text:
Sanaullah, a convicted Pakistani terrorist serving life sentence, was injured in a scuffle with another inmate in the high-security Kot Balwal jail in Jammu
Pakistani prisoner Sanaullah Ranjay, injured in a scuffle with another inmate in a Jammu jail and admitted to Post Graduate Institute of Medical Education and Research (PGIMER) at Chandiragh, died on Thursday morning.
A spokesperson from the PGIMER told reporters that "The patient was brought in a brain dead condition and died this morning due to multiple organ failure".
Meanwhile, the body of Sanaullah Ranjay will be handed over to Pakistan.Home Minister Sushilkumar Shinde said the Ministry of External Affairs is coordinating with the Pakistani authorities to complete the formalities to send back his body home. "Once the formalities are done, we will hand over the body," he said.
The 52-year-old Sanaullah was airlifted from Jammu to the PGIMER on Friday.
The PGI spokesman said that it was a case of death in custody and a post mortem will be conducted for which a medical board is being constituted.
"The post mortem will be conducted by the Medical Board in the presence of a Magistrate and videography will be done of the process," he said.
His condition had worsened yesterday following kidney failure and neurological problems.
"Patient has become extremely critical. He has developed renal failure with no urine output since morning. Hence he was put on peritoneal dialysis by Nephrologists," the bulletin issued last evening said.
A resident of Sialkot in Pakistan, Sanuallah was serving a life term after he was convicted under TADA provisions following his arrest in 1999.
He was injured in a scuffle with another inmate in the high-security Kot Balwal jail in Jammu and rushed to the PGIMER in an air ambulance on Friday last.
The assault came a day after the death of an Indian prisoner Sarabjit Singh who was brutally attacked by fellow inmates in Lahore's Kot Lakhpat Jail.
The Aditya Birla Group company saw its net profit rocket up helped by net sales and trimming of long-term debt
Aditya Birla Chemicals (India) Ltd, has reported above average net sales growth. For the quarter to end-March, the Aditya Birla Group reported total revenues, including sales, of Rs211.80 crore, a growth of 26% compared with Rs168.21 crore same period last year. During the quarter, the company's net profit rose 156.54% to end the March 2013 quarter at Rs7.85 crore from Rs3.1 crore a year ago period. Similarly, its operating profit showed a 34% y-o-y increase at Rs44.04 crore.
Moneylife wrote about the company in the Street Beat section in 27 December 2012 issue.
According to Moneylife database, the company reported above-average net sales growth rate of 26% y-o-y when compared to its three-quarter y-o-y growth rate of 22%. It has posted strong sales growth in the last four quarters. Similarly, its operating profit rose 34% y-o-y when compared to its three-quarter y-o-y growth rate of 31%.Barring a blip in the September 2012 quarter, the company posted double digits growth rates in the last four quarters. The company has significantly pruned its long-term borrowings by 10%. The company's return on networth is an abysmal 3% while its return on capital employed is much higher at 16%. The market values the company at a market capitalisation of just 1.55 times operating profit.
The company has declared a full year dividend of 10%.
Aditya Birla Chemicals is a leading manufacturer of bulk and speciality chemicals and viscose filament yarn. The business is spread across multiple manufacturing locations in three countries – India, Thailand and China.
Cobrapost has alleged that RBI is acting as a protector, and not as a regulator or enforcer. The RBI is loath to admit that there exists a massive money laundering racket in our financial institutions across the board, because if it were to admit, it would strip the RBI of its halo as a 'one of the best banking regulators in the world', Cobrapost says
Cobrapost.com continues with its exposure of money laundering taking place in banking system. In its latest episode, Cobrapost has named 10 banks, IndusInd Bank, Bank of Baroda, ING Vysya Bank, Allahabad Bank, Bank of India, Central Bank of India, Bank of Maharashtra, HDFC Bank, ICICI Bank and Axis Bank involved in money laundering.
"In this edition, codenamed Operation Red Spider Part III, we bring you 10 banking institutions and the murky world that exists within their walls, with junior to top level officials at dozens of branches admitting on camera their complicity in crime. The videos, across the large spectrum of such illegal activity, clearly prove that this is not simply the design of some criminally oriented officers, or for that matter, some young, and front office executive succumbing to performance pressures. It proves without doubt that this corruption is endemic, and spread through the Indian banking and insurance system," Cobrapost said in a release.
Earlier on 14 March 2013, Cobrapost named private sector lenders ICICI Bank, HDFC Bank and Axis Bank as facilitating money laundering. Later on 6th May, it named 23 entities including Life Insurance Corp of India (LIC), State Bank of India (SBI), Bank of Baroda (BoB), Punjab National Bank (PNB), Canara Bank, Indian Bank, IDBI Bank, Yes Bank, Federal Bank, Reliance Capital, Birla Sunlife and many others, who together manage assets worth thousands of crores of rupees and have equally staggering deposits at their disposal.
Cobrapost said today, "The RBI continues to act as a protector, and not as a regulator or enforcer. The RBI is loath to admit that there exists a massive money-laundering racket in our financial institutions across the board, flourishing under its very nose, simply because if it were to admit to this deep-rooted malaise, it would strip the RBI of its halo as a "one of the best banking regulators in the world."
All banks have been asked to strictly adhere to KYC norms. Even older account holders' antecedents are being verified. The banks involved in money laundering have punished their staff members – 50 so far – with suspension. The Finance Ministry has asked the Banking Association of India to launch a thorough probe into the functioning of all banks, while individual banks have announced their own probe, and punishing the staff involved with suspension.
"Whether such inquiries are followed up with visible and effective action on ground or not, only time will tell. However, if the RBI and the Finance Ministry are waiting for more clinching evidence to finally act on it, we here present yet more shocking videos that should normally awaken any law enforcement authority. We hope that the authorities wake up to the ground realities and start the requisite legal procedures," Cobrapost said.
Here are the snippets from Cobrapost's latest Operation Red Spider III...
IndusInd Bank
Ironically, its motto is: We make you feel richer. Now let us tell you what lessons our reporter learnt at this bank:
At one of the branches of IndusInd Bank in an upscale area of South Delhi, its personal banking head N Anand customizes the family income builder scheme, in tie-up with AVIVA – a maximum yearly deposit of Rs5 lakh – which means Rs15 lakh for three investors and Rs1.5 crore in 10 years. It also means that you are depositing less than Rs10 lakh a year in an account. Cash in lockers is okay, and you can count your money 'araam se' (in comfort) inside the branch.
At the bank's branch in Hyderabad, Branch Manager S Mishra and operations manager Amita are bolder. It is the same family income builder scheme, and when we talk about how to put in cash, they say "woh to hum baad me karenge (We will do that part later)." She has full co-operation of the bank: "Hum bank ke through kuch na kuch alternative bana sakte (We can arrange for some alternative through the bank)."
She actually solves the Rs5 lakh investment limit issue. Take out another policy a month later, without PAN card.
She throws light on the 7-year life of an Income Tax notice. She also advises the reporter on the Streedhan clause that can move a large portion of the black money out of the reach of the law for good.
At another branch of the bank in Hyderabad branch manager TJ Reddy and customer services manager Srikanth K offer great ideas.
Srikanth explains the 'pattadar passbook' or a passbook for agricultural income. The bank will do that. In this way we can get away even after we provide the PAN. When we want more accounts and more account holders, the bank offers to make these 'benami' accounts for us, with only us having the authority to withdraw. The bank even provides these faceless people for the accounts.
Bank of Baroda
The bank calls itself 'India's international bank'. Now let us tell you what lessons our reporter learnt at this bank:
For RC Verma, senior manager and S Kamran, manager at its Gurgaon branch, slow and steady wins the race. But be sure: they do win. Kamran says: "Aur toh aap humare touch mein aaoge toh aapka sub number ek mein aise hi convert kara denge … aapko pata bhi nahin chalega (If you keep in touch with us, we will convert all your money into white … you will not even know it)."
The hubris is amazing. Both managers are okay with cash in lockers. In fact, they want it that way, so they can slowly route the cash to investments.
D Singh, senior manager at BB's South Delhi branch gives us a fine idea: "Aise aadmi ke naamdaal doonga jo return file na karta ho (I will put in the name of a person who doesn't file the return)."
Or, "Joint account kar lo, pahla naam uska rakhlo jiska tax nahin hoti hai, doosra apna rakh lo jo monitoring karni hai … monitoring karenge to woh nikal hi nahin payega … paper bhi apne paas rakhna…. bagair papare ke waise hi nahin niklega (Do it through joint account. Put his name first, then for the second account holder put the name of the one who will be monitoring it. If you are monitoring then he won't be able to withdraw money … keep the papers with you … without papers it won't be withdrawn )."
RR Bhaskar, Senior Branch Manager of BoB, South Delhi, says: "Actually ikkatthe nahin karna chahenge... ekdum se light mein aa jate hain [Actually, it has not to be done in one shot… (it) may bring it to light immediately]."
Bhaskar also sees the agricultural income route: "Usme kya hai ki you can justify … agriculture land pakka hogi inki … agriculture land ka paisa is mein dekhayenge … income tax walon se bachne ka ek ye tarika hai (The thing is that you can justify … I am sure he owns agriculture land … we can show agricultural income… this is one method to stave off the Income Tax people)."
ING Vysya Bank
The following are the lessons our reporter learnt at this bank:
A Bhaskar, Deputy Manager of the bank in Chandigarh, is not exactly in the category of a serious banker, but is still conversant with most underhand dealings that the system incorporates. In bringing cash from the minister's house, he is careful to take his own car, not the bank's, and though he has not been to see the lockers, he can surely arrange for cash to be stashed there.
Manager P Saha and Relationship Manager S Das of the bank's Kolkata branch have been there, done it. Saha explains the goodness of insurance policies: "This is the only product jahan par government haat nahin laga sakta... Yeh government bond debenture me jaata hai... aur 10 saal, 12 saal baad jo maturity hota hai, government has no right to touch LICs. Because it's related to the life of an individual... (This is the only product that the government cannot touch. These are invested in government bonds and debentures that mature in 10-12 years... The government has no right to touch LICs, because it's related to the life of an individual)."
SS Sudhir, the Branch Head at the bank's another branch in Hyderabad explains when an account is tracked by authorities: "Cash aap... dal sakte.. isme tracing nahin ata. Tracing ap ko tab hoga jab agar aap ko ... kuch transactions hua. Matlab aapko transactions ka track kahin pe hua... Matlab yeh Hyderabad mein aap transaction kar rahe hain... if you do any transaction with anybody... that particular account is called for any explanation, then you check from there where it has been routed... In that sense if they to track all that then it comes... (You can put in cash. There is no tracing in this. Tracing happens when you have some transactions. That means any of your transactions are tracked anywhere. Say, this is Hyderabad, you are doing transactions...if you do any transaction with anybody... that particular account is called for any explanation, then you check from there where it has been routed... In that sense if they to track all that then it comes...)"
Basically the probability is low, she says.
Regarding transferring money out of the country, she sheds light on certain dealings: "Most of the people are doing, you know, they are taking out the money and sending it ... and then setting up the business..." And banks are complicit in this.
Allahabad Bank
This is one of the leading public sector banks of the country. Established in 1865, it prides itself in its 'Tradition of Trust'. This trust, however, is now in doubt.
Chief Manager A Sharma and Senior Manager RM Ahuja of the bank's South Delhi branch form a cozy coterie within the bank, greasing paths for all the black money available.
Even a retired officer, Khanna, is among the coterie. Says he about a benami account: "Account khologe aap ... agar uske naam pe jama karna hai toh usko mat lao ... aap khud jama kar do uske naam pe (If you want to deposit money in his name, don't bring him, you deposit)."
Central Bank of India
Another leading public sector banking behemoth, this bank has more to hide despite its glorious past.
SK Garg, Chief Manager of the bank's upscale branch in Central Delhi provides a way out to move cash into accounts. "Main aapko ek cheez sahi bata deta hoon ... jo LIC karwani hai wo toh cheque se karwaiye ... aur baaki jo main aapko jo hai ... ya toh alag alag bank pe jo hai ... pachaas pachaas hazar rupaye ki DD bana dijiye ... naam mein jismein bhi account aap kholte hain ... baad mein wo clearing mein daal ke ... wo jama ho jayega (Let me tell you one thing ... do the LIC [policy] by cheque ... and with the rest of the cash ... get several DDs for Rs. 50,000 each from other banks in the names you will open the accounts here ... later on you can put them all into clearing ... that [money] will be credited [in those accounts])."
Here are some excerpts of confessions of past criminality:
Allahabad Bank
Chief Manager A. Sharma of the bank's Defence Colony branch in New Delhi makes his apathy for the system clear: "Pehle kahte ki jisko TDS katata hai unke naam do ... ab jinka bhi tax katta hai, TDS kate ya na kate ... unka naam mangne lag gaye department ... pehle to hum log kaafi manage karte the (Earlier they [the Income Tax Department] used to ask for the names of people whose TDS is deducted ... nowadays, they have started asking names of all who are taxable, no matter if there is TDS or not ... earlier we could manage a lot)."
RM Ahuja: "Abhi to proof aur ye sab hone lag gaye hain ... humare paas ... humne ... benami humne ... kitne hi customers ka lakhon rupaya ... aaj se bees saal pehle, jis samay lakh ... aaj ka crore se bhi jyada hoga unke keemat ... wo de jaate the ... humne apne driver ke naam account khole ... yun hi sign mar diye (Now they have started asking for proofs and all that ... 20 years ago ... we had many customers who will just give us lakhs of rupees ... a lakh on those days is worth more than a crore today ... we used benami accounts ... in the name of our own drivers ... we signed for them just like that)."
National security threat
On 17 December 2012, Parliament gave its nod to a bill, which seeks to enlarge the definition of Money Laundering offences that could help the funding of terrorist operations. The Prevention of Money Laundering (Amendment) Bill was passed by the Rajya Sabha by voice vote.
It may be pointed out that the Cobrapost undercover reporter was never grilled about the origin of the money, or what it was intended for, by any banker. The bankers were simply not interested. Methods adopted in Operation Red Spider could easily be utilized by any terror organization to move huge amounts of money. This, thus, becomes a case of National Security, and the perfunctory and dismissive nature with which the RBI, the Finance Ministry and the banks have been treating this only makes the situation extremely dangerous.
Cobrapost reiterates India's responsibility as a signatory to FATF (Financial Action Task Force on Money Laundering), because the republic has treaty obligations to block precisely the kind of money laundering that the Cobrapost expose shows as rampant in the 10 banks mentioned. Operation Red Spider Part III again conclusively shows that the RBI and the Financial Intelligence Unit of the Finance Ministry have failed in stemming the systemic rot in the banks.
The gold and insurance connections
The other surprising thing observed in this huge undercover investigation by operation is the incredible urgency with which officials are pushing the sales of the banks' gold coins/ biscuits. While the coins/ biscuits present an easy option for laundering black money, this policy could have certainly helped in triggering the huge spike in retail gold prices in the country before the worldwide slump in this metal happened – not to mention, be the cause behind India's ballooning Current Account Deficit (CAD).
Yet another unholy alliance that has come unstuck is the liaison or tie-ups these banks have with the insurance companies, including the public sector behemoth Life Insurance Corporation of India. When the banks fail to provide a smooth channel to launder the cash, they quickly call upon their insurance company 'buddies' and fix a deal that suits our objective the most.
It is imperative to point out here that the financial institutions of the country, as a whole, are answerable to the people of the Republic of India. They need to explain to the average tax-payer and even to the daily wage earner, who deem these behemoth institutions as upholders of the trust they place in them, how such brazen acts of crime have been perpetuated for so long.
Our modus operandi
We used the same method as before. Our undercover reporter, Associate Editor Syed Masroor Hasan simply walks into a bank, armed with a fabricated story of a fictitious minister's ill-gotten wealth, in cash. He asks the officials for a way to 'invest' this imaginary unaccounted money – starting with amounts as large as Rs50–Rs60 lakh or even Rs1 crore, leading up to amounts often as huge as Rs37 crore – in legal schemes in the long term, with not only the sole purpose of laundering the black money but also to get handsome returns on that.
Logically, any official of any established bank should immediately distance himself/ herself from such a story and its blatantly illegal consequences. These set of bankers and insurers, as in those we exposed in our last two episodes, welcomed our reporter and offered several innovative ways to channelize the cash. They were eager to accommodate the offer at any cost.
The following were the methods suggested most frequently by the banks:
Bankers are at pains to remind you of three very important aspects of underhand dealings:
(a) The government does not have the right to touch insurance policies, because these deal with the life of an individual.
(b) If the women in a family have insurance policies bought with black money, they can sign a form that labels this as Stree dhan or Women's Wealth. As a banker helpfully put it, even if there is an Income Tax raid at your home, they will not be able to touch this particular policy.
(c) Nobody can question the source of income for gain from an insurance policy after seven years.
Be informed that the above is not even an exhaustive list.
What the Law of the Land has to Say on Money Laundering?
The videos, through Cobrapost's undercover investigation in Operation Red Spider Part III, clearly show the gross violation of rules and regulations, framed under various laws of the land, namely, the Prevention of Money Laundering Act of 2002, the Income Tax Act and the Indian Penal Code, among others.
The first exposure from Cobrapost triggered a series of muted reactions actions from banks, the finance ministry and the Reserve Bank of India (RBI), although committees have been set up to investigate the sting operation and a few bank managers and junior officials were suspended. Reacting to Cobrapost exposure on 21 March 2013, RBI's deputy governor Dr KC Chakrabarty said, "No scam has happened... Allegations don't mean that KYC norms have been flouted... If KYC is made more stringent then the opening of bank accounts for the finally excluded may become tough... Allegations do not mean flouting norms. There is not a single transaction (of money laundering as alleged by the exposé)...These are transactional issues that have nothing to do with money laundering (!)... I am not saying that there is no problem. I know there is a problem in the system... RBI's Financial Statement Reports highlight serious concerns over bank branches selling gold coins, mutual fund instruments, and insurance. RBI has undertaken thematic studies of banks that are active in selling gold coins and wealth management products… We cannot take action on allegations. We go by evidence."


