Ensure your success with this 70-338 question bank


Use authentic 70-338 dumps. brain unload high-quality and popularity does remember.

70-338 free pdf | 70-338 boot camp | 70-338 real test | 70-338 test questions | 70-338 test questions - bigdiscountsales.com



70-338 - Lync 2013 Depth Support Engineer - Dump Information

Vendor : Microsoft
Exam Code : 70-338
Exam Name : Lync 2013 Depth Support Engineer
Questions and Answers : 114 Q & A
Updated On : December 6, 2017
PDF Download Mirror : 70-338 Brain Dump
Get Full Version : Pass4sure 70-338 Full Version

Once you memorize these 70-338 Q&A, you will get 100% marks.


It is essential to assemble to the guide material on the off chance that one needs toward spare time. As you require bunches of time to search for refreshed and true investigation material for taking the IT accreditation exam. In the event that you find that at one place, what could be superior to this? It's just killexams.com that has what you require. You can spare time and avoid bother on the off chance that you purchase Adobe IT accreditation from our site.

You ought to get the most refreshed Microsoft 70-338 Braindumps with the right answers, which are set up by killexams.com experts, enabling the possibility to get a handle on learning about their 70-338 confirmation course in the greatest, you won't discover 70-338 results of such quality anyplace in the market. Our Microsoft 70-338 Practice Dumps are given to applicants at performing 100% in their exam. Our Microsoft 70-338 test dumps are most recent in the market, allowing you to get ready for your 70-338 exam in the correct way.

In the event that you are occupied with effectively finishing the Microsoft 70-338 Certification to begin procuring? killexams.com has driving edge created Microsoft exam addresses that will guarantee you pass this 70-338 exam! killexams.com conveys you the most exact, present and most recent refreshed 70-338 Certification exam questions and accessible with a 100% unconditional promise guarantee. There are many organizations that give 70-338 mind dumps yet those are not precise and most recent ones. Arrangement with killexams.com 70-338 new inquiries is a most ideal approach to pass this affirmation exam in simple way.

We are for the most part very much aware that a noteworthy issue in the IT business is that there is an absence of value ponder materials. Our exam readiness material gives you all that you should take a confirmation examination. Our Microsoft 70-338 Exam will give you exam inquiries with confirmed answers that mirror the real exam. These inquiries and answers give you the experience of taking the genuine test. High caliber and incentive for the 70-338 Exam. 100% assurance to pass your Microsoft 70-338 exam and get your Microsoft affirmation. We at killexams.com are resolved to enable you to clear your 70-338 accreditation test with high scores. The odds of you neglecting to clear your 70-338 test, in the wake of experiencing our far reaching exam dumps are practically nothing.

killexams.com top rate 70-338 Exam Testing Tool is extremely encouraging for our clients for the exam readiness. Immensely vital highlights, points and definitions are featured in mind dumps pdf. Social occasion the information in one place is a genuine help and causes you get ready for the IT accreditation exam inside a brief timeframe traverse. The 70-338 confirmation offers key focuses. The killexams.com pass4sure dumps retains the essential highlights or ideas of the 70-338 affirmation

At killexams.com, we give completely surveyed Microsoft 70-338 preparing assets which are the best to clear 70-338 test, and to get affirmed by Microsoft. It is a best decision to quicken your vocation as an expert in the Information Technology industry. We are pleased with our notoriety of helping individuals clear the 70-338 test in their first endeavors. Our prosperity rates in the previous two years have been completely great, because of our upbeat clients who are currently ready to impel their vocations in the fast track. killexams.com is the main decision among IT experts, particularly the ones who are hoping to move up the progression levels quicker in their individual associations. Microsoft is the business pioneer in data innovation, and getting affirmed by them is an ensured approach to prevail with IT vocations. We enable you to do precisely that with our superb Microsoft 70-338 preparing materials.

Killexams.com Huge Discount Coupons and Promo Codes are as under;
WC2017 : 60% Discount Coupon for all exams on website
PROF17 : 10% Discount Coupon for Orders greater than $69
DEAL17 : 15% Discount Coupon for Orders greater than $99
DECSPECIAL : 10% Special Discount Coupon for All Orders


Microsoft 70-338 is ubiquitous all around the globe, and the business and programming arrangements gave by them are being grasped by every one of the organizations. They have helped in driving a large number of organizations on the beyond any doubt shot way of achievement. Far reaching learning of Microsoft items are viewed as a critical capability, and the experts confirmed by them are exceptionally esteemed in all associations.


70-338 Discount Coupon, 70-338 Promo Code, 70-338 vce, Free 70-338 vce, Download Free 70-338 dumps, Free 70-338 braindumps, pass4sure 70-338, 70-338 practice test, 70-338 practice exam, killexams.com 70-338, 70-338 real questions, 70-338 actual test, 70-338 PDF download, Pass4sure 70-338 Download, 70-338 help, 70-338 examcollection, Passleader 70-338, exam-labs 70-338, Justcertify 70-338, certqueen 70-338, 70-338 testking


found maximum 70-338 Questions in real exam that I prepared.

The solutions are explained in short in simple language and nonetheless make pretty an impact thats clean to understand and observe. I took the help of killexams.com Q&A and exceeded my 70-338 exam with a wholesome rating of sixty nine. way to killexams.com Q&A. I would really like to indicate in favor of killexams.com Q&A for the practise of 70-338 examination

Right place to find 70-338 real question paper.

it's far exceptional! I surpassed my 70-338 examination the day prior to this with a nearly best score of 98%. thank youKillexams! The substances in the package are genuine and legitimate - that is what I were given on my other examination. I knew answers to most of the questions, and some extra questions were very similar and on the topics absolutelycovered inside the take a look at guide, so i used to be capable of solution them on my own. no longer best did i am getting an top notch getting to know device which has helped me expand my professional knowledge, but I also receivedan smooth bypass to my 70-338 certification.

Try out these real 70-338 Actual Questions.

each subject matter and vicinity, each state of affairs, Killexams 70-338 materials have been exquisite help for me even asgetting ready for this examination and actually doing it! i used to be anxious, but going again to this 70-338 Q&A and questioning that I know the whole lot due to the fact the 70-338 exam was very clean after the Killexams stuff, I got an excellent end result. Now, doing the following degree of 70-338 certifications.

Very Tough 70-338 exam questions asked in the exam.

it's miles my satisfaction to thank you very a whole lot for being here for me. I handed my 70-338 certification with flying colors. Now i'm 70-338 certified.

I feel very confident by preparing 70-338 actual test questions.

passed 70-338 examination a few days in the past and got a really perfect score. but, I can't take complete credit score for this as I used killexams.com to prepare for the 70-338 examination. two weeks after kicking off my exercise with their trying out engine, I felt like I knew the solution to any query that could come my way. and that i certainly did. each query I examine at the 70-338 examination, I had already seen it while practicing. If no longer every, then giant majority of them. the whole thing that became inside the practise % became out to be very relevant and useful, so I cant thank enough to killexams.com for making it occur for me.

That changed into remarkable! I got actual questions of 70-338 examination.

It is excellent! I passed my 70-338 exam yesterday with a nearly perfect score of 98%. Thank you Killexams! The materials in the bundle are authentic and valid - this is what I got on my other exam. I knew answers to most of the questions, and some more questions were very similar and on the subjects fully covered in the study guide, so I was able to answer them by myself. Not only did I get an excellent learning tool which has helped me expand my professional knowledge, but I also received an easy pass to my 70-338 certification.

Get these Q&As and visit holidays to put together.

Im so glad i bought 70-338 exam prep. The 70-338 examination is hard because its very large, and the questions cover the whole lot you notice in the blueprint. Killexams turned into my main preparation source, and they cowl the entirety flawlessly, and there had been lots of associated questions about the examination.

right Place to discover 70-338 brand new Braindumps paper.

I skip in my 70-338 exam and that was now not a easy bypass however a terrific one which I should inform all of us with proud steam filled in my lungs as I had got 89% marks in my 70-338 exam from studying from killexams.com.

Shortest question are protected in 70-338 question bank.

killexams.com become a blessing for 70-338 examination, for the reason that machine has masses of tiny info and configuration tricks, which may be tough in case you dont have a good deal of 70-338 experience. Killexams 70-338 questions and answers are sufficient to sit and pass the 70-338 take a look at.

70-338 questions and answers that works in the real test.

typical affect changed into superb however i failed in a single assignment but succeeded in 70-338 2nd undertaking with killexams.com group very fast. examination simulator is good.

See more Microsoft dumps

MB2-708 | 70-740 | MB4-219 | 70-511-VB | 70-552-VB | 70-475 | MB3-214 | MB7-639 | MB3-207 | 70-630 | MB2-712 | 72-640 | 77-884 | 70-487 | 70-534 | 70-765 | 70-566-CSharp | 70-357 | MB6-705 | 70-569-VB | 70-686 | 70-552-CSharp | 70-411 | 70-567-CSharp | MB6-527 | MB2-228 | MB7-255 | 70-343 | MOS-AXP | 77-601 | 70-473 | MB3-208 | 70-333 | 70-640 | 70-413 | 72-642 | MB6-890 | MOS-A2K | 70-417 | MB5-626 | 70-480 | MOS-E2E | 98-366 | MB6-892 | 70-247 | 70-342 | 70-543-CSharp | 77-887 | 70-486 | 70-528-CSharp |

Latest Exams added on bigdiscountsales

1Z0-453 | 210-250 | 300-210 | 500-205 | 500-210 | 70-765 | 9A0-409 | C2010-555 | C2090-136 | C9010-260 | C9010-262 | C9020-560 | C9020-568 | C9050-042 | C9050-548 | C9050-549 | C9510-819 | C9520-911 | C9520-923 | C9520-928 | C9520-929 | C9550-512 | CPIM-BSP | C_TADM70_73 | C_TB1200_92 | C_TBW60_74 | C_TPLM22_64 | C_TPLM50_95 | DNDNS-200 | DSDPS-200 | E20-562 | E20-624 | E_HANABW151 | E_HANAINS151 | JN0-1330 | JN0-346 | JN0-661 | MA0-104 | MB2-711 | NSE6 | OMG-OCRES-A300 | P5050-031 |

See more dumps on bigdiscountsales

VCP5-DCV | 9L0-353 | 1Z0-035 | C7010-010 | 000-M10 | M5050-716 | E20-580 | 3204 | HP2-H09 | 050-890 | 510-309 | JN0-532 | ST0-202 | 000-677 | C2090-305 | C_TPLM30_66 | E20-580 | C2040-421 | M2080-241 | 000-M246 | HP0-090 | 1Z0-531 | HP0-J19 | 000-379 | 510-405 | HP2-K21 | C9010-250 | 000-S02 | P2070-055 | HP2-E28 | LOT-986 | 9L0-509 | E20-385 | HP3-X12 | A2010-503 | A2010-023 | 132-S-900 | C2090-136 | CSSGB | 70-341 | 2V0-621 | DP-023X | 190-711 | A2010-005 | 00M-653 | FN0-202 | 250-272 | HP0-P21 | C7010-010 | A2010-652 |

70-338 Questions and Answers


Based on the network trace, you need to identify the most likely cause of the call quality issues. Which setting should you modify?

  1. Geo IP
  2. MTU
  3. DSCP value
  4. STUN port

Answer: C


QUESTION: 103

You support a customer whose Microsoft Lync Server 2013 Enterprise Pool is configured with Enterprise Voice and Dial-in Conferencing. You specify a Session Initiation Protocol (SIP) Uniform Resource Identifier (URI) for the dial-in conferencing access number. You discover that the SIP URI was configured incorrectly. You need to change the SIP URI. What should you do?

  1. Run the Set-CsDialInConferencingAccessNumber cmdlet.
  2. Delete the object and recreate the access number.
  3. Run the Set-CsSipResponseCodeTranslationRule cmdlet.
  4. Run the New-CsDialInConferencingDtmfConfiguration cmdlet.

Answer: B


QUESTION: 104

You deploy a Microsoft Lync Server 2013 Enterprise Pool that is configured with Enterprise Voice and Dial-in Conferencing. All client computers run Windows 7 or Windows 8, and all use Lync 2013. Users report that they are unable to share Microsoft PowerPoint presentations with users who are using certain mobile devices. They also report that they are unable to scroll through a PowerPoint presentation independent of the presentation itself. You need to ensure that while they are in conferences, users are able to share PowerPoint presentations with users who are using mobile devices. You also need to ensure that users are able to scroll through PowerPoint presentations. What should you do?

  1. Install an Office Web Apps Server on a server and configure Lync Server 2013 to communicate with Office Web Apps.
  2. Install an Office Web Apps Server on a server that is running Office 2013 and configure Lync Server 2013 to communicate with Office Web Apps.
  3. Install Office 2013 for all users and instruct them to use PowerPoint 2013.
  4. Configure a new conferencing policy and enable the AllowMultiView.

Answer: A


QUESTION: 105

You support a customer whose network environment includes Microsoft Lync Server 2013 Standard Edition deployed with an Edge Server that is connected to the Internet. A user reports that when he tries to share a Microsoft PowerPoint presentation in a Lync conference, external users receive the following error message: "Some sharing features are unavailable due to server connectivity issues." You ask the customer to run the Get-OfficeWebAppsFarm cmdlet on the Office Web Application Companion (WAC) server. He provides the following result:

You need to ensure that external users are able to share and view PowerPoint presentations in Lync meetings. What should you do?

  1. Install WAC on the Edge Server.
  2. Set the ExternalUrl parameter on the WAC server and publish to the Internet by using a reverse proxy.
  3. Install WAC on the Front End pool.
  4. Set the InternalUrl parameter on the WAC server and publish to the Internet by using a reverse proxy.

Answer: B


QUESTION: 106

You support a customer whose company network includes Microsoft Lync Server 2013 with web conferencing deployed at a main site named Washington and a branch site named Redmond. All users have Lync 2013 and Microsoft Outlook 2013 installed. UserA and UserB are both Lync- enabled users at your customer's main site. UserA has the Global conferencing policy and the Washington meeting configuration assigned to his account. UserB has the Redmond conferencing policy

and the Global meeting configuration assigned to his account. In online meetings that UserA schedules, users can collaborate using videoconferencing. In online meetings scheduled by UserB, videoconferencing is not available. You need to determine why videoconferencing is unavailable in UserB's meetings. Which cmdlet should you run from the Lync Server Management Shell?

  1. Get-CSConferencingPolicy -Identity Redmond
  2. Get-CSMeetingConfiguration -Identity Redmond
  3. Get-CSMeetingConfiguration -Identity Global
  4. Get-CSConferencingPolicy -Identity Global

Answer: A


QUESTION: 107

You support a customer whose Microsoft Lync Server 2013 environment includes:
  • A single Standard Edition server,
  • A single consolidated Edge server, and
  • A single Forefront Threat Management Gateway 2010 server that is acting as an HTTP(S) reverse proxy.
A user reports that his attempts to join an online meeting by clicking the Join Online Meeting link are unsuccessful. Your need to troubleshoot the Join Online Meeting functionality from this workstation without using the installed Lync 2013 client. You need to achieve this goal by using the least amount of administrative effort. What should you do?

  1. Open Internet Explorer, type the URL of the online meeting into the Address bar, and then append ?sl=l to the URL.
  2. Open the Lync Options menu and select Join meeting audio from: Lync.
  3. Uninstall the Lync 2013 client, open Internet Explorer, and then enter the URL of the online meeting into the Address bar.
D.Open the Lync Options menu and set the Logging in Lync option to Full.

Answer: A


QUESTION: 108

You support a customer whose network environment includes Microsoft Lync Server 2013 Standard deployed with an Edge Server that is connected to the Internet. Users are able to share screens remotely during conference calls. However, when they attempt to upload a Microsoft PowerPoint presentation, the

sharing attempt fails with an error. You need to ensure that users are able to share PowerPoint presentations during Lync conference calls. What should you do?

  1. Install and configure the Office Web Apps Server Components on the Lync Pool Server.
  2. Install and configure Silverlight on the user workstations.
  3. Install and configure an Office Web Apps Server in Lync Topology.
  4. Install and configure the PowerPoint 2013 Viewer on the Lync presenter workstations.

