If you're unable to download the latest version of Internet Explorer, please ensure you have compatibility view turned off. How to disable compatibility view • Open Internet Explorer. • Click on the Tools menu (you may have to press Alt to bring up the menu). • If the Compatibility View box is checked, click to remove checkmark. • Click Tools again. • Select Compatibility View Settings. • Make sure 'rogers.com' is not listed under 'Websites you've added to Compatibility View.' • Close and reopen Internet Explorer. • Go to rogers.com. Incoming calls can be automatically forwarded to a number of your choice. Activate • From your home phone, dial *72. Listen for three beeps and then a dial tone again • Dial the number to which you want your calls forwarded • When that phone is answered, you will hear an intermittent dial tone confirming that call forwarding is activated Note: If the number is busy or goes unanswered, hang up and repeat the process in 2 minutes. Deactivate • From your home phone, dial *73 • Listen for 2 beeps • Hang up Downloads. ![]() ![]() What is Call Forwarding for my mobile phone?Call Forwarding. How to use Call Forwarding on my mobile phone? Phone to another number, such as your home. With Rogers One Number, you can quickly and easily set up call forwarding. Send all your calls to one wireless number, to the cottage phone or, better yet, to someone. ![]()
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![]() Net user administrator. Using WMIC commands. To enable administrator account: wmic useraccount where name='administrator' set disabled='false'. To disable administrator account: wmic useraccount where name='administrator' set disabled='true'. In Windows XP, administrator account is not disabled as in Vista and. Make it twice as hard for a potential hacker to get into your system by renaming the Administrator account. Here's how to do so in Windows XP Home and Pro. Unhide the Administrator Account in Windows XP; Login to Windows XP with No Password Administrator Enable Administrator Account in Windows with Local. How to Login to Windows XP as the Administrator. The Administrator account is required in order to install programs and make changes to most Windows settings. If you're using your own computer, chances are that your account is already an. Note For upgrade installations, the built-in Administrator account is kept enabled when there is no other active local Administrator on the computer. However, the built-in Administrator account is disabled by default for new installations and upgrades on domain-joined computers, regardless of whether there are other active local Administrators on the domain-joined computers. In audit mode, Windows Setup will implicitly enable the built-in Administrator account as the last action in the auditSystem configuration pass if the built-in Administrator is not already enabled. The first action in the auditUser configuration pass is to disable the built-in Administrator account. This enables you to run programs and applications as an Administrator. When you complete your customizations in audit mode and log out, the built-in Administrator account will be disabled. Unless you want to explicitly leave the built-in Administrator account enabled, there’s no need to re-enable the built-in Administrator account in audit mode. Enable the Built-in Administrator Account for Windows Vista There are two ways to enable the built-in Administrator account. • Use the AutoLogon unattended Setup setting You can enable the built-in Administrator account during unattended installations by setting the AutoLogon setting to Administrator in the Microsoft-Windows-Shell-Setup component. This will enable the built-in Administrator account, even if a password is not specified in the AdministratorPassword setting. You can create an answer file by using Windows System Image Manager (Windows SIM). The following sample answer file shows how to enable the Administrator account, specify an Administrator password, and automatically log onto the system. SecurePasswd123 true Administrator true 5 SecurePasswd123 true • Use the Local Users and Groups MMC console Change the properties of the Administrator account by using the Local Users and Groups Microsoft Management Console (MMC). ![]() ![]() ![]() • Open the MMC console and select Local Users and Groups. • Right-click the Administrator account and select Properties. The Administrator Properties window appears. • On the General tab, clear the Account is Disabled check box. • Close the MMC console. Administrator access is now enabled. Enable the Built-in Administrator Account for Windows Server 2008 For Windows Server® 2008, the built-in Administrator password must be changed at first logon. This prevents the built-in Administrator account from having a blank password by default. Both Microsoft-Windows-Shell-Setup| Autologon and Microsoft-Windows-Shell-Setup| UserAccounts| AdministratorPassword sections are now needed for autologon in audit mode to work. Both of these settings should be added to the auditSystem pass. The following XML output shows how to set the appropriate values. YourBApasswordhere true true Administrator yourBApasswordhere true Microsoft-Windows-Shell-Setup| UserAccounts| AdministratorPassword can be used in oobeSystem pass to prevent having to enter a password for the built-in Administrator account after you complete the out-of-box experience. The following XML output shows how to set the appropriate values. YourBApasswordhere true Disabling the Built-in Administrator Account Original equipment manufacturers (OEMs) and system builders are required to disable the built-in Administrator account before delivering the computers to customers. • Run the sysprep /generalize command When you run the sysprep /generalize command, the next time the computer starts, the built-in Administrator account will be disabled. -or- • Use the net user command Run the following command to disable the Administrator account. Net user administrator /active:no You can run this command after configuring the computer, before delivering the computer to a customer. In the login screen in XP, one can enter the administrator account by pressing ctl-alt-del two times. That does not work in Windows 7. In fact, there doesn't appear to be the same type of built-in administrator account that exists in XP. I know that you can do certain things in any User Account by doing them as 'administrator', but that doesn't mean there is an account named 'administrator' 1) How do the administrator accounts differ between Win7 & XP. 2) Is there an actual or 'built-in' administrator account in Windows 7? Any assistance with this would be greatly appreciated. In the login screen in XP, one can enter the administrator account by pressing ctl-alt-del two times. That does not work in Windows 7. The 2 x 'clt-alt-del' only works in the PRO version of XP, and not in the Home edition. Same in Windows 7. If you have the HOME PREMIUM edition there is no 2x 'ctl-alt-del' function, but AFAIK it does exist in Professional and Ultimate. In fact, there doesn't appear to be the same type of built-in administrator account that exists in XP. I know that you can do certain things in any User Account by doing them as 'administrator', but that doesn't mean there is an account named 'administrator' 2) Is there an actual or 'built-in' administrator account in Windows 7? The built-n Administrator account IS there in Windows 7, but it is disabled by default, same as the Guest account is. Create your own administrator account and call it 'Admin' or 'Tech' or something similar, give it a strong password and leave it alone, to be used for emergency access and elevation purposes only. (Windows 7 has the 'Run as Administrator' command and UAC so there's no need to use an Admin account on a daily basis at all) If you find my response helpful, please click on the 'Vote as Helpful' button!Thank you! Adobe Acrobat Pro version 8 is not officially supported on Windows 7. These instructions describe how to work around the installation problems and get Acrobat Pro installed and patched to version 8.3.1. Note that I've successfully installed Acrobat Pro on Windows 7 64-bit, however the same approach may apply for 32-bit Windows. Additionally Adobe Acrobat Pro is compatible with Windows 10. I've downloaded the installation program from the Adobe web site here: Note the software and license keys are provided free, under the rationale of supporting existing customers now that the. Run the installer executable. When asked for the folder location to extract files, do not choose the default location (since you won't have permission to save files there). Instead you'll get an error like this. ![]() A problem occurred while extracting some files. Check available space on your computer and the write privileges on the destination folder. Follow these steps to complete the installation successfully: • Instead choose an alternate temporary location like C: Acrobat8. • After you're advised the Files are Ready, select the option to Launch Adobe Acrobat 8 and click Finish. • I've left Adobe Reader as my default PDF viewer as this is a later version. • Enter your personal details, and your serial number. • Then choose the installation type. I've chosen a Typical installation. • During installation, a dialog will complain that the file ' AdobePDF.dll' is needed. • Download the file ' AdobePDF.dll_64' from the following Adobe