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text:
Science Applications International Corporation
|This article's lead section may be too long for the length of the article. (June 2017)
|Traded as
S&P 400 Component
|information technology and engineering
|1969 (original company)
2013 (as a spin-off of Leidos, formerly SAIC)
|Reston, Fairfax, Virginia, United States
Area served
Key people
|Edward Sanderson, Jr. (Chairman)[1]
Anthony J. Moraco (Chief Executive Officer)[2]
|US$ 4.5 billion (annual)
Number of employees
|Footnotes / references
Science Applications International Corporation (SAIC) is an American company headquartered in Reston, Virginia[6] that provides government services and information technology support. The original SAIC was created in 1969 by J. Robert Beyster. Then on September 27, 2013, it spun off a $4 billion unit which retained its name,[7] while the parent company changed its name to Leidos. Following the split, Anthony J. Moraco was appointed CEO of SAIC, and John P. Jumper was appointed CEO of Leidos.[8] The primary motivation for the spinoff was the conflicts of interest provisions in the Federal Acquisition Regulations which prevented the company from bidding on some new contracts because of existing contracts.[9][10]
On May 4, 2015, SAIC acquired Scitor Holdings, Inc. for $790 million to expand their presence in the intelligence industry through classified contracts, cleared personnel, and a robust security infrastructure. [11] Scitor was previously owned by Leonard Green & Partners, L.P., a private equity firm.[12]
SAIC has adopted a matrix operating model in which different service lines collaborate to serve a given contract.[10][13]
Deborah Lee James, president of SAIC's technology and engineering sector, was sworn in as Secretary of the Air Force on December 20, 2013 after being appointed by President Barack Obama.
- ^ SAIC. "Edward Sanderson, Jr. - SAIC".
- ^ SAIC. "Anthony J. Moraco - SAIC".
- ^ Aitoro, Jill R. (27 September 2013). "What to expect from Leidos and SAIC when they start trading Sept. 30". Washington Business Journal. Retrieved 29 September 2013.
- ^ Aitoro, Jill R. (27 September 2013). "Exclusive: John Jumper explains why the Leidos-SAIC split had to happen". Washington Business Journal. Retrieved 26 September 2013.
- ^ "www.saic.com". Retrieved 29 September 2013.
- ^ SAIC. "Contact Us - SAIC".
- ^ Jayakumar, Amrita (27 September 2014). "One year later: The tale of SAIC and Leidos". The Washington Post. Retrieved 1 October 2014.
- ^ "- The Washington Post". Washington Post. Retrieved 2016-09-29.
- ^ Riekenberg, Philip (2013-09-17). "SAIC: A Profitable Solution To A Conflict Of Interest". Retrieved 2016-09-29.
- ^ a b "One year later: The tale of SAIC and Leidos". Washington Post. Retrieved 2016-09-29.
- ^ "SAIC Completes Acquisition of Scitor | SAIC". investors.saic.com. Retrieved 2016-09-29.
- ^ "Leonard Green & Partners - Past Investments". www.leonardgreen.com. Retrieved 2016-09-29.
- ^ "SAIC » Simulation & Analysis". www.saic.com. Retrieved 2016-09-29.
External links[edit]
|This United States corporation or company article is a stub. You can help Wikipedia by expanding it.


