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Get ready for a different kind of food fight!
Throw some food (or funds) our way in March as we compete in a friendly state-wide food drive competition during the annual Minnesota FoodShare March Campaign. You’ll not only be helping to stock our shelves for the busy times of need, but you’ll also be helping us secure funding distributed later in the year. Which means any donation now goes even further!
Neighbors is so grateful for the support of our community and hope you’ll join us in this important fight against hunger.
THANK YOU to the 732 Neighbors supporters – including individuals, organizations and churches – who helped us reach 98% of our goal during Minnesota FoodShare! We are so grateful for your support.


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SuperDroid Robots is an authorized reseller of Arduino products.
The Arduino Mega 2560, the successor to the Arduino Mega, is a microcontroller board based on a ATmega2560 AVR microcontroller. It has 70 digital input/output pins (of which 14 can be used as PWM outputs and 16 can be used as analog inputs), a 16 MHz resonator, a USB connection, a 2.5mm center positive barrel jack, an in-circuit serial programming (ICSP) header, and a reset button. It contains everything needed to support the microcontroller; simply connect it to a computer with a USB cable or power it with a AC-to-DC adapter or battery to get started.
The Mega 2560 differs from the preceding Mega in that it does not use the FTDI USB-to-serial driver chip. Instead, it features the Atmega8U2 programmed as a USB-to-serial converter. This auxiliary microcontroller has its own USB bootloader, which allows advanced users to reprogram it.
The Arduino Mega revision R3 adds SDA and SCL pins next to the AREF. In addition, there are two new pins placed near the RESET pin. One is the IOREF that is meant to allow shields to adapt to the voltage provided from the board. The other is a not connected and is reserved for future purposes. The Mega 2560 R3 works with all existing shields but can work with new shields that use these additional pins.
The Arduino Mega 2560 has 70 total available I/O lines; all of them can function as digital I/O lines, and sixteen of them can be used as analog inputs.
More information about the Arduino Mega 2560 is available on Arduino’s website.
The Arduino Mega 2560 does not include a USB cable, batteries, or a DC power adapter. No external programmer is required.
All 70 can used as digital I/O and 16 can be used as analog inputs.
Arduino Mega 2560 Detailed description of the Arduino Mega 2560.
How to get Arduino running on Windows A tutorial for setting up an Arduino environment on Windows.

industry_corpus2: Computing & Telecommunications


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Pebblebee | What is "Enable as Action"?
What is "Enable as Action"?
Enable as action is essentially turning on a feature. After you've set up the feature, you would enable it as an action to start using the feature. If enable as action wasn't turned on, the feature would be disabled.


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New York Giants tight end Evan Engram is reportedly expected to play in the team's Week 1 game against the Jacksonville Jaguars September 9.
On Saturday, Jordan Raanan of ESPN reported Engram has "made progress" in his recovery from a concussion and is on track to participate when the regular season gets underway.
The 23-year-old Ole Miss product suffered the head injury during the team's third preseason game against the New York Jets. He didn't appear in the exhibition finale Thursday against the New England Patriots.
Engram, the 23rd overall pick in the 2017 NFL draft, recorded 64 catches for 722 yards and six touchdowns in 15 games during a solid rookie season.
He struggled with dropped passes, however, and said in early August that head coach Pat Shurmur made avoiding those type of mistakes a "focal point" for the team during training camp.
"I have some individual goals, but I want to keep them to myself," Engram told reporters. "I just want to be the best tight end in the league and be the best tight end I can possibly be. That raises the standards for myself and for my teammates if I show up and do that every day."
Rhett Ellison would be in line to take over starting duties should the team's top tight end be unavailable for Week 1. Jerell Adams and Scott Simonson could also see some playing time.
That said, it sounds like Engram is on track to play against the Jags, barring any setbacks over the next eight days.

industry_corpus2: Sports


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Born and raised in rural Western Australia, I have lived in the United Kingdom, the U.S.A. and permanently reside in Oakville, Ontario with my husband and two children. I chose an Art Major while earning a degree in Education and later went on to pursue an Associate Diploma in Fine Arts. My painting style has evolved and changed; influenced by the places I have lived and travelled, from my early childhood in rural Australia, to my trips throughout Europe and North America.
You can find my Artwork in private collections throughout Australia, the U.S.A. and Canada.

industry_corpus2: Subject Education


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The purpose of this exhibit is to provide a foundation on the meaning of innovation, as part of engaging in the Innovation 2016 conference. Rather than accepting the dominant cultural assumptions of innovation or assuming innovation is bound in hype, the exhibit's artifacts and interactions are designed to challenge participants to rethink what constitutes innovation as a product versus innovation as a value.
Defining Innovations is located in the Napoleon Ballroom at the Sheraton New Orleans.
Dr. Rolin Moe is the Director of Educational Technology & Media at Seattle Pacific University.


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We Pray for Ukraine - Ukraine Tours, Kiev Travel Guide, Transfers, Private Guide, Ukraine Foto, Accomodation in Kiev, Walking tours, Village Life Tours, Monasteries of Ukraine, Reasonable prices - Travel to Ukraine!
may You set her in light.
keep us, Your children, enlightened.
May You, oh Lord, bring us up.
for us our Ukraine, please, keep.
May you turn to our people.
we thank you that you are a God of compassion and mercy.
We pray for the people of the Ukraine.
For those weighed down by fear and confusion we pray for peace and clarity.
For those lost in grief and sorrow bring the warmth of your care and the strength of your presence.
We pray that you may raise up peacemakers who have a positive vision of unity and hope.
We ask that you may lift the voices of understanding and reconciliation that they may be heard.
May bonds of love overcome suspicion.

industry_corpus2: Travel & Geography


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The Kid’s Clothes Week Fall 2014 theme is: storybook. There are so many ways to go with this theme! There is the more obvious costume feel (like my Princess Anna here) or a more inspired look (like these comfy reading PJ’s here). Some opted to screen print quotes or images onto tees or skirts. There are just so many wonderful ways to interpret this seasons theme.
I chose my storybook theme to center around one of my childhood favorites that I am now sharing with my children: My Father’s Dragon.
For the dragon dress, I chose the Adele Dress by Violette Fields Threads with Robert Kaufman Laguna Stretch Jersey Knit in Royal (here) with red tulle accents. I coupled the dress with DIY Leggings in Riley Blake Jersey Knit 1″ Stripes in Yellow (here). With the boots we got from Hanna Andersson, the dress could not be a more perfect dragon!
For Elmer, I chose the Rowan Tee by Titchy Threads with Riley Blake Jersey Knit 1″ Stripes in Red (here). I added a light blue knit color band and accent pocket with scrap knit I had.

industry_corpus2: Others


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Pennsylvania Sen. Arlen Specter isn't a big fan of RNC Chair Michael Steele.
Considering that Republican National Committee Chairman Michael Steele has hinted that he might find some sort of monetary punishment for people like Sen. Arlen Specter, R-Pa., who don't toe the party line, you'd expect Specter to be at least somewhat upset at Steele. But on a local radio show Turesday morning, Specter was more than just upset -- he was brutal.
And National Chairman Steele, well he’s said so many contradictory things I wouldn’t pay a whole lot of attention to him.
Actually, that about sums up Steele's troubled tenure at the RNC to this point.

industry_corpus2: Politics & Administration


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Algo Leasing and Finance Inc.
Send Inquiry - Algo Leasing and Finance Inc.
Send Inquiry - First Quadcor Finance Corp.
Send Inquiry - JK Capital Finance Inc.
Send Inquiry - Puregold Finance Inc.
Send Inquiry - Silver Finance Inc.

industry_corpus2: Finance & Economics


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When it comes to Medicare fraud, an illegal scheme usually begins with the need for qualified beneficiaries. There are a variety of methods for wooing Medicare beneficiaries to participate, and as a Department of Justice press release states, one Detroit doctor chose to prescribe controlled substances to carry out his part of a $7 million scam.
The press release states that the doctor agreed to refer Medicare beneficiaries to multiple home health care agencies in the Detroit area for medically unnecessary services. He signed medical documents, including home health care certifications and care plans that falsely certified Medicare beneficiaries to receive home health care services. (He was fully aware that the falsified documents would be used by his co-conspirators to support the false Medicare claims.) In exchange for his signature, the home health care agency owners paid him with cash and through other forms of kickbacks.
Court documents show that the doctor lured Medicare beneficiaries to his practice through patient recruiters, who were then paid in cash for their ”delivery” services. Once the patients were under his care, the doctor would bill Medicare for medically unnecessary controlled substance prescriptions, home health care and physician services. Bogus claims totaled more than $6 million and approximately $5.5 million in fraudulent claims were paid out as a result.
The 66-year-old doctor pleaded guilty to one count of conspiracy to commit health care fraud. He is awaiting sentencing for his part in the scam to bilk the government benefits program.
Everyone has a weak spot, and this doctor knew what would work for his scam. Most fraudsters also have multiple vulnerabilities that cause them to commit illegal acts. In this case, the doctor was wooed by the love of cold hard cash. Unfortunately for him, there is a good chance that selfishness and greed will land him behind bars.
Source: Today’s ”Fraud of the Day” is based on a press release titled, ”Physician Pleads Guilty for Role in Detroit-Area Medicare Fraud Scheme,” issued by the Department of Justice on July 1, 2014.
A Detroit-area physician pleaded guilty today for his role in a $7 million health care fraud scheme.
Assistant Attorney General Leslie R. Caldwell of the Justice Department’s Criminal Division, U.S. Attorney Barbara L. McQuade of the Eastern District of Michigan, Special Agent in Charge Paul M. Abbate of the FBI’s Detroit Field Office and Special Agent in Charge Lamont Pugh III of the U.S. Department of Health and Human Services Office of Inspector General (HHS-OIG) Chicago Regional Office made the announcement.

industry_corpus2: Health & Medicine


text: Prerequisite: UFDN 2000 or 3001. Considers the ethical and societal impacts of computing technology on daily life. Sample topics include invasion of privacy, computer crime, intellectual property, software theft, computer security, technology in the workplace, social networking and the global information society. Class format is a combination of lecture and discussion.


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Weekly Poll: Kirk vs. Picard; Refcardz!
This week, you get not one, but TWO weekly polls. One is displayed as usual, and will give instant results, like normal. It's at the bottom of this page and is, as you will quickly tell, not particularly development related.
The second is considerably longer than our average poll, is focused on which Refcardz topics you'd like to see in the near future, and is available by clicking either this link, or the button below. Enjoy!

industry_corpus2: Media & Journalism


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St. Mary’s Knights pitcher and point guard Jaxon Rideau is a standout two-sport athlete at the Catholic high school in Central Phoenix.
Most high school baseball teams have the benefit of playing their home games on the same property as the school. With its Central Phoenix location too small in size to accommodate a field, the St. Mary’s Knights baseball team does not get that same benefit.
Game recap of St. Mary's vs Tempe varsity baseball game on April 2.
Game recap of Shadow Mountain vs St. Mary's baseball game on March 26.
The story of St. Mary's assistant coach Michael Nunez.
St. Mary's head baseball coach Joseph Ponce breaks down his love for the job.
Preview of the St. Mary's Knights 2019 baseball season.