Answer: C


QUESTION: 109

You support a Microsoft Lync Server 2013 Enterprise pool deployed in a high availability configuration for Back End Servers named Backend1 and Backend2. You execute the Get-CSDatabaseMirrorState cmdlet and discover the following errors. WARNING: Cannot connect to database server "BackEnd2.contoso.com".Message: A network- related or instance-specific error occurred whileestablishing a connection to SQL Server. The server was not found or was notaccessible. Verify that the instance name is correct and that SQL Server isconfigured to allow remote connections. (provider: Named Pipes Provider, error: 40 - Could not open a connection to SQL Server)DatabaseName
: rtcabStateOnPrimary
: PrincipalStateOnMirror
: StatusUnavailableMirroringStatusOnPrimary
:synchronizedMirroringStatusOnMirror
:DatabaseName
: rtcxdsStateOnPrimary
: PrincipalStateOnMirror
: StatusUnavailableMirroringStatusOnPrimary
:synchronizedMirroringStatusOnMirror
You need to resolve the connectivity issue and bring up the mirror databases to the synchronized state between the Back End Servers. What should you do?

  1. From the Front End Server, create the inbound rule on the firewall.
  2. Run the Test-CsDatabase cmdlet.
  3. Run the Invoke-CsPoolFailOver cmdlet.
  4. From the mirror Back End Server, create the inbound rule on the firewall.

Answer: D


QUESTION: 110

You support a customer whose network environment includes Microsoft Lync Server 2013 Enterprise, with two Front End Servers and two Back End Servers. The Back End Servers are deployed in a high availability configuration. You create a Front End pool named pool.contoso.com and implement DNS load balancing for client access to the Lync servers. Your internal DNS records have the following configuration: _gc._tcp.contoso.com SRV priority 0, weight 100, port 3268
dc1.contoso.com_ldap._tcp.contos.com SRV priority 0, weight 100, port 389
dc1.contoso.com_kerberos._tcp.contos.com SRV priority 0, weight 100, port 88
dc1.contoso.com_sip._tls.contoso.com SRV priority 0, weight 0, port 5061, pool.contoso.com FrontEndServer1 A 192.168.1.10FrontEndServer2 A 192.168.1.11BackEndServer1 A
192.168.1.20BackEndServer2 A 192.168.1.21Pool01 A
192.168.1.10Pool01 A
192.168.1.11 Your Lync 2013 clients fail to sign in automatically. The trace log of failed sign-in attempt is as follows: INFO
:: QueryDNSSrv - DNS Name[_sipinternaltls._tcp.contoso.com]ERROR :: HRESULT failed:
80072726 = HRESULT_FROM_WIN32(::ShimWSAGetLastError()) . Failed to convert string IP to SOCKADDRERROR :: ResolveHostNameUsingGetAddrInfo - getaddrinfo(pool.contoso.com) failed WARN
:: ResolveHostName - getaddrinfo failed for pool.contoso.com ERROR :: ResolveHostNameUsingDnsQuery - DnsQuery(pool.contoso.com) failed WARN
:: ResolveHostName - DNS lookup failed for pool.contoso.com ERROR :: ResolveHostName - Name resolution for pool.contoso.com failedERROR :: ResolveHostSyncResolveHostName failed ERROR :: GetDnsResults - did not get any resultERROR :: QueryDNSSrvGetDnsResults query:
_sipinternaltls._tcp.contoso.com failed ERROR :: DNS_RESOLUTION_WORKITEM::ProcessWorkItemResolveHostName failed You need to ensure that internal Lync 2013 clients can sign in automatically. You also need to ensure that internal traffic is load balanced across the two Front End Servers. What should you do?

  1. Create the following host records:PoolA192.168.1.20PoolA192.168.1.21
  2. Create the following host records: PoolA192.168.1.10PoolA192.168.1.11
  3. Create the following service record: _sip._tls.contoso.com SRVPriority 0, weight 0, port 5061, pool01.contoso.com
  4. Create the following host records: SipA 192.168.1.20SipA 192.168.1.21
  5. Create the following service record: _sipinternal._tls.contoso.com SRVPriority 0, weight 0, port 5061, FrontEndServer1.contoso.com

Answer: B


QUESTION: 111

You support Microsoft Lync Server 2013 in your company network. Your company has four buildings on a single site. A user reports that when she calls users in another building, the call quality is poor. You receive the Quality of Experience (QoE) report: Capture device: Headset MicrophoneRender device: Headset EarphoneMicrophone timestamp error: 0.02msEcho percent microphone in: 15.21%Codec: SIRENAudio FEC: FalsePacket utilization: 32701Avg. packet loss rate: 10.21%Avg. jitter: 23msAvg. round trip: 62msAvg. network MOS: 3.71Avg. network MOS degradation (jitter):0.00%Avg. sending MOS: 2.97Avg. listening MOS: 3.17Receive noise level: -56dBoV Which of the following values is outside acceptable limits?

  1. Receive noise level: -56dBoV
  2. Packet utilization: 32701
  3. Avg. round trip: 62ms
  4. Avg. packet loss rate: 10.21%
  5. Avg. jitter: 23ms

Answer: D


QUESTION: 112

You support a customer who administers Microsoft Lync Server 2013 Enterprise servers in his company. The pool named lync.contoso.com is configured with the session initiation protocol (SIP) domain contoso.com. The internal split-DNS domain contoso.com contains the following records: _gc._tcpSRV priority 0, weight 100, port 3268 dc1.contoso.com_ldap._tcpSRV priority 0, weight 100, port
389 dc1.contoso.com_kerberos._tcpSRV priority 0, weight 100, port 88
dc1.contoso.com_sipinternal._tcpSRV priority 0, weight 0, port 5061,
lync.contoso.com _sip._tlsSRV priority 0, weight 0, port 5061, lync.contoso.com EnterpriseCA
A 192.168.10.5Admin A
192.168.10.10Lync A
192.168.10.10Lyncdiscoverinternal A
192.168.10.10Exchange2010 A
192.168.10.4OWA A

192.168.10.4DC1 A 192.168.10.3Sip A
192.168.10.10
Users who are running Lync Mobile on their mobile devices report that when they attempt to retrieve calendar information, they receive an error that references a Microsoft Exchange Web Services connectivity issue. You need to ensure that users are able to receive calendar information from Lync Mobile devices. What should you do?

  1. Reconfigure Dynamic Host Configuration Protocol (DHCP) option 120 to point to DC1.contoso.local.
  2. Create the following service record in DNS: _lyncdiscover._tcp.contoso.com SRV Priority 0, weight 0, port 5061, Lync.contoso.com
  3. Reconfigure Dynamic Host Configuration Protocol (DHCP) option 43 to point to EnterpriseCA.contoso.local.
  4. Create the following host record in DNS: Autodiscover.contoso.com A 192.168.10.4

Answer: D


QUESTION: 113

You deploy Microsoft Lync Server 2013 Enterprise Edition and create a pool named Lync2013pool.Contoso.local. You configure the Lync admin URL to be admin.contoso.local. The Front End and Back End roles are installed on servers named FE2013 and BE2013. Your DNS server hosts the following records:
_Sipinternaltls._tcp.contoso.local SRV priority 0, weight 0, port 5061, lync2013pool.contoso.local_tcp._kerberos.contoso.local SRV proiority 0, weight 100, port 88, DC.contoso.localDC
A 192.168.10.20Dialin A 192.168.10.10Admin A 192.168.10.20FE2013 A 192.168.10.10BE2013 A
192.168.10.11
You attempt to open the Lync Server Control Panel and you receive an error message. You need to be able to open the Lync Server Control Panel. Which two actions should you perform? (Each correct answer presents part of the solution. Choose two.)

  1. Create the following DNS A record:Meet.contoso.local A 192.168.10.10
  2. Create the following DNS A record: lync2013pool.contoso.local A 192.168.10.10

  3. Create the following DNS SRV record: _sipinternal._tcp.contoso.local SRV priority 0, weight 0, port 5061 lync2013pool.contoso.local
  4. Create the following DNS A record: lyncdiscoverinternal.contoso.localA 192.168.10.10
  5. Modify the following DNS A record: admin.contoso.local to point 192.168.10.10

Answer: B, E


QUESTION: 114

You deploy Microsoft Lync Server 2013 Enterprise Edition and create a pool named Lync2013pool.Contoso.local. You configure the Lync admin URL to be admin.contoso.local. The Front End and Back End roles are installed on servers named FE2013 and BE2013. You create the following DNS records:
_Sipinternaltls._tcp.contoso.local SRV priority 0, weight 0, port 5061, lync2013pool.contoso.localDialin A 192.168.10.10FE2013 A 192.168.10.10BE2013 A 192.168.10.11 You attempt to open the Lync Server Control Panel and you receive an error message. You need to be able to open the Lync Server Control Panel. Which two actions should you perform? (Each correct answer presents part of the solution. Choose two.)

  1. Create the following DNS A record:Meet.contoso.local A 192.168.10.10
  2. Create the following DNS A record: lync2013pool.contoso.local A 192.168.10.10
  3. Create the following DNS SRV record: _sipinternal._tcp.contoso.local SRV priority 0, weight 0, port 5061 lync2013pool.contoso.local
  4. Create the following DNS A record: lyncdiscoverinternal.contoso.localA 192.168.10.10
  5. Create the following DNS A record: admin.contoso.localA 192.168.10.10

Answer: B, E


Microsoft 70-338 Exam (Lync 2013 Depth Support Engineer) Detailed Information

Microsoft Certification Program benefits
Learn about the benefits of the Microsoft Certification Program. Find answers to frequently asked questions regarding program benefits and Microsoft accounts.
Program benefits
Hide all
Q. What are the benefits of achieving a Microsoft Certification?
Microsoft Certification is an industry standard that is recognised worldwide. After you earn your Microsoft Certification, you have access to a number of benefits, which can be found on your benefits and exams dashboard.
Q. What will I find on the benefits and exams dashboard and who has access?
Individuals who have passed a Microsoft exam have access to the benefits and exams dashboard.
On the site, you will find:
A downloadable version of your certificate (you can order a printed copy); MOS members will need their Certiport login information
Your official Microsoft Certification downloadable transcript and access to the transcript-sharing tool
A tool to create and download certification logos
Your contact preferences and profile
Your Certification Planner
A sign up for the MCP Flash newsletters
Promotional offers, discounts and additional services
Q. What is a Charter certificate?
Charter Members are the pioneering group of individuals who achieve a certification within six months following the retail release date of the certification. (People who pass the beta exams will receive the Charter certificate after the certification is commercially released.) Charter Members are recognised by being given the Charter version of the certificate acknowledging their early adoption of the technology solution. The Charter version of the certificate includes the word "Charter".
Q. Which certifications are eligible for Charter status?
If you have any of the following certifications, you are eligible for the Charter certificate: Microsoft Certified Solutions Associate (MCSA), Microsoft Certified Solutions Expert (MCSE), Microsoft Certified Solutions Developer (MCSD), Microsoft Certified Technology Specialist (MCTS), Microsoft Certified IT Professional (MCITP), Microsoft Certified Solutions Master (MCSM), or Microsoft Certified Professional Developer (MCPD). Exceptions include those certifications that are released when the technology covered is no longer the latest version of the technology. These include the MCSA: Windows 7, the MCSA: SQL Server 2008 and the MCSA: Windows Server 2008.
In addition, Microsoft Specialist (Specialist) certifications released in and after September 2015 are also eligible for Charter status, including all Windows 10, Big Data Analytics and Cloud Data Platform Specialist certifications.
Q. How much time do I have to order my Charter certificate?
You can digitally download or order your Charter certificate at any point after it is earned.
Q. When does the Charter certificate period start for localised exams?
The six-month "clock" start time is based on the earliest time the certification can be earned, which is typically based on English-language exam availability. For example, if an exam is available in German two months after the English version is available, the German candidate has only four more months to earn a Charter certificate.
Q. I have heard that Microsoft sponsors an Elevate America veterans initiative to help our country's veterans and their spouses acquire the skills and resources that they need to be successful in today's workplace. What is this initiative?
Through this initiative, Microsoft convenes a coalition of public, private and non-profit organisations that are interested in contributing expertise, cash and in-kind resources to help U.S. veterans and their spouses build the skills and access the resources that they need to be successful in today's workforce.
Accessing the benefits and exams dashboard
Hide all
Q. How do I qualify for access to the benefits and exams dashboard?
The benefits and exams dashboard is available if you have passed a Microsoft qualified exam. It provides customised resources specific to your achievements and certifications.
Sign in to the benefits and exams dashboard
Q. How do I access the Microsoft Certification benefits and exams dashboard for the first time?
After you pass your first Microsoft Certification exam, you will receive a welcome email message outlining the steps needed to gain access to the benefits and exams dashboard. Please check your junk folder to ensure that the automatic email message is not blocked by your spam filter. Here are the instructions you will receive in the email message.
Create a Microsoft account if you don't already have one.
On your first visit, click on the link provided in the email and use your Microsoft account to log in to the benefits and exams dashboard.
Note This must be done within 90 days of receiving the email to meet security requirements.
In some cases, you may be required to enter your Microsoft Certification ID (MC ID) and temporary access code, which will be supplied in the email message.
On future visits, log in to the benefits and exams dashboard using the same Microsoft account.
For assistance, go to Microsoft training and certification help.
Q. I do not know my Microsoft Certification ID (formerly MCP ID). What should I do?
Your MC ID is shown in your welcome email as well as in the profile information on the benefits and exams dashboard. If you are unable to access the benefits and exams dashboard, contact your Microsoft Regional Service Centre for assistance.
Q. What is the access code required for the benefits and exams dashboard?
An access code is a unique code that allows first-time access to the benefits and exams dashboard and, if required, is provided in your welcome email. You can only use the access code to sign in to the benefits and exams dashboard for the first time. If you misplace your code or it has retired, contact your Microsoft Regional Service Centre for assistance.
Q. How long does it take for my data to update after accessing the benefits and exams dashboard for the first time?
After you register, you must wait 24 hours before accessing your program benefits.
Q. I am a returning user and cannot access the benefits and exams dashboard. How do I gain access?
If you cannot remember the Microsoft account information associated with your account, or you have other difficulty accessing the benefits and exams dashboard, contact the Microsoft Regional Service Centre in your area.
Note If you have a Hotmail account, MSN email account or Microsoft Passport, it is your Microsoft account.
Q. I saved the URL to my Microsoft Certification Program benefits in my Favourites, but when I navigate there I am redirected to a sign-in page, not the page I am trying to access.
Because online benefits are available only to eligible Microsoft Certification Program members, you must enter your Microsoft account credentials to access your online benefits. After your account is authenticated, you will be redirected to the page you are trying to access.
Q. Why is my name on the benefits and exams dashboard different from what it is in my Microsoft Personal Profile?
You are required to use your legal name for the benefits and exams dashboard. The Microsoft Personal Profile is associated with your Microsoft account.
Q. How do I update my legal name?
To update your legal name, contact your Microsoft Regional Service Centre.
Q. How do I update the information in my Microsoft Personal Profile?
You can update your Microsoft Personal Profile in the Profile Centre.
To change the name that is associated with your Microsoft Certification ID (formerly, MCP ID), contact your Microsoft Regional Service Centre.
Q. I updated my email address at the Profile Centre, but it was not changed on my Microsoft Certification transcript. What is wrong?
It can take up to 48 hours for changes in your profile to appear on your Microsoft Certification transcript. Check your transcript later to verify that your email address has been updated.
Q. I entered an email address in my Microsoft Personal Profile, but the email field is still blank. What happened?
After you update your Personal Profile, a message is sent to the email address noted in your profile. For the changes to be saved, you must follow the instructions in the email message to confirm the update. Otherwise, the email address in your profile will be removed because it has not been confirmed.
Q. How do I use the Microsoft Certification logos, and what are my rights and responsibilities when I use them?
When you access the logo tool, you are required to read and accept the MCP logo guidelines to correctly promote your relationship with Microsoft and to protect the integrity of the logos. You must sign in to the benefits and exams dashboard in order to access these guidelines.
Q. How can I download electronic files of Microsoft Certification logos?
Logos are available for download from the benefits and exams dashboard.
Q. I passed a test a week ago and I want to print my logo, but I cannot select it for printing.
The test that you passed might not qualify you to print that specific logo. If you believe this is an error, contact the Microsoft Regional Service Centre.
Q. How do I report misuse of a Microsoft Certification logo?
Contact the Microsoft Regional Service Centre.
Microsoft account and Profile Centre
Hide all
Q. What is Microsoft account?
Microsoft account (formerly Windows Live ID) enables you to simply use your email address and password to sign in. After you create your Microsoft account, you can access all Microsoft account sites and services. If you already have an MSN Hotmail, MSN Messenger or Passport account, it is your Microsoft account.
Learn more about Microsoft account.
Q. If I do not have a Microsoft account, do I need to set one up to access the benefits and exams dashboard?
Yes. You can sign up for a Microsoft account now. You will not be able to access the benefits and exams dashboard without one.
Q. If I already have a Microsoft account, can I use it to access the benefits and exams dashboard?
Yes.
Q. I do not remember the Microsoft account I used to access the benefits and exams dashboard. What should I do?
If you cannot remember the Microsoft account that you used to access the benefits and exams dashboard, contact your Microsoft Regional Service Centre for assistance.
Q. Will my Microsoft account expire?
If your Microsoft account is unused for one year, your Microsoft account may expire, and you may need to re-establish a Microsoft account.
Q. How do I update my profile?
Log in to the benefits and exams dashboard and, under the Account menu in the top right corner of the page, click Profile settings.
Q. What should I regularly update in my profile?
Update the following information to ensure that you receive communications from Microsoft:
Your contact preferences: Specify whether you want to receive promotional information about Microsoft products, services and events.
Your personal information: View, add or edit your personal contact information, such as your email address, business or personal address and phone numbers.
Your business information: View, add or edit information about your job and your organisation so that we can personalise content and recommend materials that match your job, organisation or industry.
Your technology preferences: View, add or edit information about your technology interests so that we can personalise content and recommend materials that match your interests.
Q. What if I do not want to receive email messages from Microsoft?
You are under no obligation to receive email messages from Microsoft. However, if you indicate that you do not want to receive communications from Microsoft through email, you cannot receive e-newsletters, such as the MCP Flash and MCT Flash. These communications provide up-to-date information on changes in training and certification resources, such as discontinued exams, Microsoft Certified Professional (MCP) benefits and special offers.
Q. How do I ensure that I receive communications by email?
Visit the Microsoft Profile Centre to indicate that you want to receive email messages from Microsoft.
Sign in to the Microsoft Profile Center to view and update your profile
Profile Center screenshot
Click My Contact Preferences, located on either the Profile Centre home page (shown above) or on the left-hand side of the page (shown below). Select E-Mail Address, and then click Save.
Note You can always unsubscribe from selected e-newsletters. However, if you do not select the E-Mail Address option, Microsoft cannot send you any information by email, including product notifications and news.
My Contact Preferences screenshot
Sign-up for e-newsletters: Select Manage Subscriptions on the left-hand side of the page. Tick all of the subscriptions that you want to receive, and then click Subscribe. You can also unsubscribe from publications.
Accessing your transcript
Hide all
Q. Where can I find my transcript or see which exams I have passed?
Access your transcript from the benefits and exams dashboard.
Under the section called Transcript, click on the View link.
Q. How do I give my employer or others access to my transcript via a secure connection that they can trust?
Microsoft offers a tool called Transcript Sharing, which can be accessed from the benefits and exams dashboard. Under the section called Transcript, click the Share link.
You will be asked to create an Access Code (which can be changed at any time) and given a Transcript ID. You will then provide these two codes to your selected audience, along with a URL to view your transcript.
Q. I have certifications from multiple providers; can I combine all my credentials into one transcript?
Microsoft has worked with the IT Certification Council (ITCC), along with other industry certification providers, to provide MCPs with the opportunity to create one transcript across providers. Under the Transcript section of the benefits and exams dashboard, click Create a multi-vendor transcript. You will then be asked to provide your transcript sharing codes. The transcript sharing code can be found by clicking Share your transcript.
Q. What if I cannot print or access my transcript?
If you cannot print or access your transcript, contact your Microsoft Regional Service Centre.
To ensure a prompt response:
Use the email address associated with your Microsoft Certification ID (MC ID), if possible.
Have your Microsoft Certification ID number available.
If you do not know your Microsoft Certification ID, have other information available, such as your address, telephone number, exam numbers and completion dates.
Q. I viewed my transcript but it is missing data. Why can't I see all my data?
It can take up to two weeks for Microsoft to receive and process your exam records. If it has been more than two weeks since you took your exam, and the results still do not appear on your transcript, or if you notice any other problems, contact your Microsoft Regional Service Centre.
Training and certification from Microsoft
Fulfill your potential and make the most of your investment in Microsoft technology with certification, classroom training, e-learning, and books from Microsoft and Microsoft Learning Partners.
Certifications
A Microsoft Certification validates your expertise in a Microsoft technology. As a Microsoft Certified Professional, you’ll have access to community resources and tools that allow you to exchange ideas with peers, increase your knowledge and skills, and broaden your career opportunities.
Microsoft Certified Trainers
Microsoft Certified Trainers (MCTs) are classroom and e-learning instructors, training and certification consultants, authors, conference presenters, and user group leaders who combine their expertise, experience, and passion for training and leadership to help Microsoft customers and partners realize their full learning potential.
Microsoft Learning Partners
Microsoft Partners with the Learning competency (Learning Partners) are training companies that meet stringent Microsoft qualifications to train IT professionals and developers on Microsoft technologies.
Microsoft Imagine Academy
The Microsoft Imagine Academy program connects the world of education to the world of work by enabling faculty and students to acquire new technology skills in an academic setting.
Share this page
Facebook
Article by ArticleForge