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text:
South West MEP On Holidays After Brexit
Green MEP advises holidays makers of their rights post-brexit
A report has praised the Swindon Youth Offending Team for being was committed to helping children and young people stop offending, but says improvements are needed.
Inspectors found work to reduce reoffending was satisfactory, work to ensure that the sentence was served was good and the delivery and management of interventions to reduce reoffending were satisfactory.
But they were concerned work to protect the public and actual or potential victims was unsatisfactory, with the YOT often underestimating the level of harm a child or young person posed to others and the subsequent need to plan to manage this. Work to protect children and young people and reduce their vulnerability was unsatisfactory and the effectiveness of governance and partnership arrangements was unsatisfactory.
Chief Inspector of Probation Paul Wilson said: "Overall, we found an evident commitment by Swindon YOT to keep the children and young people with whom it worked safe, enhance their wellbeing and help to reduce the likelihood of them offending again. The YOT formed strong relationships with its partners and service users to provide a wide range of good interventions. However, there were a number of gaps at an operational and management level in work to protect the public and safeguarding. The YOT Management Board was interested and knowledgeable about the YOT but had yet to put the governance, scrutiny and challenge structures in place to help direct and inform its work.
"We have identified a number of areas where Swindon has cause to celebrate its performance and highlighted others here there is scope for review. We have made a small number of recommendations in specific areas for improvement and look forward to Swindon's successful implementation of these."
In response Swindon Borough Council released the following statement:
''The inspection report makes a number of recommendations about the YOT's management processes which are being acted upon, and this will give the reassurance the inspectors require about how the YOT's results are achieved. The overall message from the report is that in the areas that truly matter – reducing reoffending among young people, and making sure they properly serve their court sentences – the Swindon Youth Offending Team performs well. In fact, the rating given for the work to make sure young offenders serve their sentences places the Swindon YOT as one of the best performers in England and Wales.''
Green MEP advises holidays makers of their rights post-brexit
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An Act Relating to the Determination of Human Death
Bibliographic Citation
Missouri. General Assembly. 81st General Assembly, 2nd regular session. H.B. 1223. Pre-filed 1 December 1981
Permanent Linkhttp://hdl.handle.net/10822/791885
MetadataShow full item record
Related items
Showing items related by title, author, creator and subject.
A Bill ... Relating Generally to the Uniform Determination of Death Act; the Determination of Death; and the Definition of the Term 'Death' in the Uniform Anatomical Gift Act. S.B. 3 Unknown author (West Virginia. Legislature. Senate, 1982-01-13)
A Bill for an Act Relating to the Determination of Death Unknown author (Hawaii. Legislature. House, 1982-01-29)
Legislation: Death and Related Issues: Sweden: Regulations and General Recommendations No. 5 of 18 June 1996 of the National Board of Health and Welfare Amending Regulations and General Recommendations No. 32 of 29 October 1987 of the National Board of Health and Welfare for the Implementation of Law No. 269 of 1987 on the Criteria for the Determination of Death in Human Beings Unknown author (1996)


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Knowledge of Dynamic VLANs and VMPS is grievous in your pains to passing the BCMSN exam and take in your CCNP, and it's besides a excessive accomplishment to have for your networking work.
As a CCNA and CCNP candidate, you cognize how and why to tack together fixed VLANs. Static VLANs can be a violent contraption for reduction excess broadcast and multicast traffic, but if hosts are captive from one controller quay to another, you've got to clear those changes manually on the control. With Dynamic VLANs, the changes are made - how else? - dynamically.
The existent constellation of resurgent VLANs is out of the margin of the BCMSN exam, but as a CCNP competitor you necessitate to cognize the details of VMPS - a VLAN Membership Policy Server.Post ads:
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Using VMPS results in wharf VLAN political leanings changes person performed dynamically, because the port's VLAN rank is contracted by the root MAC computer address of the instrument united to that anchorage. (Yet different origin that the most primitive plus point a switch looks at on an elect frame is the wellspring MAC address.)
In my territory lab network, I've got a grownup related to to electrical switch left fast0/1 that resides in VLAN 12. What if we had to dislodge Host 1's seam to the electric switch to waterfront 0/6? With ageless VLANs, we'd have to bracket together to the switch, assemble the marina as an right port, and past plant the larboard into VLAN 12. With VMPS, the just entry we'd have to do is reconnect the wire to waterfront 0/6, and the VMPS would dynamically leave that waterfront into VLAN 12.
I insist you to do added language on the subject of VMPS. Use your favorite explore motor for the residence configuring vmps and you'll at the double brainwave some remarkable chief Cisco certification on this topic.Post ads:
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To review, the VLAN body of a grownup is definite by one of two factors. With monotonous VLANs, the host's VLAN membership is the VLAN to which its control port has been appointed. With projectile VLANs, it is mutually beneficial upon the host's MAC address.