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OTTAWA, Ont. /Troy Media/ – It’s not easy being young these days – especially where employment is concerned.
A recent report from Statistics Canada found that full-time employment among young people (aged 17 to 24, excluding full-time students) has declined significantly since the late 1970s. This is not just the result of a bumpy economy. Youth are more likely to be unemployed than the rest of the employable population, regardless of the business cycle.
The 15-year span between 1990 and 2015 saw an average youth unemployment rate of almost 12 per cent – well above national averages for adults. And youth under-employment is estimated at 27 to 33 per cent.
Media headlines and pundits often decry today’s youth as entitled or lazy, but these stereotypes don’t reflect the realities of employment opportunities. Increasingly, the jobs offered to youth are temporary, contract, unstable and part-time – reflecting a situation found across other Organization for Economic Co-operation and Development (OECD) countries.
The federal Liberal government has committed to doing better, promising more jobs and better opportunities for young Canadians. That’s why it launched an Expert Panel on Youth Employment, which will soon issue a final report with suggestions for tackling the issue.
So what can be done to tackle the employment obstacles facing Canada’s youth?
The good news is that we don’t have to look too far to find successful models for youth skills development and employment – the solution is in our backyard. Too often, government reports and media accounts wax poetic over our fine universities as a source of solutions to our youth employment challenges. But our equally impressive polytechnic institutions get lost in the discussion.
Many might be surprised to hear that graduates from schools that are members of Polytechnics Canada enjoyed an employment rate of 86 per cent within six to nine months of graduation (2014-2015). This number rises to 90 per cent for four-year degree programs.
Polytechnics are distinct from universities because of their close relationships with industry to design program content, to meet labour market demands, and for their research and development and innovation solutions.
Don’t be misled by those who might paint such a discussion as an either-or debate between polytechnics and other post-secondary education. In fact, an increasing number of university students are turning to polytechnics as finishing schools to ease the transition into the labour force. In the 2014-2015 academic year, 14 per cent of full-time polytechnic students had already completed a four-year university bachelor’s degree.
Polytechnics help youth, and others, launch careers by prioritizing work-integrated models of learning. Polytechnics also offer academic bridging programs and learning models that are flexible enough to accommodate our most vulnerable youth populations.
There are four key ingredients we’d like to see in a federal solution to youth employment.
First, the federal government must invest in better labour market information from Statistic Canada. Youth can’t make informed career choices without data on skills in demand, employment outcomes by education type, or the earning power of specific credentials. Let our youth make informed career decisions.
Second, new youth employment initiatives should take advantage of the resources at polytechnic and college employment centres. This would leverage public funding that has already been invested over several decades in the core mission of education for employment that is the hallmark of polytechnic education.
Third, apprenticeships need a jump-start with secure funding, public awareness and programs to connect apprentices with employers. Canada needs a culture shift that gives apprenticeships the same esteem as other credentials. Those in the skilled trades are truly some of Canada’s best and brightest. The contribution skilled tradespeople make to our economy and their necessity in growing it must be acknowledged. The federal government should promote the new Canada Apprentice Loan – a recent pilot program that gives apprentices support similar to the Canada Student Loans program.
Finally, the federal Youth Employment Strategy must be overhauled. A federal work-integrated learning strategy could enable field placements, internships and professional practice. An improved strategy should include career counselling, interview preparation, and the inclusion of part-time and newly graduated students.
Employability has always been the polytechnic measure for success. If the government wants to tackle Canada’s youth unemployment problem, looking to how polytechnics offer youth direct pathways to the labour market would be a good place to start.
Nobina Robinson is the chief executive officer of Polytechnics Canada, a national alliance of Canada’s leading polytechnics and colleges.


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The Douro Valley is located in the North of Portugal and runs from the city of Porto in the west, all the way across to the Spanish border in the east. Douro inherits it's name from the Douro River, which snakes through the valley, the region is famed for it's production of Port Fortified Wines and yet despite this Douro produces just as many table wines as it does fortified.
While the grapes responsible for the table wines are typically grown on the region's more granite based soils, those reserved for the creation of Port will likely be found in Schist. The Douro Valley produces far more red wines than white with the styles diverse, ranging from light, fruitier reds to bold, complex wines often compared to those of Bordeaux.
Douro has Portugal's highest wine classification as a Denominação de Origem Controlada (DOC).

industry_corpus2: Agriculture & Fisheries


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With a focus on leading edge technology for lifestyle products, Artison incorporates it’s patented DualMono Center Channel technology into the ultra-thin Masterpiece M- LCR-DM speakers that rest beautifully along side a flat panel TV.
The Masterpiece LCR-DM combines three, front channels (LCR) into these slim enclosures. Using their exclusive DualMono technology, the Masterpiece speakers are able to project clear dialog from the center of the video display, for the most accurate center channel placement. This replicates the kind of sound you would get by hiding speakers behind a projection screen. However, for flat panels there is usually no other placement other than above or below the screen and the sound will not be exactly “centered”.
The Masterpiece speakers are designed to be attached to a flat panel display or mounted to the wall. The side-firing tweeters widen the “sweet spot” for a more encompassing and accurate soundstage. Featuring Artison’s exclusive Curved Line Array, there are thirty-two, 25mm Artison Reference Theater drivers, which controls the vertical dispersion pattern and creates a wider horizontal soundstage. This array is combined with an Inverted Magnet Structure with two, 5.5″ long-throw Kevlar midrange/woofers that ensure a low crossover frequency of 80Hz.
Custom Grilles can be cosmetically matched for both height and color of any flat panel TV and are delivered within 48 hours after purchase. The thin mounting bracket allows attachment to newer, super thin TV’s ranging in size from .28 to 4.5″. Designed for larger video displays, the Masterpiece M-LCR=DM speakers are compatible with most displays starting at 55″ diagonal. Complementing today’s sleek and contemporary flat panel TVs, the Artison Masterpiece speakers integrate seamlessly into any decor, yet, deliver sound quality on par with high-end, floor-standing speakers.
A pair of Masterpiece M-LCR-DM speakers retail for $2,800/pair, including the custom grilles.


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In line with their ideological convictions, the Games set for July are viewed as a rightful cause to act.
“We have no inhibition to use guerrilla activity to hurt the national image and paralyze the economy however we can. Because simply, we don’t want rich tourists – we want civil war,” reads a statement on their website.
The same very statement has been used to justify a rash of recent offences currently being investigated by UK police.
Last Tuesday, railway services were severely disrupted in Bristol, a prominent city in England’s south-west, after cables were set on fire. The perpetuators had to remove protective concrete slabs to access the lines.
It followed an attack on April 11, when several police stations in the same area were forced off the air. A police radio broadcasting mast on Dundry Hill was also damaged.
The FAI statement said it “specifically chose” targets in Bristol as the Ministry of Defense has located several installations nearby. A local business park of “military industry companies” such as Raytheon, Thales and QinetiQ can also be found there.
“If they are serious about trying to disrupt the Olympics by using simple techniques such as these, they are capable of causing utter havoc,” admits a private sector security consultant.
The Olympics security bill tops £1 billion ($ 1.6 billion), with the Royal Air Force in London’s airspace promising to shoot down passenger jets if required. Troops armed with surface-to-air missiles and warships will also be in place to do battle on the Thames. Still, most of this is to prevent Al Qaeda-style terrorist attacks, not “a low-intensity war” as promised by the FAI.
Sabotaging UK transport system during July games, especially at another end of the country, has the potential of severely hampering travel during a critical period. But the FAI could go further than that, as they are also known for sending letter bombs to officials and carrying out drive-by attacks. In a recent instance, two motorcyclists shot a chief executive of a nuclear power company in the kneecaps in the Italian city of Genoa a fortnight ago.


text: Loch water rushing over rocky falls, barley malted over moorland peat, slow distillation and long maturation in oak casks; all help to shape Lagavulin’s robust and smoky character. Pungent and potent, this is the great Islay malt. With richly peaty, deep, smoky flavours, it has an inrtense, long, ambrosial finish.


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If you are experiencing an issue that causes your Samsung Galaxy Note9 to freeze, you can do a ‘soft reset’.
Doing a soft reset will not delete any personal data, files or apps on your Samsung Galaxy Note9.
To reboot the Samsung Galaxy Note9, press and hold the power button and the volume down button on the side of the Samsung Galaxy Note9 until the screen turns off.
The Samsung Galaxy Note9 will automatically reboot.
The Samsung Galaxy Note9 has been rebooted.
To completely restore the original settings and clear all data from your Samsung Galaxy Note9, you can do a ‘factory reset’.
If a reset does not resolve your issue, you can also check for software updates for your Samsung Galaxy Note9.


text: The OPC desires to plant reformed churches throughout the United States, Canada and the Caribbean. Their efforts to bring solid, bible-based churches to gospel-starved areas has yeilded much fruit and the denomination is growing. To learn more about the OPC’s Home Missions visit the OPC website page on Home Missions.


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3 BR • 2 BA • 1749 sq.ft.
Condo/Coop - Palm City, FL Discover this 3 bed, 2 bath 1st floor furnished unit in Mile Lake Village at Harbour Ridge Country Club in Palm City. Extended Gourmet kitchen with granite counters, SS appliances + upgraded cabinetry. Light n’ bright with Bamboo flooring (carpet in bedrooms). Harbour Ridge offers 2 championship golf courses, yachting & an unparalleled lifestyle. Located on the North Fork of the St. Lucie River & offering a clubhouse with award winning chefs, events & gatherings! No food or beverage minimum! Community Yacht Club (dock negotiable thru seller), $68,000 membership equity fee (refundable) + $9,000 capital contribution (non-refundable) required of all new residents! The guard gate provides 24 hour manned security.

