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Q: How to connect to any specified Wi-Fi network(SSID) using ADB command Lets assume I have Wi-Fi networks 'A' 'B' & 'C'. I want to connect the connected android device to SSID: 'A' which is password protected. Is there a way to perform this using ADB command by providing SSID and password in the same command?

A: $ am startservice \
    -n com.google.wifisetup/.WifiSetupService \
    -a WifiSetupService.Connect

Go with android developer console.


When we first released Soundproof we had to make the difficult decision to delay polishing our VoiceOver experience for accessibility users.
We had done some work on this already, but due to the nature of our custom UI elements such as the time scrubber and Count-In screen, the work was too much to complete first time around.
In hindsight, I really would like to have done this for the first release. It seems to be common that developers don't bother with this until later, if at all, but it is extremely useful as it tells you things about your visual UI that you wouldn't have considered. In short, it is a great test of your visual design.
We wanted to have a VoiceOver experience that was as polished as our visual UI. I've found very little detailed information on making a first-class VoiceOver experience, hence the detail included here.
This is going to be quite long, but it will hopefully provide useful insights into the efforts required to make your apps work really well for VoiceOver users. Those users rock by the way, they are the most helpful beta testers ever.
So, what were the problem areas?
This was a fascinating problem, with wide implications. At startup Soundproof shows a single on-boarding screen with some static text and a single "Create a Setlist" button at the bottom.
What could possibly have gone wrong here? Well some testers could not find the "Create a Setlist" button. There were at least two causes. First, the element read by VoiceOver at the top of the screen is the big "Get Started" heading. That is all it said in VoiceOver, and it was not a tappable item.
Second, the low density of interactive elements — a single button at the bottom — gets worse the bigger display you have. The fingers of a blind user have to explore much more physical space to find that one thing they can touch.
This was a very surprising discovery, after all when designing a screen what could possibly be easier to use than a screen of static text with only one button?
For now we chose to add an accessibilityHint to the "Get Started" heading indicating that they can get out of this screen using the button at the bottom, and also by using the "Magic tap" gesture.
We should probably make the "Get Started" item a button as well, as when you can't see how it is visually styled as a heading it seems like an obvious call to action, but this should do for now. The user could have spotted that VoiceOver did not say "Get Started. Button", but this is easily missed.
Right there is another lesson: font stylings are obviously invisible to VoiceOver users. However we rely on these a lot in modern UI to indicate functionality. Forget worrying about visual cues disappearing on buttons in iOS 7 and up — nobody using VoiceOver can tell one bit of text's importance relevant to another.
Our main player interface is relatively simple, but there are a number of elements that provide challenges with VoiceOver.
Our statistics view drops down from behind the navigation bar to show practice stats when the player is not active.
Being a UIView that contains an animated UILabel that rotates between different stats, we had to make the container view return YES from isAccessibilityElement and return an aggregated value for accessibilityValue so that VoiceOver could speak the full set of statistics rather than just those that were currently visible in the animated display.
Our track selector is actually a special UICollectionView derivative that supports infinite scrolling through the Setlist.
This caused a slew of problems with VoiceOver.
Problem 1: the built-in collection view VoiceOver handling would not work as page offsets would be incorrect.
To resolve the incorrect page number announcements such as "Page 2 of 7" when they are tracks and there's only 5, and have VoiceOver announce it as "Track 1 of 5", we had to add a smart implementation of the hard-to-find accessibilityScrollStatusForScrollView: method of UIScrollViewAccessibilityDelegate.
Problem 2: the single visible cell was not being used for accessibility properties.
Problem 3: tapping the visible collection view cell did not announce track information.
To solve these two related problems we had to make our custom collection view delegate its accessibilityValue and accessibilityHint properties to the first (only) visible cell, and we had to make the collection view act as an accessibility element itself with isAccessibilityElement returning YES.
Problem 4: tapping a track would not tell you which number track you were on in the setlist.
While swiping, the track selector collection view would announce the position in the setlist with for example "Track 2 of 7", but tapping on the view directly would only tell you the track information, not the position in the Setlist.
However by putting the information at the end of the value, we minimise the annoyance this could cause. It did not seem possible to change the announcement only when paging has just taken place. So if we want to have swiping announce the track number, and tapping also to convey this information, we have to live with it.
The time scrubber is based on UISlider encapsulated in a custom view. We track the start and end of drag events on the slider and perform our zoomed time display animations in response to these, and then track the changes to update the player's position when the user stops dragging. During the drag we update the "Add marker" button title to adapt to the presence of markers and the current play position so that the button doesn't read "Set end marker" when you scrub the time to before the the existing (start) marker.
Problem 1: with VoiceOver on, changes to UISlider values don't trigger drag events (well, duh), they only generate value changed events.
So the solution here was to detect if VoiceOver is running in the "value changed" action of the slider and adjust the state of the "Add marker" buttons there.
Problem 2: the VoiceOver description of the slider is not read as a time, it is just read as the float value of the slider.
To fix this a custom implementation of accessibilityValue is required on the custom view that acts as the accessibility element (parent) of the slider.
Problem 3: with the above issue solved, when the time position is at the start or end of the track, announcing "Time: 0 seconds" or "Time: 9 minutes 41 seconds" is not human enough.
I feel detail like this is important because we developers tend to go to this kind of trouble with on-screen labels for relative times and quantities.
Problem 4: the custom view containing the time slider also presents the start and end marker positions visually, however the VoiceOver description does not tell you where the markers are. Thus VoiceOver users have no idea where their markers are when they return to the app or switch tracks.
The marker interface is deliberately super-simple. Covered in a previous post, we have a one-button interface for all marker features which shows the only action you can take based on the current marker state. This means it alternates between "Add marker", "Set start marker", "Set end marker" and "Clear markers".
Any of these actions will change the state of the time slider which has the markers on it, and possibly throw the player into "Repeat markers mode" automatically. It also changes the state of the button itself which can cause VoiceOver to re-announce that button.
This causes the slider to receive VoiceOver focus and have its current values announced, including the state of the markers. In fact we do this any time anything changes the state of the markers.
This does have the side effect that the accessibility focus switches from the button the user had selected, and sometimes results in a clipped announcement of the new button title, before the focus switches to the slider. This appears to be an unavoidable problem with VoiceOver (see later notes).
When the application became active, VoiceOver would select the first accessible element, the "Settings" button. However you would not know what state the player was in, or what track was active.
For this we added the UIAccessibilityTraitSummaryElement trait to the track selector collection view. The result is that when the player becomes active, the accessibilityValue of the track selector is announced, which includes the current track information.
This problem actually highlighted a non-trivial issue. For the UI to skip to a new track and then use the normal track selector VoiceOver announcements, we would have to wait for that announcement to finish before showing the Count-In screen. The Count-In screen itself can be set to announce tracks even for non-VoiceOver users, which adds more complication.
Rather than deal with the myriad problems of delaying the skip to a new track until an announcement has finished (which seems to be hard/impossible, see later notes), we decided that when VoiceOver is running, we would pause playback when the track skip buttons are pressed.
This pragmatic solution means the user is not bombarded with conflicting announcements and does not enter some strange state where they have to wait for something to complete, which is not compatible with the goals of VoiceOver which is designed to be constantly interruptible to avoid slowing users down.
As we have our own voice announcer even for non-VoiceOver users, the solution was to set accessibilityElementsHidden = YES on the parent of the track info view, which hides all the subviews from VoiceOver. Using isAccessibilityElement = NO would not help, as the subviews have it set to YES. This makes sense, but can seem counter-intuitive at first.
We have a screen that lists all of the Setlists the user has created, and it uses a table view.
This was due to the accessibilityValue being "New setlist 1.6 tracks". We were missing a space after the full stop, so the number of tracks was running together with the end of the setlist name, which by default has a number at the end.
This might not have been noticed if we quoted the setlist name in the string, e.g: "New setlist 1".6 tracks assigned to an NSString including the quotes. I can't show the quotes escaped here due to a bug in WordPress Markdown handling.
Sadly we couldn't quote the setlist name because of a VoiceOver bug we found (see later notes).
Once we had the basics covered, we could add some nice-to-haves for VoiceOver users.
The so-called "Magic tap" is a two finger double tap anywhere on the screen. This is very handy as it is an easy gesture to perform that requires little accuracy.
So when you are on the player screen we made this gesture equivalent to the play/pause button. Just two finger double-tap anywhere with VoiceOver on and it will play or pause without talking to you.
On the Count-In screen we made this gesture perform the "Start" button action to skip the Count-In and instantly start the track.
We may extend this functionality in future releases to other screens such as to dismiss the track selector UI, or to play the selected track in Setlist view.
There is also a special two-finger "scrub" gesture for cancelling actions when VoiceOver is enabled.
We implemented this on the Count-In screen to perform the "Cancel" action which will cancel the Count-In and abandon playback.
Again, there is scope here to introduce this to other view controllers to avoid users having to hunt for the "Cancel" button.
We did hit some issues with VoiceOver that we couldn't find solutions for at this time.
The biggest problem was that it seems impossible to prevent VoiceOver default announcements when you want to control what is said. For example if you want to say something after a button is tapped, it does not seem possible to reliably do this, because the button's label is spoken as soon as the tap occurs.
If you try to use the UIAccessibilityAnnouncementNotification or UIAccessibilityLayoutChangeNotification in this situation you typically find your announcement or change does not occur, or there is a clipped announcement and then it switches back to the button label announcement, or vice versa.
This is very frustrating when a button action causes a change in UI state. We explored queuing our announcement internally and waiting for VoiceOver announcements to complete and then posting the notifications to make our announcements, but it does not seem possible to detect when VoiceOver finishes speaking its own announcements. You can hook into notifications for your own announcements, but not the default ones.
You could work around this by posting your announcement after a fixed time delay. However I find such "fixes" abhorrent as they will no doubt be unreliable or have other unforeseen side effects.
This issue causes problems all over the place. We hope Apple improves this in future. We need a way to know when VoiceOver has finished speaking, and a way to say "this layout change is all you should speak".
We want play/pause or the track selector to be active at startup, but VoiceOver insists on selecting the first accessible element, the "Settings" bar button. Yes, we tried posting a layout change notification to focus our element when the view controller appears. It didn't work.
We wanted to announce the track selector view controller when it appears, as it has no title bar. It was too hard to suppress or delay the initial segmented control announcement. We have to assume the user knew what they were doing when they pressed the button to add tracks and enter this UI.
However if the user leaves our app and returns to it later it is not clear what they are doing.
It's possible this could be worked around to some extent with a hidden UI element that is has the summary element trait.
UIStepper is bad for accessibility. Don't use it. It is hard (or impossible?) to get it to announce the current value represented by the stepper, or bind it to the relevant view that contains its current state. We changed our settings UI to push a new view controller that uses a table view that contains rows for all the possible values. This works out nicer for everyone really.
UISwitch embedded in table view cells in Settings needed to be wrapped in a custom view that handles VoiceOver properties in aggregate, so that the label and value can be announced together. Also handling a single activation (double tap) can be used to toggle the state of the switch on that row by implementing accessibilityActivate… except that is seemingly never called so we frigged it with accessibilityActivationPoint set to return the center of the switch.
Tappable labels in Settings, which are static table view cells, benefitted from adding the Button trait so the user knows they can activate that row to make something happen.
Presenting a pile of static text with multiple headings on our on-boarding screen is painful for VoiceOver, as each is announced individually as your finger travels across the screen. There's an argument to be had for presenting this as a single block of text that can be read out, using a custom container view. Referring back to the start of this post, a user doesn't know a heading from a paragraph that follows it, so you lose the natural narrative flow of text when you present items as just a set of labels.
It is tempting to try to announce all app state all the time, but this would likely be very annoying given people typically do have a good short term memory and vision impairment does not normally affect this! So we asked ourselves "Did the user instigate this UI state change?". If the answer was "Yes", don't try to announce what happened.
Custom UI elements are fairly painful for accessibility in terms of working out what the right behaviour is. Aggregating controls means aggregation VoiceOver properties.
Mispronunciations are rife in VoiceOver and there is apparently no way to correct it with specific phonetics.
Finally, there is a fun VoiceOver bug in quote handling. This "Track 01", if you include the quotes in the value, is pronounced as "Track zero-one inches". This is even with fancy unicode quotes. Yes, I radar'd it.
Soundproof is available in the App Store now. The build with these VoiceOver changes is pending review.