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28 Berger StHancock, NY 13783MLS #107334
- 3beds
- 1+1½baths
- 0.25acres
- 1,500sq ft
Est. P&I payment: $457.94/mo
Description for 28 Berger St, Hancock, NY 13783
Well built 50's ranch with addition and updates in quiet neighborhood near river.
Well built 50's ranch with addition and updates in quiet neighborhood near river.
My Notes for MLS #107334
General Features
- Property Type: Residential
- Property Subtype 1: Single Family
- Style: Ranch
- Year Built: 1958
- County: Delaware
- Acres: 0.25
Exterior Features
- Lot Dimensions: 100x95
- Lot Measurement: Acres
- Lot Size Source: (Tax Records)
- Lot Square Footage: 10,890
- House Color: Green
- Construction: Frame
- Foundation: Poured Concrete
- Garage Type: Attached
- Garage Spaces: 4
- Handicap Features: No
- Roof: Asphalt Shingle
Interior Features
- Bedrooms: 3
- Full Bathrooms: 1
- Half Bathrooms: 1
- Square Footage: 1,500
- Square Footage Source: Estimated
- Bedroom 1: 13x11 Level: 1st Floor
- Bedroom 2: 13x9 Level: 1st Floor
- Bedroom 3: 10x10 Level: 1st Floor
- Bathroom 1: 9x7 Level: 1st Floor
- Bathroom 2: 4x4 Level: 1st Floor
- Basement: Full
- Dining Room: 15x13 Level: 1st Floor
- Family Room: 15x12 Level: 1st Floor
- Flooring: Wood
- Kitchen: 18x8 Level: 1st Floor
- Living Room: 18x12 Level: 1st Floor
Utility Features
- Appliances: Dishwasher,Range,Refrigerator
- Heat/Cool Type: Forced Air Heat
- Hot Water: Electric
- Insulation: Unknown
- Sewer: Public Sewer System
- Water: Public Water Supply
Financial Features
- Assessment: 13,300
- Town/CountyTax: 967
- Village/City Tax: 1,708
- School Tax: 1,477
- Tax Id: 123601-439-006-0001-008-000
School Features
- School District: Hancock
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The listing information for 28 Berger St, Hancock, NY 13783 was graciously provided by our friends at the listing brokerage, Woodland Creek Real Estate
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Last Updated 07/25/2017 at 10:00pm
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text: The NETGEAR AirCard 762S Mobile Hotspot on the O2's 4G Network (Photo: Business Wire)O2's Pop Up Office proposition allows small business to create their own portable 4G mobile hotspot instantly. The solution allows those with a variety of devices (laptops, tablets, smartphones) to hook up to a 4G connection when they're out and about. "The NETGEAR AirCard 762S is the first mobile hotspot we're offering to our business customers, as it's easy to set up and high performing," said Paul Lawton, general manager of SMB at O2. "It makes up a crucial part of our 'Pop Up Office' proposition, as we believe that giving small businesses the opportunity to be connected at all times, particularly in non-fixed locations, will revolutionize the way some small businesses are able to work. They are no longer bound to the office, and can essentially pop up their business and be connected wherever they need to be." Next-generation office applications and services powered by 4G enable O2 business customers to be more collaborative, more efficient and more productive, as 4G networks provide faster data speeds (up to five times faster than 3G*) and a more seamless mobile experience. O2 switched on its 4G network in London, Leeds and Bradford on August 29th, with a rollout plan to have 4G available in a total of 13 cities by the end of the year.


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D2 Racing 90-93 Integra SL Series Coilovers
Part Number: D-HN-13-SL
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The All New SL Series coilover system from D2 is the first coilover specifically designed to provide your car with the lowest possible ride height while retaining the benefits of a full monotube coilover system.
The SL Series features specially designed mounts, shorter stroke shock bodies, 36-way adjustabile dampers, allowing you to slam your car more than ever before. The coilover system can lower the vehicle up to 2" more than our RS Series street coiloverFeatures:
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- Part Number: D-HN-13-SL
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| This article is about a project that is in development.
Information may change significantly as the project progresses.
The project seemed to have stopped, so the sprites were released on a Gameware thread in a zip file with other unused sprites (Other Sprites). You can download the sprites here. Not all sprites were finished.
I'm putting it on hiatus for now, but plan on finishing it at some point-Anaisa


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This isn't the Ritz but it suited our need for a good room at a decent price. The staff was friendly and the rooms were clean. The small fridge was nice and so was the breakfast. There is a coin laundry and a outdoor pool. The internet was free and worked well. We will stay here next visit. I recommend a downstairs room because there isn't an elevator.
Room Tip: ask for a downstairs room
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Semi-Private & Private Group Lessons
This is a great way to get private instruction with your own group of friends! Form a group between 2-8 people of the same level and watch your group improve as a whole. These groups work great for teams that want practice sessions between league match days. Semi-Privates are also a cost-effective way of learning the game!
Open Drills
All 3.5 & up USTA ratings or A/B level players are invited for an hour and a half of tennis drills.
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1 hour of training/drills for players that have progressed past the beginner stage. Focusing falls on slightly more advanced match tactics and game strategy in preparation towards tournaments and match play. Monthly rates are based on 4 weeks per month.


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Super Sunday Reds Case
Your Super Sunday Stock Up Reds Case includes two bottles each of
Big Cirrus Syrah
Maule is on the rise in Syrah production as this noble grape is growing in demand and popularity. Enjoy fruit flavors of blueberry, blackberry, wrapped in hints of pepper, violets, clove and thin layers of vanilla with every sip of this beautiful Reserva wine from Maule Valley located in central Chile.
Rivers End Reserve Merlot
A wonderful wine from the River's End winemakers with ripe berry fruit flavors and aromas with a nice touch of oak. An amazingly versatile choice for serving with grilled duck, turkey, chicken, braised lamb, pork or beef – and a nice partner for dishes featuring a rich tomato sauce.
Buscado Garnacha
Grenache is a varietal grown around the world and in the country of Spain, it goes by the name Garnacha. Each country where Grenache is grown, it expresses the unique terroir and shows different flavor profiles. For Spain and specifically the grapes in Buscado, Garnacha produces intense and pleasant aromas of licorice and wild berries with slight mineral undertones. Flavors of ripe red fruit, fresh thyme and hints of minerals linger on the finish of this silky and full-bodied expression of Garnacha.
Albarrada Red
Albarrada is a new and exclusive winery that makes amazing wine that showcases local grapes. This red wine (or vinho tinto, in Portuguese) features a local grape, called Castelão. Native to the soils of Portugal, Castelão. It is unusual, finding a better home in the drier and harsher climate of Portugal than the normally more grape friendly soils of temperate climates. When well cared for, the wine that results is aromatic and fruit-forward with notes of red berries and slight hints of floral notes.
Fair Oaks Zinfandel
A claret style Zinfandel with full, jammy flavors of blackberry, raspberry, black cherry and wild berry flavors with a hint of black pepper and anise. A nice touch of acidity makes a clean, food friendly finish. Our expert wine panel enjoyed it from appetizers through to dessert and wished they had more.
Vila Nova Red Duoro Blend
The Duoro region of Portugal is in the spotlight because in 2014 it produced 4 of the top ten wines in the world as rated by Wine Spectator Magazine. Now in the forefront of wine lover's minds, the wines from this region are becoming more and more sought after. A blend of Tinta Roriz (Tempranillo), Touriga Franca and Touriga Nacional, Vila Nova is a fresh red wine that has a dark, deep and intense ruby color, with a nose dominated by fruity aromas that carry through. It is well-balanced with silky tannins finishing long and balanced with silky tannins, finishing long and fruit-filled. The grapes from Vila Nova Douro Red wine are from the oldest wine region in the world. Harvest is conducted 100% by hand from vines that are 20-to- 30 years old, with the best grapes of each of the above varietals chosen for fermentation at controlled temperatures in stainless steel tanks. Fifty percent of the finished wine is then aged for six months in used 400 liter French oak barrels. This wine is ideally served at room temperature with light meat dishes, such as pork, or flavorful, hard cheeses, such as Italian cow and sheep's milk cheeses.


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The Urban Decay Shadow Box is a brand new, Ulta exclusive eyeshadow palette filled with a dozen Urban Decay Eyeshadows. You're getting both brand new shades (three never before seen shades) in this palette as well as shades that you may already own or have wanted to own.
Let's take a look!


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Thanks for calling attention to this beautiful natural area only a short drive from Cleveland. This is an excellent time of year to observe migrating birds and other wildlife at the marsh. A couple corrections, first the address of the Mentor Marsh House is 5185 Corduroy Rd. Second, the reclamation projects in the marsh continue but it is far too soon to say the invasive phragmites has been "fought off". In fact this invasive still dominates the marsh due to salt contamination from a brine dump built in the 1960's. Recently that dump has become the attention of clean-up efforts. All are encouraged to contact your elected state leaders to encourage them to expedite this.
Tom Rapini, Mentor Marsh Board


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Design the makeup and outfit for the girl next door! Choose between many casual shirts and jackets. Change the gal's eye color, and apply matching eyeshadow. Wear beaded necklaces and sparkling, dangling earrings. Complete the new look with cool sunglasses!
Make Up 22 is one of our selected Make Up Games. Also very popular on this website right now are My Dolphin Show 8, Avie Christmas Style, Christmas Pudding: Sara's Cooking Class and Papa's Cupcakeria.
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Web loses his brother in an extreme sports accident. When he learns it may have been murder he searches, from America to his ancestral home in China, pursuing answers and vengeance.
This is Urban Games and you are watching MovieZion
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Frakus Plugin - Public Beta
A real distortion plugin for serious noise musicians. Make your nice Macbook Pro and expensive audio interface sound like a broken $10 piece of garbage. More than twelve modes of audio destruction to frighten small children and woodland creatures! Find out more.
Drifter Mage: Perfect Enemy Limited Edition Cassette
The first release for Drifter Mage is now available in cassette for a short time! Go here to listen and buy Perfect Enemy.
Treefungus now on Coprolalia
The electronic duo Treefungus has joined the Coprolalia Network. Listen to their latest album Transmission 495 now!