support page (or extract that file from data1.cab as instructed). • Rename the file to ' AdobePDF.dll' and browse for that file in the installer. After installation completes you will have version 8.0.0 active. You can then proceed with applying patches to upgrade to version 8.3.1. The upgrade files are downloaded from the Adobe web site here: The specific files required are 8.1 (English), 8.1.1, 8.1.2, 8.1.3, 8.1.5, 8.1.6, 8.1.7, 8.2, 8.2.2, 8.2.3, 8.2.5, 8.2.6, 8.3 and 8.3.1. Install them in the correct order. You'll probably be requested to reboot multiple times. I ignored most of these requests when it became too time consuming and just rebooted after the last patch. I've done some basic edits in Acrobat and so far it works perfectly for me. UPDATE 4/2015: Well I finally got Adobe Acrobat Professional 8 (free) to work on my 64 bit windows 7 system. This is my OneNote file in case I ever need to reload it. Here is what worked for me. This will take some time, but I love Acrobat and its features, so it's worth it to me. On my Desktop I still use AAP7 and it works great, so I thought I'd upgrade to AAP8 on my laptop. ***During the launch process you will be asked for a registration #, use the below appropriate for you os - you can copy & paste the #. Windows Serial number: 1118-1414-1955-8737-8172-0350 Mac Serial number: 1118-0416-1130-3724-2927-7040 Note: this tutorial is for Windows 7 x64 (may be similar on other systems, use at your own risk) A. Download the now Free Adobe Acrobat Professional 8.0 for Windows or Mac B. Start the.exe program you just downloaded to start installation. You may have to let it go thru and fail once in order for the following files to be accessible to you. - you'll start it again once you have the proper file on your desktop. To install Acrobat 8 on Windows 7 (see location below in image) 1. Find folder data1.cab in installation files and go to 'AdobePDF.dll_64'. Right click and 'Extract to' desktop. Now it will be on desktop - this file will not let you rename it, so just leave it as it is. The best crack and download. Adobe acrobat xi pro crack windows 8. Adobe Acrobat XI Pro crack patch serial key free. Adobe acrobat xi pro crack windows 8. If available, uncheck/disable the option 'Check for updates automatically'. Installation notes: ******************* >> Disconnect internet during installation, and/or make sure to BLOCK internet access for the program (block unwanted traffic by using e.g. 1) Before installing Acrobat X Pro 10, apply all this steps. About TechWhirl. TechWhirl (TECHWR-L) is a resource for technical writing and technical communications professionals of all experience levels and in all industries. Start installation process again and when you get a pop-up asking for file AdobePDF.dll from Vista disk, just go to your desktop and click on AdobePDF.dll_64 file that you extracted there. It will process and open Acrobat program. BUT...go to pop-up and Cancel all the way out of pop-up (not installation process) that asked for adobepdf.dll from Vista disk as you have already installed it. The installation will continue processing, give it a moment and it will continue. Before selecting Finish, close the open AAP8 program that came up previously. Then click finish and program will open again. After installation verify version is 8.0 (which it should be) > Help >About Adobe Acrobat 4. Install patches/security updates from: Download all updates to your download folder - 8.1 thru 8.3.1 (13 updates) Go to your downloads folder: Start at 810 (8.1) and click to install, then move onto the next one. Will require several restarts/reboots (so do this when you have an extra hour or so). I did reboot after each install requesting it. Some have said they installed all & then rebooted, I didn't want to take the chance of confusing the registries. To Disable updates notifications because they really don't have any for you (v8) Go to Preferences > Updater and set a Check For Updates option to Do Not Check for download and install Updates 6. After AAP8 is open for a while it will ask you to download license. Make sure you are connected to internet. Click ok to download license. It does it automatically (nothing to download). If it doesn't work the first time, just click 'Try Again'. You may try to print to the Adobe PDF printer but it probably will not work at first, but should after all updates are installed. When printing to pdf, note that your taskbar will be have a flashing/blinking icon to 'save as' so you can name the file. ![]() BEST solution is to just 'save as' pdf - that works and it's easy. If you have any problems you can try to go to the Devices and Printers/Adobe PDF/Printer Properties/Advanced and set it to 'Print Directly to Printer'. I bought a wireless IP webcam off eBay recently. It has lots of features for the price. However, I've had problems with being unable to connect to the camera after a period of time (an hour or so). Power cycling the camera would fix the problem. I've tried upgrading to the latest (version 11.14.2.17 is supposed to fix WiFi disconnection problems), but it did not fix my problem. ![]() Adobe acrobat professional 8 free download - Adobe Acrobat Connect Professional, Adobe Acrobat Professional and Standard Update, Adobe Acrobat Pro DC, and many more. ![]() Problem analysis The problem is not related to my browser or the wireless network. I've tried disabling DHCP (router based static IP). It turns out the problem is that the webcam simply ceases to respond to requests (once in this broken state). The ARP protocol is a mechanism to translate between IP addresses and device MAC addresses. Without ARP, you cannot perform IP communications over Ethernet or WiFi (since all communications is actually between the device MAC addresses). The ARP protocol allows each network device to discover the MAC addresses of the peer dev. We recently got and found them to be noisy in the morning because they wanted to be let out of their coop. Of course, we just wanted to sleep in. What we needed was an automatic door opener. Here's our 3 hens (Edna, Dora and the bantam Dotty). Click for more photos. Here's the chicken coop, with the addition of an automatic door opener. The chicken coop was purchased from. There are some (expensive) automatic chicken doors that are available for purchase, but I decided to design and make my own. You are welcome to copy or adapt the design to suit your own requirements. Adobe Acrobat 8 Professional is quite old, but its pro features worth considering to remain in use and today can be downloaded for free with officially. Adobe Acrobat 8 Professional is useful to create PDF files and also to process (edit) the finished PDF file. PDF files are the standard format used in the exchange of documents via the internet. PDF file format can only be edited using the PDF editor software where this software is one of the famous. I don't know if you still have this problem but i found your question just the other day looking for an answer. Well now I found one! ![]() Please log in and follow the instructions to receive a new activation link. Get Help If you or someone you know needs help with a gambling problem. Aug 02, 2015 Windows 10 - How to use Draw and use WebNotes in the Edge browser - Drawing and Sharing tutorial. ![]() Money-saving coupons for adult tobacco smokers on Camel Crush, Camel Menthol and non-menthol cigarette styles. Coupons, special offers and promotions. Gifted in the prophetic and teaching ministry, the Lord uses Bruce to impart, activate, and release individuals and churches into deeper realms of the Spirit. The Father Heart of God is evidenced in Bruce's ministry as the presence and glory of the Lord is released drawing many back to the Father and causing many to draw. Without root none the less. The solution I found was in an other app called 'Settings Search'. I was tired of looking around for the right setting and installed this. Anyway, when going in from the app to the applications settings, I got in another application list apart from the one you get in the phones settings. ![]() In the cogwheel up in the right corner you can get to the 'Draw over other apps' settings. The Da Vinci Disappearance is a downloadable content addition for Assassin's Creed: Brotherhood. This is my very first tutorial.Watch and u will know how to do it. Song:Woodkid-Iron DLC Unlocker: IF THE LINK CANNOT BE OPENED. SEARCH IT IN thepiratebay.org Then Type this: Assassins Creed Brotherhood SKIDROW patch 1.01 DLC unlocker. It's the only one in there. This Dlc will automatically activated if u are at the end mission of Sequence 8 or 9 or after the long credits. ![]() Then u will see something at the map named 'Restored Memory'. Also there is 4 bonus costumes found in your inventory. AND HAVE FUN! All Credits goes to the one who post this on piratebay. And for the ubisoft. Assassin's Creed Brotherhood, DLC, PC, Ezio, Da vinci Disappearance. Halloween Special!, Download. ACB| GPS Achievement in Assassin's Creed The Ezio Collection: Complete The Da Vinci Disappearance. Xbox, Xbox Live, and all. Assassin's Creed: Brotherhood - The Da Vinci Disappearance Cheats, Codes, Action Replay Codes, Passwords, Unlockables for XBOX 360. Best archive of Assassin's Creed. A remastered version, along with Assassin's Creed II and Assassin's Creed: Revelations, was released for the PlayStation 4 and Xbox One on November 15, 2016. However, these two pre-order special