industry_corpus2: Real Estate & Construction


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India’s frontline fighter Sukhoi-30MKI is set to get a lethal edge with the Cabinet Committee on Security (CCS) tonight clearing a Rs 6,000-crore plan to acquire the air-launched version of the BrahMos missile, a joint Indo-Russian production, for the warplanes. The IAF will get 200 of these missiles.
The BrahMos, a supersonic missile with the capability of travelling at 2.8 mach, has already been inducted into the Indian Navy and the Indian Army.
Some modifications are needed to enable Sukhoi-30 carry the missile and deliver it. The missiles need some miniaturisation and Sukhoi-30 needs special under-the-wing pods to carry it.
Some of the fighter aircraft are being modified indigenously by Hindustan Aeronautics Limited (HAL) at its Nasik facility and fitted with an integrated aerial launcher developed by the BrahMos.
The first live test of the missile using a Sukhoi-30 is expected in December. Once inducted, the missile will complete a sort of triad of supersonic missiles available with India on land, in air and at sea. The IAF warplane will be able to hit a target 300 km away.
The Rs 6,000-crore outgo will involve purchase of BrahMos missiles, its integration onto the warplanes and testing, sources said.
Halwara, near Ludhiana, will be the second base of Sukhoi-30s in North India. The warplanes are currently based at Bareilly in western UP. The deployment of Sukhoi-30s at Halwara will complete in July 2013.
Sources said the CCS also cleared a Rs 2,000-crore proposal to tide over the crippling shortage of tank ammunition.
The Invar Missile, which can be fired from the barrel of the T-90 tanks, has also been okayed. The Army had projected its demand for 20,000 Invar missiles at a cost of Rs 2,000 crore.
Under the Army proposal, 10,000 Invar missiles would be procured from Russian manufacturers whereas the remaining would be license-produced by Bharat Dynamics Limited (BDL) under the transfer of technology (ToT) agreement.
Defence Minister AK Antony today joined the growing tribe of optimists who believe it was impossible to have a repeat of the humiliating military defeat of 1962 Sino-India war.
Over the past four weeks, Army Chief General Bikram Singh and Air Force Chief Air Chief Marshal NAK Browne have assured the nation of the prowess of the armed forces. General Bikram Singh had said a repeat of 1962 was not possible while the IAF boss pointed out that the outcome of the 1962 war would have been different had the IAF warplanes been used.
Antony, talking to the media on the sidelines of the Navy Commanders Conference today, said, "India of 2012 is not the India of that (1962) period. We are now capable of defending every inch of our country." He was replying to a question on India vis-a-vis China 50 years after the 1962 war in which India had suffered a defeat.
Antony quickly added a caveat to his words by talking about the abysmal infrastructure saying "infrastructure in the North-East is not up to our satisfaction, but it has improved a lot as compared to the past". Earlier, India, as per its policy, had not focused on enhancing infrastructure in border areas of northeastern states fearing the road could be used by an ingressing army.
"Now we have moved very fast. Our infrastructure, assets and manpower have substantially improved," he added. He said India would continue to build infrastructure.
In the increasingly networked world of today, cyber security is a vital national need. It is indeed heartening that the government has taken steps to enrol the private sector as an ally in its battle to defend the nation from cyber attacks — both within and from outside.
The world over, governments and the private sector work together to patrol the increasingly chaotic cyber space, and it’s high time the Government of India did so too. The ‘Recommendations of Joint Working Group on Engagement With Private Sector On Cyber Security’ prepared by the government lays down the parameters on which both sectors can work together.
The government is reacting to the misuse of cyberspace facilities that had triggered the mass movement of people from the Northeast who were working or studying in other parts of the country. The government is doing well in setting up various groups and committees that will enable it to move swiftly as and when the need arises. However, fire-fighting is only one aspect of the problem. What is needed is to train personnel and build expertise. The nation needs to develop expertise in dealing with cyber security. Private enterprise will help cut the red tape, and add the necessary verve as well as cutting-edge technology/training needed to combat cyber threats. It has been estimated that India needs over five lakh cyber security professionals, something that is simply not available right now. To fulfil this need, cyber security needs to be immediately introduced in the academic curricula of professional colleges.
The government must also take effective steps in training the police and other law enforcement agencies in the various aspects of cyber security. Only then would they be equipped to deal with cyber-crime investigation and cyber forensics. A world of caution is in order here, as in any such endeavour: the government must ensure that the democratic rights of freedom of speech and privacy of the citizens are not violated. India is a leading provider of IT talent to the world; it needs to secure itself even as it continues to embrace cyberspace.
Flaws in India's 'forward policy' of locating troops north of the disputed MacMahon line, were exposed in Kameng frontier division of Arunachal Pradesh. Within days of the attack, the well-prepared Chinese had overrun the Indian defences. Thousands of Indian Army soldiers and officers were killed, captured or wounded while some even shame-facedly took refuge in neighbouring Bhutan. Interspersed in this mayhem, some units showed flashes of military, bravado and tenacity. However, over all, the largely unhindered Chinese advance and India's retreat remains one of the saddest chapters in military history.
The debacle in the Kameng sector aptly illustrates the confusion and lack of planning. It was in August 1962 when the Chinese crossed the MacMahon line to counter the Indian moves and occupied the Thag La ridge north of Tawang. The lone Indian post at Dhola, created as part of the Forward Policy, was attacked. It signalled the Chinese were not averse to crossing the MacMahon Line -- a thing which they had not done since the 1914 Simla conference with the British and Tibetans.
The Indian leadership took a stand "Chinese must be evicted by force". The Eastern Army Command sent a message asking Indian troops "to engage Chinese patrols that came within the range of their weapons". Brig John Dalvi, Brigade Commander of the 7th Infantry Brigade, asked for logistics to support his troops. Eastern Army Commander, Lt Gen LP Sen, shared his opinion that eviction of the Chinese was not possible with existing logistics.
Brig Dalvi wanted logistics in place. The top echelons were keen to press ahead. The differences are aptly summed up by Maj Gen DK Palit in his well-documented book 'War in the Himalaya'. Palit, then the Director Military Operations, says "Senior commanders were ignorant of both the terrain and logistical difficulties." Bringing in supplies and ammunition was not easy. Only a jeep track existed only till Bomdilla located some 160 kms north of Tezpur in Assam, further north the journey was on foot and the mountains tracks were unsuitable for load carrying soldiers.
Indian troops had made a strong beginning on October 10 when the Chinese attacked Tseng Jong just north of the river Namkha Chu. A single company, about 120 men, from 9 Punjab, retaliated. Some 77 Chinese died (as reported by the Peking press at that time). The Punjabi's won three Mahavir Chakra (MVC) and two Vir Chakra (VrC). The Company Commander, Maj MS Chaudhary, died of wounds, his sepoy Kanshi Ram overpowered a Chinese with bare hands and snatched his automatic rifle. Despite a good start, the Punjabis were asked to withdraw from Tseng Jong. The Chinese stepped in and started probing around Tsangle and 'Bridge 5'.
At the first light of October 20 Chinese opened up a full attack all along the Namka Chu. Within the first few minutes Indian telephone lines broke down, leaving the units to act on their own. The Chinese had used the gaps in the Indian defences and occupied positions at the rear of the Indians, sandwiching them. Resultantly, an entire company of 2 Rajput was wiped out while the 1/9 Gorkha Rifles (GR) had been overrun within the first few hours. The enemy guns now turned towards "Tsangle" and "Bridge 5", while the Indian radio frequencies had been jammed so the gunners did not even get a signal from their superiors to fire.
The Rajputs suffered the worst. A total 282 of were killed. The 'History of the conflict with China' produced some thirty years after the war by the Historical Division of the Ministry of Defence, says "The unit fought a heroic battle, literally to the last round". All officers excepting Maj Gurdial Singh were killed. He led another charge, was taken prisoner of war (POW), and later awarded an MVC.
The Gorkhas, like the Rajputs, were also attacked from the rear. By then the 7 Infantry Brigade lost links with its units. Brig John Dalvi was taken prisoner on the afternoon of October 21 -- and entire Brigade had disintegrated. Elsewhere, the Chinese had attacked the Assam Rifles-held post at Khinzemane. A five-hour battle ensued following which Col Rattan Singh and 24 others were taken prisoner. Some miles away Second Lieutenant GVP Rao of Artillery was fighting a losing battle, Chinese had attacked in huge numbers. Rao and his men fought on till the last bullet. He died and got an MVC.
On the night of October 20, Maj Gen Niranjan Prasad, GOC, 4 Infantry Division, told his superiors that the Indian frontline had been wiped out. The next morning the Division's tactical headquarters was asked to withdraw from Zimithang and locate South, at Tawang. The Chinese broke up their force into two. One headed to Tawang and the other to get the Lumpo ridge vacated. Zimithang was taken over on the night of October 22-23 and Lumpo was under Chinese control by 9 am on October 23.
Elsewhere the Chinese targeted Bum-La before they entered Tawang, here the 1 Sikh covered itself in glory. Subedar Joginder Singh held back the invaders. With no ammunition left, Joginder Singh was asked to withdraw. He and his men responded by advancing with only the bayonets fixed to their guns. The Subedar was wounded and taken POW. He died in enemy custody. "Subedar Joginder Singh and his men mowed down the first wave, and the enemy was temporarily halted by the heavy losses it suffered. Within a few minutes, a second wave came over and was dealt with similarly", read the citation for the Param Vir Chakra - the highest war-time gallantry honour -- awarded to him. Another company of 1 Sikh led by Capt Haripal Kaushak stood firm. They "proved to be an iron curtain for the enemy", says the 'History of the Conflict with China'.
The Chinese had moved into Tawang unopposed on October 23. Lt Gen Sen, ordered withdrawal of troops to Bomdila located further south. Lt Gen BM Kaul was appointed the new IV Corps Commander on October 29 while Maj Gen AS Pathania replaced Maj Gen Niranjan Prasad as the GOC 4 Infantry Division.
Yet again the Chinese sneaked in south of the Se La pass and cut lines of communication of the 4 Infantry Division. Indian field units reporting rapid road construction and Chinese movement were admonished for 'exaggerated' reports about enemy movements. On November 17 the second attack commenced at Nuranang. The 4 Garhwal Rifles held positions with tenacity and won two MVC -- Lt Col BM Bhattacharjea and Rifleman Jaswant Singh, besides seven VrC. The 62 briagde had been withdrawn to south of the Se La. By now the Indians were virtually being pushed down the Himalayas with the Chinese in hot pursuit.
Strangely the Indian side wanted to be north of Bomdila. Strategically, the Indians could have just dug themselves in at Bomdila located at 10,000 feet and stopped the enemy. South of Bomdilla was a steep downward gradient that led to the Assam plains. The Chinese had no means to transport its guns across Se La and their supply lines were getting affected. By now the Chinese were not pursuing the retreating Indian Army, the indication was they were regrouping, a tactic they used in the 1952 Korean war. Indians had taken defensive positions at Se La (14,600 feet), Dirang and Bomdila. At Dirang the 7 cavalry was holding fort with its Stuart tanks which deterred the Chinese. The task was to stop the advance on the Bomdilla-Dirang road. The 7 cavalry provided covering fire to the troops to extricate themselves. On November 18 Bomdila had fallen without much of fightback.
On November 23, the Chinese announced a ceasefire. The Rajputs, headed by Lt Col Bhramanand Avasthy, did not know of this and were retreating with 300 odd men when they came under attack. More than a hundred Chinese were killed and the entire lot of the Rajputs was wiped out at battle now known as the 'Last stand at Lha-Gyala Gompa'. Baring a lone shepherd not a single man survived to cite the valour of Lt Col Avasthi and his men, hence no battle honours. His daughter Neeharika Naidu, who was only nine years in 1962, says, "A shrine has come up at the place where people come and worship and pay homage to my father".
Maj Gen DK Palit writes in his book, War in the High Himalay "The civilians set the pace, but the Army went along, beguiled by the conviction that the Chinese would not call our bluff."
Such was the confusion that 2 Sikh Light Infantry was withdrawing when it met with the frontline defences of 1 Sikh who did not even known about the withdrawal. With communication lines snapped, 4 Sikh LI and a section of 7 Mahar MMG unit were left to fend for themselves. When attacked these two small units were butchered. Some of them managed to save themselves by escaping into Bhutan. It was decided to pull out of Bomdila. There was little the brigade commander could do. There were no mines and just a handful of troops to defend Bomdila, which the Chinese took over on November 18. Elsewhere the 62 Brigade commander Brig Hoshiar Singh was ambushed and killed on November 27. Such was the condition that on December 1 -- a good fortnight after the withdrawal -- some 2,291 ranks were still missing.
New Delhi: The Cabinet Committee on Security (CCS), the highest decision making body for security issues headed by Prime Minister Dr Manmohan Singh, today cleared the acquisition of air-launched version of BrahMos missiles for the Russian-made Su-30 MKi air planes and Russian made Invar anti-tank missiles. The acquisition is estimated to cost the Indian exchequer Rs. 8000 crore.
Earlier, in March, 2012, in a letter to the Prime Minister and Defence Minister A K Antony, former Army Chief General V K Singh had highlighted severe shortfalls in ammunitions and missiles and had pointed out the Indian Army had tank ammunition for three-four days only.
Sources told NDTV that the CCS cleared the Rs. 6,000 crore plan to acquire the air-launched version of the BrahMos missile for India's main stay fighter plane Su-30 MKi.
BrahMos, a super-sonic missile that can travel at about 3 mach speed, is already being used by the Indian Navy and the Indian Army. It is a joint venture between India and Russia.
The Indian Air Force is expected to get about 200 of these missiles. Su-30 MKi will have to be modified to enable them to carry the air version of the BrahMos. In its current form only a single BrahMos can be fitted in the underbelly of the Su-30 MKi. There is, however, an effort to miniaturise the missiles and also modify the Sukhoi-30 under-the-wing-pods so that more than one missile can be fitted into a single aircraft. Hindustan Aeronautics Limited (HAL) is currently working on the modifications.
The first test launch of the air version of the BrahMos is expected to be conducted in December 2012. The BrahMos air version missiles will have range of about 290 kilometres and will enable the Indian Air Force to hit targets deep inside Pakistan while flying within Indian air space.
The CCS also cleared the acquisition of Invar anti-tank missiles for T-90 tanks at a total estimated cost of Rs. 2,000 crore to tide over crippling shortage of tank ammunition. The Invar Missile are fired from the barrel of the T-90 tanks. The Army wants about 20,000 Invar missiles. Source said that about 10,000 of these missiles would be brought from Russian whereas the remaining would be produced by Bharat Dynamics Limited (BDL) under Transfer of Technology (ToT).
New Delhi, Oct 18 — India's armed forces will Friday make a joint pitch before Prime Minister Manmohan Singh and members of the cabinet committee on security (CCS) on setting up three new commands to meet the threats to space assets and cyber infrastructure and for controlling commando operations.
The three new commands - special operations, aerospace and cyber security - will draw elements, assets and manpower from all the three services, as well as from other relevant government departments, a top commander said.
A presentation, laying out the modalities and feasibility of the separate formations for carrying out these specialist tasks, will be made when the commanders of the army, navy and air force meet with the prime minister and the CCS members at the combined commanders conference, the officer told IANS on condition of anonymity.
The proposals have been prepared by the Chiefs Of Staff Committee (COSC) headed by Indian Air Force (IAF) chief Air Chief Marshal N.A.K. Browne. This followed threadbare discussions with the other two COSC members - Indian Army chief General Bikram Singh and Indian Navy chief Admiral D.K. Joshi.
Browne will take lead in making the presentation. There will also be deliberations on the proposals at the daylong conference, where the heads of the sword arms of the three services will also express their views.
The idea behind the three separate commands has evolved from a suggestion made by a task force on national security headed by former cabinet secretary Naresh Chandra which submitted its report earlier this year.
The report of the task force is currently being studied by all relevant departments of the government, including the three armed forces.
The three services are of the view that each of the three new commands should be headed by a three-star officer while they synergise efforts and assets in these key areas of security.
Their opinion is that a lieutenant general should head the special operations command, which will pool in the commando forces of the army, navy and air force in a single formation, while the aerospace command should be headed by an air marshal and the cyber command by a vice admiral.
The army has been the pioneer in India in operations by special forces, the air force is the first among the three services to create a space cell and the navy is the first to raise a dedicated cyber security cadre.
At present, the Indian armed forces have two joint tri-services commands - the Port Blair-based Andaman and Nicobar Command and the New Delhi-based Strategic Forces Command that handles all nuclear weapons assets. The chiefs of these two commands are drawn by rotation from the three services.
Through the chief of integrated defence staff (IDS), the heads of these commands report to the COSC, as will the heads of the three new formations.
The decision on the three new commands will rest with the government, though the tri-services commanders will impress upon the nation's top security leadership the necessity for such formations in the wake of threats and trends in these sectors.
The special operations command has been successfully implemented by the US armed forces. Also, China has separate formations in the People's Liberation Army in the field of cyber security to carry out both offensive and defensive operations against threats to the country's information technology infrastructure.
Both these nations already have highly advanced space security programmes. China had in early 2007 demonstrated its anti-satellite capability, raising concerns among all space-faring nations on the security of their assets in space.
THE INTRODUCTION to the Henderson-Brooks report by Gen J.N. Choudhary, a former chief of the Indian army, exposes the failure of political leadership and the wrongly appointed army commanders that led to India’s defeat in the 1962 war with China.
It is claimed in the report that India had no shortage of arms and stores. But the report cannot deny that Indian soldiers without acclimatisation and adequate winter clothing were pushed into the mountain snows. Many researched and informative articles have been published in newspapers.
Recently Air Chief Marshal N.A.K. Browne stated that the use of the Indian air force, proposed by the military and disallowed by the government, would have changed the course of the conflict. He is right.
(The Henderson Brooks report is an analysis (operations review) of the Sino-Indian war of 1962. Its authors, Lt Gen Henderson Brooks and Brig P. S. Bhagat, are former officers of the Indian army). The report has been kept classified by the Indian government because its contents “are not only extremely sensitive but are of current operational value”).
The Chinese media have dismissed these claims by asserting that their air force was much too powerful then. Indian commentators have echoed the Chinese view. It is wrong. At that time even the US view was that due to logistics, the terrain and poor refueling facilities, the Chinese air force would have been ineffective.
The strident criticism in Indian parliament by Acharya Kripalani, Minoo Masani, Atal Behari Vajpayee and others has been recalled by commentators as the reason that forced the government into ill-prepared hostility to China. Implicitly a chauvinistic opposition is blamed for the government’s follies.
What the writers have omitted is the fact that there was also a sustained demand by critics to purchase arms from the West for immediate need and not to persist with the plodding, time-consuming official policy of building self-reliance in armaments inspired by the Soviet model.
The opposition leaders were not leading the charge against the government’s China policy. It was the media. The opposition was too weak at that time. Mainly it was Sri Mulgaokar, Frank Moraes and, later, Prem Bhatia who led the charge. These editors were not inspired by the politicians. The politicians were led by them.
Mulgaokar was my editor. I was around 25 and had returned after a two-year stint in Britain, dividing my time there between journalism and dish-washing in cafes. I think a flavour of those times based purely on memory would not be out of order for critics today.
The truth is that Prime Minister Jawaharlal Nehru and Defence Minister Krishna Menon were not only woefully ignorant about strategy, about realpolitik and security, but were also besotted with misplaced notions about the Soviet Union. Both were mentored by liberal Brits.
For years while Nehru foolishly kept chanting Hindi-Chini Bhai Bhai (India and China are brothers), the Chinese were steadily preparing for war. Nehru’s advisers in Britain were as silly as their Indian protégés that included an entire Indian intellectual class conditioned by the British.
VILIFICATION OF CRITICS: Those who rightly criticised India’s China policy were in those Cold War days described as CIA agents. The Economist of London in its intelligence report, restrictedly distributed to subscribers, wrote that Mulgaokar who was leading the charge against the government’s blind spot on China was inspired by the CIA.
Mulgaokar wrote a sharp letter of protest to the weekly’s editor, Donald Tyerman. The latter wrote a profuse letter of apology which I read. In those days my brother Rakshat Puri, very recently deceased, was the India correspondent of London’s New Statesman and Nation, which was a Bible for Indian intellectuals. He wrote an expose about the road being clandestinely built by the Chinese in disputed border territory.
The Indian government was aware of course, but kept parliament and the nation in the dark. The New Statesman editorially echoed the Nehru-Menon approach. After the 1962 war, the weekly’s legendary Editor, Kingsley Martin, visited India. He was god to most Indian intellectuals. He visited the India Coffee House on Delhi’s Janpath and shared a chat with a few of us. As a brash youngster I hauled him over the coals. I asked him whether or not he owed an apology to those of us in India who had been warning of the catastrophe India would suffer and had been rubbished by the likes of him.
I think he was not used to such criticism, particularly from an Indian. He mumbled, hummed and hawed and could give no reply. That was the mood of that time.
Now it is galling to read about chauvinistic critics who pushed Nehru into the Sino-Indian conflict and no recognition is given to those who accurately foresaw the defeat.
I guess it required a brash 25-year-old to say that the Emperor wore no clothes.
I subsequently had the article published in a small weekly, Thought, edited by Ram Singh. In that article I wrote: “This is therefore as good as a time as any for Nehru to resign from the prime ministership. Propriety demands it, wisdom counsels it and sympathy pleads for it.
Nehru should resign primarily because his China policy of high stakes has not succeeded. It has contained omissions which have facilitated an infringement of our sovereignty by the Chinese.
“This is no place to go into the whole depressing record of honest mistakes, dire circumstances, crass negligence and political naivete which has brought us to our present situation, where we have to sit round a table and argue with aggressors the legal validity of our claims matched against theirs on territory in their possession and control…that chunks of our territory were occupied by the Chinese and neither Parliament nor people were informed at Nehru’s sole instance, but instead led at that very time into the soporific bhai-bhainess of Panch Sheel, was an unpardonable lack of the sense of responsibility.
With India embarking on a massive defence modernisation programme, orders worth `100,000 crore is expected to be placed during the 12th Five Year plan period.
Defence and Aerospace experts participating in a panel discussion on ‘Making India the ESDM hub focus on telecom, aerospace & defence and automobiles’ at the Bangalore IT.biz on Wednesday said that with all the three armed and para-military forces set to procure latest weapons and military systems, abundant opportunities are in store for private companies.
Participating in the discussion Lt Gen AKS Chandele, former DG, Electronics and Mechanical Engineers, Indian Army said that orders worth `100,000 crore is expected to be placed during the 12th Five Year plan period (between 2012-2017) providing ample opportunities to the private companies.
He said that the Army will be embarking on big modernisation programme, which include acquiring tactical communication system, network centric warfare system, future infantry soldier as a system (F-INSAS) and procurement of radars, sensors and night vision devices.
He said the navy, apart from ordering about 40 vessels, will also be acquiring six diesel and nuclear submarines. Similarly the air force will be undertaking many upgrade and refurbishments.
NC Agarwal, chief executive officer of the Multi-Role Transport Aircraft Limited (MTAL), a joint venture between Hindustan Aeronautics Limited and Russia United Aircraft Corporation, said that many opportunities lay in the field of avionics. Apart from India developing its own aircraft, it will also be upgrading a significant number of its existing fleet. India’s First medium multi role combat aircraft (MMRCA) will roll out in four years.
Agarwal, who has been HAL’s Director (Design and Development), said the defence procurement process in India was very complicated, which led to delays in the armed forces getting the required product. Citing the example of Advanced Jet Trainer (AJT) Hawk, he said that aircraft was inducted into the airforce 21 years after India began negotiations with BAE Systems.
On the 126 MMRCA programme, he said that though the request for proposal was put out to vendors in 2005 the contract has still not been finalised and that the first aircraft will manufactured in India in about four years from now.
As per the MMRCA programme, 18 aircraft will be provided in a flyaway condition by Dassault Aviation, the remaining 108 will be produced locally by HAL.
New Delhi: An MP from Karnataka has allegedly constructed an engineering college on a field firing range in Belgaum belonging to the army, which is already facing shortage of these practice fields for training its personnel. Sources said Angadi Institute of Technology and Management has not got 'No Objection Certificate' (NOC) from civil or military authorities.
Defence Minister AK Antony has also sent two letters to the MP who has been demanding de-notification of the firing range which is available with the army till 2020, they said. Belgaum is an important military base of the army and houses the Commando school and the regimental centre of the Maratha Light Infantry regiment.
About 10 years back, the Army had 104 ranges but the numbers have come down to 66 including 12 acquired and 54 notified ranges. Thirty-eight field firing ranges have been taken away from the army and re-notified in this period, they said.
The issue of shortage of these ranges also came up for discussion during the ongoing Army Commanders' Conference. There are ranges in the Kargil and Srinagar areas also where demands have come from the local representatives for denotifying them for public use, sources said. However, the Army has been able to get a new firing range in Rewa in Madhya Pradesh which will be ready for use in next few years.