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Hide lots of fields, conditions to select one?Asked by delbuys on February 24, 2014 at 06:41 PM
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I'm making a form where people can type the make and model of their phone in and it gives them a price we will pay for it.
Basically I've got a dropdown for the make, and a dropdown of the model.
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iPhone 3GS 8GB
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Sennheiser Unveils Premium Headsets Optimized for Microsoft Lync
DW and Circle Series Portfolio Additions are Designed to Deliver Comfort and Clarity, While Leveraging the Productivity Advantages of Unified Communications
Circle Series
The Circle Series includes two Microsoft Lync-compatible models: the dual-sided SC 260 ML and the single-sided SC 230 ML. Each is designed to deliver HD voice clarity sound and comfort to busy contact center, office and Unified Communication professionals. "In increasingly busy workplaces, smart communication tools that allow users to work more efficiently can give a competitive advantage. With Sennheiser's launch of its new wired Circle Series headsets optimized for Microsoft Lync, we can help provide our mutual customers with an even wider choice of Unified Communications tools to improve their business productivity,"
DW Series
The DW Series DECT wireless headsets includes three Microsoft Lync-compatible models: the 2-in-1 DW Office ML with headband and ear hook, the DW Pro1 ML single-sided headset and the DW Pro2 ML double-sided headset. "The DW Series of headsets optimized for Microsoft Lync leverages Sennheiser's expertise in sound quality, noise cancelation and wireless audio transmission,"
"Sennheiser is very pleased to be extending our portfolio with these five new wired and wireless products, each of which are now optimized for Microsoft Lync," Seliokas concluded. "With our leadership in sound quality, comfort and hearing protection, contact center, office and Unified Communication professionals can enjoy an enriched communications experience."
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Sennheiser's Circle and DW Series headsets are now optimized for Microsoft Lync.
About Sennheiser
Sennheiser is a world-leading manufacturer of microphones, headphones and wireless transmission systems. Established in 1945 in Wedemark, Germany, Sennheiser is now a global brand represented in 60 countries around the world with U.S. headquarters in Old Lyme, Conn. Sennheiser's pioneering excellence in technology has rewarded the company with numerous awards and accolades including an Emmy, a Grammy, and the Scientific and Engineering Award of the Academy of Motion Picture Arts and Sciences.
You can find all the latest information on Sennheiser by visiting our website at www.sennheiserusa.com or by contacting:
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Get To Know About Kitten Illnesses
Kitten illnesses are to be viewed in a serious manner because the kittens are highly vulnerable to the pathogenic changes made by various pathogens or illnesses. Kitten illnesses have to be treated in a fast manner.
If you want to avoid such problems, you have to take lot of precautionary measures. The kittens get mostly affected by worm problems. In particular, the round worms are gaining more significance in this younger group of cats.
If they are larger in numbers, some worms may get vomited through the mouth. Yeah. The vomit may have some worms too, during such kitten illnesses. Getting amazed? But, it is true. Don't get panic on seeing the worms in the vomited materials.
If the kitten swallows some foreign materials, then they may make more severe attempts to vomit. Hence, the activity of vomiting is considered as a protective response and such vomiting may occur with more sounds.
However, if you see a continuous type of vomiting with pink froth, you have to consult a veterinarian immediately to get proper therapy.
Cestodiasis is another problem often encountered in the kittens. The affected animals may often squat on the ground and the rubbing of anal regions on the ground may be so severe and serrations may at times occur at the posterior regions.
Hence, they need immediate therapeutic attention. Rice beads like materials may be seen in the areas under the tail around the anal parts. Parvo viral infection may also affect the kittens severely.
The affected animals may reveal motion with blood streaks and the animals may also be dehydrated seriously. Hence, the kittens may be found depressed and with lethargic movements.
Feline infectious peritonitis has to be examined if these young cats reveal the symptoms like seizures, vomiting in frequent manner etc.
There may be signs of intense pain due to the affection by means of swelling at the peritoneal regions of the body. Feline immunodeficiency viral infections are the ones that are difficult to treat in general.
Hence, the kittens need to be protected with all types of vaccinations that are required. For this, it is essential that you have to consult a veterinarian in time.
Get some technical suggestions on the protective measures required to avoid the kitten illnesses during the meeting with veterinarian.
The gastric problems like gastritis or gastric ulcers may become the worrying kitten illnesses and the cats may never be comfortable with kitten illnesses. More incidences of vomiting and occasionally vomiting with blood streaks may occur in such occasions.
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KBC Networks, provider of industrial transmission equipment, has introduced PoE (Power over Ethernet) to its network transmission range. The first of the new products is a fully-ruggedized 10/100 Ethernet media converter with PoE+ that conforms to the higher power specification necessary for PTZ and dome cameras.
The new KBC FTLS1 is fully-compliant with the IEEE 802.3at specification and acts as a PSE (Power Sourcing Equipment) providing 25.5W @ 48Vdc. The unit supports transmission of an Ethernet channel over either one or two, multimode or singlemode optical fibres and with full-duplex operation, it enables transmission distances in excess of 30km on singlemode fiber.
Iain Deuchars, MD KBC Networks EMEA commented on the new product series, "We're seeing an increasing demand for our industrial IP products. As well as adding the PoE unit to the range, we're also concentrating on expanding our series of Ethernet switches and developing our ThruLink VPN device."