characters are also included in The Da Vinci Disappearance downloadable content as are the formerly pre-order only. 3DS DS iOS PC PS3 PS4 PSP Switch Vita Wii U Xbox 360 Xbox One More Systems. Brotherhood - The Da Vinci Disaperance? The Da Vinci Disappearance. Enter The House of Da Vinci, a new must-try 3D puzzle adventure game. Solve mechanical puzzles, discover hidden objects, escape from rooms and dive into the authentic atmosphere of the Renaissance. Use all your wits to find out what's behind your master's disappearance. You are Da Vinci’s most promising apprentice. Your master, Leonardo, has disappeared. You have no idea where he’s gone or what has happened. So you begin your quest, your search for the truth. However, Leonardo's workshop is full of puzzles, inventions, escape mechanisms and objects hidden in all corners of beautifully decorated rooms. You’ll need to use all of your brain cells to find out what’s really going on! ![]() We’ve applied our own imagination to the story of the world’s greatest inventor, Leonardo da Vinci, providing players with unique special abilities throughout gameplay. We allow you to warp back into your early days as a young apprentice, exploring the 16th century world around you, peeking into Leonardo’s mechanisms, puzzles and devices, as you eventually become powerful enough to control time itself and learn the secrets hidden in the past. Many challenging puzzles are inspired by Leonardo Da Vinci's real inventions and ideas. Mysterious locations were created based on original artworks and the wonderful Florence, Italy of 1506. Features • SOLVE MECHANICAL PUZZLES Brain twisters and baffling riddles are all based on Leonardo’s inventions. • ENJOY INTUITIVE TOUCH CONTROL Navigate Leonardo’s workshop intuitively. Playing the game couldn’t be more fun. • EXPERIENCE THE RENAISSANCE War machines, complex lockboxes, mechanical puzzles, room escapes will all test your skills. • MASTER UNIQUE MECHANICS Detect hidden objects by seeing through the surfaces around you. • PEEK INTO THE PAST An unusual gauntlet allows you to witness events that happened earlier, and learn more of the story. Compare prices and buy The House of Da Vinci Steam CD Key at the best price. Kinguin offers best deals on. Offered products are for digital download and instant delivery Compare CD Key merchants. Activate The House of Da Vinci Steam CD Key on your Steam account to download the game and play. Find the best and cheapest game on Kinguin. • Go to: and download STEAM client • Click 'Install Steam' (from the upper right corner) • Install and start application, login with your Account name and Password (create one if you don't have). • Please follow these instructions to activate a new retail purchase on Steam: • Launch Steam and log into your Steam account. • Click the Games Menu. • Choose Activate a Product on Steam. • Follow the onscreen instructions to complete the process. After successful code verification go to the 'MY GAMES' tab and start downloading. ![]() ![]() Cullis Gates were magic portals invented in the times of the Old Kingdom that allowed Heroes to travel quickly from one place to the other. In Fable, Fable: The Lost Chapters and Fable Anniversary, the teleportation through Cullis Gates was immediate and the Guild Seal allowed the. Fable II FAQ/Walkthrough| GamesRadar+ [[===========================================================================]] There are many conflicting legends as to whether or not the Hero of Oakvale killed the Guildmaster. It's said that the words, 'Your health is low' were carved into his forehead. • • • • • • » » » » Darkwood Cullis Gate - Permanent Activation Question Show::::: [] Mon, 12 December 2011 10:58 Messages: 2 Registered: December 2011 Hello. Fairly new to modding Fable, though not modding itself. Anyway, what I would like to do is once you complete the quest that sends you to Hook Coast, make the Cullis Gate stay active and usable. I looked around the site a bit, saw how to make your own Gates, but nothing about editing that one in particular. It shouldn't be a difficult thing to do, but, well, even though I have backups, I'd rather not just blindly mod my way into a problem. One last note: I have some text issues when using Fable Explorer. Long story short, I'm heavily reliant on ID numbers, though directing me to things like a weapon's CWeaponDef or other various links works just fine. Thank you for the help. EDIT: Currently using only Fable Explorer, though if additional programs are required that shouldn't be a problem. [ is a reply to ] Sat, 17 December 2011 01:21 Messages: 1 Registered: December 2011 Hi, I'm a complete noob and have just gotten into modding Fable. I have FE and have used it to remove the weapons restriction on Bowerstone South and the like. What I'm trying to do is get to Hook Coast, Knothole Glade, or Witchwood early in the game rather than mid-game. Is there any possible way if like your saying activating the Darkwood Cullis Gate would glitch the game? I've looked everywhere and can't find anything. Thanks in advance. ![]() ![]() Activating Project Activation is Microsoft’s way of verifying that you’re running a legal copy of a Microsoft program. Running Project without activating it turns the program into little more than a viewer, and you can run the program for only a short period of time. (Although Project 2013 runs with reduced functionality before you activate it, Microsoft is starting to eliminate the activation grace period. In Windows 8, you need a product key to install the software.) When you activate Project, Microsoft doesn’t ask for personal information, so there’s no harm in doing so. Note The Microsoft Office Project Activation Wizard creates a hardware identifier for your computer, which means, unfortunately, that you may have to reactivate Project if you rebuild your computer. The Activation Wizard starts automatically when you run Project for the first time. In the Activate Office dialog box, click “Microsoft account” to sign in with the account associated with your Microsoft products (SkyDrive, Windows Phone, Hotmail, or other Microsoft services). ![]() If you have a Microsoft account through work or school, click “Organizational account.” Enter your username and password, and then click “Sign in.”. Feb 02, 2017 Summary: Deactivate user accounts by using the Manage Users page in Project Web App Settings. Oct 05, 2017 I downloaded the trial version of MS Project Pro 2013, and installed it. When it tries to start the activation process it gives me and error: 'Sorry, we. This troubleshooter runs on Windows PCs and can fix activation errors for Office 365, Office 2016, and individual Office 2016 apps, like Visio, Project, Word, and Outlook. If you use your device both at home and at work, try turning off the proxy settings in Microsoft Edge or Internet Explorer before you install Office. ![]() ![]() ![]() Activate and enable 3G internet network service instantly in 15 minutes on your new or old Vodafone SIM card with pre-activated 2G edge. I bought new SIM card from Vodafone store, which was not activated with 3G service. It was pre-activated with 2G service, but I wanted to get 3G speed. So searched on various website, blog, forums, same as you did. Finally your search ends, here is some cool steps to activate 3G service instantly* on your newly. Step #1: Change your Network Mode setting in your mobile to Automatic, so that it can change to 2G or 3G which ever network available. Step #2: Restart you mobile. Step #3: Now send message activation from you new SIM. “ ACT 3G” to 111 (without double quotes) or “ ACT 3G” to 144 (same, without double quotes). Note: If above SMS is not working, please try sending message: 3G to 111 Step #4: You will receive two messages continuously. Enjoy high speed Internet and downloads, Video Calls, Live Mobile TV, HD Gaming with Vodafone 3G. We have your request for 3G service and will revert shortly After awhile, you will receive second message 3G services on your phone will be activated in 24 Hrs. Note: In-case you receive any error message for using wrong keyword, as I got, try send same message to 144 it will work out. Step #5: Now wait for 10 minutes so that your request will be accepted and changes to propagate in Vodafone server. Sep 21, 2012 - 2 min - Uploaded by Austin EvansiPhone 5: How To Set Up, Activate & Insert / Remove SIM Card iPhone 5 Review: http://www. If you want to use a new SIM in your Verizon phone, you'll have to activate it first. After that you can use it with your phone. To do this you have to make a call to. Jan 11, 2011. Are we supposed to leave these in? Are these cards worthless in the States? I'm torn whether I should leave it in my phone or take it out. After a while again change you Network Mode settings to UTMS. Step #6: Now keep you mobile with SIM card switch off for 5 minutes and start it. Now you start data connection, and you will see 3G network working in your mobile phone. Hello editor I am using 2 Vodafone simcards (both are ported from different network). My handset is moto g xt-1033 (6months old). Complaint what I am getting is 3g coverage. It is not fluent. Some time I am getting 3g coverage (H+) sometime I am getting 2g(E). But I have made my preference as 3g. I don’t know why I am getting like this. Could you clarify and help to get continuous 3G. I am just within the range of 500 meter radius circle of that tower. ![]() I am staying in tamilnadu