky: when the limb of the Earth passes between the orbiter and the satellite, line of sight is lost and no data can be sent between them. Several TDRSS satellites are in orbit, and large portions of the orbiter's possible locations are covered, but not all. When no satellites are visible from the orbiter, no communication is possible. This situation is called a Loss Of Signal (LOS). NASA announces the LOSs with high accuracy and long precognition, but they still cause headaches for experimenters. (E-mail your politicians and ask for more Tetris satellites.)
Figure 10. The plants at the end of the mission
NASA does not guarantee the delivery or correctness of data sent through their pipes. I once asked a member of their technical staff how reliable the channel is, and he replied "Oh, I think probably no more than one corrupt or dropped character in a hundred." Observations made during last year's experiment indicate that the error rate is significantly lower than that estimate.
When you rent volume on the orbiter for your experiment, you can also rent bandwidth to ground side. You must specify the number of bits per second to reserve for your payload, and you are guaranteed no less. You then get a connection to the Rack Interface Computer. The RIC presents a three-wire RS-232 connection: transmit and receive only, no handshaking.
Data generated by the experiment must be encapsulated in little packets in accordance with a specification from NASA. The fields in the header and footer of these packets are used for routing within the orbiter's communication equipment and include a checksum. If the RIC accepts your packet, NASA will do their best to deliver it to your machine at ground side, but no guarantees are made. Data sent back to the payload from ground side is encapsulated in the same packets and go over the same wires. All packets traded between the payload and the RIC contain data that is from or to the ground side support computer, wrapped in RIC packets.
There is no change at the RIC interface, no automated notification from NASA to the payload that a LOS is imminent or occurring. The obvious way to do this notification would be to use the regular RS-232 handshaking lines.
Our Remote POCC is in Boulder, Colorado. At this end of the line, NASA presents a twisted pair Ethernet interface. You connect using TCP/IP to two specified ports on two specified hosts on this network. These computers are collectively called the Virtual Remote Users Gateway (VRUG). The VRUG interface is more complex than the RIC interface.
All communication with the VRUG (in both directions) is encrypted. The computer people at MSFC asked us to identify our operating system, and then supplied us with two object files, containing compiled C-callable functions to encrypt and decrypt data. The data sent over the TCP stream between our computer and theirs is packetized, but using packets different from those used by the RIC. These packets can contain data to and from the orbiter, commands to configure the VRUG interface and "telemetry" data from the orbiter (a standard data set provided by NASA at no extra charge, describing ambient conditions within the orbiter). Checksums are dutifully computed and checked on data going over the TCP link.
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One of the best things about the UNIX environment (aside from being stable and efficient) is the vast array of software tools available to help you do your job. Traditionally, a UNIX tool does only one thing, but does that one thing very well. For example, grep is very easy to use and can search vast amounts of data quickly. The find tool can find a particular file or files based on all kinds of criteria. It's pretty easy to string these tools together to build even more powerful tools, such as a tool that finds all of the .log files in the /home directory and searches each one for a particular entry. This erector-set mentality allows UNIX system administrators to seem to always have the right tool for the job.
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With all the industry talk about the benefits of Linux on Power and all the performance advantages offered by its open architecture, you may be considering a move in that direction. If you are thinking about analytics, big data and cloud computing, you would be right to evaluate Power. The idea of using commodity x86 hardware and replacing it every three years is an outdated cost model. It doesn't consider the total cost of ownership, and it doesn't consider the advantage of real processing power, high-availability and multithreading like a demon.
This ebook takes a look at some of the practical applications of the Linux on Power platform and ways you might bring all the performance power of this open architecture to bear for your organization. There are no smoke and mirrors here—just hard, cold, empirical evidence provided by independent sources. I also consider some innovative ways Linux on Power will be used in the future.Get the Guide


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By Alex Eisman I felt like I was on Mars, yet I was only 90 minutes south of Miami. I was a 15-year-old in Havana, two years before the United Stat...
Social determinants of health (SDOH), as identified by the CDC, are conditions in the places where people live, learn, work, and play that affect a wide range of health risks and outcomes.
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In what is one of the best examples of a partnership between sharing research from a world-class university and providing valuable insights to the public, "Sea Secrets" is made possible by many whose passion is to protect our environment and preserve our oceans.
As I drove past our neighborhood elementary school, I flashed back to when I used to drive by with our younger son strapped into his car seat. Each time, he asked me when it would be his turn to go to the "big kids' school." Last week, my husband and I delivered him to the real "big kids' school," the University of Miami.
The deepest thing I took away from reading American Conspiracy Theories was how hard it is to actually define the term. Especially while conducting the interview, I was forced to reexamine and redefine what I would personally qualify as a conspiracy theory. In other words, the book made me think -- especially about my own biases and perceptions.
On October 2, the Rosenstiel School of Marine and Atmospheric Science at the University of Miami further solidified its academic and research leader...
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Skip to comments.Dubai ports takeover prompts backlash
Posted on 02/16/2006 5:21:05 PM PST by Dane
Main page content: Dubai ports takeover prompts backlash By Stephanie Kirchgaessner and Edward Alden in Washington Published: February 16 2006 23:00 | Last updated: February 16 2006 23:00
Washington lawmakers on Thursday expressed deep reservations about Dubai Ports Worlds £3.9bn acquisition of P&O, the UK-based port operator, on the grounds that the deal represented a potential national security threat, and demanded that the White House re-open a regulatory review of the deal. ADVERTISEMENT
In a letter to Treasury secretary John Snow, Senator Richard Shelby, an influential Alabama Republican, stopped short of calling for the deal to be blocked, but said the transaction merited further scrutiny, potentially raising complications for DP Worlds bid. Mr Shelby is expected to call for a hearing to discuss the issue in coming weeks.
In a separate letter to Mr Snow, New York senator Chuck Schumer and others said US ports were the most vulnerable targets for terrorist attack. They questioned whether DP World, which is owned and controlled by Dubai, should be allowed to take over P&O, charging that Dubai was a key transfer point for shipments of nuclear components bound for Iran, North Korea and Libya.
The deal, which has already been given regulatory clearance by the White House, gives DP World container terminals at six ports on the east coast of the US, including New York and New Jersey.
Although the Department of Homeland Security is ultimately charged with cargo- screening functions, the legislators said the port facility itself was responsible for securing cargo coming in and out of the port and the hiring of security personnel.
After the 9/11 attacks, your department complained of a lack of co-operation by the UAE and other Arab countries as the US was trying to track down Osama bin Ladens bank accounts, the letter stipulated.