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Diva/BclB regulates differentiation by inhibiting NDPKB/Nm23H2-mediated neuronal differentiation in PC-12 cells
Cellular and molecular mechanisms
Our results show that after differentiation, Diva/BclB expression was decreased and reciprocally, NDPKB/Nm23H2 expression was increased and it translocated into the nucleus. Overexpression of NDPKB/Nm23H2 promoted PC-12 neuronal differentiation by increasing neurite outgrowth and arresting cell cycle progression. There was a concurrent downregulation of Diva/Boo when NDPKB/Nm23H2 was overexpressed, which mirrors the effect of NGF on PC-12 cell differentiation. Overexpression of Diva/BclB did not change the expression level of NDPKB/Nm23H2, but inhibited its nuclear localization. Cells that overexpressed Diva/BclB presented a decreased percentage of differentiated cells and average neurite length was shortened. This was due to an increase in the formation of Diva/BclB and NDPKB/Nm23H2 complexes as well as Diva/BclB and β-tubulin complexes. Concomitantly, there was a decrease in formation of NDPKB/Nm23H2 and β-tubulin complexes. Overexpression of Diva/BclB also resulted in a higher percentage of S-phase cells.
Our results showed a novel role for Diva/BclB in neuronal differentiation. Its downregulation during neuronal differentiation may be necessary to allow NDPKB/Nm23H2 and β-tubulin interaction that promotes NDPKB/Nm23H2 mediated differentiation.
The PC-12 cell line has been used extensively as a model of neuronal differentiation, as the cells halt proliferation, have outgrowth of neurites and acquire properties of sympathetic neurons after exposure to nerve growth factor (NGF)
Diva/BclB is a member of the Bcl-2 family of proteins. A human ortholog has also been identified, and designated as BclB or Bcl2l10. It is also known as Boo (Bcl-2 homologue of ovary). Diva/BclB contains BH1, BH2, BH3, and BH4 regions and a carboxyl-terminal hydrophobic domain. It is distributed in the brain, liver, and heart of E15 embryonic mice. Interestingly, in adult mice the expression of Diva/BclB decreases dramatically and becomes restricted to the granulosa cells of the ovary and the seminiferous tubules of the testis
Nm23/NDPK are house keeping enzymes needed for transfer of the terminal phosphate of a nucleoside triphosphate to a nucleoside diphosphate. Out of the ten known human Nm23 family genes, NDPKA/Nm23H1 and NDPKB/Nm23H2 are the most abundant and ubiquitous isoforms, and are located both in the nuclei and cytoplasm
Thus far, the physiological role of Diva/BclB has remained elusive due to a paucity of information. Even its function in apoptosis is highly controversial with many opposing reports. Recently, there has been evidence of Diva/BclB having a direct role in development. Knockdown of the zebrafish ortholog (NRZ) resulted in disorganized somites, increased levels of Snail-1, and disorganisation of the microtubule network and F-actin depolymerisation in the yolk sac
Up-regulation of NDPKB and down-regulation of Diva during neuronal differentiation
To address whether NDPKB/Nm23H2 and Diva/BclB may play physiological roles in neuronal differentiation, we examined the expression of these proteins during PC-12 neuronal differentiation. PC-12 cells showed a steady and significant increase in NDPKB/Nm23H2 and β-tubulin mRNA levels, with a concurrent decrease in Diva/BclB mRNA levels during the course of NGF-induced differentiation (Figure
Figure 1
Reciprocal expression of Diva/BclB and NDPKB/Nm23H2 proteins in differentiated PC-12 cells
Reciprocal expression of Diva/BclB and NDPKB/Nm23H2 proteins in differentiated PC-12 cells. (A) Real time-PCR showed decreased expression of Diva/BclB mRNA and upregulation of NDPKB/Nm23H2 and β-tubulin expressions. The data was normalized using GAPDH expression. (B) Western blotting show protein levels follow the same trend after differentiation. (C) Immunocytochemistry staining with Diva/BclB (green) and NDPKB/Nm23H2 (red) antibodies show colocalization of both proteins in the cytoplasm. (D) Nuclear staining of NDPK B was observed in differentiated cells, and the percentage of cells with NDPKB/Nm23H2 in the nucleus was quantified after counting an average of 300 cells. Bar=40μm; *p<0.05, ***p<0.001.
In the cells not treated with NGF, the distribution of Diva/BclB was detected to be in the cytoplasm, and NDPKB/Nm23H2 was mostly in the cytoplasm with some cells staining positive for the nucleus as well. By 3 days after NGF-induced differentiation, the cells expressed a lesser amount of Diva/BclB and this low expression was maintained at 5 days after differentiation. In contrast, differentiated cells stained more intensely for NDPKB/Nm23H2 after 3 days of differentiation and remain elevated even at 5 days, and a greater percentage of cells showed staining for the nucleus as compared to the control (Figure
NDPKB/Nm23H2 protein promotes outgrowth of neurites
In order to investigate if the increased expression NDPKB/Nm23H2 were directly involved in the process of differentiation, we overexpressed NDPKB/Nm23H2 in the PC-12 cell line. For this purpose, PC-12 cells were transfected with 2μg of control and NDPKB/Nm23H2 plasmids. Quantification of protein levels (Figure
Figure 2
Overexpression of NDPKB/Nm23H2 increases cell differentiation and neurite length
Overexpression of NDPKB/Nm23H2 increases cell differentiation and neurite length. The PC-12 cells were harvested or fixed 24hrs post-transfection. (A) Immunoblotting showed that overexpression of NDPKB/Nm23H2 suppresses expression of Diva/BclB. (B) Immunocytochemistry staining with Diva/BclB (green) and NDPKB/Nm23H2 (red) antibodies show colocalization of both proteins in the cytoplasm. (C) The percentages of cells with NDPKB/Nm23H2 in the nucleus were greatly elevated after the protein was overexpressed. Both the percentage of differentiated cells (D) and average neurite length (E) of the differentiated cells were increased in the NDPKB/Nm23H2 overexpressing cells. An average of 150 cells per group was counted for graphical analyses, and the experiments were performed in triplicate (Bar=40μm, *p<0.05, **p<0.01, ***p<0.001).
The ectopic NDPKB/Nm23H2 protein was expressed in both nucleus and cytoplasm of the transfected cells (Figure
Diva/BclB inhibits neurite outgrowth and prevents the translocation of NDPKB/Nm23H2 into the nucleus during differentiation
Since NGF-induced differentiation of PC-12 cells as well as NDPKB/Nm23H2 overexpression resulted in downregulation of Diva/BclB expression, we next investigated if Diva/BclB might be a negative regulator of neuronal differentiation. For this purpose, PC-12 cells were transfected with control or Diva/BclB plasmid. Overexpression of Diva did not affect the protein expression level of β-tubulin and NDPKB/Nm23H2 (Figure
Figure 3
Overexpression of Diva/BclB decreases cell differentiation and neurite length
Overexpression of Diva/BclB decreases cell differentiation and neurite length. After transfection with either the empty vector (control) or Diva/BclB overexpressing plasmid(Diva/BclB), immunoblotting (A) both showed an increase in Diva/BclB expression without any change to NDPKB/Nm23H2 expression. (B) Overexpression of Diva (green) resulted in stronger immunostaining in the cytoplasm. There was no change in NDPKB/Nm23H2 (red) expression. In the Diva/BclB cells, Diva/BclB expression remained strong even after 24 hrs of differentiation. There was strong colocalization of both proteins in the cytoplasm and at the neurite. (C) Overexpression of Diva/BclB prevents translocation of NDPKB/Nm23H2 into the nucleus after differentiation. It also decreases the percentage of differentiated cells (D) and their average neurite length (E). An average of 150 cells per group was counted for graphical analyses, and the experiments were performed in triplicate (Bar =40μm, *p<0.05, **p<0.01).
While the control cells that were differentiated had translocation of NDPKB/Nm23H2 to the nucleus, cells that were overexpressing Diva/BclB had most of their nucleus unstained with the NDPKB/Nm23H2 antibody despite differentiation (Figure
Diva/BclB inhibits neurite outgrowth by preventing the association of NDPKB/Nm23H2 and β-tubulin
As it is known that β-tubulin is a component of microtubules, and that neuritogenesis requires changes to the cytoskeletal network, we studied the interactions of Diva/BclB, NDPKB/Nm23H2 and β-tubulin to further understand the mechanism through which Diva/BclB inhibited neurite outgrowth. Using Duolink kit, which is a proximity ligation assay kit, we examined the subcellular locations and amounts of Diva/BclB and NDPKB/Nm23H2, NDPKB/Nm23H2 and β-tubulin as well as Diva/BclB and β-tubulin complexes.
Overexpression of Diva/BclB led to increased amounts of Diva/BclB and NDPKB/Nm23H2, as well as Diva/BclB and β-tubulin complexes in undifferentiated cells. This was maintained at a relatively high number even after differentiation (Figure
Figure 4
Diva/BclB prevents NDPKB/Nm23H2 and β-tubulin complex formation
Diva/BclB prevents NDPKB/Nm23H2 and β-tubulin complex formation. After transfection, the cells were grown for a day under normal growth medium or differentiating medium for 24 hrs. The interaction between the proteins were analysed using the Duolink assay. Regardless of the differentiation status, there were more Diva/BclB and β-tubulin (A) and Diva/BclB and NDPKB/Nm23H2 (B) complexes, and fewer NDPKB/Nm23H2 and β tubulin (C) complexes formed in Diva-overexpressing cells. (Bar= 20; *p<0.05, **p<0.01).
Diva/BclB increases and NDPKB/Nm23H2 decreases cell proliferation
Next, we examined the effects of Diva/BclB and NDPKB/Nm23H2 on cell proliferation as cell cycle exit is one of the hallmarks of neuronal differentiation. To this end, we compared the proliferation rates of the undifferentiated and differentiated transfected cells, by staining them with Brd-U antibodies 24 hours after differentiation (Figure
Figure 5
Effect of Diva/BclB and NDPKB/Nm23H2 on the cell cycle and cellular proliferation of cells
Effect of Diva/BclB and NDPKB/Nm23H2 on the cell cycle and cellular proliferation of cells. (A) The number of cells in S phase are visualised by BrdU staining (green) in the nucleus. Diva/BclB had the highest proportion of stained nuclei, followed by control and NDPKB/Nm23H2 had the lowest proportion. (B) An average of 500 cells per group was counted, and quantitative analysis of the data showed that overexpression of NDPKB/Nm23H2 decreased the percentage of S-phase cells, while Diva/BclB increased cellular proliferation. (C) Growth curves of the control and transfected cells were plotted using MTS assay. Diva/BclB cells had a less inhibited growth curve, while NDPKB/Nm23H2 cells had a slower growth curve, when compared to control cells. (D-G) Flow cytometry data was collected after serum starvation for 24 hours (0hr), and at indicated times after serum addition. Graphical analysis using Summit software showed that overexpression of Diva/BclB resulted in a decrease in apoptotic cells and faster cell cycle progression. In contrast, overexpression of NDPKB/Nm23H2 resulted in slower cell cycle progression. (Bar=20μm, *p<0.05, **p<0.01).
Using MTS assay, growth curves for control, Diva/BclB and NDPKB/Nm23H2 were plotted over a 4 day period (Figure
In order to study the effects of both proteins on serum-induced cell cycle re-entry, the transfected cells were serum starved for 24 hours to arrest and synchronize them, before serum was added again to allow cell cycle re-entry. At stipulated time points after re-addition of serum, the cells were stained with propidium iodide and subjected to flow cytometry analysis. Both Diva/BclB and NDPKB/Nm23H2 overexpressing cells had significantly lower percentages of apoptotic cells (Figure
Thus far, reports on Diva/BclB have only covered its role in apoptosis and oocyte maturation
In this study, we showed that with NGF-induced differentiation, Diva/BclB expression decreases and NDPKB expression increases. The regulation of these 2 proteins is likely through the sustained ERK signalling by NGF, which results in phosphorylation as well as binding of CREB and AP-1 family members so as to facilitate gene transcription
Overexpression of NDPKB/Nm23H2 encouraged PC12 neuronal differentiation by promoting neurite outgrowth and retarding cell proliferation. Interestingly, there was Diva/BclB downregulation in the NDPKB/Nm23H2 overexpressing cells, which is a salient point since decreased expression of Diva/BclB was also observed during NGF-induced differentiation. Thus, this supported that Diva/BclB was a negative modulator of neuronal differentiation that had to be downregulated and suggested that the induced downregulation of Diva/BclB by NDPKB/Nm23H2 could be a positive feedback mechanism for NGF-induced neuronal differentiation.
Overexpression of Diva suppressed NDPKB/Nm23H2 function by retaining NDPKB/Nm23H2 in the cytoplasm, and attenuated the number of NDPKB/Nm23H2 and β-tubulin complexes. In turn, there was an increase in S-phase cells and shorter neurites were observed after NGF-induced neuronal differentiation.