Note: I lost the hope of Vodafone technical assistance. Thanks in advance. This is because all the service providers have tied up for provide 3G network cross circles. Let me explain you in detail: In Maharashtra, Airtel is not a 3G service provider here. So as per cross 3G network circles, Airtel either uses Idea’s or Vodafone network to provide 3G to their customers, Hence roaming is shown. In any other circle if Airtel has and Vodafone or Idea network doesn’t have 3G network circles, they use Airtel’s 3G to provide 3G internet. Hence, customers of Vodafone or Idea will be shown as Roaming. This is just a mutual 3G connection between different networks in different circles. And in India, Roaming for Internet is totally free, so no need to panic. It was originally purchased from Vodafone and. I have an account on Verizon and when I use my valid SIM card. It does not stay connected to Verizon or activate. This tutorial will provide you with the required tips if you are struggling with Sprint, Verizon, Vodafone Netherland NL or any other CDMA iPhone 4s unlocking with Gevey sim. First of all, an iPhone 4s supports both GSM and CDMA network. However, an iPhone 4S from a CDMA network like Sprint, Verizon or Vodafone NL is configured to pick CDMA network first. This is called Preferred Roaming. All CDMA iPhone 4S has few files in it’s operating system which is configured with Preferred Roaming List (PRL) that forces the iPhone 4S to look for those CDMA network first before trying to connect to any GSM network. And, that’s where the problem starts and lot of people trying to unlock a CDMA iPhone 4S with gevey sim are struggling. So, what should you do to stop your iPhone from looking for CDMA network from PRL and connect directly to the GSM network you want it to? Answer is straight forward.you need to disable CDMA modem of your phone by removing PRL files. Once you are done with disabling CDMA modem, you can proceed with the normal gevey unlock instructions. Here are the steps you should follow before you try to unlock your CDMA iPhone 4S with gevey sim: 1. Jailbreak your iPhone with Absinthe or Redsn0w, 2. Once jailbroken, connect your iPhone to your computer and open iFunbox or any other iPhone file manager of your choice, 3. Go to var/wireless/Library then delete file “carrier_310VZW_20404_11.2.plist”. Please note that this example file name is for a Verizon iPhone, in Sprint iPhone the file name will have something like SPRNT instead VZW in it, 4. Go to System/Library/Carrier Bundle/iPhone delete folder “Zeppelin_US.bundle” and “ZeppelinDAS_US.bundle”. Again, this folder example is from a Verizon iPhone, Sprint iPhone will have a different name accordingly, 5. Delete your sim card carrier bundle in the same folder (if you have Sprint iPhone, delete folder sprint_bundle. In case your iPhone does not have this folder, just skip this step). You are done with disabling CDMA modem of your iPhone and it’ll not register to any CDMA network automatically. Now you can proceed with the normal gevey sim unlock procedure. I have personally tested it with Sprint and Verizon iPhone 4S and it works perfect. Please note that if you are trying to unlock a Verizon phone, you also need to delete Vodafone NL related files along-with Verizon files. Similarly, if you are trying to unlock Vodafone NL, you need to delete Verizon files as well along-with Vodafone NL files. These are initial steps to rule out any incompatibility issue. Now once you have performed these steps. Move on to our next tutorial on. Dominick, that is what I really need – use my iPhone 4S in GSM network only. I don’t care about CDMA, even if it is locked forever. I deleted both var/wireless/Library/carrier_310VZW_20404_11.3.plist and bundles from System/Library/Carrier Bundle/iPhone (Zeppelin bundles and all Vodafone bundles). But it didn’t help yesterday. Today I started with GEVEY+white reset SIM, then GEVEY+my SIM – it showed signal, but falls into Activation and says “SIM Not Valid”. Not sure why I was not able to see it yesterday. Anyway, I have no idea how to activate it. I have an original Verizon SIM. Could you please advise how to activate? Thanks in advance! Hi Alex, the procedure mentioned above about deleting default carrier bundle is just to prevent the phone from trying to connect to default CDMA network whereas you intent to use it on a gsm network. This also helps in Gevey sim completing the unlock procedure smoothly. The activation error you are getting has nothing to do with these steps. It means either you have not followed the unlock instructions you got with your Gevey or the iOS version of your phone is not supported by the Gevey sim you are trying to use. You need to check if you are properly following the unlocking steps using Gevey Sim. Refer to the instructions you received with the sim. Please note that steps to follow differs depending on which Gevey sim you are using. I am having the same problem. My iPhone 4s has been unlocked and working fine for months now and when I connect it to itunes it always asks to update the carrier settings for my phone, I of course always decline but a few days ago I accidentally accepted. This is where I began to have the same problem Alex was having. When my phone was unlocked earlier I deleted the carrier_310VZW_20404_11.2.plist (.2 version) and the phone went through the process and unlocked. Now when I delete the carrier_310VZW_20404_11.3.plist (.3 version) it seems to always come back even after I delete it and I cant unlock the phone. What happens is I delete all the correct carrier files, restart the phone wait about 20 seconds and the phone activates. I can then access the phone and the gevy black screen comes up I click accept and it no service I wait for about 1 minute for Gevy to try and find service and then a box comes up and says phone can not be unlocked contact your carrier if this problem continues. The phone then sends me to the activation loop but I then have service bars without my service provider showing, but the phone becomes locked in activation mode. I think somehow there is another script telling the phone to reload/redownload these carrier bundles after they have been deleted which forces me back into the activation loop once Gevy picks up a signal. A Requirement Of Nuclear Factor-kappa B Activation In Fear-potentiated Startle download free1/17/2018 Abstract Previous studies have shown that biochemical changes that occur in the amygdala during fear conditioning in vivo are similar to those occur during long term potentiation (LTP) in vitro. Electrophoretic mobility shift assay of nuclear extracts from startle-potentiated rats showed a selective increase in the amygdala of nuclear factor-κB (NF-κB) DNA binding activity. Supershift experiments further indicated that p65 and p50 subunits but not c-Rel were involved in DNA binding. The protein levels of IκB-α were reduced by treatments that reliably induced LTP in this area of the brain. This was accompanied by a decrease of NF-κB in the cytoplasm concomitant with an increase in the nucleus. Quantitative analysis of IκB kinase activity demonstrated that fear training led to an increase in kinase activity, and this effect was inhibited by thalidomide. Paralleled behavioral tests revealed that thalidomide inhibited fear-potentiated startle. Intra-amygdala administration of κB decoy DNA prior to training impaired fear-potentiated startle as well as LTP induction. Similarly, NF-κB inhibitors blocked IκB-α degradation and startle response. These results provide the first evidence of a requirement of NF-κB activation in the amygdala for consolidation of fear memory. It is generally believed that consolidation of long term memory in mammalian brain and long term facilitation in Aplysiarequire new protein synthesis (). Newly synthesized proteins are thought to deposit at the synapses that have been tagged by prior activity to encode enduring changes in synaptic strength (, ). However, despite the importance of new protein synthesis for memory consolidation, little is known about signaling pathways leading to protein translation in neurons. NF-κB, originally identified as a regulator of immunoglobulin κ light chain gene expression, is a DNA-binding factor that functions as a dimer. Five mammalian members of the family have been identified; p50/NF-κB1, p65/RelA, c-Rel, RelB, and p52/NF-κB2 (). NF-κB was localized mainly to the cytoplasm in an inactive form bound to an inhibitory protein termed IκB (). Upon stimulation by extracellular inducers, IκB was rapidly phosphorylated by the IκB kinase (IKK) complex on the serine residues 32 and 36. This phosphorylation led to the ubiquitination and subsequent degradation of IκB by the proteasome followed by nuclear translocation of NF-κB (). Once translocated to the nucleus, NF-κB bound cognate DNA sequences and activated transcription of specific target genes, the majority of which encoded proteins important in immunity and inflammation (). Apart from its role in hematopoiesis, accumulated evidence indicates that NF-κB is involved in neuroprotection or neurodegeneration, depending on the particular system under investigation and the NF-κB subunits that became activated (). Furthermore, a recent study implicated that NF-κB played an important role in the synaptic plasticity, because pretreatment of hippocampal slices with κB decoy DNA prevented induction of long term depression (LTD) 1 and significantly reduced