industry_corpus2: Aerospace


text:
Nollywood actress Tonto Dikeh-Churchill has opened up on her turbulent marriage to Olakunle Churchill.
In an emotional interview with Azuka Ogujiuba, the actress revealed the challenges she’s faced thus far in her marriage.
“First of all, I’d like to say that this interview is not to discredit my husband or rather my ex-husband in anyway… It’s a bitter experience. Being here is not easy,” she said.
The actress said she suffered domestic abuse from her estranged husband and also shared with the interviewer, pictures to back her claim.
Dikeh said her love for Churchill made her endure the abuse, infidelity and his shortcomings.
“I stayed back in my marriage, not because of my child. I stayed because I was doing it for myself, because I loved my husband with all of my heart. I stayed because he begged me to stay a couple of times. I stayed because I just didn’t want to give up. I didn’t want to give up on the marriage just like that.
“People change, I changed… My husband has humiliated me so much,” she said.
The actress married Churchill, nephew of former president Olusegun Obasanjo, in 2015.
TONTO YOUR TREAS DISTURBED ME,BECAUSE I USED TO KNOW YOU WHEN YOU WERE RUFF SORT OF,I REMEMBER WHEN YOU WERE HAVING CLASHES PARTICULARLY WITH SOME IGBO MEN ON THE FACEBOOK ,BLOG OR TWEETER.I DID ACTUALLY STAND WITH AND FOR YOU TONTO,BECAUSE I BELIEVED SO MUCH IN GOD THAT HE WILL CHANGE YOU ONE DAY.NO BODY IS A WRITE OFF INCLUDING YOUR HUSBAND,GOD CAN CHANGE HIM FOR GOOD IN THE NEAR FUTURE JUST LIKE THE WAY HE HAS TRANSFORMED YOU AND AM SO GLAD GOD ANSWERED MY PRAYERS ABOUT YOU.I DON’T KNOW WHY I HAVE ALWAYS LOVED YOU AND ANYTHING CONCERNING YOU.WHEN I LEARNT YOU WERE MARRIED I WAS HAPPY,BUT SUDDENLY I AM HEARING THIS FROM YOU THAT THE MARRIAGE IS NO MORE.WELL GOD UNDERSTANDS,AND I BELIEVE THAT HE WILL SEE YOU AND YOUR SON THROUGH.HOLD ON TO HIM AND DON’T GIVE UP AND SURELY HE WILL NOT FAIL YOU.I PRAY THAT GOD WILL BY HIS SPIRIT PATH YOUR WAY.PSALM 23;91;11;ISAIAH 49 AND 62 WILL BE GOOD FOR YOU.KEEP YOUR LIGHT BUNNING MY SISTER.
You would have gotten a better man from your tribe, but I think you run to become more famous, looking to be more than you already are. Hmmmmm, of course you must understand me well? May it also sound as a warning to others. I take the opportunity to congratulate Chioma Chukwuka for respecting the Igbo tribe and upholding it’s dignity. She’s been so decent in her acts. Hmmmmm, Tonto, think twice.
Hmm..so touching.. Its high time we ladies stand up for our rights..I CNT just imagine what you are passing through.. But don’t worry.its just a phase of life.it wud pass away like every other one and all the pains, sorrow, travail, crying and everything will be a thing of the past…I love u and our Father loves u so much..You are an inspiration.. Don’t think less of yourself.. Okayy.. Always remember “THE JOY OF THE LORD IS YOUR STRENGTH” No matter the circumstances.. SHALOM!!!
May God in His infinite mercy strengthen you, and give you the grace and boldness to stand strong in this your difficult times. Chear up dear, the best is yet to come.