www.123inkjets.com.au

123inkjetsm.au

123inkjets is ranked 2,780,946 in the United States. 'Printer Ink Cartridges | Ink Toner Cartridges.'

2,780,946Rank in United States
907,216Worldwide Rank
Monthly pages viewed 52,391 Monthly visits 4,200 Value per visitor $11.95 Estimated worth $3,649.96 * External links 914 Number of pages --
*Estimated data, read disclaimer.Last Updated: 08312016
ors Country Country Rank Users % Pageviews % Other countries 24.60% 5.90% Australia 23,644 75.40% 94.10%
Most visitors are from Australia.
Content
Topics: Check Out, Hot Deals, Earn Inkjet Points, and Hot Quiz.
Category: 'PrintersSupplies'
The site has about 468 users daily, viewing on average 2.70 pages each.
Server Server Location Crystaltech Web Hosting .ArizonaPhoenixUnited States33.4481, -112.0732
The programming language environment is ASP.NET. It has 2 DNS records, ns2.webcontrolcenterm, and ns1.webcontrolcenterm. It is hosted by Crystaltech Web Hosting (Arizona, Phoenix,) using Microsoft-IIS6 web server.
IP: 63.135.116.92
Powered ASP.NET
Web Server: Microsoft-IIS6
Encoding: iso-8859-1
PING . (63.135.116.92) 56(84) bytes of data. 64 bytes from 123inkm.au (63.135.116.92): icmp_req=1 ttl=116 66.2 ms 64 bytes from .123inkm.au (63.135.116.92): icmp_req=2 ttl=116 70.4 ms 64 bytes from 123inkm.au (63.135.116.92): icmp_req=3 ttl=116 74.3 ms --- . ping statistics --- 3 packets transmitted, 3 received, 0% packet loss, time 2003ms rtt minavgmaxmdev = 66.27570.33874.3313.303 ms rtt minavgmaxmdev = 66.27570.33874.3313.303 ms
A ping to the server is timed at 66.2 ms, and the average page load time is 2490 milliseconds.
Server Setup Date: -- Server: Microsoft-IIS6.0 X-Powered-By: ASP.NET Content-Length: 86179 Content-Type: texthtml Set-Cookie: -- Set-Cookie: -- Cache-control: private
Article by ArticleForge

Quick News

NCPL offers computer classes
Natrona County Public Library offers basic computer classes to help participants navigate the computer, the Internet and basic programs like Microsoft Word.
Upcoming classes are: “Intro to Computers” at 5 p.m. Wednesday, Sept. 3; “Intro to the Internet” at 10 a.m. Thursday, Sept. 11; “Intro to PowerPoint” at 1 p.m. Monday, Sept. 15; “Intro to Genealogy” at 10 a.m. Tuesday, Sept. 16; and “Intro to Microsoft Word” at 1 p.m. Monday, Sept. 17.

Stop by the library’s Reference Desk to sign up for classes, or call 577-READ, ext. 2.
Citizenship class in need of immigrants
A free class for individuals who are within one year of being eligible to apply for citizenship will begin on Monday, Sept. 8, at Casper College.
“Anyone who knows how to speak, read, and write some English and is interested in becoming a United States citizen is urged to sign up for this free citizenship class,” said Lisa Mixer, Casper College tutor coordinator and ABEGED co-director.
The class is scheduled to run for 14 weeks, each Monday from 6:30-7:30 p.m. It is sponsored by the Casper College Adult Basic EducationGED Center.
Pre-registration is required and can be done through Sept. 12 by calling Mixer at Casper College at 268-2453.
Defensive driving courses offered
Defensive driving courses for people 55 years and older are conducted on the second Tuesday and Wednesday of each month, sponsored by the Central Wyoming Senior Citizens Center.
A course is scheduled from 2-6 p.m. on Sept. 9 and 10 at the Natrona County Senior Center, 1831 E. Fourth St. Each session is taught by AARP-trained instructors.
After successful completion, participants will receive a certificate that can qualify them for a 10 percent reduction in car insurance (liability andor collision) over two years.
The cost is $10. For reservations or more information, call Georgia at 265-4678, ext. 12.
CC Greenhouse hours announced
New fall hours for the Casper College Greenhouse, featuring a variety of plants, birds and reptiles, have been announced.
According to Evert Brown, greenhouse director and Casper College biology instructor, the facility will be open to the public from noon to 3 p.m. Monday through Friday.
Many types of plants are grown in the greenhouse, representing several different climates from desert to tropical.
ors also are likely to see several varieties of birds that live and fly within the confines of the greenhouse, and also some turtles.
The greenhouse, which is located on the west side of the Loftin Life Science Center, is free and open to the public.
Bookmobile schedule
For more information, call 577-READ, or log on to
Thursday, Sept. 4: 9:30-10 a.m., Garden Square (1950 S. Beverly); 10:15-10:45 a.m., Office Max . (441 Landmark Drive); 11-11:30 a.m., Pineview School Area II (1190 S. Forest Drive); 11:45 a.m. to 12:45 p.m., Cottonwood School (2300 Bellaire Drive); 3-3:30 p.m., Park Place (1930 E. 12th); 3:40-4:10 p.m., New Directions (LifeSteps Campus); 4:40-5:15 p.m., Evansville School.
Monday, Sept. 8: 9:30-10:10 a.m., Giggles & Wiggles (1720 S. Poplar); 10:25-10:55 a.m., Sunshine Corner (2303 E. 15th); 11:05-11:45 a.m., Sagewood School Area (1230 E. 22nd); 3:20-4 p.m., Apple Tree Learning Center (60 Magnolia); 4:15-5 p.m., Robertson Road Area (Whispering Springs & King Salmon).
Tuesday, Sept. 9: 11-11:30 a.m., Powder River School; 11:35 a.m. to noon, Powder River Post Office; 3:45-4:20 p.m., Paradise Valley I (Daffodil Street); 4:30-5 p.m., Paradise Valley II (Glendo Street).
Wednesday, Sept. 10: 10-10:30 a.m., Big Tree Area (1712 S. Oak); 10:40-11:20 a.m., Mtn. Road Christian Academy (2657 Casper Mtn. Road); 11:30 a.m. to 12:30 p.m., Willard School (129 N. Elk); 2-2:30 p.m., Prince of Peace School area (South Beverly & Eighth streets); 2:45-3:20 p.m., Sagewood School Area II (1901 E. 24th); 3:30-4:10 p.m., Leaps and Bounds Preschool (615 S. David); 4:25-5:15 p.m., Boys & Girls Club (1701 E. “K” St.).
Buffalo hunt raffle benefits CWRM
An all-inclusive guided buffalo hunt to benefit the Central Wyoming Rescue Mission has been donated by various sponsors.
The winner also will receive a Pre-64 Winchester Model 70 338 Win. Mag. rifle, 3-9 Zeiss scope, gun case, sling and ammo, as well as meat processing.
Only 250 tickets will be sold at $100 each. Tickets are available at the rescue mission at 230 N. Park St., City Service Electric, Rescued Treasures and Bar-D Signs.
Checks, written to CWRM, can be sent to PO Box 2030, Casper 82602. Be sure to write “buffalo hunt” in the memo. All proceeds will go to the mission.
For more information, call Risa or Deb at 268-4474.
Counseling association sets fall conference
The Wyoming Counseling Association has scheduled its annual fall conference for Oct. 9-11 at the Parkway Plaza in Casper.
The theme is “Wyoming Winds of Change,” and Dr. Susan McCabe will be the distinguished keynote speaker.
McCabe, a nationally known expert in psychiatric mental health nursing, will be speaking at the pre-conference on “Mind and Brain: Understanding Mood States Across the Lifespan.”
She serves as an associate professor at the Fay W. Whitney School of Nursing at the University of Wyoming and is the recipient of numerous awards, including the 2007 University of Wyoming Faculty Senate Lectureship Series Award, the 2004 Basham Faculty Fellowship Award and many more.

Mental health professionals may earn up to 15 hours of continuing education at the conference. The early-bird registration deadline is Sept. 14, and payment is by check or agency voucher.
For more information about the conference, call Becky Gurtler at 265-7545, or go to
Kids’ Grief Support Group returns
After a break for the summer, the monthly meetings of the Kids’ Grief Support Group will begin again Saturday, Sept. 13.
The group will meet from 11 a.m. to 1 p.m. at Central Wyoming Hospice, 319 S. Wilson, for water balloon volleyball and pizza. They also will make plans for upcoming meetings.
The Kids’ Grief Support Group meets the second Saturday of the month throughout the school year. Kids age 6-16 who have experienced the death of someone important in their lives are encouraged to attend. Parents also are welcome.
For more information or to RSVP, call Dama at 577-4832.
Family Fun Fest raffle tickets available
Central Wyoming Hospice & Transitions will raffle numerous quilts and gift baskets at its Fourth Annual Family Fun Fest, Saturday, Sept. 20, from 11 a.m. to 3 p.m. at the Central Wyoming Fairgrounds Industrial Building.
Each year, area quilters donate their handiwork for this event. Also, area businesses pitch in by filling gift baskets with family-friendly gifts.
Tickets for quilts and for baskets are sold separately. The cost is $3 each or two for $5. The tickets are available at CWHTP, 319 S. Wilson, and at Kalico Kat Quilt Shop, 350 W. Collins, also will be available at the event.

The Family Fun Fest is Casper’s largest indoor picnic and will feature games for all ages and prizes.
For more information, call Denise at 577-4832.
Book drive under way
Joan Anderson of Casper is conducting a book drive for the Natrona County Detention Center.
Anderson is asking for donations of paperbacks, in decent condition, of all genres: classics, action, Bibles, mysteries, romance, everything.
She noted that books in Spanish especially are needed.
Anderson plans to donate books to the detention center in September. Anyone with donations is asked to call Anderson at 472-3720.
Platte River Revival announced
The Second Annual Platte River Revival will be held on Saturday, Sept. 20, from 9 a.m. to noon, with check-in at Mike Lansing Field.
Residents are encouraged to start forming a team, perhaps with colleagues or neighbors.
“Volunteers will help remove debris and Russian olive branches, and plant trees in designated locations along the North Platte River,” said Jolene Martinez, Keep Casper Beautiful director.
In preparation for volunteers, Bureau of Land Management fire crews began cutting Russian olives, a non-native, invasive species, recently in what is expected to take a few weeks to complete.
Last year, volunteers removed 381,380 pounds of debris from the river and its banks, including appliances, tires and vehicles in the area just east of Bryan Stock Trail to the White Water Park.
Eleven native species trees were planted in Riverview Park.
“This year, the Revival’s operations committee is looking at areas along the river from Casper’s east city limit to just past Morad Park,” Martinez said.
A barbecue will follow, and there will be a group picture of all volunteers.
For more information or to sign up to volunteer, visit the Web site link at
The Platte River Revival is a Keep Casper Beautiful event in conjunction with National Public Lands Day and is organized by a partnership of public and private organizations.
Bereavement support groups in Glenrock
North Platte Home Health and Hospice are conducting Bereavement Support Group meetings in Glenrock to provide support, discussion and conversation for those who have lost a loved one.
Meetings are held the third Friday of the month at the Glenrock Senior Center, 615 W. Deer. Led by Chaplain Gayle Unruh, the meetings are held from noon to 1:30 p.m.
Lunch is provided, and the public is invited to attend these free meetings.
Hospice is a health care program that provides physical, emotional and spiritual support with life-limiting illnesses to enable the best quality of life for the patient.
It provides educational and emotional support to the patient's family to make their remaining time with the patient as meaningful as possible. Additionally, hospice provides grief support to family members following the death of the patient.
North Platte Home Health and Hospice is owned by Amedisys ., a provider of home health and hospice services at more than 480 sites across the United States and Puerto Rico.