the magnitude of LTP (). We have recently demonstrated that acquisition of fear was associated with an activation of phosphatidylinositol 3-kinase (PI 3-kinase) and its downstream target Akt in the rat amygdala (). PI 3-kinase and Akt were also activated in response to LTP-inducing tetanic stimulation (TS). In parallel, PI 3-kinase inhibitors interfered with TS-induced LTP as well as long term fear memory formation. Akt has been shown to induce the activation of NF-κB family of transcriptional factors (, ). Therefore, it is of interest to investigate whether NF-κB plays any role in synaptic plasticity and memory formation. Here we show that NF-κB is activated in the amygdala following fear-potentiated startle, and disruption of this signaling pathway impairs fear memory. Surgery Rats (4–5 weeks old) were anesthetized with sodium pentobarbital (50 mg/kg intraperitoneally) and subsequently were mounted on a stereotaxic apparatus. Two cannula made of 22-gauge stainless steel tubing (C313G; Plastic Products) were implanted bilaterally into the lateral (LA) or basolateral (BLA) amygdala (anteroposterior, −2.8 mm; mediolateral, ±4.5 mm; dorsoventral, −7.0 mm) (). A 28-gauge dummy cannula was inserted into each cannula to prevent clogging. Three jewelry screws were implanted over the skull, serving as anchors and the whole assembly was affixed on the skull with dental cement. The rats were monitored and handled daily and were given 7 days to recover. NF-κB inhibitors were administered bilaterally in a volume of 0.5–0.8 μl at a rate of 0.5 μl/min. ΚB decoy and scrambled DNA were prepared and administered based on the method of Blondeau et al. In brief, double-stranded κB decoy DNA was prepared by annealing complementary single strands with the sequences of 5′-GAGGGGACTTTCCCT-3′. Control DNA with a scrambled sequence was prepared by annealing oligonucleotides of the following sequence: 5′-GATGCGTCTGTCGCA-3′. Stocks of double-stranded DNA were prepared at a concentration of 2 m m in saline. ![]() A Requirement of Nuclear Factor-κB Activation in Fear-potentiated. B, comparison of fear-potentiated startle between thalidomide- and vehicle-treated rats. Which is required for NF-κB activation to. ΚB activation in fear-potentiated startle. 6 levels through the nuclear factor-kappa B pathway. ΚB decoy and scrambled DNA (60 μg) were infused into the amygdala at a rate of 0.5 μl/min (two injections at 24 and 2 h before conditioning). Fear Conditioning Fear conditioning was measured using the potentiated startle paradigm in a startle apparatus (San Diego Instrument, San Diego, CA) as described previously (, ). Briefly, the animal was constrained in a Plexiglas cylindrical tube (length, 20 cm; diameter, 10 cm) with an accelerometer sensor attached on the base. The whole set-up was enclosed in a ventilated, sound-attenuating cabinet (length, 38 cm; width, 38 cm; and height, 55 cm). The acoustic startle stimulus was a 50-ms white noise at an intensity of 95 dB. ![]() The visual conditioned stimulus was a 3.7-s light produced by an 8-watt fluorescent bulb attached to the back of stabilimeter. The unconditioned stimulus, which serves as the aversive component necessary for fear conditioning, was a 0.6-milliamp foot shock with a duration of 0.5 s. Startle amplitude was defined as the maximum accelerometer voltage that occurred during a 200-ms period after the onset of the startle stimulus. Seven days after surgery, each rat was exposed to habituation sessions over two consecutive days. In these sessions, rats were placed in the startle box and 3 min later presented with 10 startle stimuli at 2-min intertrial intervals. On the basis of their mean startle amplitudes in the second of these two sessions, rats were matched into groups with similar response levels. On the training day, rats were placed in the startle chamber and received 10 light-foot shock pairings. Unpaired controls received the same number of light and foot shock presentation but in a pseudorandom fashion in which the unconditioned stimulus could occur at anytime except at 3.2 s following the conditioned stimulus. Slice Preparation and Extracellular Recordings Male Sprague-Dawley 4–6-week-old rats were decapitated, and their brains were rapidly removed and placed in cold oxygenated artificial cerebrospinal fluid (ACSF) solution. Subsequently, the brain was hemisected, and transverse slices of 450-μm thickness were made. ACSF solution had the following composition: 117 m m NaCl, 4.7 m m KCl, 2.5 m m CaCl 2, 1.2 m m MgCl 2, 25 m m NaHCO 3, 1.2 m m NaH 2PO 4, and 11 m m glucose. ![]() ![]() The ACSF was bubbled continuously with 95% O 2, 5% CO 2 and had a pH of 7.4. Extracellular field potentials were made by electrical stimulation of the external capsule, which contained fibers from the auditory cortex to the lateral amygdala, with a concentric bipolar stimulating electrode (SNE-100; Kopf Instruments, Bern, Germany). Electrical stimuli (150 μs in duration) were delivered at a frequency of 0.05 Hz. Base-line field potentials were adjusted to ∼30–40% of the maximal responses. LTP was elicited by four trains of tetanus (100 Hz, 1 s at 3-min intervals) at the same stimulation intensity used for base line. For κB decoy DNA experiments, amygdala slices from each rat were divided into two groups. Slices in the treatment group were incubated with 50 μ m κB decoy DNA for at least 3 h before recordings were made. Control slices were incubated for a similar length of time with scrambled DNA or normal ACSF in different beakers. Western Blot Analysis The lateral and basolateral subregions of the amygdala were sonicated briefly in ice-cold buffer (50 m m Tris-HCl, pH 7.5, 0.3 m sucrose, 5 m m EDTA, 2 m m sodium pyrophosphate, 1 m m sodium orthovanadate, 1 m mphenylmethylsulfonyl fluoride (PMSF), 20 μg/ml leupeptin, and 4 μg/ml aprotinin). In the LTP experiments, slices were given four trains of TS or were treated with forskolin for 15 min, the protocols that reliably evoked LTP in this area of the brain. Subsequently, slices were removed from the recording chamber at various time points as indicated after the last TS or the washout of forskolin. Following sonication, the samples were centrifuged at 7500 rpm for 15 min, and the supernatant was obtained after pelleting the crude membrane fraction by centrifugation at 50,000 rpm for 1 h at 4 °C. Protein concentration in the soluble fraction was then measured using a Bradford assay, with bovine serum albumin as the standard. Equivalent amounts of protein for each sample were resolved in 8.5% SDS-polyacrylamide gels, blotted electrophoretically to Immobilon membranes, and blocked overnight in TBS buffer (50 m mTris-HCl, pH 7.5, 150 m m NaCl) containing 3% bovine serum albumin. For detection of NF-κB and IκB-α, blots were incubated with polyclonal antibodies for p50, p65, or IκB-α (Santa Cruz Biotechnology, Inc., Santa Cruz, CA). An enhanced chemiluminescence kit (Amersham Biosciences) was used for detection. Western blots were developed in the linear range used for densitometry. The density of the immunoblots was determined by an image analysis system (Vilber Lourmat). Nuclear Extract Preparation After the rats were killed, the brains were rapidly removed, and the LA and BLA were dissected out using a Dounce grinder with a loose pestle in ice-chilled buffer (15 m m HEPES, 60 m m KCl, 1 m m NaCl, 0.25 m sucrose, 5 m m EDTA, 1 m mEGTA, 1 m m PMSF, 10 μg/ml aprotinin, 15 μg/ml leupeptin, 2 m m NaF, and 1 m m sodium orthovanadate). The homogenate was centrifuged for 10 min at 2000 rpm, and pellet was resuspended in buffer (10 m m HEPES, pH 7.2, 15 m m MgCl 2, 10 m m KCl, 1 m m PMSF, 2 m m NaF, 15 μg/ml leupeptin, and 1 m m sodium orthovanadate). After a brief vortexing, they were incubated on ice for 10 min and lysed with a tight pestle. The homogenate was centrifuged at 4000 rpm for 10 min. The pelleted nuclei were resuspended in 40–60 μl of extraction buffer consisting of 100 m m HEPES, pH 7.2, 1.5 m mMgCl 2, 1 m m EDTA, 0.8 m NaCl, 15% glycerol, 2 m m NaF, 1 m m PMSF, 15 μg/ml leupeptin, 1 m m sodium orthovanadate and were incubated on ice for 2–4 h. The nuclear suspension was centrifuged at 14,000 rpm for 30 min at 4 °C, and the supernatant was saved. Electrophoretic Mobility Shift Assay (EMSA) EMSA was performed on the amygdala neurons isolated from unpaired control or conditioned animals at specific times after training. For binding reactions, 10 μg of protein was incubated in binding buffer (0.05% Nonidet P-40, 10% glycerol, 10 m m HEPES, pH 7.9, 50 m m KCl, 0.1 m m EDTA, 2.5 m mdithiothreitol, and 0.25 mg/ml poly(dI-dC)) for 15 min at room temperature. The double-stranded NF-κB consensus oligonucleotide (5′-AGTTGAGGGGACTTTCCCAGGC-3′) (Santa Cruz Biotechnology) was end-labeled with T4 polynucleotide kinase and [ 32P]ATP. Probe specificity has been thoroughly documented previously (). Radiolabeled oligonucleotide was added to the reaction mixture and incubated for 20 min. In the supershift experiments, the nuclear extracts were incubated with 1 μg of polyclonal antibodies against the p65, p50, and/or c-Rel proteins (Santa Cruz Biotechnology) for 30 min before the binding reaction. The specificity of binding was demonstrated by the addition of cold competitor or cAMP-response element-binding protein (CREB) probe (5′-AGAGATTGCCTGACGTCAGAGAGCTAG-3′). The reaction products were analyzed by electrophoresis in a 4% polyacrylamide gel with 0.5× TBE