The 'self-discovery' trip Barack Obama took in his 20s—plus bucket list places he's visiting soon "I did not have any money. I was traveling alone. In Europe, I was in these 'pensiones' [hostels] and I'd basically buy a baguette and cheese and ate that every day, and some wine on occasion."


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Luanna L. Wolff
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Luanna
Obituary of Luanna L. Wolff
Please share a memory of Luanna to include in a keepsake book for family and friends.
Luanna L. Wolff, passed peacefully on July 17th, 2021, at the age of 80. At the time of her passing, Luanna was a resident of Broken Arrow, Oklahoma where her eldest daughter Kim resides. Luanna is survived by her daughters Kim (Tim) and Wendy (Gianluca) along with her two grandchildren, Joshua and Julia, nieces, and nephews. Luanna is predeceased by her husband of 55 years, Terry. Born in Wellsville, NY she moved to Brockport, NY as a toddler, where she predominately lived and raised her family. Luanna worked as a secretary for the Brockport School District for 27 years.
Luanna was a caring and loving mother, wife, neighbor, and friend. She enjoyed tending to her garden, caring for her home, traveling, and helping family and friends. She was loved and admired by all those who knew her, and she will be greatly missed.
A memorial service for Luanna will be held at the Fowler Funeral Home in Brockport, NY on Wednesday, July 28th at 11:00 AM, followed by a graveside service at Lakeview Cemetery and a celebration of her life at the Hampton Inn in Brockport. Flower donations are welcome, or in lieu of flowers, consider donating to the Dystonia Medical Research Foundation in Luanna's honor.
To send flowers to the family or plant a tree in memory of Luanna Wolff, please visit Tribute Store
Fowler Funeral Home Inc.
Order Flowers for Luanna Wolff's Memorial Service
Lake View Cemetery
Rte 19, Lake Rd
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Luanna Wolff

redpajama_set_name: RedPajamaCommonCrawl


Dartford Borough Council has appointed national developer and urban regeneration specialist, Muse Developments, as its development partner to lead the £75 million regeneration of the town's Westgate area. Muse was appointed following an open tender by the Council to find the most suitable partner.
The mixed-use development incorporates a six-screen cinema at the heart of a new social neighbourhood with bars, restaurants and family-friendly hospitality. The scheme will see the creation of a new public square, a 109-bed hotel, a multi-storey car park and 140 new homes.
Discussions are also underway with the local Clinical Commissioning Group to collaborate on the potential creation of a new health facility providing space for GPs, family healthcare, social care providers and mental-health services.
"We are delighted to be working with Muse. So much of what happens in Dartford is developer-driven and the Council rarely has sufficient ownership to shape the use of land for the benefit of local people.
Designed by architect and urban designer tp bennett, the scheme envisages a vibrant public realm with 37,250 square feet designated for restaurant and bar space.
140 new homes – a mix of one, two and three-bedroom apartments above retail uses – will be spread among four new buildings. A range of tenures will be offered for these properties, including affordable units for leaseback to the Council.
"It is an honour to have been chosen by Dartford Borough Council to lead on a project which will have such a positive impact on the town. The site is very well-connected, being close to both the high street and train station, making it the ideal location for a lively and vibrant new community and cultural quarter.
Muse Developments is looking to submit a planning application in Summer 2018 and to start on site by Spring 2019. Before then, Dartford Borough Council and Muse Developments will be holding public consultation events in early 2018.
Gross Development Value (GDV) £75 million.
• 140 new homes (39 being affordable).
• A new public square.


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9 great things to do in Morris County this weekend
No matter what you're into--puppies; power boats; Motown; roots music; gaming or simply celebrating fall, there's plenty to make you happy in Morris this weekend.
9 great things to do in Morris County this weekend No matter what you're into--puppies; power boats; Motown; roots music; gaming or simply celebrating fall, there's plenty to make you happy in Morris this weekend. Check out this story on dailyrecord.com: http://dailyre.co/1Kl7sg1
Morristown Daily Record Published 12:20 p.m. ET Sept. 18, 2015 | Updated 12:32 p.m. ET Sept. 18, 2015
The New Jersey State Championships Power Boat Races will be held at Lake Hopatcong on Saturday and Sunday. Pictured here are speedboat races in Point Pleasant.(Photo: File photo)Buy Photo
1. "A Video Game Con" in Parsippany Saturday http://dailyre.co/1EvrCVm
A 36,000-square-foot celebration for video gamers. 60-plus vendors and 100 tables of merchandise, live music, tournaments, discussion panels, free play and collectibles, plus 60 consoles to play old-school games and new. Parsippany PAL, Saturday, Sept. 19, 10 a.m. to midnight. $20 admission. The Parsippany PAL, 33 Baldwin Road. More information: www.avideogamecon.com
2. Championship Power Boat Races at Lake Hopatcong Saturday and Sunday http://dailyre.co/1KkqrqX
"No wake" rules won't apply this weekend at the New Jersey State Championships Power Boat Races. "Inboard" and "Stock Outboard" categories; classes are based on hull size, type, weight, engine size, and modifications. Saturday and Sunday, 10 a.m. - 5 p.m. $10 donation for spectators. Hopatcong State Park, 6 Lakeside Boulevard, Landing,
3. Parsippany Fall Festival on Sunday
Thousands are expected at the 14th Annual Parsippany Fall Festival featuring entertainment, rides, and food vendors, many of whom are local. Sunday, from noon to 5:00 p.m., Lake Hiawatha, North Beverwyck Road, Parsippany. Proceeds go to Parsippany's Public Assistance Trust Fund. For more information, call 973-335-7554, ext 14.
4. 11th Annual "Puptoberfest" at Horshoe Lake in Roxbury on Saturday http://dailyre.co/1F6pYty
This family-friendly event will feature a costume contest, keepsake pet photos, games, contests, paw painting, food and live entertainment, live music by "Frantic! Band," 72 Eyland Ave, Roxbury. The cost is $5 per person or $10 per family. This fee is waived for previous Eleventh Hour adopters. For more information, call 973-664-0865 or email to MainOffice@ehrdogs.org.
5. Tribute performers honor legacy of Motown http://dailyre.co/1Kl28t9
David Revels, formerly of the Drifters, and his band will perform such classics as "I Can't Help Myself," "Reach Out I'll Be There," "Standing in the Shadows of Love," "Bernadette" and more of the Four Tops and other Motown Bands. 8 p.m. Saturday, David and Carol Lackland Center, Centenary College, 715 Grand Ave., Hackettstown. $20 to $30 (advance); $25 to $35 (day of show). More information call 908-979-0900 or visit www.centenarystageco.org
6. Geoff Muldaur to play the Minstrel Sept. 18 http://dailyre.co/1F6FllQ
Muldaur specializes in roots music, or Americana and will perform at 8 p.m. Friday, at the Morristown Unitarian Fellowship, 21 Normandy Heights Road, Morristown $9 plus a free-will offering. For more information call 973-335-9489 or visit www.folkproject.org.
7. 'Equivocation' play at Shakespeare Theatre of NJ http://dailyre.co/1Kl4fgv
The play centers on the notorious 1605 Gunpowder Plot, in which Guy Fawkes and his followers attempted to blow up Parliament in an incident that may have been backed by the government. A key figure was Robert Garnet, a Jesuit priest who was hanged, drawn, and quartered for his complicity.Through Oct. 4.F.M. Kirby Shakespeare Theatre, 36 Madison Ave., Madison. $15 to $70. Student rush tickets available half-hour before performance. For performance schedule and tickets, call 973-408-5600 or visit www.ShakespeareNJ.org.
8. Mary Fahl to play in Whippany on Sunday http://dailyre.co/1Kl4jga
Singer-songwriter Mary Fahl singles out the inspiration of such artists as the Beatles, British singer-songwriter Richard Thompson, and Roger Waters of Pink Floyd in this solo acoustic concert. 7 p.m. Sunday. Doors open at 6:30 p.m. Ukrainian American Cultural Center, 60 N. Jefferson Rd., Whippany. $26 in advance, $28 day of show. $5 for ages 13 to 17, free for ages 12 and under. For more information call 973-585-7175 or www.splatterconcerts.com.
9. The Great Pumpkin Festival at Heaven Hill Farm in Vernon
Billed as the largest fall festival in Northern New Jersey, the event will feature hayrides, a pumpkin patch, corn maze, farm animals, spider web crawl, bee barn, vortex tunnel, dancing robot show and obstacle course. Through November 8. $8; $10 on weekends. More information visit: heavenhillfarm.com/special-events/great-pumpkin-festival.
Julia Martin, 973-428-6653, @JuliaMartinDR
Read or Share this story: http://dailyre.co/1Kl7sg1
Morris Parks Summer events and activities catalog
Morris Parks Spring events and activities catalog
Shakespeare Theatre unveils 2019 season
April 4, 2019, 6 a.m.
'Snooki' of 'Jersey Shore' opening NJ shop
Lots of winter activities offered in Morris County
Jan. 20, 2018, 9:42 a.m.
Things to do in Morris County this week