The growing congressional backlash against the deal is likely to reignite a debate in Washington on the effectiveness of the committee on foreign investments in the US, or Cfius, an inter-agency panel that vets foreign takeovers of US assets on national security grounds.
Cfius, which is chaired by the Treasury Department, approved Dubais bid for P&O after reviewing it for a standard 30-day-period, although the committee could have called for the deal to be examined more thoroughly over an additional 45 days.
Stewart Baker, assistant secretary for policy in the Department of Homeland Security, said that Cfius had completed its review after it was notified last year of the pending takeover, and had concluded there was no national security basis for blocking the transaction.
He said the US government had worked cooperatively with DP World in the past as part of the global container security initiative launched by the US after the September 11 attacks.
People familiar with the deal said administration officials were yesterday briefing lawmakers on the deal to assuage their concerns.
A spokesperson for the Treasury Department said the administration would not re-open its review unless evidence emerged that DP World had given the committee false information.
Cfius came under harsh scrutiny by lawmakers last year following a failed attempt by CNOOC, the Chinese oil company, to takeover California-based Unocal. Although the deal was quashed before Cfius reviewed the transaction, the widespread opposition to the deal in Congress prompted the Treasury Department and other agencies to promise to increase transparency at Cfius by communicating more with Congress and to be more willing to conduct full investigations of deals.
Basically, we have one foreign company operating port services being sold to another foreign company.
Now without the usual hyoerbole that seems to go along with these threads, maybe we can figure this out. Notice that the article says the port facility itself is responsible for security. I would surmise that the foreign port company is supervised by the municipal port authority and in one case that would be the NY/NJ port authority, so that means the local authorites are charged with security(i.e the Dubai company is not in charge with security).
BTW, notice how chuckie schumer mentioned Libya. I guess he missed the news that Libya has given up it's WMD program, due to US pressure and is now cooperating with the US.
I know a lot of people are queasy about this, but it should not be looked through the prism of hyperbole from the press and a Senator who falls in love with every TV camera there is(chuckie schumer).
And also what are other "solutions". Should there be a govt. takeover. Should the govt. block a sale of one foreign owned company to another foreign company.
So instead of the usual knee jerk hyperbole, how about intelligent discussion.
"Reality Based
Since Feb 17, 2006"
Welcome to Free Republic
great info in this thread..
I think that is the misnomer, they are not taking over the ports, they are operating from the ports which are under the local municipal port authority.
Like I said before it is one foreign company buying the port facilities from another foreign company(which was British). And I wonder where schumer's outrage was over that foreign company that was overseen by the local port authority.
In actuality, this is a business deal, with a UK firm that has contracts in 6 US ports being taken over by another firm.
It's not like that DP world is taking over the ports. They will have a contract to load and unload ships from all a round the world from that port(which is under US control and overseen by local, state, and federal security).
Now there can be a debate about if that is good or not, but let's get rid of the chuckie schumer misnomer that DP World is "taking" over the ports, which they are not.
There are no muslim countries that are staunch U.S. allies. Their loyalty is to their religion which is in direct conflict with the security of the U.S. There is no such thing as a tolerant branch of islam. This is a business deal that would give a potentially hostile foreign power all information about the security of vital ports. What possible reason would we have to allow such a potentially suicidal transaction?
This is just plain stupid.
Maybe he doesn't trust Muslims.
Here are the new players: left Pres.UAE.... Right VP UAE
Rep. Mark Foley said Sec. John Snow told him he didn't know anything about the deliberations, yet he heads the Department which oversees this very decision. Additionally, when John Snow ran CSX Transporation, he sold his entire port operations to a Dubai-based corporation. So it appears Secretary Snow is either a fool, or a damned liar.
Hoooo boy!
Actually that happened one year after Snow had left CSX.
Snow is a former chairman of freight rail company CSX Corp. (CSX.N: Quote, Profile, Research), which sold its global port assets to Dubai Ports World for $1.15 billion in 2004 -- the year after Snow had left the company for the Bush administration.
So who is being the liar now.
Will someone please tell me why there is not an American firm that can handle this acquisition? Why is it not even talked about? How about the cities where the ports are located having ownership, control and management?
You are right about that part and I stand corrected. However his claims of not knowing about the CFIUS deliberations were not credible.
JMO, chuckie schumer would be screaming 100 times louder if Halliburton took over.
I have no idea why there is not an American company that could take over.
What can be done to stop this? If we let 6 major ports be controlled by a terrorist-sponsoring nation, we're not serious about the War on Terror.
Allowing this to transpire is utter foolishness.
Are they holding hands or is one of them slipping a little something to the other?
Come right in with nukes,we have your prey rugs on the
dock so you can prey to allah before setting off the
nuke oh we also have some tea incase your thirsty.Oh
do you want some signs to show you the best place to set
your nuke off.We aim to please.
"If we let 6 major ports be controlled by a terrorist-sponsoring nation, we're not serious about the War on Terror."
Good job that's not happening then. The control of the ports isn't changing and P+O cannot really be considered a 'terrorist-sponsering nation'.
Err Dubai. P+O of course would not be a nation at all. Oh, it's way to early!
Do you ACTUALLY believe that Multi-Billion dollar deals with Multi-National Companies MAGICALLY occur AFTER certain players are bought by one of the interested parties? That they ONLY take one year?
Just how long does a Billion dollar deal take, Dane?
Because I'd bet that this buyout was proposed, started, researched, and negotiated when Snow was still head of CSX!
But not in your Kool-Aid addled world! Multi-National deals and Billion dollar buyouts ONLY occur when GWB and his sock-puppets say they do!
Wow, you really do NOT have a clue of what you speak...
January 13, 2003, John Snow, CEO of CSX leaves and Bush nominates John Snow to be Secretary of the treasury
December 9, 2004, CSX World Terminals is purchased by DPW
January 24, 2006, David Sanborn leaves DPW and joins Whitehouse as new administrator of the Maritime Administration in the Transportation Department
Present, the fight whether to allow DPW into 6 US Ports
It's hard to be logical with the Kool Aid cultists..
And I have concluded there is little difference between the Treasury Department and Foggy Bottom when it comes to national security and protecting America's best interests. What a sorry assed collection of useless bureaucrats.
"Why can't I QUIT you?!"
Correct. But then that's just Dane being Dane.
For once, Senator Charles Schumer (D-NY) is right about something. I guess it just goes to show that even a blind bird catches a worm once in a while.