Article by ArticleForge

Discovery of immunodominant T-cell epitopes reveals penton protein as a second immunodominant target in human adenovirus infection

Human adenovirus (HAdV) infection constitutes a major cause of morbidity and mortality in patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). The incidence of HAdV infection ranges from 5 to 30 %, with pediatric recipients showing the highest rates of infection with up to 83 % lethality [1–6]. Monitoring for HAdV infection and therapeutic intervention (reduction of immunosuppression, antiviral treatment) may reduce mortality due to HAdV in pediatric HSCT recipients [7]. However, antiviral treatments for HAdV infection with agents like cidofovir and ribavirin are associated with toxicity and may result in delayed immune reconstitution. Previous studies clearly indicate that T cells, the most potent effectors of the human immune system, are crucial for HAdV clearance [2]. It was demonstrated that children with HAdV-associated mortality had no HAdV-specific T cells, whereas patients who cleared HAdV infection showed HAdV-specific T-cell responses [2, 8]. Adoptive transfer of HAdV-specific T cells offers an effective and non-toxic immunotherapeutic strategy to reduce or prevent the clinical manifestation of HAdV in HSCT recipients with no or low numbers of HAdV-specific T cells [2, 8–12]. Monitoring HAdV-specific T-cell immunity may improve risk assessment in HSCT recipients and enhance treatment efficacy by determining the optimal time point for adoptive T-cell transfer. The median time between the first detection of HAdV DNA in the blood and the onset of symptoms is 3 weeks, which therefore seems to be the optimal time point for adoptive T-cell transfer [2, 13, 14]. Since the generation of short-term in vitro generated virus-specific T-cell lines takes about 3 weeks including quality controls, the production should start even earlier at the time of high viral load in stool (>106 copies) [12, 15].
The 70 different human HAdV types identified to date are divided into seven species (A to G) [16, 17]. Type 31 (of species HAdV-A) and HAdV 1, 2, and 5 (of species HAdV-C) are the most prevalent types in HSCT recipients [4–7]. Occasionally, types of species HAdV-B can be observed in adult HSCT recipients [18]. The major capsid protein hexon serves as an immunodominant target antigen across the different HAdV types, but few hexon-derived epitopes have been identified as immunodominant so far [13, 19–23]. Most of these epitopes are highly conserved, demonstrating that HAdV-specific T cells can cross-react across HAdV species and may therefore provide protection against a wide range of HAdV types [20]. HAdV-specific T-cell responses to the recombinant hexon protein, the overlapping peptide pool covering the complete hexon sequence, HLA-restricted peptides, and whole viral lysates have been investigated. A study by Feuchtinger et al. revealed that 10.5 % of donors had a specific T-cell response to the whole adenovirus but no response to the hexon protein, while 17 % of donors had no detectable T-cell response to HAdV [11]. Moreover, Zandvliet et al. detected specific CD8+ T cells in 616 healthy donors (37.5 %) after stimulation with the 15-mer hexon peptide pool, but only 316 donors (18.8 %) had specific T cells for known CD8+ hexon epitopes [24]. Sukdolak et al. observed a specific T-cell response to the 15-mer hexon peptide pool in 73 % of HAdV seropositive healthy donors, while 30 % were classified as high responders and 43 % as low responders [25]. Interestingly, 27 % of all HAdV seropositive healthy donors tested showed no response to the hexon peptide pool. These results underline the need to identify more immunogenic T-cell epitopes to improve the selection of HAdV-specific T cells for adoptive transfer and the immunomonitoring of high-risk patients.
T-cell epitopes can be identified by direct or reverse immunology. Various computer algorithms have been developed over the past years that allow for the prediction of peptide binding to MHC class I and II molecules, proteasome cleavage patterns and transporter associated with antigen processing translocation [26]. Naturally presented CD8+ T-cell epitopes are usually among the top-scoring 2 in 80 % of all predictions, whereas the reliability of CD4+ T-cell epitope prediction is much lower due to the more variable pocket binding behavior of MHC class II molecules [27]. SYFPEITHI [26, 28, 29], BIMAS [26, 30] and NetChop [31] are the most widely used algorithms to identify cytotoxic T lymphocyte (CTL) epitopes in viral, microbial, and tumor antigens. These well-established algorithms, which have been validated and compared [26], were employed in this study to predict new HAdV epitopes.
The major focus of this study was to identify and evaluate novel immunodominant HAdV-specific T-cell epitopes by analyzing the main structural proteins, hexon and penton. HLA-A*01-, A*02-, A*03- and B*08-restricted peptide epitopes within conserved protein regions (Table 1) were pre-selected based on the predictions of several established computer algorithms. Immunogenicity of the top-ranked epitopes was investigated by established methods: IFN-γ-based EliSpot, cytokine secretion assay (CSA), peptide MHC (pMHC) multimer staining and multicolor flow cytometry. Four of the selected peptide candidates were classified as low immunodominant and two as high immunodominant according to the number of responders in the healthy donors and HAdV-infected HSCT recipients. This paper describes for the first time the immunogenic potential of penton-derived epitopes and demonstrates that the penton, as an immunological target, it is not secondary to the hexon. Expanding the repertoire of immunodominant HAdV-specific T-cell epitopes will enable more precise immunomonitoring and more effective multi-epitope-based T-cell therapy by targeting epitopes presented in a broader array of HLA molecules. Table 1
Predicted peptide candidates used for HAdV-specific T-cell screening in healthy donors

Article by ArticleForge

Award-Winning Chef Elizabeth Falkner Reveals Her Struggle with Atopic Dermatitis to Highlight the Physical and Psychological Impact of the Disease

CAMBRIDGE, Mass. and TARRYTOWN, N.Y., July 12, 2016 PRNewswire -- Celebrity chef, restaurateur, and media personality Elizabeth Falkner has teamed up with Sanofi Genzyme, Regeneron Pharmaceuticals, ., the National Eczema Association, and the Dermatology Nurses Association to launch Understand AD, a national awareness campaign focused on educating people about moderate-to-severe atopic dermatitis (AD), a potentially serious, chronic inflammatory skin disease.1 Falkner is speaking out for the first time about her own struggle with the disease to drive awareness about the physical impact and effects on quality of life for people living with atopic dermatitis, and to encourage others to speak up about their experience.
"I have been living with the challenges of atopic dermatitis for more than 20 years. At its worst, my atopic dermatitis causes constant, unbearable itching, scabbing, visible rashes on my body and even bleeding, and that's only the physical part," says Elizabeth Falkner. "Having atopic dermatitis can affect many aspects of a person's life – physically and emotionally – and yet many people don't understand the severity and impact. I joined Understand AD to empower people to have more open conversations with their doctors and loved ones about the impact this disease has on their lives."
Atopic dermatitis is a chronic, systemic inflammatory disease characterized by rashes and can include intense itching, skin dryness, cracking, redness, crusting and oozing.1,2,3 Though symptoms can appear on the surface of the skin all over the body,4 advances in research have provided new insights on the cause of atopic dermatitis.5 Scientists now believe AD is caused in part by systemic allergic inflammation that results from a malfunctioning immune system.4,6 The physical symptoms are challenging and impact people's sleep and daily lives and the disease can also make people feel self-conscious and embarrassed about their appearance.7,8,9
"Understand AD aligns with our mission to educate the public and support patients impacted by atopic dermatitis," says Julie Block, President and CEO, National Eczema Association. "Unfortunately, there's a misperception that atopic dermatitis is just a 'skin condition' that people can deal with on their own, but in reality, it's an immunological disease that has a huge impact on patients' lives. We want people living with this disease to know that they're not alone and that we're committed to advocating for better care and treatments, providing support and raising the level of awareness about this serious, and often overlooked, disease."
An estimated 1.6 million adults in the United States live with uncontrolled moderate-to-severe atopic dermatitis.10 Researchers continue to discover more about atopic dermatitis and there is still a need for additional treatment options for atopic dermatitis.
"Our community of nurses on the front lines see people every day who are suffering with atopic dermatitis," says Donna Beyer, MSN, RN, DNC, President of the Dermatology Nurses Association. "But there is still a gap in public awareness about this disease and a clear need for continued education and supportive resources for patients. We're excited to join Understand AD to help educate about the disease and to drive the dialogue that atopic dermatitis is more than skin deep."
.UnderstandADm to learn more about moderate-to-severe atopic dermatitis, get connected with advocates such as the National Eczema Association and Dermatology Nurses Association, and hear from award-winning chef, media personality and restaurateur Elizabeth Falkner who has lived with atopic dermatitis for the past 20 years.
About SanofiSanofi, a global healthcare leader, discovers, develops and distributes therapeutic solutions focused on patients' needs. Sanofi is organized into five global business units: Diabetes and Cardiovascular, General Medicines and Emerging Markets, Sanofi Genzyme, Sanofi Pasteur and Merial.
Sanofi Genzyme focuses on developing specialty treatments for debilitating diseases that are often difficult to diagnose and treat, providing hope to patients and their families.
Genzyme® is a registered trademark of Genzyme Corporation. Sanofi® is a registered trademark of Sanofi. .


About Regeneron Pharmaceuticals, .Regeneron is a leading science-based biopharmaceutical company based in Tarrytown, New York that discovers, invents, develops, manufactures, and commercializes medicines for the treatment of serious medical conditions. Regeneron commercializes medicines for eye diseases, high LDL cholesterol and a rare inflammatory condition and has product candidates in development in other areas of high unmet medical need, including rheumatoid arthritis, asthma, atopic dermatitis, pain, cancer, and infectious diseases. For additional information about the company, please visit .regeneronm  or follow Regeneron on Twitter.
About the National Eczema AssociationThe National Eczema Association (NEA) is a non-profit 501 (3) patient advocacy organization whose mission is to improve the health and quality of life for individuals with eczema through research, support, and education. In the United States alone, over 10% of the population has some form of atopic dermatitiseczema. NEA was founded in 1988 by a group of patients, medical professionals, and parents to help individuals and families living with this skin disease live healthier lives. Through a variety of educational materials, including a quarterly patient-oriented magazine, a monthly electronic newsletter, and trustworthy website, the NEA reaches out to a diverse audience that includes eczema patients, caregivers, medical professionals, and other stakeholders. NEA also conducts patient conferences and participates in a wide-variety of medical symposiums. NEA is active year round to promote eczema awareness, break through stereotypes and address issues critical to patient care. Advocacy efforts include advancing increases in skin disease research funding through the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) of the National Institutes of Health, as well as increasing public understanding regarding the burden of eczema. NEA provides a network of support groups, an up-to-date website with the latest research and treatment information, a Seal of Acceptance program for over-the-counter products to help eczema patients navigate the myriad of products necessary for their daily skin care regimen, and a research program to advance scientific knowledge and care.  All NEA programs and services result in benefits for eczema patients and their families. NEA does not endorse specific products. For more information about the National Eczema Association, visit .nationaleczema, contact at infonationaleczema, or call 1-800-818-7546.
About the Dermatology Nurses AssociationThe Dermatology Nurses Association (DNA) is a professional nursing organization comprised of a diverse group of individuals committed to quality care through sharing knowledge and expertise. The core purpose of the DNA is to promote excellence in dermatologic care. Members include nurse practitioners, registered nurses, licensed practical and vocational nurses, medical assistants and others associated with dermatology nursing, who work in a variety of settings including clinics, academic institutions, private practice, public health centers, and government facilities. DNA offers education and training in fundamental and cutting-edge dermatology care and treatment through its annual convention, local chapter meetings, dermatology nurse and nurse practitioner certification review courses and expert workshops. Members of the DNA's Nurse Practitioner Society are afforded tools, resources and education focused on the needs of the advanced nurse practitioner. The DNA Focus Newsletter and official journal, the Journal of Dermatology Nurses Association, extend the DNA's informational and education presence with association and practice news, learner-paced continuing education and timely resources.
s Sanofi:
Media Relations                                                     Carrie Brown                                                           Tel: (908) 981-6486                                           carrie.brownsanofim                                           
s Regeneron:
Media Relations                                                     Ilana Tabak                                                              Tel: (914) 847-3836Mobile: (914) 450-6677                                                ilana.tabakregeneronm
1 World Allergy Association 2004:
 
To view the original version on ,

Article by ArticleForge

NF-κB Regulates Mesenchymal Transition for the Induction of Non-Small Cell Lung Cancer Initiating Cells

Abstract The epithelial-to-mesenchymal transition (EMT) is a de-differentiation process that has been implicated in metastasis and the generation of cancer initiating cells (CICs) in solid tumors. To examine EMT in non-small cell lung cancer (NSCLC), we utilized a three dimensional (3D) cell culture system in which cells were co-stimulated with tumor necrosis factor alpha (TNF) and transforming growth factor beta (TGFβ). NSCLC spheroid cultures display elevated expression of EMT master-switch transcription factors, TWIST1, SNAI1Snail1, SNAI2Slug and ZEB2Sip1, and are highly invasive. Mesenchymal NSCLC cultures show CIC characteristics, displaying elevated expression of transcription factors KLF4, SOX2, POU5F1Oct4, MYCN, and KIT. As a result, these putative CIC display a cancer “stem-like” phenotype by forming lung metastases under limiting cell dilution. The pleiotropic transcription factor, NF-κB, has been implicated in EMT and metastasis. Thus, we set out to develop a NSCLC model to further characterize the role of NF-κB activation in the development of CICs. Here, we demonstrate that induction of EMT in 3D cultures results in constitutive NF-κB activity. Furthermore, inhibition of NF-κB resulted in the loss of TWIST1, SNAI2, and ZEB2 induction, and a failure of cells to invade and metastasize. Our work indicates that NF-κB is required for NSCLC metastasis, in part, by transcriptionally upregulating master-switch transcription factors required for EMT.

Citation: Kumar M, Allison DF, Baranova NN, Wamsley JJ, Katz AJ, Bekiranov S, et al. (2013) NF-κB Regulates Mesenchymal Transition for the Induction of Non-Small Cell Lung Cancer Initiating Cells. PLoS ONE 8(7): e68597. doi:10.1371journal.pone.0068597
Editor: Srikumar P. Chellappan, H. Lee Moffitt Cancer Center & Research Institute, United States of America
Received: January 14, 2013; Accepted: May 30, 2013; July 30, 2013
: 2013 Kumar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: Work was supported by National Institutes of Health grants R01CA132580, R01CA104397 (to M.W.M.), and R01CA136705 (to D.R.J.). All funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Introduction
Cancer development from early pre-malignant neoplasm to full metastatic disease is a multistep process that involves tumor epithelial-stromal interactions, angiogenesis, and infiltration of tumor-associated pro-inflammatory cells [1], [2]. An emerging hypothesis proposes that this milieu of cell-cell interactions, growth factors, and cytokines known as the tumor microenvironment, stimulates morphogenesis within tumor cells referred to as the epithelial-to-mesenchymal transition (EMT) [3]–[5]. EMT induces a redistribution of intracellular architecture, decreased cell-cell adhesion, and loss of cellular polarization. Carcinoma cells that have undergone EMT are characteristically motile, invasive and highly metastatic. Over the past several years, EMT has also been recognized as a de-differentiation program attributed to generation of tumor-initiating or cancer-initiating cells (CICs) that are important in the maintenance of cancer “stemness” [6]–[9].