Webinse CRM allows you to split tasks and reminders among different employees and groups of your store.
Webinse СRM (Customer Relationship Management) allows you to create reminders for orders and customers. That would be rather handy for your store administration. Our extension will help you to split tasks and reminders among different employees and groups of your Magento store. It also can help you not to forget about client or order. You can't track all of your clients and their associated tasks and events. You can simply forget about your client, and such situation can be a blow to your reputation. With CRM extension you can store and manage hundreds of clients and let a computer system sent you reminders.
To create reminders you just need to appropriate order tab or customer profile in the CRM tab, and click on the "Add New Reminder" button. CRM also maintains attachments that are not reminders.
All reminders include start and due dates and all reminders are visible on the top of back-end pages at all the times until they are marked as being completed.
CRM Extension also allows you to create groups of employees (e.g., sellers or account managers) in System->Permissions->Groups, and create task not only for a particular employee, but for all department. The system also sends notifications to the group, if task has been assigned or changed.
Also you may customize reminders by using configuration in the System -> Configuration -> Webinse -> CRM tab.
Note: In the top of the pages only reminders which have status 'Running' or 'Failed' will be outputted.
System->Configuration->General->Content Management ->Use Static URLs for Media Content in WYSIWYG for Catalog field set to 'Yes'. And after this, please clean cache.
Ability to send notifications when creating or editing (reminder itself or automatic change of reminder's status) a task.
All features and user interface are fully configurable, installation is fast, and all module settings can be controlled from the Magento Administration Panel.


The scarcity of charging points in cities is a major challenging factor for growth of the global electric scooter market. These electric scooters does not have any other alternative source in place of battery. Thus, complete drain of the battery can stall the scooter and can put the traveller at risk. Moreover, prices of electric scooter are also very high, which in turn decreases the adoption of these scooters.
Increasing prices of gasoline and diesel fuel is expected to be major factor for increasing adoption of vehicles that are environment-friendly. Thus, increasing demand for electric vehicles is one of the major factors for high demand for lithium-ion battery. For instance, according to Coherent Market Insights' analysis, the annual sales of electric scooters as 542,000 Units in 2015, which witnessed an increase to 850,000 Units in 2016. Moreover, another factor leading to increasing demand for lithium-ion batteries is that the weight of the battery is very low as compare to the other alternatives. This is expected to aid in increasing adoption of lithium-ion batteries, which is in turn supporting growth of the market during the forecasted period.


Yesterday one off the hectic days . No one in the building to atleast talk.
Mohun is in Channai now, So no new Films,Suresh went to Spectramind, and I get on to my singing sting with my small FM radio which makes stero effects very well. I enjoyed the "Return Ticket " which is talked on the 1970's best 10 movies. According to that the top song on 1970 is "ye jo mohabat hai" frm the film "kati Patang" .
Read on the National Geographic old books and the update my diary.
before going to my room I had a talk with Sarwar in Kashmir .


Below is a throwback video of Ex-President, Olusegun Obasanjo attacking his former VP now Presidential candidate of the Peoples Democratic Party, PDP, Atiku Abubakar.
SAVAGE!!! Festus Keyamo Shades "Man Of God" Bishop Oyedepo In New Tweet… And It's Causing A Lot Of Reactions Online!


The first Thanksgiving in the New World was celebrated in 1621, nearly a year after the Pilgrims landed at Plymouth in the Massachusetts Bay Colony.
I do recommend and assign Thursday, the 26th day of November next, to be devoted by the people of these States to the service of that great and glorious Being who is the beneficent author of all the good that was, that is, or that will be; that we may then all unite in rendering unto Him our sincere and humble thanks for His kind care and protection of the people of this country previous to their becoming a nation; for the signal and manifold mercies and the favorable interpositions of His providence in the course and conclusion of the late war; for the great degree of tranquillity, union, and plenty which we have since enjoyed; for the peaceable and rational manner in which we have been enabled to establish constitutions of government for our safety and happiness, and particularly the national one now lately instituted; for the civil and religious liberty with which we are blessed, and the means we have of acquiring and diffusing useful knowledge; and, in general, for all the great and various favors which He has been pleased to confer upon us.
And so it was until President Franklin Roosevelt, in 1939, temporarily moved the celebration back to the third Thursday in November to stimulate Depression-era Christmas sales.
Rooted in a story of generosity and partnership, Thanksgiving offers an opportunity for us to express our gratitude for the gifts we have and to show our appreciation for all we hold dear. Today, as we give of ourselves in service to others and spend cherished time with family and friends, we give thanks for the many blessings bestowed upon us.
We also honor the men and women in uniform who fight to safeguard our country and our freedoms so we can share occasions like this with loved ones, and we thank our selfless military families who stand beside and support them each and every day. . .
In the same spirit of togetherness and thanksgiving that inspired the Pilgrims and the Wampanoag, we pay tribute to people of every background and belief who contribute in their own unique ways to our country's story.
Each of us brings our own traditions, cultures, and recipes to this quintessential American holiday — whether around dinner tables, in soup kitchens, or at home cheering on our favorite sports teams — but we are all united in appreciation of the bounty of our Nation.
Let us express our gratitude by welcoming others to our celebrations and recognize those who volunteer today to ensure a dinner is possible for those who might have gone without. Together, we can secure our founding ideals as the birthright of all future generations of Americans.
NOW, THEREFORE, I, BARACK OBAMA, President of the United States of America, by virtue of the authority vested in me by the Constitution and the laws of the United States, do hereby proclaim Nov. 26, 2015, as a National Day of Thanksgiving.
I encourage the people of the United States to join together — whether in our homes, places of worship, community centers, or any place of fellowship for friends and neighbors — and give thanks for all we have received in the past year, express appreciation to those whose lives enrich our own, and share our bounty with others.
IN WITNESS WHEREOF, I have hereunto set my hand this twentieth day of November, in the year of our Lord two thousand fifteen, and of the Independence of the United States of America the two hundred and fortieth.


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How University of Sask. researchers hope cannabis could one day help pets
Published Thursday, November 28, 2019 4:16PM CST Last Updated Thursday, November 28, 2019 7:03PM CST
SASKATOON -- With growing interest in potential therapeutic uses for cannabis-derived products, researchers at the University of Saskatchewan's Western College of Veterinary Medicine are examining the effects of cannabis extracts in pets.
"What we want to do is administer various doses of cannabis extracts to dogs currently and eventually cats, to see what we find in terms of changes in behaviour or neurological signs or gastral intestinal signs," Dr. Al Chicoine said in an interview.
The department of veterinary biomedical sciences assistant professor is leading the project and a team of three research students and three other veterinarians.
The preliminary portion of the research project started in October.
"Right now we are in the preliminary stages of generating that safety data."
A total of 13 dogs belonging to the college are taking part in the initial testing.
"We start with administering very low doses, we are very protective of our teaching colony of dogs," Chicoine said.
The researchers then monitor the animals, particularly the level of cannabis in the bloodstream.
"Once we generate the data from our experimental animals then we are more sure we have an appropriate dose and will have sufficient safety data to move into client-owned animals from our vet center."
According to Chicoine, cannabis-related testing in animals is difficult to conduct at many universities across Canada.
"The approval process is very rigorous for these trials in veterinary medicine," Chicone said.
"Because people think cannabis is legal now for recreational use and legal for medicinal use for people, so they assume vets can prescribe it as well, but that is not the case."
Currently, Health Canada has not approved any cannabis products intended for animals.
However, Chicoine said Health Canada has expressed interest in possibly approving products if sufficient evidence of safety is generated by his team.
"It's something we are trying to get done through this conducted research."
Chicoine's team has received approval from both Health Canada and the university's animal research ethics board.
"The approval from Health Canada and the university animal research ethics board actually allowed us to acquire the cannabis extract. Without that, we couldn't do the study or use the dogs, and we needed both of those to start the study," said Chicoine.
Chicoine said he is hoping the research could one day lead to treatments for ailments found in older dogs such as osteoporosis, nausea, and vomiting.
"We are excited for a number of reasons, we don't know how it will be effective but either way we think it will be valuable information for veterinarians and their clients," said Chicoine.
Chicoine said he and his team are hopeful clinical research that will include cats along with dogs will begin in January
Dr. Al Chicoine is leading a team of researchers hoping to learn more about the effects of cannabis derived products in pets.
Video surfaces of group beating girl, Saskatoon police believe victim under 13