What Schumer is right about is his opposition to the deal that would allow Dubai Ports World, a company based in the United Arab Emirates (UAE), to gain control of six major American ports through its purchase of London-based Peninsular and Oriental Staem Navigation Company, which currently runs the ports: New York, Newark, Philadelphia, Baltimore, New Orleans, and Miami.
The obvious question is, aren't there any American companies capable of running these ports? And the more important question is, Are we serious about the War on Terror? We have left our borders wide open for anyone (including terrorists) to come through and now we are turning control of six major ports over to a country that sponsors terrorism.
The deal has been approved by the Committee on Foreign Investment in the United States, but Schumer, along with Senators Tom Coburn (R-Okla.) and Frank Lautenberg (D-NJ) and Representaties Vito Fossella (R-NY), Christopher Shays (R-Ct.), and Mark Foley (R-Fla.), is trying to stop the transaction. REpresentaigve Peter King (R-NY), chairman of the House Homeland Security Committee, has urged the Bush Administration to reconsider.
New York City Council members of both parties have spoken out against the deal. "On its face, this looks like insanity to me," the Republican minority leader of the City Council, James Oddo, said. The chairman of the Council's Public Safety Committee, Democrat Peter Vallone, said that the deal "raises some legitimate concerns."
The Bush administration considers the UAE a key ally in the War on Terror, but the UAE has been a sponsor of terrorism. It was the home of Marwan al-Shehi, one of the 19 hijackers who killed 3,000 people on September 11, 2001. Other hijackers travelled through the UAE. The UAE was an important financial base for the September 11 hijackers. The country is a transit point for Al Qaeda. It has been a financier of Al Qaeda. It continues to regard the Taliban as the legitimate government of Afghanistan and is militantly anti-Israel. The CIA has identified the UAE as a major center for drug running and money laundering. It also has close ties to Iran.
The UAE is a federation of seven Arab emirates on the Arabian peninsula.
As Rep. Foley has said, "If our ports are the most vulnerable targets for terrorism and if we are at war, as the president says, we should be overly critical of handing over the management of our ports to any foreign countries, post 9/11."
What happens if nuclear materials pass through one of these ports? How will the UAE direct its employees to deal with that? The threat to our national security is clear.
That is why these Senators and Representatives are right to speak out against the transfer of control of our ports to UAE control, just as Red Chinese control of the Panama Canal poses a threat to our national security.
Can we afford to turn control of key ports over to a company controlled by a terrorist-sponsoring government?
I am sorry, but you simply incorrect on all of your points. With all due respect, you are uninformed, nativistic, and prejudiced. Thousands of Muslims have died as our allies resisting Islamofascist extremism. Thousands more died resisting Communism. The kind of attitude you exemplify will simply drive more Muslims into the camp of the extremists.
The ignorance and childish naivete of people like you threatens my security. Kumbaya is not a strategy against a ruthless enemy.
What flavor kool-aide do I need to drink to make this Arab control of our ports sound like a good idea?
Bush is so freaking stupid on this. Who in the XXXX is in control in the White House on policy and politics.
It appears that he is batting 50% instead of in the high 90s right now. I cannot believe the his instincts are in this or Karl Rove's. He has badly, badly misjudged the populace on this issue. It is literal idiocy.
We even have Senator Schumer suggesting Halliburton, which is beyond sensible, but maybe only a Dem could do it.
Maybe what should be printed and discussed is HOW MANY COMPANIES in the world can handle this job?
Bush is even allowing Hillary Clinton to get to his right for crying out loud...why would he allow that to happen.
Novak's report says today that the impression that he is seeing people get, is that control in the White House is off balance.
Who's on first? What's on Second? and I don't know is on Third?
If all this weren't so important with the mid terms coming up this year, you would have to laugh and cry.
From one of my favorite Star Trek films:
The lady says, "Is this that dip s**t stuff?"
and Captain Kirk says, "No Mam, no dip s**t.
The problem? This has dip s**t written all over it, just like Mier's for SCOTUS!!!!!!!!!!!!!!
Where has Bush's game gone??????
I understand the line of reasoning...
but unequivocally disagree!!!!!!!!!!!!!
Disclaimer: Opinions posted on Free Republic are those of the individual posters and do not necessarily represent the opinion of Free Republic or its management. All materials posted herein are protected by copyright law and the exemption for fair use of copyrighted works.


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To work as a doctor in another EU country, you must apply to the authority that oversees the doctor profession in that country, providing proof of your qualifications. The authority must:
- acknowledge your application within 1 month of receiving it, and ask you missing but necessary documents to process the application
- assess your qualifications and decide whether to grant your application within 3 months (or 4 months for certain complicated cases in the area of non automatic recognition)
If you do not accept their decision, you can appeal to the relevant court in that country. As to the type of documents an authority may ask, please see also the Code of conduct
Which qualifications are recognised?
Automatic recognition
Basic medical training and general practitioner qualifications
Basic medical training and general practitioner training are automatically recognised throughout the EU, entitling you to work in any EU country, if they are listed in respectively Annex V, points 5.1.1 and 5.1.4 of the Directive 2005/36/EC, which are conform to Art. 24 or 28 of the Directive, or benefit from acquired rights (Art. 23 and 30 of the Directive).
Specialist doctors' qualifications
Some specialist doctors' qualifications are automatically recognised in certain EU countries. These are listed in:
- Annexes V, point 5.1 of the Directive on recognition of professional qualifications
All of the countries listed for each given field will automatically recognise each others' qualifications in that field.
Non-automatic recognition
Even if your qualification does not meet the automatic criteria for recognition, it may still be recognised in another EU country, under the general system for recognition of qualifications. See article 10 to 15 of the Directive.
If there are major differences between the qualifications you have from your home country and the qualifications required to work in the EU country where you want to work, you may be asked to:
- sit an aptitude test to check that you have the skills required in your chosen country
- complete an adaptation period
Links – detailed rules
- Directive on recognition of professional qualifications (2005/36/EC)
- Automatically recognised qualifications – recent additions
Read more:
- Updates list of diplomas
- Archives
- Other documents:


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Diversified Financial Services
Company Overview of Säk I AB
Company Overview
As of July 6, 2011, Säk I AB was acquired by Investment AB Latour (publ). Säk I AB is a public investment firm listed on the Stockholm Stock Exchange O-list. The firm mainly invests in listed companies but also invests in private growth companies. Säk I AB is based in Stockholm, Sweden.