buffer (22.3 m m Tris, 22.2 m m borate, and 0.5 m m EDTA). The dried gels were analyzed by autoradiography after an overnight exposure. IKK Kinase Assay IKK activity was quantitated using an immune complex kinase assay as described previously (). Cells were prepared from unpaired conditioned or conditioned rats given thalidomide (10 μg dissolved in 1.0 μl of saline, 0.5 μl per side) 30 min before training. Equal amounts of whole cell extracts were immunoprecipitated using an antibody against the β subunit of IKK. Kinase activity was determined in kinase buffer (20 m mHepes (pH 7.7), 2 m m MgCl 2, 2 m mMnCl 2, 10 μ m ATP, 1–3 μCi of [γ- 32P]ATP, 10 m m β-glycerophosphate, 10 m m NaF, 10 m m PNPP, 300 μ mNa 3VO 4, 1 m m benzamidine, 2 μ m PMSF, aprotinin at 10 μg/ml, leupeptin at 1 μg/ml, pepstatin at 1 μg/ml, and 1 m m dithiothreitol) at 30 °C for 30–60 min in the presence of the indicated substrate. The kinase reaction was stopped by the addition of 5× SDS-PAGE sample buffer, subjected to SDS-PAGE analysis, and visualized by autoradiography. IκB-α-(1–317) substrate (Santa Cruz Biotechnology) was expressed and purified from Escherichia coli with glutathione-agarose affinity chromatography. Figure 1 Fear-potentiated startle results in a selective activation of DNA binding of NF-κB in the amygdala. A, gel shift analysis of nuclear extracts of the amygdala from paired and unpaired rats using consensus sequence of NF-κB at different time points after training. B, rats were given 10 pairings of light and foot shock, and DNA binding activity of NF-κB was analyzed using EMSA from nuclear extracts of the amygdala, hippocampus, and cerebellum. NF-κB was activated selectively in the amygdala but not in the hippocampus or cerebellum. The data shown are representative of six independent experiments (one rat per experiment in each time point). Regulation of NF-κB may be a general phenomenon across the entire brain. To determine whether it was specific for the amygdala, additional experiments were performed to measure NF-κB DNA binding activity in the hippocampus and cerebellum. As illustrated in Fig. B, training of visual conditioned fear did not lead to an increase in NF-κB binding activity in the hippocampus and cerebellum. These results indicate that regulation of NF-κB activity occurs in the region of the brain involved in the formation of fear memory. Previous studies have shown that biochemical changes that occur in areas LA and BLA during fear conditioning in vivo are similar to those occur during LTP in vitro (). We next determined whether forskolin or TS, the protocols that reliably evoked LTP in this area of the brain, produced a similar effect as fear training. Slices were treated with forskolin (50 μ m) for 15 min, and nuclear extracts prepared from the LA and BLA were assessed for NF-κB activation using EMSA. A shows that NF-κB binding activity was eliminated in the presence of a 100-fold molar excess of unlabeled κB DNA. In contrast, unlabeled CREB probe had no effect demonstrating the specificity of binding. To identify which NF-κB family members that were binding, we performed supershift experiments. A revealed that both p65 and p50 antibodies retarded the migration of proteins interacting with the NF-κB oligonucleotide. Similar experiments were performed following TS. As shown in Fig. B, supershift assay indicated that the major NF-κB complex induced by TS was p65/p50 heterodimer. Interestingly, CREB antibody also slightly retarded the migration, suggesting the possible interaction between NF-κB and CREB (see below). Figure 2 Supershift analysis of DNA binding activity of NF-κB. A, slices were treated with forskolin (50 μ m) for 15 min, and nuclear extracts from the LA and BLA were analyzed with EMSA using radiolabeled NF-κB probe. DNA binding activity of NF-κB was eliminated in the presence of a 100-fold excess of unlabeled NF-κB probe but was not affected by unlabeled CREB probe. Supershift experiments with anti-p65 and anti-p50 demonstrated that the protein complex interacting with the NF-κB oligonucleotide contains a p65/p50 heterodimer. B, slices were given four trains of TS, and nuclear extracts were preincubated with antibodies raised against various NF-κB subunits and CREB antibody. The data shown are representative of five independent experiments. TS or Forskolin Treatment Induces Degradation of IκB-α The induction of the active state of NF-κB involves phosphorylation and subsequent degradation of IκB-α (). We performed Western blot analysis to test what effects TS or forskolin treatment have on the regulation of IκB-α. A shows that IκB-α protein level was degraded by 25.7 ± 1.8% ( n = 6, p. Figure 3 Induction of IκB-α degradation by TS or forskolin. Slices were given four trains of TS ( A) or were treated with forskolin (50 μ m) for 15 min ( B). Cytoplasmic extract proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride membrane. The membrane was analyzed for degradation of IκB-α with an anti-IκB-α antibody. C, suppression of IκB-α degradation by wortmannin and TLCK. Slices were treated with wortmannin (100 n m) or TLCK (50 μ m) 30 min prior to application of forskolin. Cytoplasmic extracts were analyzed for degradation of IκB-α as stated in B. We further examined the effect of TLCK on the NF-κB DNA binding activity. Pretreatment of TLCK before training blocked conditioning-induced binding of NF-κB (Fig. However, the direct addition of TLCK to the nuclear extracts from the conditioned rats did not affect the binding of NF-κB to the DNA. These data indicate that suppression of NF-κB binding by TLCK is not due to interference between the transcription factor and the DNA but rather that TLCK acts specifically to inhibit conditioning-induced NF-κB activity. Fear Training Activates IKK Activity IKK is the enzyme responsible for the phosphorylation of IκB-α; therefore, we examined the regulation of IKK activity in behaviorally trained rats. By using agarose-conjugated antibody against the β subunit of IKK complex, IKK was immunoprecipitated from whole cell extracts of rats that had been conditioned. Immunoprecipitates were assayed for kinase activity by incubating with IκB-α (amino acids 1–317) fusion protein in the presence of [γ- 32P]ATP, electrophoresed on a polyacrylamide gel, and analyzed by autoradiography. Shows that fear training led to an increase in kinase activity by 10 min that continued through the 30-min time point. It has been shown that anti-inflammatory and antiangiogenic effects of thalidomide were based on its ability to block NF-κB activation through suppression of IKK activity (). We tested whether thalidomide produces a similar effect on the central neurons by administration of this drug (10 μg dissolved in 1.0 μl of saline, 0.5 μl per side) 30 min before training. Quantitative analysis of conditioning-induced IKK activity demonstrated that thalidomide inhibited the increase at 10 ( p. Figure 4 Fear conditioning activates IKK activity. Rats were given light and foot shock in a paired or unpaired manner. Thalidomide (10 μg dissolved in 1.0 μl of saline, 0.5 μl per side) was injected bilaterally into the amygdala 30 min before training. A, whole cell extracts were prepared and immunoprecipitated with an anti-IKKβ antibody. Immunoprecipitates were incubated with IκB-α-(1–317) fusion protein in the presence of [γ- 32P]ATP. The immunoprecipitates were run on a 8.5% SDS-PAGE gel, dried, and exposed to film. Translocation of NF-κB to the Nucleus Amygdala slices were given four trains of TS or were incubated with 50 μ mforskolin for 15 min, and extracts of treated cells were separated into cytoplasmic and nuclear fractions and analyzed by immunoblotting assay. A shows that NF-κB p50 protein was reduced to 48.0 ± 3.9 ( n = 6, p. NF-κB Is Required for Fear-potentiated Startle To investigate whether the activation of NF-κB was not only correlated but also obligatory for conditioned fear, we microinjected TLCK (0.4, 20, or 50 m m, 0.5 μl per side) bilaterally into the LA or BLA 30 min before training. Fear retention was tested 24 h after training. A shows that TLCK dose-dependently impaired fear memory. The analysis of variance for startle scores revealed a significant effect for dose ( F (4,25) = 8.10; p 0.05). Similarly, intra-amygdala infusion of another NF-κB inhibitor, N-tosyl- l-phenylalanine chloromethyl ketone (TPCK; 100 m m, 0.5 μl per side) blocked the acquisition of fear memory ( n = 6, p. Figure 6 Effect of intra-amygdala administration of NF-κB inhibitors on fear conditioning. A, rats were microinjected with TLCK (0.4, 20, or 50 m m, 0.5 μl per side) or TPCK (100 m m, 0.5 μl per side) bilaterally into the LA or BLA 30 min before training. Fear retention was tested 24 h after training. Fear-potentiated Startle and LTP Were Markedly Reduced in Rats Treated with κB Decoy DNA We administered double-stranded κB decoy DNA or control double-stranded DNA with a scrambled sequence bilaterally into the amygdala based on the method of Yu et al. After conditioning, the rats were sacrificed, and nuclear extracts from the amygdala were incubated with radiolabeled NF-κB consensus DNA sequence. As shown in Fig. A, the conditioning-induced increase in NF-κB DNA binding activity was not seen in rats that were given κB decoy DNA. Furthermore, quantification of startle potentiation 24 h after conditioning revealed a marked depression of the degree of potentiation (48.3 ± 14.8% of potentiation, n = 6) as