DJ Software - VirtualDJ - Oh my.....Roland DJ-808!
....and it's just been added to the latest version of Traktor, along with a sequencer for the Remix Decks.
I do hope Atomix are paying attention and considering these features.
Our Canadian Dollar is very low too.
For the1500.00 US price I had to pay 2000.00 CDN + 100.00 tax.
It's totally worth it though.
That's how good the soundcard is!
Plus the other bells and whistles, I would say it's a very good start for Roland in DJ controller line indeed!!!
I think it was a waste of time.
For somebody who said this "Old School around the way DJ...Started DJing in 1985..." I won't be surprised. LOL!
Some people just want to stay within their comfort zones.
Some, want to explore and adapt the changes.
And of course there are some who are in between.
It's our choice where to waste our time!
First its my opinion. Also, Old school meaning im over 40, around my way I dj for CNN and Akinyele(before RobSwift). So dont think my around the way is equivalent to your around the way. Im from Lefrak City, Queens. I dj every signing party, I djed Kenny Aderson signing party also along side spinderella (his girl at the time). Andre Harrell from around my way did all his uptown records parties. So dont judge my around the way cause around my way we had a lot of successful HipHop artist. So I got a lot of experience from being an around the way DJ, been on a lot of stages being an around the way dj. My Niche now underground speak easy after hour spots in the south.
I would have rather had an onboard effects processor Like the echoplex. especially using VDJ. So dont think I dont embrace technology, but if its not practical it makes no sense to me. With an Ableton link and the pads is a much better integration Idea. With much more capabilities.
I tried LINK in different ways on Ableton, Traktor, Ipad Apps with Push, 2x F1s, keyboard-synth, and even with RMX1000 plugin. I made it work to the max that I needed on my workflow. I too, said to myself that I don't need the 808 because of all the possibilities in using LINK.
What changed my mind is when I came accross the hassles in... switching between displays if only using one laptop, un-sync'ing and re-sync'ing when going in and out on manual beatching, using an extra soundcard if the mixer/controller cannot route the audio signal of Ableton to a specific channel that I need to use, lots of wiring, etc.
Big selling point for me is the midi-clock-out on 2 USBs and 5pin midi, the TR-S blends well with the mix way, way, better than using 3rd/4th deck or the sampler, and of course the sound quality.
And I didn't mean any disrespect on my previous post. Born on 1967, I am turning 50 myself next year. I still mix with Tech1200s every once in a while but for compact set ups, I could say this the best controller I've ever had.
As soon as this get native support with VDJ, the more people will get it. Not just the producers, musicians (like me) but the regular DJs as well.
I could sense that lots of VDJ users will switch into this unit especially the folks who uses other external midi devices.
So can anybody say how well this works with VDJ??
I mean the sequencer and all??
Don't think I've seen anyone here who actually has one.
938MyDJ has one but I'm pretty sure he's a heavy SDJ user.


IN JANUARY 1998, about halfway through my first year out of graduate school, where I'd just completed a PhD in engineering, my housemate handed me a copy of New Scientist containing a book review by the physicist and science writer, John Gribbin. The book was Tricks of the Trade by Chicago sociologist, Howard Becker, mostly a collection of Becker's musings on how to do productive social science research.
Gribbin clearly hated it, judging Becker's insights to be the kind of self-evident checks that "real scientists learn in the cradle". But Gribbin didn't stop there, noting that the book had merely reinforced his opinion that all social science was "something of an oxymoron" and that "any physicist threatened by cuts in funding ought to consider a career in the social sciences, where it ought to be possible to solve the problems the social scientists are worked up about in a trice".
Twelve years later, and now a sociologist myself, I've learned that Gribbin is not alone in his scepticism about what social science offers. To many people, and I suspect to many readers of this magazine, it is deeply unclear what sociology has to say about the world that an intelligent person couldn't have figured out on their own. It's a reasonable question, and as a former "hard" scientist, it's one that I admit to having asked myself. But as Paul Lazarsfeld, one of the giants of 20th-century American sociology, pointed out nearly 60 years ago, it also reveals a common misconception about the nature of social science.
"Aha," says Lazarsfeld's imagined reader, "that makes perfect sense. Rural men in the 1940s were accustomed to harsher living standards and more physical labour than city men, so naturally they had an easier time adjusting. Why did we need such a vast and expensive study to tell me what I already knew?" Why indeed.
But Lazarsfeld then reveals the truth: all six of the "findings" were in fact the exact opposite of what the study found. It was city men, not rural men, who were happier during their army life. Of course, had the reader been told the real answers in the first place, they could just as easily have reconciled them with other things they already thought they knew: "City men are more used to working in crowded conditions and in corporations, with chains of command, strict standards of clothing, etiquette, and so on. That's obvious!" But this is exactly the point Lazarsfeld was making. When every answer and its opposite appears equally obvious then, as he put it, "something is wrong with the entire argument of 'obviousness'".
Lazarsfeld was talking about social science, but as I argue in my new book, his point is just as relevant to any activity that involves understanding, predicting, changing or responding to human behaviour, from marketing to politics to policy-making.
Politicians dealing with urban poverty feel they already have a good idea why people are poor. Marketers planning campaigns already feel that they have a decent sense of what consumers want and how to make them want more of it. Economic policy-makers believe that they can do a reasonable job of getting the incentives right, whatever end they are trying to achieve. They don't expect to get them right all the time, and they would be the first to admit that it's complicated. Nevertheless, they do think the problems they are contemplating are mostly within their grasp, that they are "not rocket science".
What's puzzling about this attitude is that if we compare recent progress in the physical versus the social sciences, it should be clear that we're actually much better at rocket science than managing the economy, merging corporations, or even predicting how many copies of a book will be sold.
So why does rocket science seem hard, while problems to do with people – which in some respects are clearly much harder – seem like they ought to be just a matter of common sense?
As it turns out, the key is common sense itself. Common sense is exquisitely adapted to handling the kind of complexity that arises in everyday situations, such as how to behave at work versus in front of your children versus in the pub with your mates. And because it works so well in these situations, we're inclined to trust it.
But situations involving corporations, cultures, markets, nations and global institutions exhibit a very different kind of complexity. Large-scale social problems necessarily involve anticipating or managing the behaviour of many individuals in diverse contexts over extended periods of time. Under these circumstances, the ability that Lazarsfeld highlighted of common sense to rationalise equally one behaviour and also its opposite causes us to commit all manner of prediction errors.
Yet because of the way we learn from experiences – even ones that are never repeated – the failings of common sense reasoning are rarely apparent to us. Rather, they manifest simply as "things we didn't know at the time" but which seem obvious in hindsight.
The paradox of common sense, then, is that even as it helps us make sense of the world, it can actively undermine our ability to understand it.


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The NVBA has published newsletters for many years. The schedule has changed from 6 times a year to 4 times a year. Information about players, the unit, upcoming events such as the annual meeting, the holiday party and the LM/NLM game are in the newsletter. You can use the matrix below to access and read any historical newsletter that we have in electronic format. They're in PDF format and may be printed on your home printer.


text:
SDSU student takes honors with calf research
News | April 21, 2016
Nirosh Dias Senevirathne, a South Dakota State University (SDSU) graduate student, was awarded second place in the Alltech Young Scientist North American regional graduate division. The title of the award winning paper was "Calf nutrition and growth performance."
Senevirathne was nominated to participate in the Alltech Young Scientist program by his mentor, Jill Anderson, assistant professor in the SDSU dairy science department. For individuals to compete in this year's competition, the student had to be nominated by a professor from their university.
The Alltech Young Scientist program is an opportunity for students to participate with the company in pursuit of natural solutions in animal health, plant technology, environmental sciences and other biotechnology-related sectors, while also gaining valuable experience with Alltech, a global leader in the feed and food industry.
"I feel amazed and am delighted to have achieved this award through my education. I am proud to be an Alltech Young Scientist winner," said Senevirathne. "I someday hope to help the agriculture industry through my research and in the future obtain a position in research and development."
For Senevirathne's second place award, he received $1,000 cash prize, a certificate of achievement, an Alltech Young Scientist Medal and a gift basket.
Senevirathne is working toward his doctorate through South Dakota State University. He completed his undergraduate degree in Sri Lanka and his master's degree in Japan.