The regulatory molecules that control the rate of neurite growth and the signals that determine when and where the axons and dendrites have to grow are still largely unknown. Overexpression of negative regulators of neuronal differentiation have been shown to block neurite outgrowth and branching by inhibiting the Erk/MAPK and Rac1 but not Akt signaling pathway in response to NGF
In differentiated cells, the overexpression of Diva/BclB in PC-12 cells resulted in intense immunostaining in the cytoplasm, indicating the colocalisation of the 2 proteins, and significantly decreased the number of cells with NDPKB/Nm23H2 in their nucleus. This suggests that overexpression of Diva/BclB was able to sequester NDPKB/Nm23H2 in the cytoplasm and prevent its translocation into the cytoplasm after receiving the signal for differentiation. This is interesting because overexpression of NDPKB/Nm23H2 results in nuclear expression and decreased cellular proliferation. In contrast, the effect of overexpressing Diva/BclB is a higher proliferation rate and decreased nuclear expression of NDPKB/Nm23H2. Hence, this suggests that Diva/BclB is able to regulate proliferation by controlling the nuclear translocation of NDPKB/Nm23H2. NDPKs have been reported to translocate to the nucleus, although there is controversy whether NDPKA/Nm23H1 and NDPKB/Nm23H2 visit the nucleus together or separately
In conclusion, our results present a model in which downregulation of Diva/BclB during differentiation is necessary to allow increased formation of NDPKB/Nm23H2 and β-tubulin complexes for neurite outgrowth, and for NDPKB/Nm23H2 to enter the nucleus to inhibit proliferation. Hence this clarifies the developmental observation for the increased NDPKB/Nm23H2 expression seen during the onset of organogenesis in the brain
Cell culture
The PC12 cell line is established from rat adrenal medullary pheochromocytoma has been used as a model for neuronal differentiation. PC12 cells were routinely maintained at 37°C and 5% CO2 in DMEM (Sigma Aldrich, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Hyclone, Germany). To differentiate the cells, 50ng/ml of NGF (Invitrogen, USA) was added to DMEM with 1% FBS (differentiating medium). The differentiating medium was then either added to 1x106 cells in a flask (for real-time PCR and immunoprecipitation studies), or 5x105 cells in a well (for immunocytochemistry studies). The media was replaced with fresh differentiating medium every other day.
Real-time polymerase chain reaction (RT-PCR)
After differentiation, total RNA was isolated from the cells in each well of a 6-well plate using the RNeasy Mini kit according to the manufacturer's protocol (Qiagen). The qRT-PCR was performed using a LightCycler instrument (Roche, Germany) and QuantiTect SYBR Green (Qiagen) according to manufacturer's instructions. The sequences for the primers used are as follows: GAPDH-F (51-AGTCTACTGGCGTCTTCACCA-31), GAPDH-R (51-AGTTGTCATGGATGACCTTGG-31), Diva-F (51-ACGGCTGCTGACTGACTACAT-31), Diva-R (51-ACTCTTGGTCATTGGAGAGCA-31), NDPKB-F (51-GAGATCATCAAGCGATTCGAG-31), NDPKB-R (51-CCTGTTTTCACCACATTGAGC-31), β-tubulin-F (51-CAAGATGTCGTCCACCTTCAT-31) and β-tubulin-R (51-CTCAGACACCAGGTCGTTCAT-31).
The cells were seeded onto sterilized cover slips placed in a 6-well plate. After differentiation, the cells were fixed with 4% paraformaldehyde. Next, the cells were blocked with 5 % horse serum for 1 hour at room temperature and incubated with Diva (sc8740; Santa Cruz), Bcl-B (sc101875; Santa Cruz, USA) and NDPK B (L-14) (sc14789; Santa Cruz, USA), and β-tubulin (sc9104; Santa Cruz, USA) antibodies (All diluted at 1:200) overnight at 4°C. The cells were washed before incubation with secondary antibodies, Alexa Fluor® 488 or Alexa Fluor® 555 (1:200, Invitrogen, USA), for 1 hour at room temperature. The cells were then washed, before they were mounted onto glass slides with Vectashield Mounting Medium with DAPI (Vector Labs, USA). All images were captured using a confocal microscope (Fluoview1000, Olympus, Japan).
Transfection of PC12 cells
The plasmids from the company Blue Heron Biotechnology (USA) were used. The full length Diva and NDPK B were cloned into pcDNA6A (Invitrogen, USA) with a 6xHis tag. 1x106 cells were seeded into a six well plate and incubated at 37°C and 5% CO2 for 24 hours. 4μl of Lipofectamine 2000; 2μg of pcDNA6A, pcDNA6A-Diva and pcDNA6A-NDPK B were diluted in 250μl of OPTI-MEM for 5 minutes respectively. The diluted Lipofectamine 2000 reagent was then added to the diluted plasmids, and the mixture was incubated at room temperature for 25 minutes. The cells were then incubated with the plasmids for 5 hours at 37°C and 5% CO2. Thereafter, they were rinsed with 1xPBS once, before fresh media was added.
Western blot
Equal amounts of protein were separated electrophoretically using SDS-PAGE and then the gel was transferred to 0.45 μm polyvinylidene fluoride (PVDF: Millipore, USA). The membranes were soaked in blocking buffer (5% skimmed milk) for 1 hour at room temperature and then incubated overnight at 4°C with primary antibodies. This was followed by washing with TBS-Tween and incubation with secondary antibodies. For IP studies, Clean-blot reagent (Thermoscientific, USA) was used as it allows clear Western blot detection without IgG bands. This was followed by visualization by the enhanced chemiluminescence (ECL) detection system according to the recommended procedure (Pierce, USA).
The Duolink Kit I (Olink Bioscience, Sweden) was used to study the direct interaction of the proteins. In brief, the wild type, pcDNA6A, pcDNA6A-Diva and pcDNA6A-NDPK B cells were seeded onto 24 well cover slips at 1 x 104 cell density, and treated with NGF for a day. The cells were then fixed in 4% PF for 10 minutes, The cells were then incubated with 1X Duolink blocking stock for 1 hour, before overnight incubation at 4°C with the primary antibodies. Both the anti-goat-plus and anti-rabbit-minus PLA probes were added to the diluent in 1:10 ratio. Next, the cells were washed with 1X TBS-T and the diluted PLA probes were incubated with the samples in a humidity chamber for 2 hours at 37°C. The coverslips were washed and hybridisation solution was added and the samples were incubated for 15 minutes at 37°C. The coverslips were washed with 1X TBS-T and Duolink Ligase was added to the ligation solution at 1:40 dilution, and incubated for 15 minutes at 37°C. The coverslips were washed with 1X TBS-T and Duolink Polymerase was added to the diluted amplification solution at 1:80 dilution, before addition to samples and incubation for 90 minutes at 37°C. The slides were then washed with 1X TBS-T. The detection solution was added to the samples and incubated for 60 minutes at 37°C. The slides were then washed in 2xSSC for 2minutes, 1x SSC for 2 minutes, 0.2x SSC for 2 minutes, 0.02x SSC for 2 minutes and 70% ethanol for 1 minutes before the samples were left to dry. The coverslips were then mounted on slides using Vectashield Mounting Medium with DAPI (Vector Labs, USA) and examined using confocal microscopy (Fluoview1000, Olympus, Japan).
Brd-U(5-bromo-2'-deoxy-uridine) staining
The cells were transfected as mentioned above, before they were reseeded at 50% confluency. The number of proliferating cells was visualized and counted using the BrdU Labeling and Detection Kit I by Roche. In brief, the cell culture medium was aspirated and the cells were incubated with BrdU labeling medium for 30 minutes at 37°C and 5% CO2. The cells were washed before they were fixed in ethanol for half an hour at −20°C, washed and then incubated with the Anti-BrdU working solution for 30 minutes at 37°C. After washing, Anti-mouse-Ig-fluorescein working solution was added to the cells and incubated for 30 min at 37°C. After a final 3 washes, the coverslips were mounted onto glass slides with Vectashield Mounting Medium with DAPI (Vector Labs, USA) and photos were taken using the confocal microscope.
MTS assay
In order to study if the NGF treatment had decreased the proliferation rate of the cells, cell numbers were assayed using CellTiter 96® Aqueous Non-Radioactive Cell Proliferation Assay (Promega), a colorimetric based technique that exploits the ability of viable cells to convert tetrazolium salt into formazan. In brief, cells were plated at a fixed number in 100 μl of cell medium into a 96-well plate and allowed to equilibrate for 2 hours. Thereafter, 100 μl of PMS solution was added to 2 ml of MTS solution, and 20 μl of the PMS/MTS solution was added to each well. The plate was then incubated for 2.5 hours in an incubator at 37°C and 5% CO2. The absorbance for the plate was then read at 490nm in the GENios plate reader (Tecan, Switzerland).
Flow cytometry
The cells were serum starved so that they would be synchronised. At the designated time point, the cells were washed with 1xPBS, trypsinized and spun down at 1500 rpm, 4°C for 5 minutes. The cell pellet was then resuspended in 70% ethanol and left on ice for a minimum of 2 hours, or incubated at 4°C overnight so as to fix the cells. Before flow cytometry analysis using Beckman Coulter Epics Altra, USA), the ethanol was removed, and the cells washed with 1xPBS once. 1ml of the propidium iodide mixture (1xPBS, 0.1% triton X-100, 0.2mg/ml RNase A, 20ug/ml PI) was then added to the cells, and incubated at 37°C for 15 minutes.
Statistical analyses
All the experiments were repeated at least in triplicate for statistical purpose, and where appropriate, the data were presented as mean and standard deviation (SD). When comparing only two groups, the quantitative data was statistically evaluated using the Student's
Apaf-1: Apoptosis-activating factor-1; Boo: Bcl-2 homologue of ovary; Diva: Death Inducer binding to vBcl-2 and Apaf-1; H1P1R: Hungtington-interacting Protein 1-related; NDPK: Nucleoside Diphosphate Kinase; NHE: Nuclease-hypersensitive element; TCTP: Translationally Controlled Tumor-associated Protein.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
LQRJ was involved in performing and analysing all experiments. HB conceived of the study. LJ participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.
This work is supported by the Singapore National Research Foundation (NRF-CRP 002–082) under its Competitive Research Programme Funding Scheme.
Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor
Signaling Pathways for PC12 Cell Differentiation: Making the Right Connections
Sprouty4 Is an Endogenous Negative Modulator of TrkA Signaling and Neuronal Differentiation Induced by NGF
Up-regulation of Idh3 α causes reduction of neuronal differentiation in PC12 cells
Diva, a Bcl-2 homologue that binds directly to Apaf-1 and induces BH3-independent cell death
Boo a novel negative regulator of cell death, interacts with Apaf-1
Nm23–H2 involves in negative regulation of Diva and Bcl2L10 in apoptosis signalling
The mammalian Nm23/NDPK family: from metastasis control to cilia movement
Nm23: unraveling its biological function in cell differentiation
DR-nm23 expression affects neuroblastoma cell differentiation, integrin expression, and adhesion characteristics
Nm23/nucleoside diphosphate kinase and signal transduction
Nm23 influences proliferation and differentiation of PC12 cells in response to nerve growth factor
The zebrafish bcl-2 homologue Nrz controls development during somitogenesis and gastrulation via apoptosis-dependent and -independent mechanisms
The Apoptotic Regulator Nrz Controls Cytoskeletal Dynamics via the Regulation of Ca(2+) Trafficking in the Zebrafish Blastula
Oocytes and early embryos selectively express the survival factor BCL2L10
Role of Bcl2-like 10 (Bcl2l10) in Regulating Mouse Oocyte Maturation
Global Expression Analysis Identified a Preferentially Nerve Growth Factor-induced Transcriptional Program Regulated by Sustained Mitogen-activated Protein Kinase/Extracellular Signal-regulated Kinase (ERK) and AP-1 Protein Activation during PC12 Cell Differentiation
Nerve Growth Factor (NGF) Down-regulates the Bcl-2 Homology 3 (BH3) Domain-only Protein Bim and Suppresses Its Proapoptotic Activity by Phosphorylation
Sprouty2 inhibits BDNF-induced signaling and modulates neuronal differentiation and survival
The association of the nm23-Ml and oc-tubulin correlates with cell differentiation
Interplay between the antimetastatic nm23 and the retinoblastoma-related Rb2/p130 genes in promoting neuronal differentiation of PC12 cells
Cytoskeletal association of the A and B nucleoside diphosphate kinases of interphasic but not mitotic human carcinoma cell lines: specific nuclear localization of the B subunit
Receptor activation regulates cortical, but not vesicular localization of NDP kinase
Cytoskeletal alterations during neurogenesis in cultures of avian neural crest cells
HIP1R interacts with a member of Bcl-2 family, BCL2L10, and induces BAK-dependent cell death
Subcellular localization of A and B Nm23/NDPK subunits
Nuclear localization of nucleoside diphosphate kinase type B (nm23-H2) in cultured cells
Human c-myc transcription factor PuF identified as nm23-H2 nucleoside diphosphate kinase, a candidate suppressor of tumor metastasis
The transcription factor, Nm23-H2, binds to and activates the translocated c-myc allele in Burkitt's lymphoma
Multiple biochemical activities of Nm23/NDP kinase in gene regulation
Embryonic expression of nm23 during mouse organogenesis