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11th Circuit gets new chief judge
Posted: 06/02/2009 - The federal appeals court in Atlanta has a new chief judge.
Posted: 06/02/2009 - The federal appeals court in Atlanta has a new chief judge.
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Posted: 05/28/2009 - A U.S. health official said a swine flu vaccine could be available as early as October, but only if vaccine production and testing run smoothly this summer.
Posted: 05/28/2009 - Documents from a police investigation into the former University of Georgia professor who fatally shot his wife and two others show he had secretly recorded a conversation between he and his wife about her affair with one of the men killed.


text:
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Conceived and designed the experiments: HK MK. Performed the experiments: HK OT YT EJW MK. Analyzed the data: HK OT YT MK. Contributed reagents/materials/analysis tools: HCL JWR KMK HY. Wrote the paper: HK JRW MK.
Aberrantly activated Wnt/β-catenin signaling is important in hepatocellular carcinoma (HCC) development. Downstream gene expressions involving the Wnt/β-catenin cascade occur through T-cell factor (TCF) proteins. Here, we show the oncogenic potential of human TCF-4 isoforms based on the expression of a single conserved SxxSS motif.
We investigated the TCF-4J and K isoform pair characterized by the presence (K) or absence (J) of the SxxSS motif. The mRNA expression profiles were examined in 47 pairs of human HCCs and adjacent non-cancerous liver tissues by RT-PCR. Proliferation, sphere assays and immunoblot analysis were performed under normoxia and hypoxia conditions. The ability of HCC cells overexpressing TCF-4J (J cells) and K (K cells) to grow as solid tumors in nude mice was explored.
TCF-4J expression was significantly upregulated in HCC tumors compared to corresponding peritumor and normal liver and was preferentially expressed in poorly differentiated HCCs. In contrast, TCF-4K was downregulated in those same HCC tumors. TCF-4J-overexpressing HCC cells (J cells) revealed a survival advantage under hypoxic conditions, high proliferation rate and formation of aggregates/spheres compared to overexpression of TCF-4K (K cells). The hypoxic J cells had high expression levels of HIF-2α and EGFR as possible mechanisms to promote tumorigenesis. Increased stability of HIF-2α under hypoxia in J cells was associated with a decreased level of von Hippel-Lindau (VHL) protein, a known E3 ligase for HIF-αs. In a xenograft model, the J cells rapidly developed tumors compared to K cells. Tumor tissues derived from J cells exhibited high expression levels of HIF-2α and EGFR compared to the slow developing and small K cell derived tumors.
Our results suggest that the specific TCF-4J isoform, which lacks a regulatory SxxSS motif, has robust tumor-initiating potential under hypoxic conditions.
The Wnt/β-catenin signaling pathway plays a crucial role in cell fate determination and stem cell renewal in adult tissue [1], [2]. Genetic and/or epigenetic deregulation of this pathway leads to aberrant nuclear accumulation of β-catenin where it binds with T-cell factor-4 (TCF-4) to form a transcriptional complex [3]; this event drives gene expression such as c-Myc, cyclin D1, and EPCAM [4], [5], [6] that contributes to a malignant phenotype.
Hepatocellular carcinoma (HCC) is the third common cause of cancer mortality worldwide [7]. Aberrantly activated Wnt/β-catenin signaling due to overexpression of upstream components of this pathway such as Frizzled (FZD) receptors and Wnt ligands is a common early event in the molecular pathogenesis of this disease [8], [9], [10]. However, the involvement of the canonical Wnt TCF-4 effector proteins in this process has yet to be examined.
The human TCF-4 gene (TCF7L2) consists of 17 exons and has several alternative splicing sites in exons 13–17 and exon 4 [11]. The alternative splicing sites in the central domain of TCF-4 can also generate isoforms with or without the conserved LVPQ and SxxSS motifs located at the end of exon 7 and the beginning of exon 9, respectively [11], [12]. Recently, we identified and characterized 14 (12 of which are unique) TCF-4 isoforms derived from human HCC cell lines [13]. Among such isoforms, three structurally identical pairs (TCF-4J and K; TCF-4A and B; TCF-4G and H) were observed that differed only by the presence (K, A, and H) or absence (J, B, and G) of a SxxSS motif. In this context, two pairs of isoforms (TCF-4A and B, and TCF-4H and G) are short forms, whereas TCF-4K and J are long forms due to the inclusion of a C-terminal tail (exon 13–17) [13]. Previous studies suggest that the SxxSS motif may modulate transcriptional activity of TCF-4 [12]; however, both the cell-based and in vivo functional consequences of the SxxSS motif expression as well as subsequent gene regulatory activity have not been determined.
Emerging evidence indicates that both embryonic (ES), adult and inducible pluripotent stem (iPS) cells [14] prefer hypoxic conditions for growth and survival [15]. Hypoxia generates diverse cellular signals through the stabilization of hypoxia-inducible factors (HIF-αs) such as HIF-1α and HIF-2α. Recent studies reveal that HIF-αs interact with β-catenin and, therefore, may regulate TCF-4-driven gene expression not only in stem/progenitors but also tumor cells experiencing hypoxic conditions during rapid growth [16], [17]. These findings imply that Wnt/β-catenin/TCF-4 signaling may be directly regulated by the professional oxygen-sensing system in the nucleus. For instance, stabilized HIF-2α, a partner of β-catenin and often found in the hypoxic core of the tumor, upregulates the expression of epidermal growth factor receptor (EGFR) and may contribute to tumor growth [18]. In human HCC, the HIF-αs are involved in the multi-step process of tumor dedifferentiation via promotion of angiogenesis [19]. Accordingly, we determined if different functional properties of TCF-4 isoforms associated with the HCC malignant phenotype were regulated in the context of a SxxSS motif-dependent mechanisms under conditions of oxygen deprivation.
Full ethical approval was obtained for all human sample collections from either the Asan Medical Center Ethics Committee or the Kurume University Ethics Committee. All samples were obtained with written consent. All animal experiments were conducted in accordance with the NIH Guidelines for the Care and Use of Laboratory Animals and were approved by the Lifespan Animal Welfare Committee of Rhode Island Hospital, Providence, RI (permit number A3922-01).
Preparations of human TCF-4A, B, J and K-myc plasmids have been previously described [13]. Two independent RT-PCR analyses were performed using 47 pairs of human HCCs (Tables 1 and and2)2) as previously described [13].