text:
This Weekend's Top Six Events
November 13, 14, & 15
Thank Based God it's Friday!
Here are six ways to have a based weekend in the East Bay.
Silence Dem All
Maya Songbird.
Yoko O.K., Maya Songbird, and Jon Jon Cassagnol
To celebrate painter Laurie Shapiro's latest solo show at Grease Diner, the North Oakland art gallery and print shop is hosting an opening reception that features three musical guests: Yoko O.K., Maya Songbird, and Jon Jon Cassagnol. Maya Songbird is a San Francisco native who creates danceable, disco-inspired tracks that evoke post-riot grrrl electro pop artists, such as Le Tigre. While Yoko O.K.'s left-field folk music departs from Songbird's sonically, both artists champion a similar ethos of self-acceptance and quirkiness. O.K.'s album, Menahan Tree, is a collection of mumblecore, bedroom acoustic songs with a slightly off-beat sense of humor. Jon Jon Cassagnol, co-owner of Grease Diner, creates music in a similar vein, except with a rougher sound that has a noticeable punk influence. This show, which takes place on November 13, will be the first time Cassagnol performs his new, electronic material. — Nastia Voynovskaya
Fri., Nov. 13, 6-10 p.m.TheGreaseDiner.com.
Oakland Art Market
Next up in East Bay alternative holiday shopping experiences: The Museum of Children's Arts (1625 Clay St., Suite 100, Oakland) will be hosting an Oakland Art Market pop-up on Friday, November 13 (5–8 p.m.) and Saturday, November 14 (noon–5 p.m.). Spearheaded by MOCHA "Teaching Artist" Kaya Fortune, the fair will offer a sampling of gifts made by more than 25 local artists and artisans. Many of the artists will be MOCHA's own Teaching Artists — those who lead the museum's many youth art classes. MOCHA has been offering hands-on programming intended to inspire children's creativity in Oakland for more than 27 years. Now, you can buy photography, clothing, jewelry, and paintings from the community of artists that facilitate that education. — Sarah Burke
Fri., Nov. 13, 5-9 p.m. and Sat., Nov. 14, 12-5 p.m. 510.987.8560. Free. Mocha.org
Pure One.
San Francisco producer Pure One (Alexandra Crotta) utilizes her voice as an instrument in her experimental electronic compositions. On many of her tracks, she manipulates her soft, cooing vocals beyond recognition, chopping them up and layering them until they take on fuzzy textures and warped shapes. Throughout "Melchedezek, My Love," one of her latest tracks, she murmurs lyrics that evoke spontaneous word associations: Let's go on/Let's go to war/Let's go to her. Her repetitive, whispered verses become a haunting mantra that washes over a slow, ambient beat, foregrounding the warbled, ebbing and flowing of her synth. Pure One's tracks are subtle and atmospheric, and while many of them use hip-hop and trip-hop beat structures as a starting point, they often unravel into avant-garde, electronic sound collages. Pure One is a relatively new music project, and she is releasing her debut EP later this month. She performs at Stork Club on November 14 with producer LoWGritt and indie band Arrows, both from Santa Cruz, as well as Oakland noise rockers Knowles Hand. — N.V.
Sat., Nov. 14, 9 p.m. $TBA. StorkClubOakland.com
Tom Manning
Welcome to Range, a small Washington town you can never leave. It's a land inhabited by talking animals, a cute floating ghost thing, an undead lunatic, a pirate (of course), and a whole cast of other characters that are both familiar and strange. It's also the setting of Oaklander Tom Manning's comic, Runoff, an epic three-part tale that is finally being published as one collection after eight years of production and self-publishing. Manning will be at Umami Mart (815 Broadway, Oakland) on Friday, November 13 — appropriately timed for this spooky storyline — to officially release the compilation and sign copies. The series, which Manning started in 1999 and completed in 2007, reportedly caught the eye of Guillermo Del Toro (Hellboy, Pan's Labyrinth, etc.) who had set his sights on turning the part mystery, part drama, part comedy into a movie. Whatever came of that isn't clear, but it is certain that this comic offers more than a dystopian view of humanity with a few laughs. The characters are vivid, poignant, and compelling. — Erin Baldassari
Fri., Nov. 13, 6-9 p.m. Free. Umamimart.com
Bert Johnson/File
Krystle Ahmadyar.
Krystle Ahmadyar
Oakland singer-songwriter Krystle Ahmadyar is known for her vocal-driven solo project, Ruby Mountain, which pairs her soulful singing with ukulele and, occasionally, guitar and minimal electronic production. Since August, Ahmadyar has been the artist-in-residence at Studio Grand, where she has been developing new songs that fuse elements of her background in composing experimental electronic music at Mills College and playing Afghan folk songs with her father. Earlier this year, Ahmadyar visited Afghanistan for the first time, where she spent the majority of her visit with her extended family, listening to elders' stories, singing traditional music, and taking field recordings. The next performance of her residency, Crescent Moon, pays homage to her heritage with a night of Afghan music and storytelling with an experimental bend. Her father, Fazal Ahmadyar, who is a singer and harmonium player, and percussionist Said Hashemi will join Ahmadyar on stage. — N.V.
Sun., Nov. 15, 6-8 p.m. $7-$15. StudioGrandOakland.org
Ayodele Nzinga.
Zara's Faith: Somebody Has Got to Stand Up
Zara's Faith follows a woman whose two sons were stopped by police. One was murdered and the other wounded. As Zara attempts to work through her anger and grief, she receives advice from her ancestors that leads her to explore her own conception of faith and community amid the plot's unraveling social conspiracy. The play, which was originally written by Marc Sapir before the Black Lives Matter movement arose, was performed as a staged reading at the Flight Deck by The Lower Bottom Playaz back in June. Now, the Lower Bottom Playaz, led by Ayodele Nzinga, will be presenting a newly revised and fully produced version of the play starring Black Lives Matter organizer Cat Brooks as Zara and Reginald Wilkins as supporting lead, Pastor Simms. The performance will take place at La Peña Cultural Center (3105 Shattuck Ave., Berkeley) on November 13 at 8 p.m. — S. B.
Fri., Nov. 13, 8-10 p.m. $8, $12. LaPena.org
If your pockets are feelin' light and you're still yearning for more suggestions, we've got a ton, and these ones are all FREE! We're Hungry: Got any East Bay news, events, video, or miscellany we should know about? Feed us at Sarah.Burke@EastBayExpress.com.