industry_corpus2: Film & Entertainment


text:
Prolactinoma is the most frequent pituitary tumor in humans. The dopamine D2 receptor agonist bromocriptine has been widely used clinically to treat human breast tumor and prolactinoma through inhibition of hyperprolactinemia and induction of tumor cell apoptosis, respectively, but the molecular mechanism of bromocriptine induction of pituitary tumor apoptosis remains unclear. Caveolin-1 is a membrane-anchored protein enriched on caveolae, inverted flask-shaped invaginations on plasma membranes where signal transduction molecules are concentrated. Currently, caveolin-1 is thought to be a negative regulator of cellular proliferation and an enhancer of apoptosis by blocking signal transduction between cell surface membrane receptors and intracellular signaling protein cascades. Rat pituitary adenoma GH3 cells, which express endogenous caveolin-1, exhibit increased apoptosis and shrinkage after exposure to bromocriptine. Hence, the GH3 cell line is an ideal model for studying the molecular action of bromocriptine on prolactinoma.
The expression of endogenous caveolin-1 in GH3 cells was elevated after bromocriptine treatment. Transiently expressed mouse recombinant caveolin-1 induced apoptosis in GH3 cells by enhancing the activity of caspase 8. Significantly, caveolin-1 induction of GH3 cell apoptosis was sensitized by the administration of bromocriptine. Phosphorylation of caveolin-1 at tyrosine 14 was enhanced after bromocriptine treatment, suggesting that bromocriptine-induced phosphorylation of caveolin-1 may contribute to sensitization of apoptosis in GH3 cells exposed to bromocriptine.
Our results reveal that caveolin-1 increases sensitivity for apoptosis induction in pituitary adenoma GH3 cells and may contribute to tumor shrinkage after clinical bromocriptine treatment.
Prolactinomas, prolactin-secreting pituitary adenomas, are the most frequently found functional pituitary tumors in humans. The pathology of prolactinomas involves dysfunction of lactotrophic cells in the anterior pituitary gland which in turn leads to hyperprolactinemia . First line therapy for prolactinomas includes management with dopamine agonists. Bromocriptine is commonly chosen as a therapeutic agent for patients with prolactinomas or other pituitary adenomas; bromocriptine binds to the dopamine D2 receptor on pituitary epithelial cells to inhibit prolactin secretion . Treatment with bromocriptine causes tumor shrinkage . A rat pituitary prolactinoma cell line, GH3 is commonly used as a cellular model for studying prolactinoma formation. Long-term incubation of GH3 with bromocriptine induces cell apoptosis .
Caveolin-1 is a 21–24 kDa major integral membrane protein on caveolae, an invaginated structure on cellular membranes enriched with high numbers of cholesterol, glycosphingolipid and signaling molecules . Caveolin-1 has been suggested to negatively regulate many different signaling molecules located on caveolae via mutual interactions that compartmentalize the signaling molecules and suppress cell growth . Caveolin-1 is functionally involved in endocytosis, transcytosis, cholesterol transport, homeostasis, negative regulation of Ras-, NO-, and G-protein-coupled-receptors, and growth-factor-mediated protein kinase signaling cascades [7–9].
There is growing evidence that loss of caveolin-1 expression is associated with tumorigenesis [10–12]. Down-regulation or absence of caveolin-1 expression has been found in many human cancers, including primary breast, prostate, and colon cancers [13–15]. Furthermore, caveolin-1 null mice are more susceptible to carcinogen-induced tumorigenesis , suggesting that caveolin-1 may be a tumor suppressor.
There is accumulating experimental evidence in vivo and in vitro that caveolin-1 expression sensitizes cells to apoptotic stimulation. Elevated expression of endogenous caveolin-1 is associated with induction of apoptosis in mouse peritoneal macrophages . Ectopic expression of caveolin-1 in NIH3T3 cells and T24 human bladder carcinoma cells sensitizes cells to staurosporine-induced apoptosis . These data demonstrate that an up-regulation of caveolin-1 may be involved in promoting cell apoptosis.
In the present study, we investigated the effects of caveolin-1 on pituitary adenoma shrinkage in response to bromocriptine treatment at clinically-relevant concentrations in GH3 cells. Here we show that caveolin-1 in GH3 cells was up-regulated after bromocriptine treatment. Our data show that increased caveolin-1 expression sensitizes pituitary adenoma GH3 cells to apoptosis induced by bromocriptine treatment and clarifies the molecular mechanism of bromocriptine therapy of pituitary adenoma.
Caveolin-1 is associated with apoptosis and has been detected in GH3 cells . As bromocriptine stimulates GH3 cell shrinkage and apoptosis, we hypothesized that bromocriptine treatment would induce GH3 cell apoptosis via caveolin-1. Semi-quantitative RT-PCR was used to detect the amount of caveolin-1 mRNA in rat GH3 cells before and after bromocriptine administration at different dosages according previous report . Caveolin-1 mRNA was elevated after 24 hours of bromocriptine treatment in a dose-dependent manner (Fig. 1A,a). To explore the function of caveolin-1 in GH3 cells, a pcDNA4-Caveolin-1 plasmid containing Myc-tagged mouse caveolin-1 under the control of the CMV promoter was constructed and successfully transfected into GH3 cells. A predicted 30 kDa recombinant protein was recognized by an anti-caveolin-1 antibody (Fig. 1A,b) and by an anti-Myc antibody (Fig. 1A,c). Twenty-four kDa endogenous caveolin-1 expressed in the A431 cell line, and recognized by an anti-caveolin-1 antibody, was used as the positive control in Western blotting (Fig. 1A,b). We next clarified whether the recombinant caveolin-1 was localized in cells in the same manner as endogenous caveolin-1. The distribution of endogenous caveolin-1 in A431 cells was determined by immunofluorescent staining with an anti-caveolin-1 polyclonal antibody. Exogenous Myc-tagged caveolin-1 distribution in A431 cells was detected by double immunofluorescent staining with anti-caveolin-1 and anti-Myc polyclonal antibodies. Both endogenous and exogenous caveolin-1 had the same punctate distribution in the perinuclear region (Fig. 1B,a and 1B,b). We then transiently expressed the Myc-tagged caveolin-1 in GH3 cells to examine the effect of caveolin-1 on GH3 cells. By 48 hours after caveolin-1 expression in GH3 cells, apoptosis-like nuclear condensation was visible upon staining with Hoechst 33342 (bisbenzimide, Fig. 2E). In the control experiment, transient expression of enhanced-green fluorescent protein (EGFP) did not alter nuclear morphology (Fig. 2F). These data indicate that transient expression of Caveolin-1 induces apoptosis in GH3 cells. To test this, the TUNEL assay, which discriminates apoptosis from necrosis and from primary DNA strand breaks, was used to measure DNA fragmentation 48 hours after transfection . Recombinant caveolin-1-expressing cells appeared shrunken with positive TUNEL labeling (Fig. 3A,d). In the control experiments, all cells were positively TUNEL labeled after exposure to DNase I (Fig. 3A,f), whereas no cell was TUNEL labeled after vehicle treatment (Fig. 3A,h). We quantified the proportion of apoptotic cells in ectopic caveolin-1 or EGFP-expressing cells by counting the number of cells exhibiting nuclear condensations per 100 exogenous caveolin-1- or EGFP-expressing cells, stained with Hoechst 33342. Significant numbers of caveolin-1 expressing cells exhibited apoptosis after transfection (13% at 24 and 53% at 48 hours), whereas only 2% and 3% of EGFP-expressing cells and 1.6–2% vehicle-treated cells exhibited apoptosis 24 and 48 hours after transfection (Fig. 3B). This result revealed that even transient caveolin-1 expression increased GH3 cell apoptosis.
Bromocriptine enhances endogenous caveolin-1 mRNA expression in GH3 cells. (A) Panel a: Expression of caveolin-1 mRNA was enhanced after bromocriptine treatment. Panel b and c: Cellular proteins were extracted and Western blots performed 48 hours after recombinant caveolin-1 was transfected into GH3 cells (lane 2 and 5). Vehicle transfections (lane 1 and 4) were used as controls. Panel b is an immunoblot using rabbit anti-caveolin-1 antibody and panel c is an immunoblot using monoclonal anti-Myc antibody against Myc-tagged caveolin-1. Arrowhead indicates Myc-tagged caveolin-1. Arrow indicates endogenous caveolin-1. Protein extracted from A431 cells was used as an immunoblotting positive control. (B) Recombinant caveolin-1 expressed in A431 cells has identical perinuclear localization with endogenous caveolin-1. Myc-tagged caveolin-1 was transfected into A431 cells (a, c) for 48 hours. Non-transfected A431 cells were used to examine endogenous caveolin-1 localization (b, d). Cells were fixed and immunocytochemically stained with anti-caveolin-1 antibody then visualized by anti-rabbit IgG conjugated-FITC secondary antibody to detect exogenous (a) and endogenous (b) caveolin-1. Monoclonal anti-Myc antibody combined with anti-mouse IgG conjugated-Texas-Red was used to recognize recombinant caveolin-1 (c). Filamentary actin was stained with phalloidin conjugated-Texas-Red to observe cell morphology (d). Scale bar = 20 μm.
Overexpression of caveolin-1 induces nuclear condensation in GH3 cells. GH3 cells were transfected with either pcDNA4-caveolin-1 (A, C and E) or pcDNA4-EGFP (B, D and F). Immunocytochemical staining was carried out 48 hours after transfection. Phalloidin conjugated-Texas-Red (A and B) and Hoechst 33342 (E and F) dyes were used to stain F-actin and nuclei respectively. Cells expressing caveolin-1 (arrowhead) were visualized by anti-rabbit IgG conjugated-FITC secondary antibody (C). Caveolin-1 overexpressing cells exhibit nuclear condensation (E), while normal nuclei (arrow) were detected in cells over-expressing EGFP (F). Scale bar = 20 μm.
Over-expression of caveolin-1 induces apoptosis of GH3 cells. (A) GH3 cells were transfected with pcDNA4-Caveolin-1 for 48 hours then subjected to TUNEL assays. Cells transiently expressing exogenous caveolin-1 (indicated by arrowheads) showed shrunken morphologies (a and b) and nuclear fragmentation when stained with Hoechst 33342 dye (c) and were positively fluorescence-labeled by TUNEL assay (d). Cells directly treated with DNase I (e, f) or vehicle (g, h) were TUNEL positive (f) or negative (h), respectively. Nuclei stained with Hoechst 33342 dye are shown in c, e and g. (B) Caveolin-1 elicited apoptosis of GH3 cells. GH3 cells were transfected with pcDNA4-caveolin-1, pcDNA4-EGFP or vehicle (null treatment) and subjected to immunocytochemical staining 24 and 48 hours after transfection. Three hundred cells were randomly chosen and counted in each experiment (vehicle, caveolin-1 or EGFP) to determine the percentage with fragmented nuclei after Hoechst 33342 dye labeling. Apoptotic cells were measured by fluorescence labeling and data were expressed as mean ± standard deviation from n = 3 independent experiments (angular transformed for analysis, back-transformed for presentation). The standard deviations are too small to observe in the 48 hours data. **P < 0.01 and *P < 0.05 versus EGFP or vehicle experiment.
The activation of caspases plays a pivotal role in the execution of apoptosis by various signaling pathways [20, 21]. To examine the role of caspases in apoptotic GH3 cells after transient caveolin-1 expression (Fig. 4), we separately treated the ectopic caveolin-1 and DsRed-N1 expressing GH3 cells with a general caspase inhibitor (Z-VAD-fmk), as well as specific caspase inhibitors, for caspase 3 (Z-DEVE-fmk), caspase 8 (Z-IETD-fmk) or caspase 9 (Z-LEHO-fmk) before determining the number of apoptotic cells by TUNEL assay. Over-expression of caveolin-1 resulted in 62% of the transiently-transfected cells becoming apoptotic. This effect was inhibited by treating GH3 cells with the general caspase inhibitor Z-VAD-fmk (24% apoptotic caveolin-1 expressing cells) and with the caspase 8 specific inhibitor, Z-IETD-fmk, (20% apoptosis). Treatment with caspase 3 or caspase 9 specific inhibitors did not inhibit caveolin-1 induced apoptosis (apoptosis decreased by 8 and 13%, respectively; P > 0.05). In negative control experiments, cells expressing the red fluorescent protein, DsRed-N1 had no significant increase in apoptosis compared to untreated GH3 cells (8% versus 2%, respectively). Treatment of GH3 cells with Z-FA-fmk, a negative control caspase inhibitor reagent, however, did not significantly inhibit caveolin-1 induced apoptosis (59% of cell apoptosis), indicating that activation of caspases plays a role in caveolin-1 induced GH3 cell apoptosis, at least in part through caspase 8 signaling.
Caveolin-1 induces GH3 apoptosis via caspase 8 activity. Caspase inhibitors (50 μM) were added to either pcDNA4-Caveolin-1 or pcDNA4-DsRed-N1 (negative control) transfected GH3 cells and the proportion of apoptotic cells quantified 48 hours after transfection by immunocytochemical and TUNEL assays. Data were expressed as mean ± standard deviation from 3 independent experiments (angular transformed for analysis, back-transformed for presentation). Abbreviations on the x-axis: General caspase inhibitor (Z-VAD-fmk); a negative control inhibitor (Z-FA-fmk); specific caspase inhibitors for caspase 3 (Z-DEVE-fmk), caspase 8 (Z-IETD-fmk) or caspase 9 (Z-LEHO-fmk).
Bromocriptine induces activation of p38 MAP kinase in GH3 cells . Activation of the p38 MAP kinase signaling pathway in NIH3T3 cells causes phosphorylation of caveolin-1 on Tyr14 . We examined if there was any relationship between bromocriptine and caveolin-1 that affected GH3 cell apoptosis. GH3 cells were allowed to transiently express caveolin-1 for 24 hours, then 30 μM bromocriptine was added for another 12 hours. The proportion of apoptotic cells was determined by estimating the number of cells containing condensed nuclear DNA. After 12 hours of bromocriptine treatment, the number of apoptotic cells was 38% of the total caveolin-1 expressing cell population compared to 24% without bromocriptine (Fig. 5A). Only 14% and 6% of the total cell population underwent apoptosis when cells were treated with bromocriptine or vehicle for 12 hours (Fig. 5A). The data indicate that there is an interaction between caveolin-1 and bromocriptine in the induction of GH3 apoptosis.