Human cell lines Hep3B, Huh7, HepG2, and HEK293 were obtained from the American Type Culture Collection (ATCC, Manassas, VA). FOCUS cell line was obtained from Dr. Wands (Liver Research Center, Rhode Island Hospital, RI). HAK-1A and HAK-1B cells were provided by Dr. Yano (Kurume University School of Medicine, Japan). The immortalized hepatocyte derived cell line OUMS-29 was a kind gift from Drs. Namba and Kobayashi (Okayama University, Japan). Hep3B, Huh7, HepG2, and FOCUS cell lines are hepatitis B virus (HBV)-related HCC cells. HAK-1A and HAK-1B cells are hepatitis C virus (HCV)-related HCC cell lines. All cells have been described and employed in previous studies (Hep3B [20], Huh7 [21], HepG2 [20], HEK293 [22], FOCUS [23], and HAK-1A and HAK-1B [24]). To generate stable transfectants, HAK-1A cells were transfected with the TCF-4 or empty vector plasmid using TransIT-LT1 Reagent (Mirus Bio Co., Madison, WI) and selected by G418.
Hypoxic conditions (1% oxygen) were produced by an incubator equipped with an oxygen concentration regulator system (ASTEC, Fukuoka, Japan). Alternatively, cells were exposed to hypoxia-mimic chemical cobalt chloride (CoCl2, Sigma-Aldrich, St. Louis, MO) [25].
Cells, grown on 35 mm diameter Glass Bottom Dishes (MatTek, Ashland, MA), were fixed with cold acetone/methanol (11) for 10 min, and then washed in PBS containing 0.05% Tween 20 (PBS-T). Nonspecific reactions were blocked with Protein Block Serum-Free (DAKO North America, Carpinteria, CA) and then incubated with a mouse anti-Myc-tag antibody (Santa Cruz Biotechnology, Santa Cruz, CA) or a rabbit anti-HIF-2α antibody (Abcam, Cambridge, UK) at 4°C overnight. After washing in PBS-T, the specimens were treated with Alexa Fluor goat anti-mouse or goat anti-rabbit IgG (H+L) antibody (Molecular Probes, Eugene, OR) for 40 min at RT, and then counterstained by VECTASHIELD Mounting Medium with 4′,6-diamino-2-phenylindole (DAPI) (Burlingame, CA). The Zeiss LSM510 Confocal Laser Scanning Microscope (Carl Zeiss MicroImaging, Inc., Thornwood, NY) equipped with the user interface software Zen 2008 was used to visualize the immunofluorescence staining for Myc-tag or HIF-2α and nuclear localization was provided by DAPI. The negative controls were obtained by incubating cells with non-specific mouse IgG or rabbit IgG as described above.
The primary antibodies used were against for Myc-tag, β-catenin, actin, γ-tubulin, lamin A/C, HIF-1α, c-myc, and ubiquitin (Santa Cruz Bio, Santa Cruz, CA), TCF-4, EpCAM, and von Hipple-Lindau (VHL) (Cell Signaling Technology, Beverly, MA), HIF-2α (Abcam, Cambridge, UK), and keratin-19 (K-19) (DAKO, Carpinteria, CA). A TCF-4 rabbit derived mAb (Cell Signaling Technology, Cat# 2569) recognizes surrounding the Leu330 of human TCF-4 and detects endogenous TCF-4 proteins including both the short and long isoforms. For immunoprecipitation, cell extracts were prepared as previously described [26]. Total cell lysate or nuclear extract were incubated with antibodies against HIF-1α, HIF-2α, and VHL followed by Recombinant Protein G Agarose (Invitrogen, Carlsbad, CA). Immunoblots were probed with antibodies to ubiquitin, HIF-1α, HIF-2α, and VHL and detected with HRP-labeled anti-mouse or anti-rabbit IgG (Amersham Pharmacia Biotech, Buckinghamshire, UK) using ECL Advanced kit (Amersham). Positive signals were captured by the Image analyzer LAS-1000plus (Fujifilm, Tokyo, Japan), and the band intensity of protein was determined using Image Gauge version 3.45 (Fujifilm).
Cells were seeded into 24-well plates at a density of 2.5×103/well. A colorimetric assay (CellTiter 96® Aqueous One Solution Cell Proliferation Assay; Promega, Madison, WI) was performed at days 0, 1, 3, 5, and 7, and the signals were measured using Spectra Max M5 (Molecular Devices, Sunnyvale, CA).
Single-cell suspensions were put into Ultra Low Attachment 6-well plates (Corning, Lowell, MA). After 27 days, the number of cell aggregates/spheres per well was quantified and photographed under the Zeiss Axiovert 200 M Fluorescence Microscope (Carl Zeiss MicroImaging). The area of the cell aggregates was measured using the MetaMorph 6.0 software (Molecular Devices).
Cells were exposed to 5 µg/mL cycloheximide (CHX; Sigma-Aldrich) for 0 or 60 min at the last phase of the 48 h CoCl2 treatment. Total cell lystes were used for immunoblot analysis to evaluate HIF-1α and HIF-2α expression levels after protein synthesis was inhibited by CHX.
HAK-1A-derived stable clones were used; parental HAK-1A cells do not form tumors in nude mice, and are therefore suitable for assessment of malignant transformation following stable transfection with TCF-4 isoforms [24]. Cells (1×106) were sub-cutaneously injected into the back of 5-week old male BALB/c nude mice (n =12) (Taconic Farms, Cranbury, NJ). The tumor size was measured twice per week, and the tumor volume (mm3) was estimated using the equation length × (width)2×0.5. When the longer diameter reached 10 mm, the mice were sacrificed and the tumors were prepared for subsequent analysis.
Paraffin-embedded tissue sections were deparaffinized and heated in 10 mM citrate buffer at 120°C for 5 min for antigen retrieval. The sections were pre-blocked by Protein Block Serum-Free (DAKO) and incubated with primary antibodies for HIF-2α (Abcam) and EGFR (D38B1 XP™) (Cell Signaling Technology). After washing, the sections were incubated with EnVision secondary antibodies labeled with HRP-polymer complexes (DAKO) and visualized by 0.1% 3–3′′-diamino-benzidine-tetrahydrochloride. The cell nuclei were counterstained with hematoxylin. Specimen incubated with mouse or rabbit IgG were set as the negative controls.
Statistical significance was performed by the Mann-Whitney U test using GraphPad Prism 4.0 software (San Diego, CA). P<0.05 was considered statistically significant. The Pearson's correlation coefficient was used to calculate correlation between expression of TCF-4J and K.