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The dollar and Euro fell sharply in trading in Tokyo
In Tokyo is influenced by the outcome of the referendum on the UK's membership of the EU dramatically aplicarse the dollar and the Euro. Investors are shifting their assets in Japanese currency, considering it a more reliable asset after a referendum in the UK.
As of 11:00 local time (05:00 GMT) for one dollar gave 101.8 yen 1.2 yen less. Even more fell sharply, the Euro – he is listed at a level below 112,1 yen.Related posts:


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Target: San Mateo County Sheriff's Office
Goal: Thank the San Mateo County Sheriff's Office for setting up a regular patrol the Caltrain tracks and saving half a dozen lives
The Caltrain commuter train runs between San Francisco, San Mateo and Santa Clara counties. Running at nearly 80 miles per hour, the train is a fast and reliable source of transportation for commuters and tourists alike. Unfortunately, the train has gained a reputation for a much sadder reason as well. This year alone, 11 people have died on the train tracks; five of those deaths were conclusively ruled as suicides. This figure puts California in the top 10 states for suicide by train.
In order to combat the alarmingly high number of suicides, the San Mateo County Sheriff's Office has set up regular patrols of the Caltrain station and surrounding tracks. So far the office has prevented half a dozen suicides in the past couple of months, and hopes continued patrols will keep saving lives.
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Sign the petition below, and thank the San Mateo Sheriff's Office for keeping people safe on the train tracks. It is a difficult and often heartbreaking job, and the office should be commended for saving lives and getting people the help they need.
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I would like to thank you for your efforts to prevent suicide on the Caltrain tracks. The high suicide rate on these tracks has put California in the top 10 states where people are most likely to commit suicide by train. This is a statistic that is deeply troubling to me, and I'm glad your office is taking steps to prevent more deaths.
In the past couple of months, your officers have saved half a dozen lives. That is half a dozen people who now have a second chance at life. Together with Caltrain's efforts, you are lowering the suicide rate and giving California a happier reputation.
[Your Name Here]
Photo Credit: Flickr


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The Salado boys soccer team advanced into the second round of postseason play after a 2-0 blanking of Mexia on Tuesday at Waco ISD Stadium. Salado scored both its goals in the first half.
Fifteen minutes into the contest, center-midfielder Daniel Lemus assisted Brad Cosper who booted home an 18-yarder.
"It was a fantastic pass by Daniel," said Salado head coach Michael Goos. "It was a very clutch play all the way around."
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seminary — mid 15c., plot where plants are raised from seeds, from L. seminarium plant nursery, figuratively, breeding ground, from seminarius of seed, from semen (gen. seminis) seed (see SEMEN (Cf. semen)). Meaning school for training priests first… … Etymology dictionary
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seminary — [sem′ə ner΄ē] n. pl. seminaries [ME, seed plot < L seminarium, seed plot, nursery, neut. of seminarius, of seed < semen, a SEED] 1. Now Rare a place where something develops, grows, or is bred 2. Old fashioned a school, esp. a private… … English World dictionary
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How to Give other cyclists a safety cushion
Bicycle riders in groups often don't leave each other enough room. This how-to video demonstrates good safety cushion technique for bikes. Space is important to allow for maneuvering around pot holes or traffic. Watch this video cycling tutorial and learn how to maintain a safety cushion between bike riders.


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