A single correlate of effective vaccine protection against chronic HCV infection has yet to be defined. In this study, we analyzed T-cell responses in four chimpanzees, immunized with core-E1-E2-NS3 and subsequently infected with HCV1b. Viral clearance was observed in one animal, while the other three became chronically infected. In the animal that cleared infection, NS3-specific CD8 T-cell responses were observed to be more potent in terms of frequency and polyfunctionality of cytokine producing cells. Unique to this animal was the presence of killing-competent CD8 T-cells, specific for NS31258–1272, being presented by the chimpanzee MHC class I molecule Patr-A*03∶01, and a high affinity recognition of this epitope. In the animals that became chronically infected, T-cells were able to produce cytokines against the same peptide but no cytolysis could be detected. In conclusion, in the animal that was able to clear HCV infection not only cytokine production was observed but also cytolytic potential against specific MHC class I/peptide-combinations.
Copyright: © 2014 Verstrepen 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: This research was initiated by EC grant number QLK2-CT-1999-00356 and supported by ZonMw number 114024014. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Hepatitis C virus (HCV) infection is characterized by a high propensity for development of chronic infection, which typically manifests as asymptomatic for a long period of time. However, over decades the virus causes subtle, but cumulatively irreversible, hepatic damage. A vaccine that could prevent development of persistent HCV infection would therefore be of great clinical benefit.
The chimpanzee is the only validated animal model of HCV infection , . The dilemma that biomedical research in non-human primates, and chimpanzees in particular, is inevitably associated with low animal numbers and limiting statistical analysis, has been discussed elsewhere . Nevertheless, we believe that results from in depth immune-profiling of immunized chimpanzees may provide new insights into immune mechanisms operating in the early phase after infection and as such is important for optimal vaccine development in the future.
Spontaneous HCV clearance has been associated with the presence of broad and strong T-cell responses in both humans and chimpanzees –. Moreover, specific memory T-cell responses often correlate with early clearance after HCV reinfection –. These data imply that T-cell-based vaccines may facilitate HCV clearance, protect from viral persistence and thus from HCV-related disease progression.
For this reason, several HCV T-cell vaccine-candidates have been evaluated. Some of these vaccine-strategies induced strong T-cell responses however, no correlation was observed between either the magnitude or the breadth of vaccine-induced T-cell responses, and viral clearance. The lytic capacity of vaccine-induced CD8 T-cells, in the context of MHC class-I presentation was not previously evaluated in relation to viral clearance.
Previously, we reported partial control of an HCV 1b challenge after a DNA prime-MVA-boost vaccine strategy, targeting HCV core, E1, E2 and NS3 in chimpanzees . In these animals, neither (neutralizing) antibody responses nor ex vivo cytokine production or proliferative responses were uniquely associated with control of HCV infection. Therefore, the current study aimed to identify potential mechanisms of protection via detailed functional characterization of the T-cell responses against the most dominantly recognized antigen, NS3 .
We report that the animal that cleared infection had a high percentage of polyfunctional cytokine producing CD4 and CD8 T-cells against NS3-peptides. Moreover, a strong cytolytic T-cell (CTL) response against epitope NS31258–1272 was identified, which was uniquely presented in the context of the chimpanzee MHC class I molecule Patr-A*03∶01.
This retrospective study was performed on cryopreserved PBMCs, isolated from animals that were part of an HCV vaccine-efficacy study . As previously described, the preceding study had been performed in six purpose bred, naive mature chimpanzees (Pan troglodytes) that were housed at the Biomedical Primate Research Centre (BPRC), Rijswijk, The Netherlands, according to international guidelines for non-human primate care and use (The European Council Directive 86/609/EEC, and Convention ETS 123, including the revised Appendix A). The Institutional Animals Care and Use Committee (DEC-BPRC) approved study protocols (#426) according to strict international ethical and scientific standards and guidelines. The qualification of the members of this committee, including their independence from a research institute, is stated in the "Wet op de Dierproeven" (1996). The experiment can only be performed after a positive recommendation from this ethical committee. Permission for projects on non-human primates is requested in an animal ethical committee form. This form is discussed in the ethical committee and, if approved, a written permission is sent to the project leader and a copy to the director of the BPRC, who holds licence to perform animal experimentation. The project was monitored by a qualified independent veterinarian, specifically regarding the ethical issues of the projects.
The animals were in good physical health with normal biochemical and hematological values. The animals were socially housed in a BSL3-facility with spacious cages, and were provided with commercial food pellets supplemented with appropriate treats. Drinking water was provided ad libitum. Environmental enrichment was provided daily and suffering of the animals was alleviated wherever possible. In 2002, the European Council banned the use of apes for biomedical research in Europe. Yet, this was decided after the initiation of the original vaccine-study described by Rollier et. al. . For this reason, the European Council approved continuation of this particular study. The following years, all chimpanzees that were housed at the BPRC were outplaced to public zoos or animal sanctuaries. Before 2001, the use of chimpanzees for biomedical research was highly restricted and regulated, it was not allowed to perform terminal experiments on these animals .
For stimulation and expansion of HCV-specific cells, two different NS3-peptide-pools were compiled: NS3vaccine was a mixture of 15-mer peptides with 11 amino acids overlap, covering the NS3-sequence of the vaccine constructs; NS3challenge was a mixture of 15-mer peptides with 11 amino acids overlap covering the NS3-variants observed in the inoculum. Identification of NS3 epitopes within NS3 was first performed via the matrix setup  and confirmed in a second assay using individual peptides (peptide-sequences are listed in Table S1).
Cells isolated before infection were expanded and tested for response against peptide-pool NS3vaccine, whereas cells isolated after challenge were tested against peptide-pool NS3challenge. PBMCs isolated at the day of challenge were stimulated in two separate cultures with either NS3vaccine or NS3challenge.
After thawing, PBMCs were washed and resuspended in culture medium, R20 (RPMI, Invitrogen, CA, USA) supplemented with 20% FCS (MP medicals, Solon, OH, USA), pen/strep (100 U/ml/100 µg/ml) (Invitrogen) and L-glutamine (2 mM) (Invitrogen)) and cultured at a final density of 10 million cells per ml in a 24 well culture plate (Greiner bio-one). The cultures were initiated in R20 containing 5 ng/ml IL-7 (Peprotech) and NS3-peptide-pool (5 µg/ml per peptide). After overnight incubation, 1 ml R20 containing IL-7 (10 ng/ml) and IL-2 (Proleukin Chiron, final concentration 10 IU/ml) was added per well. Cultures were checked daily for general condition and depending on cell density, fresh R20 IL-2/IL-7 was added or cultures were split. After 12 days, the expanded cells were collected and resuspended in fresh R20 for further analysis.
The cytokine production profile of HCV-specific T-cell lines was determined by intracellular cytokine staining (ICS) after restimulation with NS3-peptides. For T-cell epitope mapping 0.2×106 expanded cells were either restimulated with NS3-peptide-pool (1 µg/ml/peptide) or without NS3-peptide, in medium containing co-stimulatory αCD28 and αCD49d molecules (2 µg each, BD-Biosciences) and 5% FCS (MP-Biomedicals). After 2 hrs, Brefeldin A (Golgiplug 1∶1000, BD-Biosciences) was added and 16 hours later the surface markers were stained with a panel of fluorochrome labeled antibodies, containing CD3-PacificBlue (clone SP-34-2, (BD-Pharmingen), CD14-PE-TexasRed (clone RMO52, BeckmanCoulter), CD20-PE-TexasRed (clone B9E9, BeckmanCoulter), CD4-PE-Cy7 (clone SK3, BD-Pharmingen) and CD8-APC-H7 (clone SK1, BD-Pharmingen). Stained cells were fixed and permeabilized (Cytofix/Cytoperm, BD Biosciences), followed by staining of accumulated intracellular cytokines with IFNγ-APC (clone B27, BD Pharmingen), IL-2-PE (clone MQ1-17H12, BD Pharmingen) and TNFα-FITC (clone MAb11, BD Pharmingen). Cell staining was analyzed on a FACSAria (BD Bioscience) and DIVA software Version 6.1.1.
To evaluate antigen sensitivity, T-cell lines were restimulated with increasing concentrations of individual peptide ranging from 0.05 to 10 µg/ml before intracellular cytokine production was measured.
Animals were genotyped for their MHC-class-I repertoire as previously described  (Table 1).
Table 1. MHC class I genotypes of the chimpanzees.
NS3-peptide-specific T-cell lines were tested for their capacity to kill peptide-loaded target cells, using a new non-radioactive cytotoxicity assay. In this assay, a panel of matched Patr MHC-class-I expressing transfectants, either constructed in 721.221 cells (kindly provided by C. Walker, Center for Vaccines and Immunity, The Ohio State University) or K562  were used as target cells. In brief, 3×106 MHC-matched transfected target cells were labeled with 1 µM CFSE (CFSElow) (Fluka) and another 3×106 MHC-matched transfected target cells, expressing the same class-I molecule were labeled with 8 µM CFSE (CFSEhigh). Subsequently, CFSElow cells were incubated with medium alone, whereas, CFSEhigh target cells were pulsed with HCV-peptide (8 µg/ml) for 2 hrs at 37°C. Excess peptide was removed by washing before mixing CFSEhigh and CFSElow with 1×106 NS3-specific expanded T-cells (ET ratio 10∶1). The mixed target and effector cells were cocultured for 16 hrs, after which CFSE staining was analyzed using a FACSAria (BD Bioscience) and DIVA software Version 6.1.1.
and lysis was considered positive when exceeding 25%.
A cell-based peptide-binding-competition-assay was performed as previously described . In this assay, peptides are tested for binding to transfected cell lines expressing chimpanzee class-I molecules, while competing with a biotin-labeled reference-peptide. In a 96 wells plate, 100.000 MHC class-I transfected K562 or 721.221 cells were incubated with 500nM of the biotinylated-indicator-peptide (in case of A*03∶01 ATALECVYK was used, and in case of B*01∶01 LSDMHLCSI, in which C indicates the position of the biotin-labeled cysteine) and 0.05 to 100 nM of the peptide of interest, for instance p59 (NS31258–1272), LGFGAYMSK or AATLGFGAY. Peptide KGGLRPRAG, predicted to have no or a very low binding affinity to MHC class I molecules based on the incorporation of glycines on the anchor positions, was used as control. After overnight incubation, excess peptide was removed by washing and indicator-peptide bound to cells was quantified using the DELFIA-system and the Victor3 1420 multilabel counter (Perkin Elmer). The binding affinity of the indicator peptide was verified using its unlabeled peptide as competitor, this is referred to as standardized competitor .
We previously reported  an HCV-vaccine evaluation-study in chimpanzees, in which 4 out of 4 immunized animals became viremic shortly after challenge with HCV 1b J4. However, at a later stage, animal Vac1 cleared HCV infection while the three other animals, Vac2, Vac3 and Vac4 became chronically infected. As reported previously, IFNγ release, ex vivo lymphoproliferative responses, Th1/Th2 cytokine release patterns, and HCV-specific antibodies were comparable amongst the four animals . NS3 was the immunodominant antigen in all four animals, while the responses against core, E1 and E2 were not only less intense, but also largely absent in Vac1, the animal which was protected against chronic infection. Therefore, functional analysis of the cellular responses was focused on the immunodominant NS3 antigen .
After 12 days of culturing in the presence of a pool of NS3 peptides, T-cell lines were restimulated with individual peptides and analyzed for intracellular expression of IFNγ, IL-2 and TNFα using the gating strategy shown in Figure S1. Cells within the lymphocyte gate that were CD3 positive, and CD14 and CD20 negative were selected and subsequently IL-2, IFNγ and TNFα single, dual or triple cytokine expression was analyzed in the CD4 and CD8 T-cell subsets within the T-cell lines. Cytokine expression profiles varied per peptide as illustrated in Figure 1. For instance, only moderate cytokine induction was observed after peptide specific restimulation of expanded CD4 cells in Vac1. In contract, strong dual IFNγ and IL-2 expression was seen in expanded CD8 T-cells restimulated with peptide 59 (NS31258–1272), while medium alone or peptide 64 (IRTGVRTITTGGPIT) did not induce cytokine production. In comparison, in Vac2 p59 only gave a marginal cytokine induction in expanded CD8 T-cells, while p64 induced moderate dual IFNγ/IL-2 and IFNγ/TNFa cytokine production (Figure 1). Instead, expanded CD4 T-cells in Vac2, showed stronger response towards p59.
Figure 1. Evaluation of peptide-specific IL-2, IFNγ and TNFα expression in CD4 and CD8 T-cells.
A representative example of the analysis performed in Vac1 (left panel) and Vac2 (right panel), two weeks after the final vaccine boost, is shown. PBMC were first expanded for a 12 day period using a pool of all NS3 peptides, restimulated with individual peptides (medium alone, p59 or p64) and analyzed for induction of IFNγ, IL-2 and TNFα cytokine expression by expanded CD4 and expanded CD8 cells, using the gating strategy described in Figure S1. The numbers in the quadrants, represent the percentage of positive cells calculated from the parent-population.
In this manner the responses against each individual NS3 peptide was investigated and the results are summarized in Figure 2 in the form of heat maps; showing the percentages of IFNγ producing CD4 (Figure 2(A)) and IFNγ/IL-2 dual cytokine producing CD8 (Figure 2(B)) T-cells in all four animals at all time points tested. In general, analysis of peptide responses by dual cytokine expression gives a clear-cut distinction between positive and negative responses, because of low background responses. However, for CD4 T-cells the number of dual cytokine expressing cells was often rather low, making it necessary to consider the total number of IFNγ cytokine producing cells. The peptide recognition profiles shown in Figures 2(A and B) indicate that some regions within the NS3 peptide sequence, indicated by red boxes, are broadly recognized amongst the animals. Other peptide specific responses were more restricted and only gave positive responses at some time points in one to two animals. No unique peptide response-pattern could be discerned for Vac1 that could explain vaccine induced viral clearance as compared to Vac2, 3, and 4. Moreover, no relation was observed between the number of peptides recognized and the course of infection (Table 2).
Figure 2. Heat map of NS3-peptide-specific T-cell responses in four immunized chimpanzees at different study time points.
Table 2. Number of NS3-peptides that induce IFNγ production by CD4 T-cells or IL-2/IFNγ dual cytokine production by CD8 T-cells.
Next, the sum of the % of cytokine producing cells induced by all 156 individual peptides was calculated for expanded CD4 and CD8 lines. This total frequency of peptide-specific IFNγ-producing expanded CD4 T-cells was comparable between Vac1 and Vac2, but somewhat lower in Vac3 and Vac4 (Figure 2(C)). After the last immunization (week -4), total peptide specific CD8 T-cell responses were higher in Vac1 as compared to the other animals (Figure 2(D)). Triple cytokine production, i.e. simultaneous expression of IFNγ, IL-2 and TNFα by expanded CD8 cells, was also higher in Vac1 and for instance observed in 12.3% of the p59-specific expanded CD8 T-cells 4 weeks prior to HCV infection, whereas this was less than 7% in Vac2, Vac3 and Vac4 (Figure. 2(E)).
In conclusion, we observed increased number of NS3 specific cytokine producing expanded CD8 T-cells after the last immunization, and especially more triple IFNγ, IL-2 and TNFα cytokine producing cells in the animal that cleared the virus, relative to those animals that became chronically infected.
Next, NS3-peptide-specific cells were evaluated for their capacity to kill peptide-loaded target cells. Similar to standard 51chromium release assays, T-cells were expanded prior to analysis , . The expanded T-cells were tested for their lytic capacity for all individual peptides that were found to induce IL-2/IFNγ dual expression in expanded CD8 cells (Figure 2(B)) and was tested against a panel of MHC matched target cells  (Table 1).
Figure 3(A) illustrates specific killing of p59-pulsed CFSEhigh relative to unpulsed CFSElow Patr-A*03∶01 target cells, using expanded T-cells from Vac1, isolated 10 weeks after HCV challenge. As a control, no difference in ratio of the area under the curve was observed if both the CFSEhigh and CFSE low Patr-A*03∶01 targets were unpulsed (Figure 3(C)), precluding non-specific toxicity due to labeling with higher CFSE concentration. Killing of target cells was only observed for particular peptide-MHC class I combinations. As shown in Figures 3(B and D), neither unpulsed nor p59-pulsed Patr-B*02∶01 targets were lysed by expanded cells from Vac1.
Figure 3. Cytolytic killing of CFSE-loaded target cells.