compared with those receiving scrambled DNA (205.1 ± 29.6%, n = 6, p. Figure 7 Blockade of fear-potentiated startle and amygdala LTP by κB decoy DNA. A, nuclear extracts from conditioned or conditioned rats given κB decoy or scrambled DNA 24 and 2 h prior to training were analyzed by EMSA for NF-κB DNA binding activity. B, rats were either not administered (sham-operated) or were given κB decoy DNA or scrambled DNA 24 and 2 h before training and tested 24 h later. Administration of κB decoy DNA impaired fear memory, which could be reversed when the rats were retrained 3 days after the initial testing. C, LTP induced by TS in the cortical-LA synapses. All slices were incubated in the κB decoy DNA (50 μ m, ○), scrambled DNA (50 μ m, ▾), or normal ACSF (●) for at least 3 h. LTP was evoked by delivery of four sets of TS (100 Hz, 1 s) at an interstimulus interval of 3 min. ΚB decoy DNA blocked LTP without affecting basal synaptic transmission (▿). D, sample traces taken from experiments in C at the times indicated. It has been shown that many hippocampal pyramidal neurons exhibited intense fluorescence when slices were incubated for 3 h in the presence of 50 μ m κB decoy DNA, indicating penetration of the κB decoy DNA into these cells (). Amygdala slices were incubated with 50 μ m κB decoy DNA for at least 3 h before recordings were made. Control slices were incubated for a similar length of time with scrambled DNA in a different beaker. C shows that delivery of four sets of TS at an interstimulus interval of 3 min produced a long lasting, robust enhancement of synaptic responses in control and scrambled DNA-treated slices. The slopes of field excitatory postsynaptic potential (fEPSP) were 213 ± 18, 201 ± 15, 189 ± 16, and 178 ± 17% ( n = 6), at 1, 2, 3, and 4 h after the stimulation. By contrast, the same amount and pattern of stimulation elicited a gradually decaying long-term potentiation (LTP) in κB decoy DNA-treated slices. The fEPSP slopes at 1, 2, 3, and 4 h after the stimulation were 134 ± 23, 108 ± 26, 86 ± 14, and 88 ± 14% ( n = 6) of control, respectively. The difference between control and κB decoy DNA-treated slices was statistically significant at 1 ( p. Increase in Co-immunoprecipitation of CREB and NF-κB after TS or Treatment with Forskolin Since CREB antibody slightly retarded the migration of proteins interacting with the NF-κB oligonucleotide (Fig. ), we investigated possible interaction between CREB and NF-κB with co-immunoprecipitation. In the first set of experiments, slices were given four trains of TS, and nuclear extracts from the LA and BLA were immunoprecipitated with CREB antibody and immunoblotted with NF-κB antibody. As shown in Fig. A, TS resulted in a significant increase in NF-κB (177.0 ± 6.3% of control, n = 6, p. Figure 8 Effects of TS and forskolin on the interaction between NF-κB and CREB. A, slices were given four trains of TS and nuclear extracts from the LA and BLA were immunoprecipitated with CREB antibody and then blotted with NF-κB antibody. Nuclear extracts were also immunoprecipitated with NF-κB antibody and then blotted with CREB antibody. TS resulted in a significant increase in NF-κB, which bound to CREB, and the effect was attenuated by d-APV (50 μ m). B, slices were incubated with forskolin (50 μ m) for 15 min, and nuclear extracts from the LA and BLA were immunoprecipitated with CREB antibody and then blotted with NF-κB antibody. Nuclear extracts were also immunoprecipitated with NF-κB antibody and then blotted with CREB antibody. Forskolin caused a significant increase in NF-κB, which bound to CREB, and the effect was blocked by KT 5720 (1 μ m). Second, slices were incubated with forskolin (50 μ m) for 15 min, and nuclear extracts from the LA and BLA were immunoprecipitated with CREB antibody and then blotted with NF-κB antibody. As shown in Fig. B, activation of adenylyl cyclase by forskolin caused a significant increase in NF-κB (180.3 ± 13.2, n = 6, p. DISCUSSION We have examined the potential role of NF-κB in amygdala synaptic plasticity. Here we show for the first time that fear-potentiated startle results in an activation of NF-κB and intra-amygdala administration of NF-κB inhibitors, or κB decoy DNA blocks memory consolidation and LTP. Since activated NF-κB binds to cognate DNA sequences to initiate transcription of specific target genes, the present results are consistent with pharmacological studies demonstrating a requirement of new protein synthesis during fear conditioning and the late phase of LTP in the amygdala (, ). The protein levels as well as the DNA binding activity of NF-κB were significantly increased in the conditioned animals. By contrast, the control group that received conditioned stimulus and unconditioned stimulus in an unpaired fashion did not present any increase, suggesting that NF-κB activation is specific to the learning component of the task. In addition, activation was restricted to the amygdala but not the hippocampus or cerebellum, supporting the notion that amygdala but not hippocampus or cerebellum is critically involved in auditory fear-potentiated startle. Previous reports also showed that auditory fear conditioning was associated with an activation of CREB () and cAMP-response element-mediated transcription () selectively in the amygdala. Furthermore, overexpression of CREB in the BLA using viral vectors has been shown to facilitate long term memory of fear-potentiated startle (). Conversely, training for contextual conditioning or passive avoidance led to significant increase in cAMP-response element-dependent gene expression in areas CA1 and CA3 of the hippocampus (). The central nucleus of the amygdala, which receives inputs from LA and BLA and in turn synapses on autonomic control cells in the hypothalamus and brainstem, is generally thought to mediate behavioral, autonomic, and emotional responses to stressful and fearful stimuli (). Although we did not screen NF-κB activity in the central nucleus of the amygdala, it is very likely that its activity could be increased after training. In this respect, it has been demonstrated that auditory fear conditioning resulted in an increased in phosphorylated mitogen-activated protein kinase and phosphorylated CREB-immunoreactive cells not only in the LA and BLA but also in the central nucleus of the amygdala (, ). LTP at sensory input synapses to the LA and BLA is a candidate mechanism for memory storage during fear conditioning (, ). Synaptic responses in the LA could be persistently enhanced after repetitive stimulation of central auditory pathways or in animals given auditory fear conditioning. In the present study, we found that TS or forskolin that reliably induced LTP in this area of the brain (, ) also enhanced NF-κB binding activity. In the same vein, administration of κB decoy DNA or NF-κB inhibitors resulted in an impairment of LTP and memory formation. These results reinforce the idea that memory consolidation of auditory fear and LTP in the LA may share a common molecular substrate. Supershift assays using antibodies against p65 or p50 subunits further indicated that the major NF-κB complex induced by TS or forskolin was the p65/p50 heterodimer. This finding is compatible with previous reports showing that both constitutively active and inducible NF-κB activities in the central nervous system predominantly consist of p50/p65 complexes (, ). Using antibody against IκB-α, we found that protein levels of IκB-α were reduced by 20–40% at 10 min after TS or application of forskolin followed by its reappearance at a later time point (∼30 min). This was accompanied by a decrease of NF-κB level in the cytoplasm concomitant with an increase in the nucleus. The rapid resynthesis of IκB-α could be due to the presence of three NF-κB binding sites in the promoter region of IκB-α gene, which was activated by NF-κB (, ). In the present study, forskolin-induced degradation of IκB-α was prevented by wortmannin, suggesting that PI 3-kinase is necessary for forskolin activation of NF-κB. A role for PI 3-kinase in the mediation of tumor necrosis factor and platelet-derived growth factor activation of NF-κB has been reported in embryonic kidney and primary fibroblast cells, respectively (, ). Thalidomide, once used as a sedative and an anti-nausea medication for first trimester pregnancy, produced several developmental defects to the human fetus, leading to its removal from the market. It possesses anti-inflammatory and antioncogenic properties and has attracted considerable attention due to its possible clinical potential for the treatment of some intractable diseases such as rheumatoid arthritis and several forms of cancer (). NF-κB is a key regulator of inflammatory genes, and it has been shown that thalidomide inhibited tumor necrosis factor-α-induced NF-κB DNA binding activity and transcriptional up-regulation of interleukin-8 in Jurkat T lymphocytes and EA.hy926 endothelial cells through a mechanism involving the inhibition of IKK (). In the present study, we found that fear training induced an increase in IKK activity, resulting in the activation of NF-κB in the amygdala. Consistent with the observation in lymphocytes and endothelial cells, thalidomide inhibited NF-κB activation probably via an inhibition of IKK. More importantly, behavioral study revealed that thalidomide blocked fear-potentiated startle, which reinforces the obligatory role of NF-κB in the acquisition of fear memory. The