A combination of bromocriptine treatment and overexpression of Caveolin-1 enhances apoptosis of GH3 cells. (A) GH3 cells were transfected with caveolin-1 for 24 hours and then treated with or without 30 μM bromocriptine for another 12 hours. Non-transfected cells were treated for 12 hours with ethanol alone (vehicle) or bromocriptine. Cells expressing exogenous Caveolin-1 were immunostained with anti-Myc antibody and then detected with anti-mouse IgG Texas Red-conjugated secondary antibody. Three hundred Caveolin-1 expressing cells were counted in each experiment and apoptotic cells were determined by nuclear fragmentation after staining with Hoechst 33342 dye. Data were expressed as mean ± standard deviation from 3 independent experiments (angular transformed for analysis, back-transformed for presentation). (B) Phosphorylation of Tyr14 was enhanced when GH3 cells were exposed to bromocriptine. Cells were administered 30 μM bromocriptine as indicated, 24 hours after pcDNA4-caveolin-1 transfection. Cellular protein was extracted and separated on 12% SDS-PAGE then subjected to Western blotting. Anti-caveolin-1 or anti-phosphorylated caveolin-1 (Tyr14) specific antibodies were used to measure total and phosphorylated caveolin-1 respectively. Cellular protein extracts from NIH3T3 cells exposed to H2O2 were used as a positive control to detect phosphorylated caveolin-1. Dash (-) indicates vehicle treatment. The arrowhead points to endogenous caveolin-1 and the arrow marks recombinant caveolin-1. Abbreviation: Br, bromocriptine. *P < 0.05 versus experiment for caveolin-1 transient expression only or bromocriptine treatment only.
Tyrosine14 phosphorylation of caveolin-1 was found to be associated with apoptosis in the human promyelocytic leukemia cell line HL-60 after etoposide induction , indicating phosphorylation of caveolin-1 tyrosine may play a role in apoptosis. To explore whether bromocriptine could induce caveolin-1 phosphorylation, GH3 cells were transfected with pcDNA4-caveolin-1 for 24 hours, then exposed to bromocriptine as indicated in figure 5B. Total proteins were extracted, separated using SDS-PAGE and examined by Western blotting using an antibody specific for caveolin-1 phosphorylated at Tyr14. After 12 hours of bromocriptine treatment the amount of caveolin-1 phosphorylation was 3.75 times higher than with vehicle (ethanol) treatment (Fig. 5B). Cellular protein from H2O2-exposed NIH3T3 cells was used as a positive control for phosphorylated caveolin-1. These data demonstrated that bromocriptine enhanced phosphorylation of caveolin-1 in GH3 cells.
In the present study, we demonstrated GH3 cells overexpressing caveolin-1 underwent apoptosis. Caveolin-1 has previously been reported to be associated with enhancement of apoptotic sensitivity. For example, NIH3T3 cells treated with anti-sense caveolin-1 were resistant to staurosporine-induced apoptosis, and T24 bladder carcinoma cells were sensitized to caspase 3 activation when stably expressing recombinant caveolin-1 . Large increases in caveolin-1 expression have also been observed in apoptotic-agent-treated mouse peritoneal macrophages . In addition, retrovirus insertion-mediated random mutagenesis applied to L292 cells revealed caveolin-1 is necessary for apoptosis induced by TNF-α . These reports highlight a growing body of evidence indicating that up-regulated expression of caveolin-1 is associated with cell apoptosis. This study has now shown, for the first time, that caveolin-1 itself induces apoptosis of rat anterior pituitary epithelial cells of the GH3 cell line.
We have shown that caveolin-1 mRNA expression was further enhanced in GH3 cells after 24 hours of bromocriptine stimulation. It is known that with bromocriptine stimulation of GH3 cells leads to accumulation of wild type p53 . There are two consensus half sites (-292 to -283 bp and -273 and -264 bp) in the caveolin-1 promoter that are predicted to be wild type p53 binding sites. Previous experiments have also shown that transfection of wild type p53 into human skin fibroblasts is accompanied by a six-fold increase in caveolin-1 promoter transcription activity . Therefore, and supported by evidence from this study, we speculate that bromocriptine enhances caveolin-1 expression via an increase in wild-type p53 expression in GH3 cells.
Caspases play critical roles in the initiation and execution of the apoptotic process . We found that, at least in part, caspase 8 might be required for caveolin-1 induction of GH3 cell apoptosis. In contrast, up-regulation of caveolin-1 sensitizes NIH3T3 fibroblasts and T24 bladder carcinoma epithelial cells to apoptotic stimuli via increased activation of caspase 3 . Caveolin-1 up-regulation is also associated with simvastatin-induced apoptosis of thioglycollate-elicited mouse peritoneal macrophages; this effect was independent of caspase activity, because the general caspase inhibitor Z-VAD-Fmk failed to block cell death . Together these results indicate caveolin-1 mediates cellular apoptosis through variant signaling pathways in different cell types.
Treatment of lymphocytes and NIH3T3 cells with H2O2 or UV light induces phosphorylation of caveolin-1 Tyr14 [27, 17]. In addition, caveolin-1 phosphorylated at Tyr14 specifically co-localizes with paxillin at focal adhesion complexes after epidermal growth factor, insulin or H2O2 treatment, suggesting phosphorylation of caveolin-1 is associated with cellular morphological changes and shrinkage when cells adapt to external cellular stresses [22, 27, 28]. In addition, etoposide, a therapeutic agent for leukemia, in inducing tumor cell apoptosis, increases phosphorylation of caveolin-1 Tyr14 in HL-60 cells . We demonstrated that phosphorylation of caveolin-1 Tyr14 was increased after bromocriptine treatment and this accelerated GH3 cells apoptosis, indicating phosphorylation of caveolin-1 Tyr14 may be important for bromocriptine mediation of cellular apoptosis or morphological changes in cells encountering apoptotic stress.
In summary, the present results demonstrate that caveolin-1 expression is induced after bromocriptine treatment in rat pituitary adenoma cells. Moreover, exogenous overexpression of caveolin-1 increases apoptosis of pituitary adenoma cells and enhances bromocriptine-induced cell apoptosis. Interestingly, caveolin-1 was phosphorylated at Tyr14 when GH3 cells responded to bromocriptine treatment. Phosphorylation of caveolin-1 Tyr14 is predicted to be associated with cellular apoptosis. These data suggest that bromocriptine-induced pituitary adenoma cell apoptosis may result from enhanced expression and activation of caveolin-1 via increased caveolin-1 phosphorylation. Our result explains the therapeutic effect of bromocriptine in curing pituitary adenoma.
Dulbecco's modified Eagle medium, penicillin, streptomycin, L-glutamine, fetal bovine serum and horse serum were purchased from Life Technologies (Gaithersburg, MD). Rabbit anti-caveolin-1 and rabbit anti-phosphorylated caveolin-1 (Tyr14) antibody were bought from Chemicon Internal Inc. (Temecula, CA). Mouse anti-c-Myc-antibody (9E10) was obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). Texas-Red and FITC conjugated secondary antibodies and normal goat serum were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA).
Total RNA was extracted from adult C57Bl/6 mouse brain with TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Caveolin-1 cDNA was amplified from total RNA by RT-PCR and was subcloned into pGEM T-easy vector by TA cloning (Promega, Madison, WI). The paired primer sequences used for RT-PCR were as follows: forward primer, 5'-CTCGAGATGTCTGGGGGCAAATACGTG-3'; reverse primer, 5'-TCTAGATATCTCTTTCTGCGTGCTGATGCG-3'. The DNA sequence of caveolin-1 was confirmed by auto-sequencing using an ABI 3730 autosequenser. Caveolin-1 in the pGEM T-easy vector was digested with Xho I and Xba I restriction enzymes and then subcloned into pcDNA4 (Invitrogen, Carlsbad, CA) mammalian expression vector; this plasmid was termed pcDNA4-caveolin-1. pcDNA4-EGFP was cloned as follows: the clone pEGFP-N1 plasmid containing enhanced green fluorescent protein (EGFP), obtained from Clontech (Palo Alto, CA), was digested with Pst I and Not I restriction enzymes, then subcloned into pcDNA4 vector; this plasmid was designated pcDNA4-EGFP. pDsRed-N1 containing red fluorescent protein was purchased from Clontech Laboratories. For quantifying the level of caveolin-1 cDNA expressed in GH3 cells modulated after bromocriptine treatment, the cells were treated with either bromocriptine (Sigma) at different concentrations of 5, 10, 20 μM or with vehicle for 24 hours, when total RNA was extracted and the transcripts quantified by RT-PCR, as described above.
The rat GH3 pituitary adenoma and human A431 epithelial cell lines were obtained from the American Type Cell Collection. The GH3 cells were propagated in F12K nutrient mix medium (Invitrogen, Carlsbad, CA) supplemented with 2.5% fetal bovine serum, 15% horse serum, 2 mM L-glutamine, 100 units/ml penicillin, and 100 units/ml streptomycin. The A431 cells were maintained in DMEM medium containing 10% fetal bovine serum, 2 mM L-glutamine, 100 units/ml penicillin, and 100 units/ml streptomycin.
Transfection of GH3 and A431 cells was performed using Lipofectamine Plus™ reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's protocol. Briefly, the day before transfection, 6 × 105 cells were plated on a 6-well cell culture grade Petri dish. One μg DNA and 6 μl Plus™ reagent were diluted into 100 μl serum-free medium and 4 μl lipofectamine was added to 100 μl serum-free medium; these two pre-complexes were then mixed and incubated for 15 min at room temperature. The DNA-Plus™-lipofectamine reagent complex was added to each well containing GH3 or A431 cells in fresh serum-free medium. Cells were incubated at 37°C in 5% CO2 in air for 3 hours, then the old medium was replaced with fresh complete medium after incubation. The times after transfection for immunocytochemical staining or TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling) analyses are indicated in the results.
For the analysis of Myc-tagged caveolin-1 expression in GH3 cells, cells were briefly washed with PBS and fixed with 4% paraformaldehyde in PBS for 15 min at room temperature. Cells were permeabilized by incubating with PBS containing 0.5% Triton X-100 for 10 min. The permeabilized cells were immersed in blocking solution containing 10% normal goat serum in PBS for 1 hour. The cells were then incubated over night at 4°C with either anti-caveolin-1 (diluted 1:100 in PBS) or Myc (diluted 1:100 in PBS) primary antibody. After three washes with PBS, cells were incubated with the secondary antibody (anti-mouse IgG conjugated with FITC, diluted 1:300 in PBS; anti-rabbit IgG conjugated with Texas-Red or FITC, diluted 1:300 in PBS) for 2 hours at room temperature. Slides were mounted with Mowiol 4–88 (Calbiochem, La Jolla, CA) and visualized by confocal laser scanning microscopy (Ziess, LSM510) before being digitally photographed.
The TUNEL assays were conducted as previously described  with some modifications. Briefly, DNase I-treated GH3 cells or cells ectopically expressing caveolin-1 were washed twice with PBS and fixed with 4% paraformaldehyde in PBS, pH7.4, for 10 min at room temperature. Cells were permeabilized with 0.1% Triton X-100 in 0.1% sodium citrate for 2 min on ice. Cell ectopically expressing caveolin-1 were labeled with monoclonal anti-Myc antibody, then visualized by Texas-Red conjugated anti-mouse IgG antibody. Cells were TUNEL labeled using the In situ Cell Death Detection Kit (Roche, Alcaloid, Mannheim, Germany) according to the manufacturer's instruction.
Treatment with caspase inhibitors and quantification of cell apoptosis were conducted as follows: GH3 cells were seeded in a 24 well dish one day before transfection. Cells were transfected with pcDNA4-caveolin-1 or pDsRed-N1 by Lipofectamine Plus™ reagent. Transfected cells were treated with caspase inhibitors at 50 mM final concentration for 48 hours, then immunocytochemical and TUNEL assays were used to quantify apoptotic cells. Anti-c-Myc monoclonal antibody was used as the first antibody to recognize caveolin-1 expressing cells, followed by Texas Red-conjugated anti-mouse IgG. Cells expressing DsRed-N1 were directly detected by fluorescent microscopy. The caspase inhibitors Z-VAD-fmk, Z-DEVE-fmk, Z-IETD-fmk, and Z-LEHO-fmk, and the negative control Z-FA-fmk were purchased from Calbiochem Inc. One hundred Myc-tagged and DsRed-N1 expressing cells were examined in each experiment for positive TUNEL labeling. Each experiment was repeated three times. Data were analyzed using the GLM procedure of SAS (v9.1) and significant differences between means were determined by a Duncan's New Multiple Range Test (P < .05).
For detecting caveolin-1 and phosphorylated caveolin-1 protein expression in GH3 cells, cellular proteins were extracted with RIPA buffer (50 mM Tris-HCl, pH 7.0, 1% NP40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM PMSF, 1 μg/ml aprotinin, 1 μg/ml leupeptine, 1 μg/ml peptasin A, 1 mM Na3VO4, 1 mM NaF, 10 mM Na4P2O7) 36 hours after pcDNA4-caveolin-1 transfection. Cell extracts were then centrifuged at 12000 rpm at 4°C for 10 min, and the supernatants collected. Protein samples were quantified by the Bio-Rad protein assay kit (cat. no. #500-0006; Bio-Rad, Inc., Hercules, CA). Each 10 μg sample was denatured for 5 min at 95°C in Laemmli sample buffer (2% SDS, 10% glycerol, 100 mM DTT, 60 mM Tris HCl, pH 6.8, 0.01% bromophenol blue). The proteins were then separated by 12% SDS-PAGE and transferred to PVDF membranes. Blots were washed with TBST buffer (120 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Tween 20) then placed in TBST buffer supplemented with 5% skimmed milk powder for blocking non-specific interactions for 1 hour at room temperature. Blots were then incubated with rabbit anti-caveolin-1 antibody (1:1000 in TBST), mouse anti-Myc antibody (1:3000 in TBST) or anti-phosphorylated caveolin-1 (Tyr14) polyclonal antibody (1:1000 in TBST) overnight at 4°C. After washing the blots with TBST, membranes were incubated with horse-radish peroxidase-conjugated secondary antibodies (1:3000, Amersham Biosciences, Arlington Heights, IL) for 2 hours and washed again as described previously. Membrane-bound secondary antibodies were detected using the ECL procedure developed by Amersham Biosciences. To ensure equal protein loading membranes were rehybridized with a mouse anti-actin antibody (1:3000, Chemicon, Inc., Temecula, CA) and developed using X-ray film (Kodak).
Bar charts of the proportion of apoptotic cells of each treatment group were drawn with the sample mean plus and minus one standard deviation from three independent experiments. The angular transformation of observed proportion data were used for statistical analysis. Analysis of variance of the group factor in blocks (treating each experiment as a block) and Tukey's Studentized range test for group means were performed with SAS v9.1 statistical analysis package (SAS Institute Inc., Cary, North Carolina).
This work was supported by a grant from the National Science Council of Taiwan, Republic of China (NSC 92-2313-B-002-145 and NSC 94-2317-B-002-020) and Council of Agriculture of Taiwan, Republic of China [93AD-4.1.3-AS-U1 (4)]. We are grateful to Drs. Kenrick Deen and Harry Wilson of Academia Sinica for help and advice in reviewing this manuscript.
YNJ and YTJ conceived the study, participated in its design and helped to draft the manuscript. YHL carried out most of experiments. YNJ and YHL contributed equally toward this work. MWK and TYT performed TUNEL and apoptosis analysis. MCH, YBT, WTKC and YTJ participated in its design and helped to draft the manuscript. All authors read and approved the final manuscript.