(A) Specific lysis of NS31258–1272-pulsed Patr-A*03∶01 target cells (red peak) in comparison with unpulsed Patr-A*03∶01 target cells (yellow peak) by NS3-peptide-pool expanded PBMC from Vac1 isolated 10 weeks after HCV 1bJ4 challenge (blue peak). (B) No specific lysis of NS31258–1272-pulsed Patr-B*02∶01 target cells. (C) and (D) Control experiments showing no killing of unpulsed Patr-A*03∶01 or Patr-B*02∶01 target cells. (E,F,G) Percentage of lysis for all MHC class-I-peptide-combinations tested in Vac1, Vac2, Vac3 respectively.
Amongst all peptides tested, i.e. 24, 22 and 35 in Vac1, Vac2 and Vac3 respectively, highly efficient killing was only observed against p59 in Vac1 and only when this peptide was presented by Patr-A*03∶01 (Figure 3(E to G)). Only the peptides partly overlapping with p59 gave some lysis, while none of the other MHC class-I-peptide-combinations exceeded 25% of specific killing. Due to the limited number of cells after expansion, Vac4 could not be tested.
Collectively, the data show that although stimulation with p59 can induce some level of cytokine production in all vaccinees, a p59-mediated cytolytic capacity was only observed in Vac1.
Peptide binding to MHC-class I is determined by the anchor-residues fitting into the B- and the F-pocket of the relevant allotype for most MHC class-I-molecules. Usually MHC class I molecules bind nonamer peptides, in which the p2-position of the peptide anchors into the B-pocket and the p9-position of the peptide anchors into the F-pocket. Peptide 59, which was observed to initiate cytolysis when presented in the context of Patr-A*03∶01, was scanned using the NetMHCpan version 2.8 prediction algorithm  against the MHC class I molecules present in the studied cohort (Table 1). It is known that peptides that bind, or are predicted to bind to MHC class I molecules with an affinity <500 nm are potential T-cell epitopes –. In p59 four potential nonamer peptides are present, AATLGFGAY, ATLGFGAYM, TLGFGAYMS, and LGFGAYMSK. The only combination that was predicted to bind with an affinity <500 nm was LGFGAYMSK in the context of Patr-A*03∶01. A cell-based peptide binding competition assay was performed to test the binding-affinity of p59, and the nonamer peptides AATLGFGAY and LGFGAYMSK to Patr-A*03∶01 and as control to Patr-B*01∶01. The peptide binding motif of Patr-A*03∶01 is defined as X(STA)XXXXXX(RK) and for Patr-B*01∶01 as (X(ST)XXXXXX(IL) , . Figure 4 shows that all three NS3 peptides have a high binding affinity to Patr-A*03∶01, with IC50 values between 0.03 to 0.2 µM, which is higher compared to the control peptide (IC50 = 8.2 µM) and the standardized competitor peptide of Patr-A*03∶01 with IC50 = 1.5 µM . Peptide LGFGAYMSK was found to have the highest binding affinity to Patr-A*03∶01. In contrast, only low binding affinity was observed for p59 in the context of Patr-B*01∶01 as compared to its standardized competitor (IC50 = 1.0 µM), whereas for the two nonamer peptides, no amenable regression curves could be drawn suggesting low or no binding affinity (not shown).
Figure 4. Dose-inhibition curves for selected NS3-peptides to Patr-A*03∶01.
Binding affinity of selected NS3-peptides to Patr-A*03∶01, established by peptide binding competition assay. Patr-A*0301 cells were incubated with increasing concentrations of NS3 p59 (top left), LGFGAYMSK (LGF, top right), AATLGFGAY (AAT, bottom left), or KGGLRPRAG (control, bottom right) in the presence of biotin-labeled indicator peptide. Binding of indicator peptide was quantified by incubation with europium labeled streptavidin. Indicated is reduction in percentage of europium positive cells relative to cells incubated with indicator-peptide only. The indicated IC50 values (µM) are derived from the regression curves of three independent experiments in the case of p59, LGF and AAT, and of two individual experiments in the case of KGG. The IC50 values were estimated using non-linear least-squares regression with the "R" platform for statistical computing.
Antigen specific responses not only depend on the presentation of peptides by MHC class I molecules but also on the binding of the peptide/MHC complex to the T-cell-receptor (TCR). Therefore, we tested whether the response-threshold of CD4 and CD8 T-cell lines from Vac1 differed from Vac2 and Vac3. To this end, NS3-specific expanded T-cells, isolated 4 weeks prior to HCV infection, were expanded from PBMCs, restimulated with different concentrations of NS31258–1272 ranging from 0.05 to 10 µg/ml, and stained for intracellular IL-2 and IFNγ. Expanded CD8 T-cells from Vac1 were found to respond at a peptide concentration as low as 0.05 µg/ml whereas, at least, 20 times more peptide was required for CD8 T-cells from Vac2 and Vac3 (Figure 5). For IFNγ production by expanded CD4 T-cells the difference is even more striking. Here, we could hardly detect p59-specific IFNγ producing CD4 T-cells in Vac2 and Vac3 while these cells are abundantly present in Vac1.
Figure 5. Antigen sensitivity of p59 specific cytokine induction.
NS3-expanded T-cells from Vac1, Vac2 and Vac3, from 4 weeks prior to HCV challenge, were restimulated with a concentration range of p59 and evaluated for induction of IL-2/IFNγ expressing CD8 T-cells (left graph) and IFNγ expressing CD4 T-cells (right graph).
The chimpanzee is the closest living relative of man and the only validated animal model for HCV infection. The evolutionary proximity of both species is reflected by close genetic and immunological similarity. For these reasons the chimpanzee provides an exquisite animal model, in which immune responses against HCV vaccines and the effect on deliberate infection can be investigated in an experimental setting . Here, we report of an in depth profiling of CD4 and CD8 T-cell responses in four chimpanzees, which were immunized with an experimental vaccine for HCV and subsequently challenged with HCV 1b J4 .
While all four animals became infected, only the Patr-A*03∶01 positive animal was able to clear HCV infection. Detailed mapping of CD4 and CD8 T-cell cytokine responses, and the analysis of their cytolytic potential, revealed that expanded T-cells of the vaccinee which effectively cleared HCV infection displayed: a) a higher frequency of peptide-specific IL-2/IFNγ dual cytokine producing CD8 T-cells; b) peptide-specific TNFα production in CD8 T-cells; c) cytolytic capacity against peptide-loaded target cells; d) cytokine induction at a low antigen concentration for the cytolysis-inducing peptide. Remarkably, the cytolysis-inducing peptide p59 was found to bind strongly to Patr-A*03∶01, which was only present in Vac1. Using the NetMHCpan algorithm, it was predicted that peptide LGFGAYMSK, one of the nonamer peptides present in the 15-mer p59, would show the strongest binding to Patr-A*03∶01. Peptide LGFGAYMSK was also predicted to bind to the MHC class I molecules Patr-A*01∶01, A*04∶01, A*04∶04, and A*09∶01, present in all of the studied animals, but only with intermediate affinity between 0.9 and 5 µM. These data are in agreement with the differences in cytokine responses and cytolyses reported in expanded T-cells here and could indicate a particular affinity threshold is necessary for a peptide to induce adequate immune responses for combating HCV. The strong binding of the 15-mer p59 to Patr-A*03∶01 is somewhat unexpected, as generally nonamer peptides bind to MHC class I molecules. Nonetheless, in humans it has been shown that particular MHC class I molecules are capable of binding longer peptides with bulged structures , .
Stimulation assays would give information about the biological relevance of the binding experiments. However, the limited number of cells stored from these animals, preclude performing such assays.
Unlike ex vivo detection of CTL in humans, detection of these cells in chimpanzees can often only be achieved after amplification of antigen specific responses , . Obviously, in vitro expansion of T-cells from frozen PBMC has its limitations. Antigen presenting cells may be affected by the freezing/thawing process resulting in –partly hampered- antigen presentation. Furthermore, the addition of cytokines and peptides may skew the immune system differently as compared to the in vivo situation.
To determine the degranulating capacity of T-cells, CD107a stainings were performed on T-cell lines. However, the results of these assays were unclear and no conclusion could be drawn from it, basically because of the lack of a proper negative control.
Unlike in the intracellular cytokine staining assays, where the difference between cells restimulated with peptide or medium alone was very clear, no clear difference could be observed when CD107a was used. Probably the expansion-protocol used here does agree with antigen specific CD107a staining. Unfortunately, the number of stored cells from these animals is limited and assays like this require too many cells to repeat the assays.
And although we cannot exclude that upon further amplification of antigen specific responses, CTL responses might also be detectable in the other animals, our data suggest that they are of lower magnitude.
To study the possibility of viral escape from immune pressure as a cause of development of chronicity in Vac2, Vac3 and Vac4, analysis of viral sequences was performed and compared with the inoculum sequence (data not shown). Only a limited number of amino acid substitutions were detected in the areas that overlap with cytokine-inducing peptides. There was however no evidence for mutational escape in the immunodominant p59 in any of the animals.
This study highlights a potential role for CD8 mediated cytolytic responses as a discriminatory factor between viral clearance and the development of chronic infection in HCV vaccinated subjects and underscores that this parameter is to be considered as an important immunological readout in future vaccine trials. Indeed, studying peptide binding and CD8 cytolytic responses could complement the other contributing immune mechanisms previously described , , .
It should be noted that the reported data were obtained from peripheral blood and not from the site of HCV-induced inflammation, the liver. For ethical reasons and animal welfare, the number of liver biopsies taken during the study, was limited and not sufficient for the comprehensive analysis as presented in this manuscript. Nevertheless, a direct comparison of CD8 T-cell function between liver and peripheral blood in chimpanzees during HCV infection, showed in essence that the cells in the circulation reflect the situation in the liver , .
The data presented here, are in line with the finding that self-limiting HCV infection in humans is associated with the presence of HLA-B*57 and HLA-B*27 restricted CD8 T-cell responses , . Chimpanzee Patr-A*03∶01 and Patr-B*01∶01 have almost identical B-pocket residues as compared to HLA-B*57. Moreover, peptide binding studies have shown that both Patr-molecules may potentially bind HLA-B*57 or HLA-B*27 .
In conclusion, we observed that in contrast to peptide-specific cytokine production, the induction of an effective CD8 T-cell effector function elicited in the context of peptide presentation by appropriate class-I molecules appeared to be associated with clearance of infection.
Gating strategy for the evaluation of cytokine production by CD4 and CD8 cells. Cells from Vac1, 2 weeks after the last vaccine boost were stimulated with NS3vaccine and restimulated with either NS31258–1272 (p59) or medium alone. Cells within the lymphogate were selected, followed by selection of the CD3 positive, CD20/CD14 negative population, subsequently CD4 and CD8 positive cells were selected. Expression of IFNγ as a function of IL-2 or TNFα was plotted.
Peptide numbering of Vaccine- and Challenge-sequence relative to HCV reference strain H77.
The authors would like to thank Thea de Koning for editing the manuscript and Henk van Westbroek for preparing the graphics. The NS3-peptides were provided by BeiResources from ATCC.
Conceived and designed the experiments: BV EV PM NdG RB WB JH GK. Performed the experiments: BV NdG ZF. Analyzed the data: BV NdG EV ZF PM GK. Contributed reagents/materials/analysis tools: BV NdG EV. Wrote the paper: BV GK NdG PM.
1. Bettauer RH (2010) Chimpanzees in hepatitis C virus research: 1998–2007. J Med Primatol 39: 9–23.
2. Bukh J (2012) Animal models for the study of hepatitis C virus infection and related liver disease. Gastroenterology 142: : 1279–1287 e1273.
3. Bacchetti P, Deeks SG, McCune JM (2011) Breaking free of sample size dogma to perform innovative translational research. Sci Transl Med 3: 87ps24.
4. Thimme R, Bukh J, Spangenberg HC, Wieland S, Pemberton J, et al. (2002) Viral and immunological determinants of hepatitis C virus clearance, persistence, and disease. Proc Natl Acad Sci U S A 99: 15661–15668.
5. Diepolder HM, Gruener NH, Heeney J (2003) Of chimps and men—finding the path to an HCV vaccine. Hepatology 37: 1472–1474.
6. Cooper S, Erickson AL, Adams EJ, Kansopon J, Weiner AJ, et al. (1999) Analysis of a successful immune response against hepatitis C virus. Immunity 10: 439–449.
7. Watanabe H, Wells F, Major ME (2010) Clearance of hepatitis C in chimpanzees is associated with intrahepatic T-cell perforin expression during the late acute phase. J Viral Hepat 17: 245–253.
8. Osburn WO, Fisher BE, Dowd KA, Urban G, Liu L, et al. (2010) Spontaneous control of primary hepatitis C virus infection and immunity against persistent reinfection. Gastroenterology 138: 315–324.
9. Prince AM, Brotman B, Lee DH, Pfahler W, Tricoche N, et al. (2005) Protection against chronic hepatitis C virus infection after rechallenge with homologous, but not heterologous, genotypes in a chimpanzee model. J Infect Dis 192: 1701–1709.
10. Nascimbeni M, Mizukoshi E, Bosmann M, Major ME, Mihalik K, et al. (2003) Kinetics of CD4+ and CD8+ memory T-cell responses during hepatitis C virus rechallenge of previously recovered chimpanzees. J Virol 77: 4781–4793.
11. Rollier CS, Paranhos-Baccala G, Verschoor EJ, Verstrepen BE, Drexhage JA, et al. (2007) Vaccine-induced early control of hepatitis C virus infection in chimpanzees fails to impact on hepatic PD-1 and chronicity. Hepatology 45: 602–613.
12. Haaijman JJ, Heeney JL (1995) European centralized chimpanzee facility 'preclinical studies for evaluation of effective vaccines and therapeutic strtegies for AIDS'. AIDS research at EC level. IOS press. pp. 139–148.
13. Hoffmeister B, Kiecker F, Tesfa L, Volk HD, Picker LJ, et al. (2003) Mapping T cell epitopes by flow cytometry. Methods 29: 270–281.
14. de Groot NG, Otting N, Arguello R, Watkins DI, Doxiadis GG, et al. (2000) Major histocompatibility complex class I diversity in a West African chimpanzee population: implications for HIV research. Immunogenetics 51: 398–409.
15. de Groot NG, Heijmans CM, Zoet YM, de Ru AH, Verreck FA, et al. (2010) AIDS-protective HLA-B*27/B*57 and chimpanzee MHC class I molecules target analogous conserved areas of HIV-1/SIVcpz. Proc Natl Acad Sci U S A 107: 15175–15180.
16. Balla-Jhagjhoorsingh SS, Koopman G, Mooij P, Haaksma TG, Teeuwsen VJ, et al. (1999) Conserved CTL epitopes shared between HIV-infected human long-term survivors and chimpanzees. J Immunol 162: 2308–2314.
17. Hoof I, Peters B, Sidney J, Pedersen LE, Sette A, et al. (2009) NetMHCpan, a method for MHC class I binding prediction beyond humans. Immunogenetics 61: 1–13.
18. Allen TM, Mothe BR, Sidney J, Jing P, Dzuris JL, et al. (2001) CD8(+) lymphocytes from simian immunodeficiency virus-infected rhesus macaques recognize 14 different epitopes bound by the major histocompatibility complex class I molecule mamu-A*01: implications for vaccine design and testing. J Virol 75: 738–749.
19. Loffredo JT, Sidney J, Wojewoda C, Dodds E, Reynolds MR, et al. (2004) Identification of seventeen new simian immunodeficiency virus-derived CD8+ T cell epitopes restricted by the high frequency molecule, Mamu-A*02, and potential escape from CTL recognition. J Immunol 173: 5064–5076.
20. Sette A, Vitiello A, Reherman B, Fowler P, Nayersina R, et al. (1994) The relationship between class I binding affinity and immunogenicity of potential cytotoxic T cell epitopes. J Immunol 153: 5586–5592.
21. Sidney J, Peters B, Moore C, Pencille TJ, Ngo S, et al. (2007) Characterization of the peptide-binding specificity of the chimpanzee class I alleles A 0301 and A 0401 using a combinatorial peptide library. Immunogenetics 59: 745–751.
22. Paul S, Weiskopf D, Angelo MA, Sidney J, Peters B, et al. (2013) HLA class I alleles are associated with peptide-binding repertoires of different size, affinity, and immunogenicity. J Immunol 191: 5831–5839.
23. Bell MJ, Burrows JM, Brennan R, Miles JJ, Tellam J, et al. (2009) The peptide length specificity of some HLA class I alleles is very broad and includes peptides of up to 25 amino acids in length. Mol Immunol 46: 1911–1917.
24. Fattori E, Zampaglione I, Arcuri M, Meola A, Ercole BB, et al. (2006) Efficient immunization of rhesus macaques with an HCV candidate vaccine by heterologous priming-boosting with novel adenoviral vectors based on different serotypes. Gene Ther 13: 1088–1096.
25. Zubkova I, Choi YH, Chang E, Pirollo K, Uren T, et al. (2009) T-cell vaccines that elicit effective immune responses against HCV in chimpanzees may create greater immune pressure for viral mutation. Vaccine 27: 2594–2602.
26. Shin EC, Capone S, Cortese R, Colloca S, Nicosia A, et al. (2008) The kinetics of hepatitis C virus-specific CD8 T-cell responses in the blood mirror those in the liver in acute hepatitis C virus infection. J Virol 82: 9782–9788.
27. Ramalingam RK, Meyer-Olson D, Shoukry NH, Bowen DG, Walker CM, et al. (2008) Kinetic analysis by real-time PCR of hepatitis C virus (HCV)-specific T cells in peripheral blood and liver after challenge with HCV. Journal of virology 82: 10487–10492.
28. Kim AY, Kuntzen T, Timm J, Nolan BE, Baca MA, et al.. (2011) Spontaneous control of HCV is associated with expression of HLA-B 57 and preservation of targeted epitopes. Gastroenterology 140: : 686–696 e681.
29. Neumann-Haefelin C, Timm J, Schmidt J, Kersting N, Fitzmaurice K, et al. (2010) Protective effect of human leukocyte antigen B27 in hepatitis C virus infection requires the presence of a genotype-specific immunodominant CD8+ T-cell epitope. Hepatology 51: 54–62.