Expression of TCF-4 isoforms with or without the SxxSS motif may influence their transcriptional activity [12], [13], [27]. Therefore, we determined if three pairs of isoforms (TCF-4A and B; TCF-4G and H; and TCF-4J and K) exhibited different expression profiles that may suggest SxxSS-dependent regulation in human HCC tumors. The relative mRNA levels of these TCF-4 isoforms in 27 pairs of HCC tumors and corresponding adjacent liver tissue obtained from chronic HBV-related (26/27) disease were measured. Comparisons were made to three normal liver specimens by semi-quantitative RT-PCR (Fig. 1A, Fig. S1A and C, and Table 1) as previously described [13]. TCF-4J expression was significantly upregulated in HCC tumors compared to peritumor tissue and normal liver. Among the 27-paired samples, 85% had increased TCF-4J expression in HCC compared to matched-peritumor tissue. In contrast, TCF-4K was significantly downregulated in tumors compared to normal liver and peritumor tissue (Fig. 1A). Furthermore, a similar expression profile of these TCF-4 isoforms was observed in a second cohort of HCC-adjacent uninvolved tissue pairs from individuals where 85% (17/20) of the tumors were related to chronic HCV infection (Fig. 1B, Table 2). However, this differential expression pattern was not observed in the two other pairs of short isoforms (TCF-4A and B, G and H) (Fig. S1). TCF-4A, a short form of K was significantly downregulated in HBV-related HCC tumor compared to normal liver as well as peritumor tissue. In addition, the expression level of TCF-4B, a short form of J, was found to be similar among normal, tumor, and peritumor tissues (Fig. S1A). Furthermore, the other pair of TCF-4 isoforms (G and H) also revealed no differences of their expression levels among normal, HCC tumor, and peri-tumor tissues (Fig. S1C). In HCV-related HCC tissues, however, all four isoforms (TCF-4A and B, G and H) were significantly downregulated in both peritumor and tumor tissues compared to normal liver; in addition no difference was found between tumor and peritumor samples (Fig. S1B and D). These results indicated that the influence of SxxSS motif on generation of a HCC malignant phenotype may be associated with the long (TCF-4J and K), not short isoforms (TCF-4A and B, G and H).
Next, we analyzed the correlation between clinicopathological features and expression of these TCF-4 isoforms (Table 3, Table S1). The results suggest that only the degree(s) of tumor differentiation significantly correlated with TCF-4J and K expression levels (Table 3). Indeed, the TCF-4K level was significantly decreased in poorly differentiated (PD) HCC tumors, which was in contrast to high level of TCF-4J found in those same HCC tumors (Fig. 1A and B, Table 3). Such studies suggest that the upregulated expression of TCF-4J may be a common feature of PD HCC. In support of this hypothesis, TCF-4J was highly expressed in both HBV-related (HepG2, Hep3B, Huh7, and FOCUS) and HCV-related (HAK-1A and HAK-1B) cell lines (Fig. 1C). This isoform was also expressed in OUMS-29 and HEK293. In contrast, TCF-4K expression level was very low in PD HCC tumors and cell lines except for HAK-1B and HEK293 (Fig. 1A, B, and C). In contrast, the two pairs of short isoforms (TCF-4A and B, or G and H) did not demonstrate such a correlation between clinicopathological features and their expression of the SxxSS motif (Table S1).
Further analysis demonstrated that TCF-4J expression in HCC was significantly inversely correlated with TCF-4K (Fig. 1D, left panel; p =0.0031, correlation coefficient r =0.297). The inverse correlation between TCF-4J and K expression levels was more evident in PD HCC (Fig. 1D, right panel; p =0.0077, r =0.437). Thus, loss of the SxxSS motif in the long isoforms of TCF-4 due to a splicing event was associated with a PD HCC phenotype.
To better understand how expression of the SxxSS motif may promote the malignant phenotype of HCC in vitro, we first examined the localization of TCF-4J and K in HAK-1A HCC cell lines after transient transfection. As shown in Fig. 2, cellular expression was predominantly observed in the nucleus as assessed by confocal microscopy and nuclear/cytoplasmic fractionation studies, suggesting that exogenous expression of TCF-4J and K proteins will localize to the nucleus compartment.
During the multi-step process of hepatocarcinogenesis, less differentiated HCC cells arise from the center of HCC tumor nodules revealing a "nodule-in-nodule" histologic appearance on pathologic examination of tissue sections [28]. The central tumor area generates hypoxic stress to tumor cells and promotes apoptosis [18], [29]. However, there may be a hypoxia-resistant population of HCC cells that survive under these severe conditions to promote tumor growth through accelerated angiogenesis and vascular invasion. These two phenomena are regulated by HIF-1α and HIF-2α [30], [31]. Therefore, we hypothesized that the high expression of TCF-4J in PD HCC was a possible reflection of an induced hypoxia-resistant HCC phenotype. To test this idea, stable cell clones overexpressing TCF-4J (J cells) and TCF-4K (K cells) were established, and comparisons were made to empty vector-transfected cell clones (EV cells), as well as to the parental HAK-1A cells. Under phase-contrast microscopy, the morphologic appearance of these clones was similar and comparable to the parental cells (Fig. 3A), which was in contrast to the HAK-1A-derived dedifferentiated cell clone HAK-1B [24].
Wnt/β-catenin signaling often correlates with stem cell-like molecular alterations [32], [33]. In this regard, the expression of EpCAM, a putative liver cancer stem cell (CSC) marker and a β-catenin/TCF-4 downstream target gene product [6], was not upregulated in these cells. However, the expression of K-19, a biliary lineage marker for an aggressive HCC phenotype [34], was increased in J cells but not in K, EV, or parental HAK-1A cells (Fig. 3B).
Consistent with our previous report in Huh7 cells [13], J cells had an increased basal growth rate compared to K and EV control cells. The J cells were resistant under chemically induced, severe hypoxic conditions (150 µM CoCl2), proliferated significantly faster than K or EV cells (Fig. 3C). This finding was further supported when cells were exposed to 1% oxygen for 5 days (Fig. 3D). Cell growth in hypoxia (1% oxygen) was reduced in EV and K cells compared to cell proliferation in normoxia (20% oxygen) by 22 and 27%, respectively, while J cells demostrated only a 13% reduction in cell growth. Moreover, robust resistance to severe hypoxia in the J cells was also exhibited in a sphere assay. In this context, the J cells formed the largest cell aggregates at all the CoCl2 concentrations employed. The J cells' potential for anchorage-independent growth was most evident with severe hypoxia induced by 150 µM CoCl2 (Fig. 3E).