Although multiple cytokines and growth factors induce EMT, one of the best studied factors is transforming growth factor beta (TGFβ) [2], [3], [10]–[13]. Stimulation of cells with TGFβ results in expression of the EMT master-switch transcription factors, TWIST1, SNAI1Snail, SNAI2Slug, and ZEB2Sip1 that together differentially regulate genes to promote the mesenchymal phenotype [10], [12]. While extensive research details the ability for TGFβ to induce EMT, evidence indicates that tumor necrosis factor (TNF) further potentiates the transition [14], [15]. During cancer progression, secretion of TGFβ within the tumor microenvironment occurs through many different cell types, including tumor-associated fibroblasts, while secretion of TNF originates from tumor-associated M2 macrophages [3], [16], [17]. A prevailing hypothesis in the field is that exposure of cancer cells to these cytokines within the tumor microenvironment promotes EMT, de-differentiation, and the formation of CICs [2], [5], [17].

TNF is a powerful pro-inflammatory cytokine that stimulates signaling cascades to activate nuclear factor kappa B (NF-κB). As a transcription factor, NF-κB plays a key role in the expression of genes involved in cancer initiation and progression. Upregulation of NF-κB activity often occurs in primary solid and hematological tumors, directly correlating with de-differentiated morphology, advanced tumor stage, and poor clinical prognosis [18]. Importantly, NF-κB has been linked to mammary CICs [19], [20]. NF-κB induces and maintains EMT in model systems through two mechanisms, upregulation of EMT master-switch transcription factors [21]–[24] and stabilization of Snail [25]. NF-κB is composed of five Rel family members: RelAp65, RelB, cRel, p50 and p52. In unstimulated cells, inhibitory IκB subunits associate with NF-κB dimers and sequester them in the cytoplasm. Upon cellular stimulation by pro-inflammatory cytokines, IκBα is phosphorylated by the IκB kinase (IKK) complex, ubiquitinated by the SCF-type E3 ligase, E3RSIκBβ-TrCP and degraded by the 26S proteasome [26]. Liberated NF-κB then translocates to the nucleus to activate gene expression by recruiting transcriptional coactivators [27]. Our laboratory has shown that posttranslational modifications on RelA are required for full NF-κB transcriptional activity [27]–[30].

Although EMT in breast cancer models requires NF-κB activity [31], the role of this transcription factor in stimulating EMT and developing CICs in NCSLC has not been thoroughly examined. However, strong evidence exists for the presence of NSCLC stemprogenitor cells in primary adenocarcinomas and established cell lines [32]–[35]. Here, we demonstrate that coordinated activation of TNF and TGFβ signaling cascades effectively induces EMT and the expression of genes related to de-differentiation and stemness. Further, we show that mesenchymal NSCLC cells possess constitutively active NF-κB, and that inhibition of NF-κB decreases EMT, CIC formation, and metastatic potential.
Materials and Methods Cell culture and reagents
NSCLC lines A549, H359, H1299, and H157 were obtained from ATCC and maintained as 2D cultures in DMEM (CellGro), 10% FBS (Invitrogen) and penicillinstreptomycin (Invitrogen). The antibodies used include: E-cadherin (BD Pharmingen- 610404), N-cadherin (BD Pharmingen- 610920), Vimentin (V6630), Fibronectin (BD Pharmingen- 610078), α-Tubulin (Sigma T6793), HMGA2 (Biocheck 59170AP), Twist1 (Cell Signaling 4119), Snail1 (Cell Signaling 4719), Sip1 (SCBT sc-48789), Slug (Abcam ab27568), IκBα (pS32, Cell Signaling 2859), IκBα (SCBT sc-371), RelA (pS536, Cell Signaling 3031), RelA (SCBT sc-372), and M2-Flag (Sigma F1804). Baculogold protease inhibitors were obtained from BD Biosciences. TGFβ (PHG 9204) and TNF (PHG 3015) were purchased from InvitrogenLife technologies. All other chemicals were from Sigma.
Three-dimensional multicellular spheroid cultures
Three-dimensional multicellular spheroid cultures were created using a modified hanging droplet method [36]. Cells were grown to approximately 80% confluence on standard tissue-culture plates. The cells were subsequently trypsinized, resuspended in DMEM10% FBS, and counted. To create 25,000 cell spheroids, the cell suspension was diluted to a concentration of 1,000,000 cellsml, and 25 µl of the cell suspension were pipetted onto the underside of a sterile 10 cm tissue-culture plate lid. Each lid holds approximately fifty droplets. After loading the droplets, the lid was placed onto a tissue culture plate containing 6 mL of sterile PBS and incubated for 48 hours to facilitate cellular aggregation and spheroid formation. The freshly formed spheroids were then transferred into 10 cm suspension plates containing DMEM and 2% FBS to prevent cell attachment to the dish. Suspension plates were made by adding 8 ml of poly-HEMA solution (Sigma-Aldrich P3932, 10 mgml) in 95% ethanol to sterile polystyrene petri dish plates (Fisher Scientific). The plates were then incubated for 24 hours in a sterile environment to allow the ethanol to evaporate. Prior to use, plates were washed with sterile PBS to remove any residual ethanol or other contaminants. Each suspension plate holds up to 100 spheroids. After transfer, the spheroids were treated with vehicle or with 10 ngml TNF and 2 ngml TGFβ, and incubated for 48 hours. After incubation, cells were subjected to a second treatment of vehicle or TNF and TGFβ, and incubated an additional 48 hours. The spheroids were then collected and analyzed by various assays.
Immunofluorescence Microscopy
A549 cells were seeded on glass coverslips and subjected to EMT induction or left untreated. After induction, the cells were fixed in 100% methanol and subsequently incubated with primary antibodies to the extracellular domain of E-Cadherin (SCBT, sc-7870). An AlexaFlour-conjugated, goat anti-rabbit antibody (Invitrogen) was used as a secondary antibody, and indirect immunofluorescence of E-Cadherin was imaged using a Nikon E3800 fluorescence microscope.
Migration and Invasion
In vitro migration and invasion assays were carried out according to the manufacturer's protocol (BD Biosciences). 2D and 3D cultures were disaggregated by trypsin and subsequently counted. 1×105 cells (migration) or 1×104 cells (invasion) were seeded in plain DMEM in the top well of a transwell control plate (BD 354578) or Matrigel invasion plate (BD 354480). The bottom well was loaded with DMEM containing 10% FBS as a chemoattractant, and the plates were incubated for eight hours (migration) or twenty-four hours (invasion) at 37°C and 5% CO2. Afterwards, cells on the upper side of the membrane were removed, and the remaining cells were fixed in 100% methanol and stained with 0.1% crystal violet. The stained cells were imaged and quantified using Adobe® Photoshop.
Tumor model
Monolayer (2D) and 3D A549 cultures that had been left untreated or treated with TNF and TGFβ were trypsinized, resuspended in DMEM0.5% FBS, and carefully counted and diluted in the appropriate volume for injection. Cells were subcutaneously (SC) injected into female outbred Crl:NUNU nude mice (Charles River). Five mice were injected per experimental condition. All animal studies were performed as three independent experiments. Mice were sacrificed forty days post-injection. The primary SC tumors were removed and weighed. Additionally, the lungs were removed, fixed in formalin, and surface lung metastases were counted. To quantify the amount of total tumor burden in the formalin fixed lung tissue, genomic DNA was extracted [37] and assayed for the presence of human genomic material as described using quantitative real time-polymerase chain reaction (QRT-PCR) primers specific to human endogenous retrovirus-3 (ERV3, Table S1) [38], [39].

This study was carried out in strict accordance with recommendation from the Animal Care and Use Committee (ACUC) of the University of Virginia. The protocol was approved by ACUC Number 3914. All experiments were terminated after 40 days at which time SC tumors were less than 1.0 cm3 in size; thus, restricting tumor burden. All efforts were made to minimize pain and suffering.
QRT-PCR, Immunoblots, and Electrophoretic mobility shift assays (EMSAs)
QRT-PCR and immunoblot experiments were carried out as previously described [28]. PCR primers are shown in Table S1. Nuclear extracts were prepared using spheroids from A549.V and A549.I cell lines treated with or without TNF and TGFβ. EMSAs and supershift assays were performed as described previously [40].
Statistics
Where appropriate, comparisons between experimental groups were carried out by performing a one-tailed Student's t test in Microsoft excel. Data for all experiments was considered statistically significant when p<0.05.
Results A model to study EMT in NSCLC
TNF has been shown to potentiate TGFβ-mediated EMT through the activation of co-stimulatory pathways [15]. To confirm this observation in our three-dimensional (3D) model, a timecourse was performed using both cytokines in tandem and alone. Multicellular spheroid cultures were created using a modified hanging droplet method [36]. After two days, spheroids were suspended in poly-HEMA coated plates and treated every two days with the indicated cytokines to induce EMT (Figure 1A). Samples were collected from untreated (0 days) and cytokine-treated cultures (1–8 days). Epithelial (E-cadherin) and mesenchymal (N-cadherin, Vimentin, and Fibronectin) markers were measured by immunoblot. Treatment with TNF resulted in a modest increase in N-cadherin and Fibronectin, but failed to show differences in other markers (Figure 1B). Consistent with the induction of EMT, TGFβ treatment resulted in a loss of E-cadherin expression and an increase in N-cadherin, Vimentin, and Fibronectin. Moreover, co-stimulation with TNF and TGFβ yielded a more mesenchymal phenotype and persisted throughout the eight day time course (Figure 1B). Importantly, stimulation with TNF and TGFβ effectively induced EMT in both A549 and H358 cell lines within four days of treatment, compared to H1299, which already shows changes in E-cadherin and vimentin (Figure 1C). Based on results in Figure 1, we used the four day timeframe throughout our remaining experiments.

Figure 1. Establishment of three-dimensional multicellular culture model for EMT studies.

(A) A timeline illustrates the procedure used to create a three-dimensional mesenchymal cell population from confluent monolayers. (B) Spheroid cultures of A549 cells were treated, with TNF, TGFβ, or both cytokines every forty-eight hours for the indicated times. Immunoblot analysis measured changes in epithelial (E-cadherin) and mesenchymal (N-cadherin, Vimentin, and fibronectin) markers over an eight day timecourse. (C) 3D cultures of multiple NSCLC cell lines (A549, H358, H1299) were incubated for ninety-six hours in the absence or presence of TNF and TGFβ. Epithelial and mesenchymal markers were subsequently measured by immunoblot. Results from Figure 1B and 1C are representative examples from at least three independent experiments; α-tubulin acts as a protein loading control.
3D cultures undergo EMT more efficiently than 2D cultures
To determine whether 3D A549 cultures undergo EMT more efficiently than two-dimensional (2D) monolayer cultures, we measured expression of epithelial and mesenchymal markers in response to stimulation with TNF and TGFβ as described in Figure 1. Following cytokine treatment, 3D cultures show significant loss of CDH1E-cadherin expression when compared to 2D cultures (Figure 2A). Moreover, the spheroids also possess increased expression of mesenchymal markers VIM, HMGA2, and the EMT master-switch transcription factors, TWIST1, SNAI1Snail1, SNAI2Slug and ZEB2Sip1 (Figures 2A and 2B). Immunoblot analysis of spheroid cultures confirm that the differential mRNA expression resulted in a corresponding change in protein levels (Figure 2C). Additionally, we examined changes in cellular morphology and E-cadherin localization by microscopy. Both 2D and 3D cultures were treated with cytokines as described, trypsinized, re-plated on glass coverslips, and indirect immunofluorescent staining was carried out eighteen hours later. As expected, untreated monolayer and spheroid A549 samples showed robust E-cadherin expression, though the junctional localization appeared diminished in cells from the 3D cultures (Figure S1). Furthermore, cells derived from cytokine-treated spheroids displayed enhanced loss of E-cadherin when compared to 2D treated samples, suggesting that 3D cultures underwent more efficient EMT. Results shown in Figure 2 and Figure S1 illustrate significant EMT induction in 3D cultures as measured by changes in mesenchymal markers, EMT master-switch transcription factor expression, and cellular morphology.

Figure 2. Three-dimensional cultures show enhanced sensitivity to cytokine treatment.

(A and B) Monolayer (2D) and 3D cultures of A549 cells were left alone (No Add) or treated with TNF and TGFβ (TNFTGF) for ninety-six hours. Expression of epithelial markers (CDH1), mesenchymal markers (VIM, HMGA2), and EMT master-switch transcription factors (TWIST1, SNAI1, ZEB2, SNAI2) were measured by QRT-PCR. (C) Immunoblot analysis of 3D A549 cultures, left alone (No Add) or treated with TNF and TGFβ (TNFTGF), was performed on E-cadherin, Vimentin, HMGA2, Twist1, Snail1, Sip1, Slug, and α-tubulin. Results in Figure 2A and 2B were normalized to GAPDH, and are calculated mean ± S.D, *p<0.05, N = 3. Immunoblots in Figure 2C are representative example from at least three independent experiments.
Mesenchymal NSCLC cells are invasive and endogenously express genes known to promote stem-like properties
Phenotypically, mesenchymal cells have high migration rates and secrete enzymes that degrade extracellular matrix to facilitate cellular invasion. Using in vitro transwell assays, we measured the migration and invasion characteristics of A549 cells grown as either 2D or 3D cultures. Interestingly, untreated 3D spheroid cultures showed higher migration rates than 2D monolayer cultures (Figure 3A, left). However, treatment of 3D cultures with TNF and TGFβ further potentiated migration when compared to untreated 3D cultures. Spheroids treated with cytokines invaded through Matrigel more effectively than any other condition (Figure 3A, right). Additionally, cytokine treated A549 spheroids displayed upregulated expression of MMP9, LOX, and COL22A1 (Figure 3B), genes known to potentiate invasion [41], [42]. These results demonstrate that culturing 3D spheroids in the presence of TNF and TGFβ establishes a highly invasive mesenchymal population. Finally, cytokine-treated spheroids showed endogenous upregulation of markers associated with de-differentiation and maintenance of CICs [43]–[48], including KLF4, SOX2, POU5F1Oct4, MYCN, and KIT (Figure 3C). Data shown in Figure 3 indicate that co-stimulation of spheroids with TNF and TGFβ promotes phenotypic changes in A549 cells that result in increased invasion and expression of gene products associated with stem-like properties.

Figure 3. Efficient induction of EMT promotes invasion and the expression of genes required to maintain CICs.

Monolayer and 3D A549 cultures were left alone (No Add) or treated with TNF and TGFβ (TNFTGF) for ninety-six hours. (A) Cells were disaggregated and subsequently subjected to migration and invasion assays. (B and C) Expression of invasion (MMP-9, LOX, COL22A1) and stem cell markers (KLF4, Sox2, POU5F1, MYCN, and KIT) was measured by QRT-PCR. Results in Figure 3 are calculated mean ± S.D, *p<0.05, N = 3. Results from 3B and 3C were normalized to GAPDH.
Mesenchymal cells are highly metastatic and display cancer initiating phenotypes
To examine whether induction of EMT promotes the development of CICs in vivo, we utilized a xenograft tumor model in nude mice. TNF and TGFβ treated 2D and 3D cultures were disaggregated and cell suspensions were SC injected into the right flank of nude mice. Forty days later, animals were sacrificed and SC tumors were resected and weighed while the lungs were excised and scored for surface metastases. To our surprise, TNF and TGFβ-treated cells did not form SC tumors to the same extent as cytokine-treated 2D cultures (Figure 4A, left). However, examination of the lung surface in these mice revealed extensive metastasis (Figure 4A, right). The only plausible explanation for these results is that mesenchymal cells from 3D cultures invaded and metastasized to the lung without developing SC tumors.

Figure 4. Cytokine-treated 3D cultures contain CICs with increased metastatic potential.