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Any suggestion that fraternities be reintroduced at Old Nassau conjures up in the minds of most Princetonians forebodings of “Animal Houses” further fragmenting loyalties that are already divided among classes, eating clubs, residential colleges, dormitories, athletic teams, and other extracurricular activities. “Fraternities are not Princeton,” hooted one alumnus at the idea. Nonetheless, it can be argued that Princeton – not William and Mary, where Phi Beta Kappa was founded in I 776 – was the true birthplace of the Greek-letter societies that spread to most American college campuses during the 19th century. This is so despite the fact that Princeton’s fraternities, with one exception, were childless and reached their zenith without residential frat houses.
The example of the Halls soon led to the formation of similar literary and debating societies at other campuses. It also paved the way for a third secret society, known as the Princeton Order of Chi Phi, which was founded on Dec. 24, 1824, by Robert Baird, then a tutor at the college, in association with other faculty members and students of both Princeton and the Princeton Theological Seminary. Chi Phi ceased to be active the following year, and Princeton's next Greek-letter fraternity, Beta Theta Pi, did not arrive until 1843. But in 1854, when President John Maclean Jr. 1816 was launching his campaign to ban fraternities from the college, his nephew and namesake in the Class of 1858 found among his papers the old constitution, minute book, and ritual of the Chi Phi Society of 1824. With these as a guide, the younger Maclean united with Charles Smith DeGraw 1857 and Gustave W. Mayer 1857 in reviving Chi Phi at Princeton. Mayer went onto found a second chapter at Franklin and Marshall College, one that exists to this day, along with 50-odd other chapters from coast to coast.
By some accounts, Princeton Reunions began as Halls Reunions, when alumni returned to witness the debating and oratorical performances of the graduating seniors at their Commencement exercises. Beam noted, “Inter-Hall rivalry [was] the oldest, most prominent, and most lasting characteristic of Whig and Clio down to the time of the World War. Every [classroom] performance could be looked upon as a preliminary, a training for an open contest with Clio. Under penalty of a fine, members were forbidden to speak a piece on the College stage without having rehearsed it in Hall and received Hall’s criticism.” Furthermore, the Halls had passwords, robes, insignia, initiation ceremonies, and other rituals. In short, they functioned as fraternities.
According to Baird’s Manual of American College Fraternities, Princeton’s Beta Theta Pi chapter (1843-45) was initially composed of five members: one Whig, one Clio, two unaffiliated undergraduates, and one seminarian. Then came Delta Kappa Epsilon (1845-57), Zeta Psi (1850-82), Chi Psi (1851-57), Delta Psi (also known as St. Anthony's Hall, 1851-52), Kappa Alpha Society (1852-56), Phi Kappa Sigma (1853-81), Sigma Phi (1853-58), Delta Phi (1854-77), Theta Delta Chi (1863-67), Sigma Chi (1869-82), Delta Upsilon (187071), and Alpha Sigma Chi (1876-79).
More important, the experience of Yale after 40 years of running its residential college system proves that though frat-house living went into decline, the secret societies and fraternities-which meet as often as twice weekly in their own “tombs” or more informally as “undergrounds” – continue as important centers of congeniality, personal growth, and intellectual inquiry.
Every president of Yale who was himself an Eli has belonged to one of these groups, with none other than A. Bartlett Giamatti (Y’60) authoring Scroll and Key’s most recent history. And while Princeton’s Triangle Club shows have long poked fun at Skull and Bones (founded 1832) – whose members are reportedly supposed to leave the room whenever its name is mentioned – the Yale secret-society format is ascertainable: Typically, after dinner together on Sunday evening, a faculty member or alumnus or invited public figure discourses on a subject of interest, followed by a question-and-answer period. Then, on a week night, an hour or two is set aside for each of the 15 or so undergraduate members, in turn, to present a paper or talk on a topical or autobiographical subject of his own choosing and to be questioned by his peers.
A freshman is entitled at once to not only the friendship but also the social and academic encouragement of upperclassmen which membership in a fraternity affords. Princeton undergraduates drawn from all four classes and all walks of campus life, in turn, would benefit from contact not only with the idealism embodied in their rituals but also the hospitality of their chapters scattered across the United States and Canada.
With flexibility on the part of national fraternity officials – who should sigh with relief that Princeton would not be asking them to bankroll any new frat houses – the restoration of fraternities to the Princeton campus could be accomplished in a variety of nonexclusive ways: The practice discontinued in the 1890s of individual Princetonians joining chapters at Rutgers, Pennsylvania, and Columbia could be resumed. Neighboring undergraduate Greeks could colonize interested students into campus groups meeting in their rooms or, say, Whig Hall or the eating clubs. Or the Masonic Lodge on Nassau Street, which dates back to 1765 and probably inspired the initiation ceremonies of Whig and Clio, might be prevailed upon to open its doors. The 1979 Report of the Committee on Undergraduate Residential Life came to the conclusion that “in addition to establishing general opportunities and programs for all students, we believe that specific groups of students from all four classes who have strong interests in common must continue to be able to find appropriate means of advice and support within the university, and effective ways of pursuing their special concerns.” The residential colleges and eating clubs are not sufficient to accomplish this. At least for some students, fraternities could provide a valuable environment now absent from the Princeton scene. In furtherance of the return of four-year Greek-letter societies to Princeton, a group of interested undergraduates formed an exploratory “Council for Fraternities” last June. It should be given every encouragement.