encoding of new memories is thought to require long lasting changes in the strength of specific synaptic connections between neurons mediated by specific alterations in gene expression. Accumulated evidence indicates that CREB is one of the core components in the molecular switch that converts short term to long term synaptic plasticity (, ) including the consolidation of fear memory (,,, ). The present co-immunoprecipitation experiments revealed that TS or forskolin stimulated the interaction between CREB and NF-κB, and the effect was blocked by d-APV and KT 5720, respectively. It is speculated that the physical association between NF-κB and CREB may endow both transcriptional factors with enhanced affinity for the cAMP-response element and κB sites, thus playing the role of a positive feedback mechanism. Taken together, it is conceivable that TS, forskolin, and fear training may activate NF-κB through the signal-induced activation of PI 3-kinase, IKK, and subsequent proteolytic degradation of IκB-α in the cytoplasm. This allows NF-κB to translocate into the nucleus, where it binds to specific κB DNA consensus sequences in the enhancer region of κB-responsive genes to initiate transcription and translation of new protein necessary for memory consolidation (Fig.). Figure 9 A model for activation of NF-κB by fear conditioning. Fear conditioning or heightened synaptic activities in amygdala neurons induce Ca 2+ influx through N-methyl- d-aspartate receptors. The synaptic activity in the amygdala also induces cAMP signaling through modulatory input such as β-adrenergic and dopamine receptors. Ca 2+and cAMP activate PI 3-kinase and its downstream target Akt, which in turn phosphorylates IKK resulting in the phosphorylation and degradation of IκB followed by nuclear translocation of NF-κB. PI 3-kinase and Akt could also induce MAPK phosphorylation, and subsequent translocation of mitogen-activated protein kinase ( MAPK) into the nucleus activates CREB. In the nucleus, NF-κB and CREB may interact to fully transactivate effector genes for memory formation. In conclusion, in an intriguing parallel, the present results show stimuli that generate LTP in slices or elicit fear-potentiated startle in animals activate NF-κB in the amygdala. NF-κB inhibitors, on the other hand, block IκB-α degradation as well as LTP and fear memory. These results provide the first evidence of a requirement of NF-κB activation in the amygdala for synaptic plasticity and memory consolidation. Furthermore, the identification of NF-κB as a molecular substrate in the formation of fear-potentiated startle suggests a potential new target for the treatment of anxiety and posttraumatic stress disorders. Footnotes • * This work was supported by National Science Council Grant NSC89-2320-B006-011 and by the Academic Excellence Program of the Ministry of Education of Taiwan, Republic of China, Grant 89-B-FA08-1-4.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “ advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. • ‡ To whom correspondence should be addressed: Dept. Of Pharmacology, College of Medicine, National Cheng-Kung University, Tainan, Taiwan 701. Tel.: 886-6-2353535 (ext. 5507); Fax: 886-6-2749296; E-mail: [email protected]. • Published, JBC Papers in Press, September 25, 2002, DOI 10.1074/jbc.M206258200. LTD long term depression CREB cAMP-response element-binding protein EMSA electrophoretic mobility shift assay IKK IκB kinase LTP long term potentiation D-APV D-2-amino-phosphonovalerate PI 3-kinase phosphatidylinositol 3-kinase TPCK N-tosyl- l-phenylalanine chloromethyl ketone TLCK N α- p-tosyl- l-lysine chloromethyl ketone TS tetanic stimulation LA lateral BLA basolateral ACSF artificial cerebrospinal fluid PMSF phenylmethylsulfonyl fluoride • Received June 24, 2002. • Revision received August 26, 2002. • The American Society for Biochemistry and Molecular Biology, Inc. How to make Lion’s Dashboard overlay the desktop. First introduced with Mac OS X Tiger back in 2005. Now when you activate Dashboard it will show over the. How to make Lion’s Dashboard overlay the desktop. First introduced with Mac OS X Tiger back in 2005. Now when you activate Dashboard it will show over the. Some people have claimed memory or performance problems with Dashboard. In my case, I just don't use it, and dislike having applications running that I don't use. Activate and Use Xdebug. Xdebug is a powerful open source debugger and profiler for PHP. It is included with XAMPP and can be used to display stack traces. Remember this? Introduction & Evolution Introduced with Mac OS X 10.4 Tiger, Dashboard was Dashboard is home to widgets: mini-applications that let you perform common tasks and provide you with fast access to information. With a single click, Dashboard appears, complete with widgets that bring you a world of information — real-time weather, stock tickers, flight information and more — instantly. Dashboard disappears just as easily, so you can get back to what you were doing. While not as flashy or important as Tiger’s keystone feature, Spotlight, Dashboard still enjoyed. These widgets would auto-update, so it was a great way to keep tabs on headlines or a sports score without having to leave it open in a web browser window. Dashcode In Leopard, Apple shipped an application named Dashcode. It made creating widgets simple, giving developers a visual workflow for putting assets together and bundling everything up as a.wdgt file. Dashcode.app was last updated in July 2012. It can still be downloaded from Apple’s developer site, but doesn’t run on macOS Sierra. Many of its features have been rolled into Xcode, and re-used in tools to build webapps for Safari on the desktop and iOS. ![]() Concerning Konfabulator So, uhhhh, Dashboard wasn’t an original idea. Konfabulator was an existing third-party application that allowed users Konfabulator is a JavaScript runtime engine for Windows and Mac OS X that lets you run little files called Widgets that can do pretty much whatever you want them to. Widgets can be alarm clocks, calculators, can tell you your WiFi signal strength, will fetch the latest stock quotes for your preferred symbols, and even give your current local weather. What sets Konfabulator apart from other scripting applications is that it takes full advantage of today’s advanced graphics. This allows Widgets to blend fluidly into your desktop without the constraints of traditional window borders. ![]() Toss in some sliding and fading, and these little guys are right at home in Windows XP and Mac OS X. Apple this application, and Sherlocked it hard. Arlo Rose, the developer behind Konfabulator Arlo Rose “It’s insulting, is what it is,” Rose said in a telephone interview. “They could have at least offered to work with us or to buy it.” Apple, for its part, maintains that Dashboard is the company’s own creation, noting that Widgets have long been a part of Mac OS X and the NextStep OS. “The goal isn’t to be like anything else,” Apple Senior Vice President Phil Schiller said in an interview. “It’s not his stuff. What we’ve done is ours.” Not everyone thought Apple ripped it off, however. Dashboard is not a rip-off of Konfabulator. Yes, they are doing very much the same thing. But what it is that they’re doing was not an original idea to Konfabulator. The scope of a “widget” is very much the modern-day equivalent of a desk accessory. The classic Mac OS shipped with which were small, single-purpose programs that could run alongside a user’s main application, even before the Mac could truly multitask. Regardless of where the original idea came from, many thought Dashboard was a pretty clear clone of Konfabulator. The app’s website certainly said as much after Dashboard had been unveiled: I can only assume Apple was aware of Konfabulator’s existence when developing Dashboard. While Sherlocking is something that happens, it’s always a little icky to consider. Konfabulator was purchased by Yahoo in July 2005 and powered Yahoo Widgets until 2012 when the platform was shut down. Dashboard’s Present & Future For a couple of years, I used Dashboard pretty heavily. I had all sorts of little utilities handy, but over time, it slowly faded from my workflow. My guess is that’s the case for most Mac users. Slowly but surely, stuff that widgets were great at has been replaced by things like simple iOS apps or even quick Google searches. Dashboard itself has faded, too. Starting with OS X Lion, Dashboard could be configured to act as a space to the left of the first desktop, accessible via Mission Control or a swipe. I remember the change, and thought at the time that it felt like a demotion for the once-impressive user interface and placement Dashboard had previously enjoyed. As of OS X Yosemite, Dashboard is off by default on fresh installations. Notification Center has become the new target for Mac developers wanting to build widgets, and Dashboard’s dedicated media key on MacBooks and external keyboard has been replaced with one for Launchpad. (You can add it back if your Mac has a Touch Bar, though.) Every year, I think to myself that Dashboard has reached the end of the line, but Apple keeps surprising me. Maybe someone important on the macOS team still really love Dashboard, or maybe everyone’s forgotten it’s still in there, deep in the codebase somewhere. Whatever the reason, Dashboard lives on, for now at least. |
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