(A) Monolayer and 3D A549 cultures were treated with TNF and TGFβ for ninety-six hours. Cells were disaggregated and SC injected into nude mice (1×106 cellsanimal). Forty days later, the primary SC tumors were resected and weighed. Additionally, the lungs were excised and the number of surface metastases were determine. (B) Monolayer and 3D A549 cultures were either left untreated or treated with TNF and TGFβ and limiting cell numbers (1×103animal) were SC injected into nude mice to evaluate the presence of CICs. Metastasis was evaluated by surface lung tumor count and lung tumor burden was evaluated using genomic QRT-PCR to detect human DNA in total lung tissue. Weight and lung metastases data from Figure 4 are mean ± S.D. of five mice per condition, *p<0.05, N = 3 independent experiments. Genomic QRT-PCR data from Figure 4B are normalized to total lung tissue (mg).

Measuring the extent of metastasis under limiting cell dilution proves a reliable test for the presence of enriched CICs in epithelial-derived tumors [49]. Therefore, experiments were repeated using one-thousand cells per SC injection. Cell suspensions, derived from TNF and TGFβ treated spheroids, produced more surface lung metastases under limiting cell dilution than cytokine-treated monolayers or untreated 3D cultures (Figure 4B left). Limiting cell dilution assays indicate that induction of EMT in 3D cultures produces a CIC population that effectively metastasizes to lung. As expected, analysis of DNA isolated from mouse lungs confirmed the presence of metastatic burden and verified that the lesions were of human origin (Figure 4B right). We conclude from the experiments in Figure 4 that de-differentiation, CIC formation, and metastatic potential are all significantly enhanced in EMT-induced spheroid cultures.
NF-κB is constitutively active in 3D cultures and is required for induction of EMT
TNF, a potent NF-κB activator, enhances induction of EMT in NSCLC cell lines. Therefore, we assessed whether EMT induction results in activation of NF-κB signaling by immunoblot. Interestingly, mesenchymal A549 spheroids displayed constitutive IKK activity as measured by phospho-specific antibodies that detect IκBα pS32 and RelA pS536 (Figure 5A and Figure S2A). Change in E-cadherin and Vimentin levels confirmed efficient EMT in the cytokine-treated spheroids. Moreover, QRT-PCR experiments demonstrated increased expression of NF-κB-regulated genes IL8 and BIRC3cIAP2 in mesenchymal 3D cultures (Figure 5B). Collectively, these data indicate that cytokine-treatment of 3D A549 cultures results in the increased phosphorylation of IKK-regulated substrates and constitutive NF-κB transcriptional activation.

Figure 5. Mesenchymal cells display constitutive NF-κB activity.

Monolayer and 3D cultures of A549 cells were incubated with cytokines for ninety-six hours. (A) Mesenchymal A549 cells display constitutive NF-κB activated pathways, as determined using phospho-specific antibodies to IκBα and RelA. (B) Untreated and TNF and TGFβ stimulated 2D and 3D cultures of A549 cells were harvested and analyzed for expression of NF-κB regulated genes by QRT-PCR. (C and D) Three dimensional cultures of A549.V (vector control) and A549.I (SR-IκB) were incubated for ninety-six hours in the absence or presence of TNF and TGFβ. (C) Immunoblots confirm the expression of the Flag-tagged SR-IκBα in the A549.I line, which successfully blocked nuclear translocation and DNA binding, as measured by EMSA. (D) QRT-PCR confirmed the inability of A549.I cell to upregulate NF-κB-regulated genes following TNF and TGFβ treatment. Immunoblots in Figure 5A are a representative example from three independent experiments. Results in Figure 5B and 5D are calculated mean ± S.D, *p<0.05, N = 3. RNA values were normalized to GAPDH.

To determine the importance of NF-κB activity during induction of EMT in NSCLC cell lines, stable clonal pools expressing the super-repressor IκBα (SR-IκBα) were generated. The SR-IκBα is resistant to proteasomal degradation, and consequently sequesters NF-κB in the cytosol. Cells expressing the SR-IκBα protein therefore display an inhibition of NF-κB-mediated transcription [50]. Figure 5C (top) confirms expression of Flag-tagged SR-IκBα in A549 stable cells (A549.I) compared to empty vector control cells (A549.V). Furthermore, nuclear protein extracts from A549.I spheroid cultures, treated with TNF and TGFβ, lacked NF-κB DNA binding activity as compared to A549.V extracts (Figure 5C, bottom). Supershift experiments confirm that the NF-κB activity is composed predominantly of a RelA-p50 heterodimer complex (Figure S2B). QRT-PCR assays show repressed cytokine-mediated induction of IL8 and BIRC3cIAP2 in A549.I cells when compared to control cells A549.V (Figure 5D). In contrast to high doses of TNF (100 ngml), low doses (10 ngml) did not result in a loss of cell viability in A549.I lines, since expression of the house keeping gene, HPRT, did not change and was used for normalization in Figure 5D. These data verify that SR-IκBα expression in the A549.I cell line effectively blocks NF-κB transcriptional activity.
Characterization of NF-κB in potentiating the mesenchymal phenotype
NF-κB has been shown to regulate the expression of EMT master-switch transcription factors in multiple model systems [21]–[24]. Therefore, we hypothesized that inhibiting NF-κB activity in the A549.I cell line would dampen EMT induction. Immunoblot analysis confirmed that A549.I cells fail to down regulate E-cadherin expression or upregulate mesenchymal markers (Vimentin, N-cadherin and Fibronectin) compared to control cells (Figure 6A). Moreover, cytokine-treated A549.I cells showed only minimal upregulation of TWIST1, ZEB2 and SNAI2 gene expression following TNF and TGFβ treatment (Figure 6B). These results indicate that NF-κB is required to upregulate TWIST1, ZEB2 and SNAI2, while expression of SNAI1 appears independent of NF-κB-dependent transcription in the A549.I cell line. These results suggest that the expression of critical EMT master-switch transcription factors requires NF-κB activity.

Figure 6. NF-κB is required for the maintenance of CICs and lung metastasis.

(A) A549.I cells fail to show changes in mesenchymal markers, as determined by immunoblot analysis. (B) NF-κB is required to upregulate mRNA expression of master-switch transcription factors. (C) Spheroid cultures of A549 and H157 cell lines, expressing empty vector or the Flag-IκB super-repressor, were left alone (No Add) or treated with TNF and TGFβ (TNFTGF) for ninety-six hours. The cells were disaggregated and subjected to invasion assays. (D) A549.V and A549.I 3D cultures were left alone (No Add) or treated with TNF and TGFβ (TNFTGF) for ninety-six hours. The cells were disaggregated and SC injected into nude mice (1×106animal). Forty days later, animals were sacrificed and the number of surface lung metastasis were determined. In addition, SC tumors were excised and wet tumor weight determined. Weight and lung metastases data from Figure 6 are mean ± S.D. of five mice per condition, *p<0.05, N = 3 independent experiments. The graphs in Figure 6 are mean ± S.D., *p<0.05, of three independent experiments. Data with P values greater than 0.05 were considered not significant (ns). QRT-PCR experiments are normalized to GAPDH expression.

Next, we assessed whether NSCLC required NF-κB for invasion using transwell assays. Inhibited NF-κB activity in A549.I cells abolished invasion through Matrigel when compared to the control lines (Figure 6C). This effect was not cell-line specific since another NSCLC line expressing the SR-IκB (H157.I) showed similar results as A549.I cells. Because data shown in Figure 6 indicate that NF-κB is required for NSCLC to undergo EMT, we tested the A549.V and A549.I cell for their ability to metastasize to lung using a nude mouse model. As expected, cytokine-treated A549.I cells failed to form lung metastases (Figure 6D, left). The inability of these cells to metastasize to lung was not due to a loss of cell viability or an inability to form primary tumors, since untreated A549.I formed SC tumors with similar growth rates as A549.V cells (Figure 6D, right). Thus, data shown in Figure 6 indicates that TNF and TGFβ treated 3D NSCLC cultures require NF-κB to upregulate master-switch transcription factors, induce EMT, and promote invasive properties. Moreover, without NF-κB transcriptional activity A549 cells lose their ability to metastasize to lung without impacting primary tumor growth.
Discusson NF-κB regulates EMT to potentiate metastatic progression of NSCLC
We implemented a simple and relatively quick 3D culture system to examine the importance of NF-κB signaling during EMT induction and CIC propagation within NSCLC cell lines. In response to TNF and TGFβ exposure, A549 spheroid cultures displayed a loss of E-Cadherin and elevated expression of mesenchymal markers, N-Cadherin, Vimentin, and Fibronectin. The increased expression of mesenchymal protein markers likely occurs due to induction of the EMT master-switch transcription factors, TWIST1, SNAI1, SNAI2 and ZEB2. Furthermore, spheroid populations of mesenchymal A549 cells show elevated expression of endogenous transcription factors known to potentiate dedifferentiation, including KLF4, SOX2, POU5F1, MYCN and KIT. Interestingly, mesenchymal A549 cells from spheroid cultures failed to generate large SC tumors, compared to 2D cultures. Despite this effect, cytokine-treated 3D A549 cells displayed elevated lung surface metastatic lesions. These results support the hypothesis that CICs extravasated into the circulatory system and metastasize to the lung without forming SC tumors. We further demonstrated that EMT-induced A549 3D cultures effectively metastasize to lung under limiting cell dilutions, confirming the presence of an enriched “stem-like” CIC population. Thus, our results suggest that EMT induction effectively selects for self-renewing CIC with metastatic potential, a phenotype described by Dieter and colleagues as self-renewing long-term tumor initiating cells responsible for color cancer metastasis [51]. Since IKK and NF-κB pathways have been linked to EMT and development of CICs [21]–[25], [31], we examined whether mesenchymal A549 cells upregulate NF-κB transcriptional activity. Surprisingly, EMT-induced spheroid A549 cultures displayed chronic IKK activity as measured by phosphorylation of IκBα(pS32) and RelA(pS536), and by constitutive expression of IL8 and BIRC3 transcripts. Moreover, cytokine-treated spheroid A549 cultures maintained the activation of IKK signaling pathways well beyond the half-life of the TNF and TGFβ cytokines added to the culture media. These results suggest that mesenchymal A549 spheroid cultures must produce autocrine factors capable of maintaining the EMT phenotype. Importantly, constitutive NF-κB activity proves essential for effective EMT partially through its ability to upregulate the master-switch transcription factors TWIST1, ZEB2 and SNAI2. As a result, the loss of NF-κB activity prohibited cytokine-treated spheroid A549 cells from becoming invasive and also abolished lung metastasis in the mouse xenograft model. This work firmly establishes a role for NF-κB in the induction of EMT and for the development of NSCLC CICs that promote metastasis.
Spheroid models and the propagation of CICs
Various 3D culture models have been developed that more accurately mimic tumor biology, such as cell-cell contacts, extracellular matrix composition, and nutrient accessgradients [52]–[54]. The advantage to using the hanging drop technique, over other techniques, is that 2D cultures can be quickly expanded to form multicellular aggregates that share similar size and shape, and mesenchymal populations are generated within six days. Data provided in Figure 1C demonstrate that multiple NSCLC cell lines form compact spheroids that undergo highly reproducible EMT when exposed to TNF and TGFβ. Moreover, these spheroids possess increased sensitivity to TNF and TGFβ compared to monolayer cultures (Figures 2, 3, and 4). Therefore, we utilized this 3D system to examine EMT and CIC formation in NSCLC cell lines. Surprisingly, A549 spheroids show increased migration without requiring exposure to TNF and TGFβ and despite expressing epithelial markers (Figure 1B, 2A, and 3A). This indicates that phenotypic changes occur in 3D cultures prior to exposure to EMT-inducing cytokines. However, increased invasion is restricted to cytokine-induced A549 spheroid cultures and corresponds with the upregulation of matrix and extracellular remodeling enzymes known to induce invasive properties [41], [42]. Therefore, spheroid cultures are poised to respond to TNF and TGFβ cytokines and are able to sustain EMT reprogramming. Together, we establish that CIC populations formed from EMT induction of 3D NSCLC cell lines provide a useful tool for further characterization of cancer progression in the lung.
Mesenchymal A549 cells show constitutively active NF-κB signaling pathways
Constitutive NF-κB activation occurs in many different types of hematopoietic and epithelial-derived carcinomas. However, mutations that result in chronic activation of NF-κB signaling are extremely rare in epithelial cancers [19]. Thus, activation of NF-κB most likely results from autocrine and paracrine signaling within the tumor microenvironment rather than genetic alterations [2], [19]. Our data support this hypothesis by showing that TNF- and TGFβ-treated A549 spheroid populations both undergo EMT and maintain constitutive NF-κB signaling (Figure 5A and 5B). Rather than using co-culture systems, which introduce contaminating cell types other than NSCLC cells, we chose to treat A549 spheroids with EMT-inducing cytokines. TNF and TGFβ were selected because within the tumor microenvironment, TNF is believed to be produced predominantly by tumor-associated macrophages, while TGFβ is secreted by fibroblast and endothelial cells. This combination of cytokines not only effectively and reproducibly induces EMT, but also facilitates a reprogramming event that results in chronic NF-κB signaling. The molecular mechanism by which this occurs is currently unknown, but most likely is due to an increase in NF-κB-regulated gene products that function in an autocrine-dependent manner to maintain active NF-κB.
Inflammatory regulatory circuits that drive constitutive NF-κB activation
In the past two years, evidence has emerged that an epigenetic switch occurs during breast cancer transformation in which inflammatory circuits involving IL6 and IL8 mediate self-renewal of CICs [55]–[57]. Ginesteir and colleagues showed that breast CICs upregulate the IL8 receptor CXCR1 to potentiate self-renewal, tumorigenicity and metastasis [57]. Additional studies indicate that oncogenic transformation of breast cancer cells leads to chronic activation of NF-κB required to upregulate Lin-28B and downregulate the negative microRNA regulator of IL6, Let-7a [56]. As a result, IL6 provides an inflammatory feedback loop that further activates NF-κB as well as the STAT3 signaling pathway [56], [58]. Interestingly, this pro-inflammatory feedback loop also exists in some prostate and hepatocellular carcinomas, but only a subset of lung cancers showed increased IL6 expression [56]. In addition to the autocrine feedback mechanism, IL6 signaling pathways downregulate mir200c in a chemically-induced transformed breast cancer cell line. Loss of mir200c subsequently results in constitutively activated NF-κB through an inflammatory feedforward signaling circuit [59]. In these papers [55], [56], [59], IL6 was found to be required for the maintenance of breast CICs.