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Returning to college from a long lazy winter break is difficult. Many college students have well over a month off before they are expected to return for spring semester. During this time students enjoy a variety of pastimes, from earning extra cash for next semester to spending time with family.
Even though school is just around the corner, many students put off preparing for the upcoming semester. When the first day arrives, these students find that they are overwhelmed and unprepared to begin class. Time seems to move more slowly for students: Classes feel longer, homework becomes an increasingly tedious task and it feels as though there is an eternity stretched out between now and summer break. With a little preparation beforehand, however, students can avoid feeling doomed before the semester starts.
If you go home during winter break, plan on returning to campus at least two or three days before spring semester begins. Many students wait to take the very last plane out before classes start, allowing no time for unexpected delays. As a result, these students often miss the first few classes of the semester or the chance to purchase textbooks for class. Allow time for the surprises, inconveniences and circumstances that you can’t anticipate so that they don’t interfere with your college success spring semester.
By now, you’ve received your grades from fall semester. If they aren’t what you expected, it may be time to reprioritize your personal schedule and goals so that you can get organized and raise your GPA over the spring semester. Identify where you fell short in the past few months. Did you skip class? Fail to complete homework assignments? Arrive late on a daily basis? Whatever the excuse, as soon as you can identify why you fell short of your goals, you can begin making new ones.
Clean out your folders and start fresh. Unlike high school, college starts twice a year. If you wouldn’t use a tattered notebook that has already been all but destroyed to begin a new year, avoid using a notebook like this for second semester as well. College classes are full of information; if possible make sure you have a notebook for each class. Organize your new materials to fit your new schedule, and store all of your old papers in a safe place that is out of your way. Old class notes are useful reference materials, but don’t allow them get mixed in with the current semester’s work. Know where all your classes are located and what your weekly schedule looks like.
You’re only half way to summer break. Don’t check your brain at the door when you return to campus for spring semester. Focus on achieving your goals for the semester and hold fast to the schedule you’ve outlined. The best way to break into second semester is to proceed full steam ahead - don’t let a little exhaustion stand in the way of your success.
It’s always tempting to blow off a couple of classes when school first resumes. Resist this urge. Establishing a consistent routine at the beginning of each semester is one of the best ways to ensure that your schedule is manageable. Skipping a class or two can unnecessarily put you behind before the semester is underway. Before you know it, you’ll have missed a quiz, homework assignment or study session and making up the points will require extra effort. It also helps to take at least one elective or "fun class" if your schedule permits...that alone will keep you thirsty for knowledge!