Based on the hypoxia-resistant features exhibited by J cells, we determined if these cells expressed HIF-αs under hypoxia. Both HIF-1α and HIF-2α expression in J and K cells were explored since HIF-2α has recently been shown to be a key molecule that promotes survival of CSCs under hypoxic conditions [35], [36], [37]. As expected, HIF-1α and HIF-2α expression were induced with moderate hypoxia (75 µM CoCl2) in EV, J and K cells; however, under severe hypoxic conditions (150 µM CoCl2), only J cells maintained these high protein levels (Fig. 4A). Stabilized HIF-2α, often found in the hypoxic core of the tumor, upregulated the expression of EGFR which may contribute to tumor growth [18], and the activation of EGFR signaling pathway was associated with the development of K-19-positive HCC [38]. In J cells, expression of EGFR and K-19 was associated with an increased level of HIF-2α (Fig. 4A). The consistent HIF response was also observed in 1% oxygen-exposed J cells (Fig. 4B). Furthermore, the 150 µM CoCl2-induced HIF-2α expression was localized in the nuclei of J cells as assessed by immunofluorescence staining (Fig. 4C).
We also determined if HIF-αs upregulation under severe hypoxia in J cells was linked to loss of the SxxSS motif. If that were the case, a similar result would be found in the TCF-4B overexpressing cells (B cells), the so-called "short isoform" of the TCF-4J (Fig. 2A). In this regard, total cell lysates were subjected to immunoblot analysis to compare B with A cells where TCF-4A is overexpressed as the "short form" counterpart of TCF-4K (Fig. 2A). It is important to note that an increase in total cell lysate HIF-α expression was evident in B cells (Fig. 4D) and suggests that the SxxSS motif may have a regulatory role in modulating HIF-αs expression under severe hypoxic conditions generated by 150 µM CoCl2 in two pairs of TCF-4 isoforms.
Because HIF-αs proteins are highly degraded by the ubiquitin-dependent proteasomal pathway, it was hypothesized that a differential HIF-αs degradation mechanism may exist under severe hypoxia in J and K cells. The levels of HIF-1α and HIF-2α were substantially increased in K cells treated with a proteasomal inhibitor (MG-132) compared to non-treated K cells; in contrast we observed less of an increase in these proteins in EV and J cells (Fig. 5A). Accumulation of HIF-αs in the MG-132-treated K cells strongly suggests that SxxSS motif-harboring K cells may degrade HIF-αs under hypoxic conditions in a proteasomal-dependent manner. Indeed, protein stability of HIF-αs was found to be decreased in K cells (Fig. 5B). We found that the level of VHL protein, a known E3 ligase for HIF-αs, in K cells was higher compared to J cells in the nucleus, but not cytoplasm, which coincided with intense nuclear accumulation of HIF-1α and HIF-2α in J cells under hypoxic conditions (Fig. 5C). We further examined interaction between HIF-2α and VHL on nuclear extracts and found that nuclear VHL in K cells strongly interacted with HIF-2α compared to J cells to promote the HIF-2α ubiquitination in the nucleus (Fig. 5D). Moreover, K cells revealed poly-ubiquitination of HIF-2α compared to J cells which supports the concept of accelerated degradation of HIF-α in K cells (Fig. 5E). This finding implies a possible role for nuclear VHL in regulating stability of HIF-αs in the TCF-4 isoform-overexpressing cells.
The tumorigenic potential of EV, J, and K cells was assessed in nude mice. As might be predicted from the in vitro findings, J cells were highly tumorigenic. Although K cells generated small tumors, they appeared later (about 40 days) after tumor cell injection and grew very slowly (Fig. 6A). Control (EV) cells did not produce tumors as reported previously [24].
Immunoblot analysis was applied to confirm whether the xenograft tumors had the same protein phenotype as exhibited in the cell clones. The Myc-tag detected exogenous TCF-4J and K isoforms derived from tumor tissues; in addition, RT-PCR was performed to verify overexpression of the TCF-4J or K without cross-tissue contamination during these experiments (Fig. 6B; lower panel). Consistent with previous report that HIF-2α upregulated EGFR expression in the hypoxic core of solid tumors [18], higher expression levels of EGFR in concert with HIF-2α were found in J compared to K cell-derived tumors (Fig. 6B).
We also assessed HIF-2α and EGFR protein expression in xenograft tumors by immunohistochemistry. The histological features of the tumor tissue as well as cellular morphology did not differ among tumors induced by either J or K cells using hematoxylin and eosin staining. However, there was increased expression of both HIF-2α and EGFR predominantly in the J cell-derived tumor tissue (Fig. 6C). Since tumors frequently contain central necrotic areas that form a boundary between the areas of living and hypoxic dead tumor cells [29], we focused on the immunoreactive signals at this interface. Striking nuclear and cytoplasmic expression of HIF-2α and membranous EGFR were predominantly found at the hypoxic boundary in J cell derived tumor tissues (Fig. 6C and D).
In the present study, we demonstrated the following: (1) in hepatocyte derived cells, overexpression of the TCF-4J isoform lacking the SxxSS motif, exhibited robust tumorigenic potential; (2) the TCF-4J isoform was highly expressed in rapidly growing and presumably in the hypoxic milieu of human PD HCC, and (3) TCF-4J expression contributed to nuclear accumulation of HIF-2α and was due to, in part, less ubiquitin-dependent degradation of HIF-αs in J cells compared to K cells under severe hypoxia. This observation was linked to reduced expression of VHL.
There is evidence that genetic and/or epigenetic deregulation of the Wnt/β-catenin signaling pathway leads to tumor formation [2]. Indeed, genetic mutations in components of this pathway such as APC, AXIN1/2 and CTNNB1 are well-established molecular events in colorectal, as well as gastric carcinomas and HCC [39], [40]. Epigenetic disruptions in the Wnt/β-catenin signaling are also involved in oncogenesis through inappropriate stabilization of β-catenin and aberrant activation of upstream components of this signal transduction cascade [9], [10], [41]. There is little information, however, on the potential role of TCF-4 transcription factor isoforms in the oncogenic process. We demonstrated that overexpression of human TCF-4 isoforms may promote hepatic oncogenesis through actions of their intrinsic motifs which suggest that the Wnt/β-catenin signaling cascade may be augmented and further able to contribute to the malignant phenotype due to mechanisms that are operative in the nucleus. Indeed, expression analysis in human HCC tumors revealed that both TCF-4J and TCF-4K were present in non-cancerous dysplastic and normal liver, although both isoforms were initially identified and cloned from HCC cell lines. This finding is consistent with the previous report that emphasized the physiological distribution of TCF-4 isoforms in mice [42]. In human HCC tissues, however, PD HCCs demonstrated reduced expression of TCF-4K and exhibited very high levels of TCF-4J. The differential expression pattern of this TCF-4 isoform pair was independent of viral etiology and more directly related to the hypoxic microenvironment characteristic of PD HCC.