Additional work is needed to determine the importance of IL8 and IL6 as feedforward mediators of NF-κB activation in mesenchymal NSCLC cell lines. As shown in Figure 5B, IL8 is highly upregulated and maintained in mesenchymal A549 cultures; however, IL6 transcripts do not significantly change between untreated 3D and cytokine-treated 3D cultures (Figure S2C). Thus, in agreement with IIiopoulos and colleagues [59], IL6 may not be a common requirement for CICs in lung cancer. However, since CXCR1 is highly expressed in A549 cells following exposure to DNA methytransferase inhibitors [60], inflammatory circuits that regulate promoter demethylation, as observed for IL6 signaling [59], may play an important role for controlling the IL8CXCR1 responsiveness in lung cancers. Future work is needed to explore the importance of IL8CXCR1 in the maintenance of constitutive NF-κB activation and development of NSCLC CICs.
Supporting Information Figure S1.
Cytokine-treated 3D A549 cells show increased fibroid and mesenchymal morphology. Monolayer (2D) and 3D A549 cultures were left alone or treated with TNF and TGFβ for ninety-six hours. Cells were subsequently disaggregated, replated on glass coverslips, and cultured for an additional eighteen hours in 2% FBS. The cells were then fixed in methanol, and indirect immunofluorescence was used to detect the presence of junctional E-cadherin. Images are a representative field from three independent experiments.
doi:10.1371journal.pone.0068597.s001

(TIF)
Figure S2.
TNF and TGFβ-treated 3D A549 cells show increased RelA phosphorylation and nuclear DNA binding activity. (A) Immunoblot analysis of 3D A549 cells indicates that cells display constitutive RelA phosphorylation upon co-stimulation with both TNF and TGFβ over the three day period. (B) Nuclear extracts from cytokine-treated 3D control A549.V cells show elevated NF-κB binding activity by EMSA, compared to unstimulated cell extracts. The NF-κB DNA-protein complex is composed of both RelA and p50 proteins as detected by antibody super shift (SS) assays. (C) In contrast to IL8 expression shown in Figure 5B, cytokine-treated 3D cultures fail to upregulate IL6 transcripts as measured by QRT-PCR.
doi:10.1371journal.pone.0068597.s002

(TIF)
Table S1.
QRT-PCR Primers.
doi:10.1371journal.pone.0068597.s003

(DOC)
Contributions
Conceived and designed the experiments: MWM MK DFA NNB JJW. Performed the experiments: MK DFA NNB JJW. Analyzed the data: MWM MK DFA NNB JJW. Contributed reagentsmaterialsanalysis tools: AJK SB DRJ. Wrote the paper: MWM MK DFA JJW.
References
  • 1. Cirri P, Chiarugi P (2012) Cancer-associated-fibroblasts and tumour cells: A diabolic liaison driving cancer progression. Cancer Metastasis Rev 31: 195–208 10.1007s10555-011-9340-x.
  • 2. Korkaya H, Liu S, Wicha MS (2011) Breast cancer stem cells, cytokine networks, and the tumor microenvironment. J Clin Invest 121: 3804–3809 10.1172JCI57099.
  • 3. Fuxe J, Karlsson MC (2012) TGF-beta-induced epithelial-mesenchymal transition: A link between cancer and inflammation. Semin Cancer Biol 22: 455–461 10.1016j.semcancer.2012.05.004.
  • 4. Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139: 871–890.
  • 5. Yang J, Weinberg RA (2008) Epithelial-mesenchymal transition: At the crossroads of development and tumor metastasis. Dev Cell 14: 818–829 10.1016j.devcel.2008.05.009.
  • 6. Floor S, van Staveren WC, Larsimont D, Dumont JE, Maenhaut C (2011) Cancer cells in epithelial-to-mesenchymal transition and tumor-propagating-cancer stem cells: Distinct, overlapping or same populations. Oncogene 30: 4609–4621 10.1038onc.2011.184.
  • 7. Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, et al. (2008) The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 133: 704–715.
  • 8. Morel AP, Lievre M, Thomas C, Hinkal G, Ansieau S, et al. (2008) Generation of breast cancer stem cells through epithelial-mesenchymal transition. PLoS One 3: e2888 10.1371journal.pone.0002888.
  • 9. Scheel C, Weinberg RA (2011) Phenotypic plasticity and epithelial-mesenchymal transitions in cancer and normal stem cells? Int J Cancer 129: 2310–2314 10.1002ijc.26311.
  • 10. Heldin CH, Vanlandewijck M, Moustakas A (2012) Regulation of EMT by TGFbeta in cancer. FEBS Lett 586: 1959–1970 10.1016j.febslet.2012.02.037.
  • 11. Wendt MK, Tian M, Schiemann WP (2012) Deconstructing the mechanisms and consequences of TGF-beta-induced EMT during cancer progression. Cell Tissue Res 347: 85–101 10.1007s00441-011-1199-1.
  • 12. Xu J, Lamouille S, Derynck R (2009) TGF-beta-induced epithelial to mesenchymal transition. Cell Res 19: 156–172 10.1038cr.2009.5.
  • 13. Zu X, Zhang Q, Cao R, Liu J, Zhong J, et al. (2012) Transforming growth factor-beta signaling in tumor initiation, progression and therapy in breast cancer: An update. Cell Tissue Res 347: 73–84 10.1007s00441-011-1225-3.
  • 14. Bates RC, Mercurio AM (2003) Tumor necrosis factor-alpha stimulates the epithelial-to-mesenchymal transition of human colonic organoids. Mol Biol Cell 14: 1790–1800 10.1091mbc.E02-09-0583.
  • 15. Kawata M, Koinuma D, Ogami T, Umezawa K, Iwata C, et al. (2012) TGF-beta-induced epithelial-mesenchymal transition of A549 lung adenocarcinoma cells is enhanced by pro-inflammatory cytokines derived from RAW 264.7 macrophage cells. J Biochem 151: 205–216 10.1093jbmvr136.
  • 16. Gao D, Vahdat LT, Wong S, Chang JC, Mittal V (2012) Microenvironmental regulation of epithelial-mesenchymal transitions in cancer. Cancer Res 72: 4883–4889 10.11580008-5472.CAN-12-1223.
  • 17. Scheel C, Eaton EN, Li SH, Chaffer CL, Reinhardt F, et al. (2011) Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast. Cell 145: 926–940 10.1016j.cell.2011.04.029.
  • 18. Basseres DS, Baldwin AS (2006) Nuclear factor-kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. Oncogene 25: 6817–6830 10.1038sj.onc.1209942.
  • 19. Ben-Neriah Y, Karin M (2011) Inflammation meets cancer, with NF-kappaB as the matchmaker. Nat Immunol 12: 715–723 10.1038ni.2060.
  • 20. Shostak K, Chariot A (2011) NF-kappaB, stem cells and breast cancer: The links get stronger. Breast Cancer Res 13: 214 10.1186bcr2886.
  • 21. Barbera MJ, Puig I, Dominguez D, Julien-Grille S, Guaita-Esteruelas S, et al. (2004) Regulation of snail transcription during epithelial to mesenchymal transition of tumor cells. Oncogene 23: 7345–7354 10.1038sj.onc.1207990.
  • 22. Chua HL, Bhat-Nakshatri P, Clare SE, Morimiya A, Badve S, et al. (2007) NF-kappaB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells: Potential involvement of ZEB-1 and ZEB-2. Oncogene 26: 711–724 10.1038sj.onc.1209808.
  • 23. Julien S, Puig I, Caretti E, Bonaventure J, Nelles L, et al. (2007) Activation of NF-kappaB by akt upregulates snail expression and induces epithelium mesenchyme transition. Oncogene 26: 7445–7456 10.1038sj.onc.1210546.
  • 24. Pham CG, Bubici C, Zazzeroni F, Knabb JR, Papa S, et al. (2007) Upregulation of twist-1 by NF-kappaB blocks cytotoxicity induced by chemotherapeutic drugs. Mol Cell Biol 27: 3920–3935 10.1128MCB.01219-06.
  • 25. Wu Y, Deng J, Rychahou PG, Qiu S, Evers BM, et al. (2009) Stabilization of snail by NF-kappaB is required for inflammation-induced cell migration and invasion. Cancer Cell 15: 416–428 10.1016j.ccr.2009.03.016.
  • 26. Hayden MS, Ghosh S (2008) Shared principles in NF-kappaB signaling. Cell 132: 344–362.
  • 27. Hoberg JE, Popko AE, Ramsey CS, Mayo MW (2006) IkappaB kinase alpha-mediated derepression of SMRT potentiates acetylation of RelAp65 by p300. Mol Cell Biol 26: 457–471.
  • 28. Allison DF, Wamsley JJ, Kumar M, Li D, Gray LG, et al. (2012) Modification of RelA by O-linked N-acetylglucosamine links glucose metabolism to NF-kappaB acetylation and transcription. Proc Natl Acad Sci U S A 109: 16888–16893 10.1073pnas.1208468109.
  • 29. Hoberg JE, Yeung F, Mayo MW (2004) SMRT derepression by the IkappaB kinase alpha: A prerequisite to NF-kappaB transcription and survival. Mol Cell 16: 245–255.
  • 30. Yeung F, Hoberg JE, Ramsey CS, Keller MD, Jones DR, et al. (2004) Modulation of NF-kappaB-dependent transcription and cell survival by the SIRT1 deacetylase. EMBO J 23: 2369–2380.
  • 31. Huber MA, Azoitei N, Baumann B, Grunert S, Sommer A, et al. (2004) NF-kappaB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression. J Clin Invest 114: 569–581.
  • 32. Akunuru S, James Zhai Q, Zheng Y (2012) Non-small cell lung cancer stemprogenitor cells are enriched in multiple distinct phenotypic subpopulations and exhibit plasticity. Cell Death Dis 3: e352 10.1038cddis.2012.93.
  • 33. Ho MM, Ng AV, Lam S, Hung JY (2007) Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res 67: 4827–4833 10.11580008-5472.CAN-06-3557.
  • 34. Leung EL, Fiscus RR, Tung JW, Tin VP, Cheng LC, et al. (2010) Non-small cell lung cancer cells expressing CD44 are enriched for stem cell-like properties. PLoS One 5: e14062 10.1371journal.pone.0014062.
  • 35. Sung JM, Cho HJ, Yi H, Lee CH, Kim HS, et al. (2008) Characterization of a stem cell population in lung cancer A549 cells. Biochem Biophys Res Commun 371: 163–167 10.1016j.bbrc.2008.04.038.
  • 36. Kelm JM, Timmins NE, Brown CJ, Fussenegger M, Nielsen LK (2003) Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng 83: 173–180 10.1002bit.10655.
  • 37. Gilbert MT, Haselkorn T, Bunce M, Sanchez JJ, Lucas SB, et al. (2007) The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when? PLoS One 2: e537 10.1371journal.pone.0000537.
  • 38. Nitz MD, Harding MA, Theodorescu D (2008) Invasion and metastasis models for studying RhoGDI2 in bladder cancer. Methods Enzymol 439: 219–233 10.1016S0076-6879(07)00417-X.
  • 39. Thulke S, Radonic A, Siegert W, Nitsche A (2003) Highly sensitive quantification of human cells in chimeric NODSCID mice by real-time PCR. Haematologica 88: ELT18.
  • 40. Mayo MW, Norris JL, Baldwin AS (2001) Ras regulation of NF-kappa B and apoptosis. Methods Enzymol 333: 73–87.
  • 41. Kirschmann DA, Seftor EA, Fong SF, Nieva DR, Sullivan CM, et al. (2002) A molecular role for lysyl oxidase in breast cancer invasion. Cancer Res 62: 4478–4483.
  • 42. Yu Q, Stamenkovic I (2000) Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumor invasion and angiogenesis. Genes Dev 14: 163–176.
  • 43. Chen YC, Hsu HS, Chen YW, Tsai TH, How CK, et al. (2008) Oct-4 expression maintained cancer stem-like properties in lung cancer-derived CD133-positive cells. PLoS One 3: e2637 10.1371journal.pone.0002637.
  • 44. Leis O, Eguiara A, Lopez-Arribillaga E, Alberdi MJ, Hernandez-Garcia S, et al. (2012) Sox2 expression in breast tumours and activation in breast cancer stem cells. Oncogene 31: 1354–1365 10.1038onc.2011.338.
  • 45. Levina V, Marrangoni A, Wang T, Parikh S, Su Y, et al. (2010) Elimination of human lung cancer stem cells through targeting of the stem cell factor-c-kit autocrine signaling loop. Cancer Res 70: 338–346 10.11580008-5472.CAN-09-1102.
  • 46. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, et al. (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131: 861–872 10.1016j.cell.2007.11.019.
  • 47. Yu F, Li J, Chen H, Fu J, Ray S, et al. (2011) Kruppel-like factor 4 (KLF4) is required for maintenance of breast cancer stem cells and for cell migration and invasion. Oncogene 30: 2161–2172 10.1038onc.2010.591.
  • 48. Singh S, Trevino J, Bora-Singhal N, Coppola D, Haura E, et al. (2012) EGFRSrcAkt signaling modulates Sox2 expression and self-renewal of stem-like side-population cells in non-small cell lung cancer. Mol Cancer 11: 73–4598-11-73 10.11861476-4598-11-73.
  • 49. Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A 100: 3983–3988.
  • 50. Mayo MW, Wang CY, Cogswell PC, Rogers-Graham KS, Lowe SW, et al. (1997) Requirement of NF-kappaB activation to suppress p53-independent apoptosis induced by oncogenic ras. Science 278: 1812–1815.
  • 51. Dieter SM, Ball CR, Hoffmann CM, Nowrouzi A, Herbst F, et al. (2011) Distinct types of tumor-initiating cells form human colon cancer tumors and metastases. Cell Stem Cell 9: 357–365 10.1016j.stem.2011.08.010.
  • 52. Kunz-Schughart LA, Kreutz M, Knuechel R (1998) Multicellular spheroids: A three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol 79: 1–23.
  • 53. Li Q, Chen C, Kapadia A, Zhou Q, Harper MK, et al. (2011) 3D models of epithelial-mesenchymal transition in breast cancer metastasis: High-throughput screening assay development, validation, and pilot screen. J Biomol Screen 16: 141–154 10.11771087057110392995.
  • 54. Pampaloni F, Reynaud EG, Stelzer EH (2007) The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol 8: 839–845 10.1038nrm2236.
  • 55. Xie G, Yao Q, Liu Y, Du S, Liu A, et al. (2012) IL-6-induced epithelial-mesenchymal transition promotes the generation of breast cancer stem-like cells analogous to mammosphere cultures. Int J Oncol 40: 1171–1179 10.3892ijo.2011.1275.
  • 56. Iliopoulos D, Hirsch HA, Struhl K (2009) An epigenetic switch involving NF-kappaB, Lin28, let-7 MicroRNA, and IL6 links inflammation to cell transformation. Cell 139: 693–706 10.1016j.cell.2009.10.014.
  • 57. Ginestier C, Liu S, Diebel ME, Korkaya H, Luo M, et al. (2010) CXCR1 blockade selectively targets human breast cancer stem cells in vitro and in xenografts. J Clin Invest 120: 485–497 10.1172JCI39397.
  • 58. Grivennikov S, Karin E, Terzic J, Mucida D, Yu GY, et al. (2009) IL-6 and Stat3 are required for survival of intestinal epithelial cells and development of colitis-associated cancer. Cancer Cell 15: 103–113.
  • 59. Rokavec M, Wu W, Luo JL (2012) IL6-mediated suppression of miR-200c directs constitutive activation of inflammatory signaling circuit driving transformation and tumorigenesis. Mol Cell 45: 777–789 10.1016j.molcel.2012.01.015.
  • 60. Baird AM, Gray SG, O'Byrne KJ (2011) Epigenetics underpinning the regulation of the CXC (ELR+) chemokines in non-small cell lung cancer. PLoS One 6: e14593 10.1371journal.pone.0014593.


  • References:


    Pass4sure Certification Exam Questions and Answers - www.founco.com
    Killexams Exam Study Notes | study guides - www.founco.com
    Pass4sure Certification Exam Questions and Answers - st.edu.ge
    Killexams Exam Study Notes | study guides - st.edu.ge
    Pass4sure Certification Exam Questions and Answers - www.jabbat.com
    Killexams Exam Study Notes | study guides - www.jabbat.com
    Pass4sure Certification Exam Questions and Answers - www.jorgefrazao.esy.es
    Killexams Exam Study Notes | study guides - www.jorgefrazao.esy.es
    Pass4sure Certification Exam Questions and Answers and Study Notes - www.makkesoft.com
    Killexams Exam Study Notes | study guides | QA - www.makkesoft.com
    Pass4sure Exam Study Notes - maipu.gob.ar
    Pass4sure Certification Exam Study Notes - idprod.esy.es
    Download Hottest Pass4sure Certification Exams - cscpk.org
    Killexams Study Guides and Exam Simulator - www.simepe.com.br
    Comprehensive Questions and Answers for Certification Exams - www.ynb.no
    Exam Questions and Answers | Brain Dumps - www.4seasonrentacar.com
    Certification Training Questions and Answers - www.interactiveforum.com.mx
    Pass4sure Training Questions and Answers - www.menchinidesign.com
    Real exam Questions and Answers with Exam Simulators - www.pastoriaborgofuro.it
    Real Questions and accurate answers for exam - playmagem.com.br
    Certification Questions and Answers | Exam Simulator | Study Guides - www.rafflesdesignltd.com
    Kill exams certification Training Exams - www.sitespin.co.za
    Latest Certification Exams with Exam Simulator - www.philreeve.com
    Latest and Updated Certification Exams with Exam Simulator - www.tmicon.com.au
    Pass you exam at first attempt with Pass4sure Questions and Answers - tractaricurteadearges.ro
    Latest Certification Exams with Exam Simulator - addscrave.net
    Pass you exam at first attempt with Pass4sure Questions and Answers - alessaconsulting.com
    Get Great Success with Pass4sure Exam Questions/Answers - alchemiawellness.com
    Best Exam Simulator and brain dumps for the exam - andracarmina.com
    Real exam Questions and Answers with Exam Simulators - empoweredbeliefs.com
    Real Questions and accurate answers for exam - www.alexanndre.com
    Certification Questions and Answers | Exam Simulator | Study Guides - allsoulsholidayclub.co.uk

    Comments

    Popular posts from this blog

    Pass4sure SY0-501 Practice Tests with Real Questions

    Just memorize these CTFA questions before you go for test.

    WTF! questions were exactly the same in exam that I prepared! A2010-501 reading practice test | A2010-501 free prep | A2010-501 quest bars | A2010-501 exam prep | A2010-501 past exams - bigdiscountsales.com A2010-501 - Accessment: IBM Maximo Asset Management V7.5 Infrastructure Implementation - Dump Information Vendor : IBM Exam Code : A2010-501 Exam Name : Accessment: IBM Maximo Asset Management V7.5 Infrastructure Implementation Questions and Answers : 167 Q & A Updated On : December 8, 2017 PDF Download Mirror : A2010-501 Brain Dump Get Full Version : Pass4sure A2010-501 Full Version Looking for A2010-501 exam dumps that works in real exam? At killexams.com, we provide thoroughly reviewed IBM A2010-501 training resources which are the best for clearing A2010-501 test, and to get certified by IBM. It is a best choice to accelerate your career as a professional in the Information Technology industry. We are proud of our reputation of helping people clear the A2010-501 test in their very first attempts. Our success rates in the past two years have been absolutely impressive, thanks to our happy customers who are now able to propel their careers in the fast lane. killexams.com is the number one choice among IT professionals, especially the ones who are looking to climb up the hierarchy levels faster in their respective organizations. IBM is the industry leader in information technology, and getting certified by them is a guaranteed way to succeed with IT careers. We help you do exactly that with our high quality IBM A2010-501 training materials. IBM A2010-501 is omnipresent all around the world, and the business and software solutions provided by them are being embraced by almost all the companies. They have helped in driving thousands of companies on the sure-shot path of success. Comprehensive knowledge of IBM products are considered a very important qualification, and the professionals certified by them are highly valued in all organizations. We provide real A2010-501 pdf exam questions and answers braindumps in two formats. Download PDF & Practice Tests. Pass IBM A2010-501 book Exam quickly & easily. The A2010-501 syllabus PDF type is available for reading and printing. You can print more and practice many times. Our pass rate is high to 98.9% and the similarity percentage between our A2010-501 syllabus study guide and real exam is 90% based on our seven-year educating experience. Do you want achievements in the A2010-501 exam in just one try? I am currently studying for the IBM A2010-501 syllabus exam. Cause all that matters here is passing the IBM A2010-501 exam. Cause all that you need is a high score of IBM A2010-501 exam. The only one thing you need to do is downloading Examcollection A2010-501 exam study guides now. We will not let you down with our money-back guarantee. The professionals also keep pace with the most up-to-date exam in order to present with the the majority of updated materials. One year free access to be able to them through the date of buy. Every candidates may afford the IBM exam dumps via killexams.com at a low price. Often there is a discount for anyone all. In the presence of the authentic exam content of the brain dumps at killexams.com you can easily develop your niche. For the IT professionals, it is vital to enhance their skills according to their career requirement. We make it easy for our customers to take certification exam with the help of killexams.com verified and authentic exam material. For a bright future in the world of IT, our brain dumps are the best option. Killexams.com Huge Discount Coupons and Promo Codes are as under; WC2017 : 60% Discount Coupon for all exams on website PROF17 : 10% Discount Coupon for Orders greater than $69 DEAL17 : 15% Discount Coupon for Orders greater than $99 DECSPECIAL : 10% Special Discount Coupon for All Orders A top dumps writing is a very important feature that makes it easy for you to take IBM certifications. But IBM braindumps PDF offers convenience for candidates. The IT certification is quite a difficult task if one does not find proper guidance in the form of authentic resource material. Thus, we have authentic and updated content for the preparation of certification exam. A2010-501 Discount Coupon, A2010-501 Promo Code, A2010-501 vce, Free A2010-501 vce, Download Free A2010-501 dumps, Free A2010-501 braindumps, pass4sure A2010-501, A2010-501 practice test, A2010-501 practice exam, killexams.com A2010-501, A2010-501 real questions, A2010-501 actual test, A2010-501 PDF download, Pass4sure A2010-501 Download, A2010-501 help, A2010-501 examcollection, Passleader A2010-501, exam-labs A2010-501, Justcertify A2010-501, certqueen A2010-501, A2010-501 testking View Full Exam » I got extraordinary Questions bank for my A2010-501 examination. The A2010-501 exam is supposed to be a totally diffcult exam to clear however I cleared it remaining week in my first try. The killexams.com Q&As guided me properly and i used to be properly organized. recommendation to other students - dont take this examination gently and observe very well. Unbelieveable! but proper source of A2010-501 real take a look at questions. candidates spend months seeking to get themselves organized for his or her A2010-501 assessments however for me it changed into all just a days paintings. you would wonder how a person could be able to finish this type of awesome venture in only a day let me let you know, all I needed to do turned into sign up my Feel confident by preparing A2010-501 dumps. This preparation kit has helped me skip the exam and emerge as A2010-501 certified. I couldn't be extra excited and thankful to Killexams for such an clean and reliable education tool. i'm able to confirm that the questions within the bundle are actual, this is not a fake. I chose it for being a dependable (recommended by way of a chum) manner to streamline the exam practise. Like many others, I couldn't have the funds for studying full time for weeks or maybe months, and Killexams has allowed me to squeeze down my preparation time and nonetheless get a extremely good end result. top notch answer for busy IT specialists. Do you need real test qustions of A2010-501 exam? They price me for A2010-501 exam simulator and QA file however first i did now not got the A2010-501 QA fabric. there was a few report mistakes, later they fixed the mistake. i prepared with the exam simulator and it become properly. Do you know the fastest way to pass A2010-501 exam? I've got it. just surpassed the A2010-501 exam with this braindump. i can affirm that it is 99% valid and includes all this years updates. I handiest got 2 question wrong, so very excited and relieved. Get A2010-501 licensed with actual test exam bank. killexams.com had enabled a pleasurable revel in the whole while I used A2010-501 prep resource from it. I observed the study publications, exam engine and, the A2010-501 to each tiniest little detail. It was due to such excellent way that I became talented in the A2010-501 examination curriculum in count of days and were given the A2010-501 certification with an excellent rating. i'm so thankful to every unmarried man or woman in the back of the killexams.com platform. Easy way to pass A2010-501 exam with these q&a and Exam Simulator. Killexams.com became very refreshing access in my life, specifically due to the fact the material that I used thru this killexams.coms help became the one that got me to clean my A2010-501 examination. Passing A2010-501 exam isn't clean however it become for me because I had get admission to to the great studying fabric and i am immensely thankful for that. wherein am i able to locate A2010-501 trendy and updated dumps questions? even though i've enough heritage and enjoy in IT, I predicted the A2010-501 examination to be simpler. Killexams has saved my time and money, with out these QAs i would have failed the A2010-501 examination. I got burdened for few questions, so I almost needed to wager, but that is my fault. I should have memorized well and concentrate the questions better. Its correct to realize that I surpassed the A2010-501 exam. Dont waste your time on searching internet, just cross for those A2010-501 Questions and solutions. I am very much happy with your test papers particularly with the solved problems. Your test papers gave me courage to appear in the A2010-501 paper with confidence. The result is 77.25%. Once again I whole heartedly thank the killexams.com institution. No other way to pass the A2010-501 exam other than killexams.com model papers. I personally cleared other exams with the help of killexams.com question bank. I recommend it to every one. If you want to pass the A2010-501 exam then take killexamss help. Surprised to see A2010-501 Actual Questions! I am Aggarwal and I work for Smart Corp. I had applied to appear for the A2010-501 exam and was very apprehensive about it as it contained difficult case studies etc. I then applied for your question bank. My many doubts got cleared due to the explainations provided for the answers. I also got the case studies in my email which were properly solved. I appeared for the exam and am happy to say that I got 73.75% and I give you the whole credit. Further I congratulate you and look further to clear more exams with the help of your site. See more IBM dumps A2010-503 | A2180-188 | 00M-222 | C2010-655 | C2010-568 | 000-258 | 000-106 | 000-276 | 000-748 | C2040-417 | 00M-642 | 000-035 | 000-733 | C2090-180 | 00M-605 | 000-868 | COG-320 | LOT-829 | P9050-005 | 000-302 | 000-905 | 000-070 | 000-181 | 000-712 | 000-289 | 000-859 | 00M-220 | 000-M17 | 000-N24 | 000-939 | 000-914 | 000-N25 | C2010-570 | 000-M605 | 000-703 | 000-934 | 00M-665 | 000-M35 | A2040-922 | C2040-929 | C9520-929 | 000-M226 | C9020-568 | P2065-016 | 000-972 | LOT-801 | M9560-670 | LOT-985 | C9560-023 | 00M-646 | Latest Exams added on bigdiscountsales 1Z0-453 | 210-250 | 300-210 | 500-205 | 500-210 | 70-765 | 9A0-409 | C2010-555 | C2090-136 | C9010-260 | C9010-262 | C9020-560 | C9020-568 | C9050-042 | C9050-548 | C9050-549 | C9510-819 | C9520-911 | C9520-923 | C9520-928 | C9520-929 | C9550-512 | CPIM-BSP | C_TADM70_73 | C_TB1200_92 | C_TBW60_74 | C_TPLM22_64 | C_TPLM50_95 | DNDNS-200 | DSDPS-200 | E20-562 | E20-624 | E_HANABW151 | E_HANAINS151 | JN0-1330 | JN0-346 | JN0-661 | MA0-104 | MB2-711 | NSE6 | OMG-OCRES-A300 | P5050-031 | See more dumps on bigdiscountsales C_TSCM44_65 | HP0-753 | COG-205 | 9A0-146 | A2090-421 | 642-584 | 000-296 | 98-366 | 4H0-100 | 922-020 | ISEB-ITILV3F | 000-452 | E22-275 | 1Z0-338 | HP0-090 | HP0-Y32 | 000-340 | 000-607 | A2010-578 | 000-424 | 000-M40 | HP0-J73 | C2010-657 | 250-307 | 050-694 | E20-554 | 000-712 | 000-434 | E20-533 | A2090-611 | 1Y0-259 | 1Z1-522 | C2020-180 | NPTE | HP0-M74 | 000-M06 | LOT-952 | 646-206 | HP2-E56 | HPE0-S37 | HP0-M33 | 000-386 | 1Z0-430 | 000-118 | 9L0-007 | C9560-510 | EE0-511 | A2070-581 | BAS-011 | 000-219 | A2010-501 Questions and Answers QUESTION: 104 What is the recommended method to build a Maximo EAR file? runthebuiidear.bat from a command prompt runthebuiidmaximoear.cmd from a command prompt double-click on buiidear.bat from the IBM\SMP\maximo\deployment directory right-click on buildmaximoear.cmd from the IBM\SMP\maximo\deployment\default directory and run as administrator Answer: B QUESTION: 105 When configuring a LDAP query for the LDAPSYNC cron task, it is important to understand the content of the LDAP structure and how it relates to information in IBM Maximo Asset Management V7.5 (Maximo). What happens when users imported from a LDAP capable server do not have the required data intheir record? The person record is created but the associated Maximo user record is not. The user record is imported into the Maximo users table. An error is logged to the bulletin board group for user security updates. The user record is not imported into the Maximo users table. There is an error which can be captured if cron task logging is enabled to do so. The user record is imported into a temporary user table where the security admin role user can review and change any data to allow the user full access. Answer: C QUESTION: 106 An IBM Maximo Asset Management V7.5 administrator completes the configuration of the Maximo Integration Framework and attempts to execute an import from the EXTSYS1 external system. The import appears tocomplete without error but when the database is checked the data is not there. The administrator checks the J2EE JMS sequential inbound queue and can see that the message is there, but it is not being processed. What is the most likely cause the message isnot getting delivered to the database? The external system EXTSYS1 is not active. The Maximo Java Virtual Machine is not started. The JMSQSEQCONSUMER.SEQQOUT is not active. The JMSQSEQCONSUMER.SEQQIN cron is not active. Answer: D QUESTION: 107 The IBMMaximo Asset Management updatedb process includes the execution of scripts in numeric order for each product in which folder location? ibm\smp\maximo\tools\maximo ibm\smp\maximo\applications\maximo ibm\smp\maximo\tools\maximo\\script ibm\smp\maximo\applications\maximo\\script Answer: C QUESTION: 108 An IBM Maximo Asset Management V7.5 environment has a multi-EAR configuration. Each EAR file needs to be uniquely named based on its function. The multi-EAR configuration consists of a User Interface, Maximo Integration Framework, and CRON. Which option controls the name of the EAR file built with the buildmaximoear.cmd? MAXIMO_HOME= EAR_FILENAME= BUILD_EAR_NAME= MAXIMO PROPERTIES= Answer: B QUESTION: 109 The customer plans to use a Novell directory server for user authentication with IBM Maximo Asset Management (Maximo). Which statement provides a reason to change the customer's Maximo deployment plan based on security integration? The customer's environment has an unsupported LDAP capable server. The customer needs an additional user to be supported by user interface JVMs. The customer has a single sign-on solution that can be integrated with the selected J2EE server. The customer wantstheir groups to be managed by Maximo and the users to be managed by a LDAP capable server. Answer: A QUESTION: 110 Which operating system(s) are supported for the IBM Maximo Asset Management V7.5 installation on the administrative workstation? Windows Windows and AIX Windows and Linux Windows, AIX, and Linux Answer: A IBM A2010-501 Exam (Accessment: IBM Maximo Asset Management V7.5 Infrastructure Implementation) Detailed Information C2010-501 Test Information / Examination Information Number of questions : 57 Time allowed in minutes: 90 Required passing score : 71% Languages : English C2010-501 Objectives References: Pass4sure Certification Exam Questions and Answers - www.founco.com Killexams Exam Study Notes | study guides - www.founco.com Pass4sure Certification Exam Questions and Answers - st.edu.ge Killexams Exam Study Notes | study guides - st.edu.ge Pass4sure Certification Exam Questions and Answers - www.jabbat.com Killexams Exam Study Notes | study guides - www.jabbat.com Pass4sure Certification Exam Questions and Answers - www.jorgefrazao.esy.es Killexams Exam Study Notes | study guides - www.jorgefrazao.esy.es Pass4sure Certification Exam Questions and Answers and Study Notes - www.makkesoft.com Killexams Exam Study Notes | study guides | QA - www.makkesoft.com Pass4sure Exam Study Notes - maipu.gob.ar Pass4sure Certification Exam Study Notes - idprod.esy.es Download Hottest Pass4sure Certification Exams - cscpk.org Killexams Study Guides and Exam Simulator - www.simepe.com.br Comprehensive Questions and Answers for Certification Exams - www.ynb.no Exam Questions and Answers | Brain Dumps - www.4seasonrentacar.com Certification Training Questions and Answers - www.interactiveforum.com.mx Pass4sure Training Questions and Answers - www.menchinidesign.com Real exam Questions and Answers with Exam Simulators - www.pastoriaborgofuro.it Real Questions and accurate answers for exam - playmagem.com.br Certification Questions and Answers | Exam Simulator | Study Guides - www.rafflesdesignltd.com Kill exams certification Training Exams - www.sitespin.co.za Latest Certification Exams with Exam Simulator - www.philreeve.com Latest and Updated Certification Exams with Exam Simulator - www.tmicon.com.au Pass you exam at first attempt with Pass4sure Questions and Answers - tractaricurteadearges.ro Latest Certification Exams with Exam Simulator - addscrave.net Pass you exam at first attempt with Pass4sure Questions and Answers - alessaconsulting.com Get Great Success with Pass4sure Exam Questions/Answers - alchemiawellness.com Best Exam Simulator and brain dumps for the exam - andracarmina.com Real exam Questions and Answers with Exam Simulators - empoweredbeliefs.com Real Questions and accurate answers for exam - www.alexanndre.com Certification Questions and Answers | Exam Simulator | Study Guides - allsoulsholidayclub.co.uk © Search4Exams.com 2017. All Rights Reserved !