Although the precise interactions between HIF-αs and the β-catenin/TCF-4 complex are under evaluation, it has been demonstrated that HIF-1α [16] and HIF-2α [17] directly interact with β-catenin and thereby modulate TCF-4-mediated transcriptional activity. These observations suggest an important role for tissue hypoxia in regulating the Wnt/β-catenin/TCF-4 signaling cascade as a mechanism, which contributes to hepatic oncogenesis. Furthermore, the role of the SxxSS motif in mediating TCF-4 effects on HIF-αs stability under hypoxic conditions has been emphasized here. We found that TCF-4J and TCF-4B, which lack this motif, are associated with enhanced expression of HIF-1α and HIF-2α under severe hypoxia, whereas TCF-4K and TCF-4A, which express the SxxSS sequence, do not exhibit this property. As a potential mechanism(s), we found less ubiquitin-dependent degradation of HIF-αs in J than K cells under severe hypoxia; less degradation in J cells was linked to reduced expression of VHL. In contrast, the HIF-αs were highly degraded in K cells, raising the possibility that HIF-αs were subjected to VHL-mediated degradation due to the presence of the SxxSS motif. These findings emphasize the possible involvement of TCF-4 isoforms in mediating HIF-α stability via this SxxSS motif; however, it is unclear whether each S of the SxxSS plays a distinct role in the regulation of HIF-α stability.
The biological roles of HIF-1α and HIF-2α under hypoxia is controversial; however, HIF-2α, but not HIF-1α, is considered to be important in tumor formation [43], by promoting Myc activity [44]. In addition, HIF-2α is known to upregulate EGFR expression in the hypoxic core of solid tumors [18]. Emerging evidence has been presented that HIF-2α should be recognized as a CSC marker [45]. It is of interest that HIF-2α expression was significantly associated with poor prognosis in aggressive gliomas [37]. Since TCF-4J expression in our study was a potential biomarker for an aggressive HCC phenotype [34], we focused on the possible relevance of SxxSS as a mechanism of HIF-2α stabilization under hypoxic conditions. Since K cells showed weak and little tumorigenic ability in xenograft model, it was suggested that mechanism of drastic HIF-2α degradation in those cells even under hypoxia worked also may occur in vivo, possibly through affecting the EGFR expression in a SxxSS-dependent manner. It was observed in xenograft tissues that the driving force (HIF-2α) for EGFR expression was active in J cells as compared to K cells, especially in tumor cells under hypoxic conditions as observed in necrotic area. This HIF-2α-induced upregulation of EGFR in J cells may be associated with activation of EGFR signaling pathways and further studies will be needed.
The robust tumorigenic potential of J cells may be predicted by their significant anchorage-independent growth ability in vitro under hypoxic conditions. The oncogenic potential of J cells grown as xenografts is likely to be triggered by chronic hypoxic stress derived from subcutaneous inoculation which has previously been shown to have a poor neovascularization and thus, would generate a hypoxic environment during rapid tumor growth. The expression levels of both HIF-2α and EGFR were much higher in J tumors than K derived tumors and suggests that the J tumors may have grown faster through activating the HIF-2α/EGFR system under a hypoxic environment as summarized by the illustration presented in Fig. 7. This hypoxia-resistant property of J cells may be the biologic basis for the finding that TCF-4J was the predominant isoform expressed in human PD HCCs, which are known to have a hypoxic core in the central region of the tumor [18], [29].
Expression levels of TCF-4A, B, G and H isoforms in human HCCs. (A and C) Comparative analysis of TCF-4A/B (A) and G/H (C) mRNA expression levels in 27 HBV-related HCC tumors (T) adjacent peritumor tissue (pT) and histologic normal liver (N) by RT-PCR. (B and D) Comparative analysis of TCF-4A/B (B) and G/H (D) mRNA expression levels in another 20 HCC tumors including five WD HCCs from a different clinical site. Seventeen individuals had HCV-related chronic liver disease, and the remainder had chronic HBV infection. Values are normalized to GAPDH. Statistical results from all tissues are expressed as mean + SD (right panel). MD, moderately differentiated; PD, poorly differentiated; WD, well differentiated; *p<0.01; **p<0.05
Relation between clinicopathological factors and TCF-4A and B, and G and H expression in patients.
The authors would like to thank Drs. Osamu Nakashima and Sachiko Ogasawara for providing human tissue samples and the corresponding pathological information for the 2nd TCF-4 isoform analysis.
Competing Interests: The authors have declared that no competing interests exist.
Funding: This work was supported by the National Institutes of Health (CA-123544 to JRW). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Steve Forbes to give Landon Lecture March 9
Steve Forbes, chairman and editor in chief of Forbes Media, will give the first Landon Lecture of the year at Kansas State.
Steve Forbes, chairman and editor in chief of Forbes Media, will give the first Landon Lecture of the year at Kansas State.
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Published on Monday, July 18, 2011
Amadeus has launched in Asia Pacific the Amadeus Printmytrip, a solution that enables travel quotations, itineraries and electronic tickets to be customised according to the travellers' needs.
Travel agents can modify the design layout and include airline contact details, weather forecast at the travellers' destination, emergency contact numbers and other important information.
Amadeus said Printmytrip provides travellers "with a more informative, professional travel document to meet individual travel needsâ€.
Bruno des Fontaines, vice president, business solutions for Amadeus Asia Pacific said, "This new solution…. has the potential to positively impact a travel agency's brand. Travellers will also benefit from the added convenience of having additional booking information on their itineraries that previously was not possible.â€
Amadeus Printmytrip, which is fully integrated with the Amadeus Selling Platform, enables travel agents to brand the itinerary and upsell products and services to provide an alternative source of revenue through advertising.
Amadeus Printmytrip will be rolled out across Asia Pacific from August 2011.
As part of the launch promotion, Printmytrip will be offered free of charge to interested travel agencies until the end of October 2011.
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Holidaymakers admit to exaggerating bad experiences and threatening negative reviews
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Texture and colour looks good, Red, would love to pull out that top left corner a bit to straighten that side
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