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Patient Safety Blog - Lubin & Meyer PC
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FDA Advisory Panel Recommends Banning Shock Therapy at Judge Rotenberg Center
Video of Electronic Shock Treatment Played at Trial Helps Turn the Tide on Judge Rotenberg Center
As described by an article in New York magazine in April of 2012:
"For years, the state of New York has been sending troubled kids to a notorious 'school' in Massachusetts known as the Judge Rotenberg Center, where students who don't follow the rules receive painful electric shocks. The 'treatment' at the center has nothing to do with electroconvulsive therapy, where people with severe depression are induced into having seizures under anesthesia — it's behavior modification for troubled children, and the Rotenberg Center is the only place in America that uses this approach."
Now, after two years of in-depth reporting and a significant medical malpractice settlement between the Center and Cheryl McCollins (the mother of a former patient who filed a lawsuit when her 18-year-old autistic child, Andre, received 31 skin shocks, while strapped to a four-point restraint board for 7 hours resulting in PTSD), change has finally come.
As the New York article points out, she was not the first parent to sue.
"But McCollins and her lawyer have accomplished something that no one else has managed to do: show the public exactly what it looks like when a student is being shocked."
The disturbing video played a big part in illuminating the inhumane practice, reaching a settlement in the trial, turning public opinion and moving an FDA advisory panel to hold a hearing this week and make a recommendation to ban the practice. See the investigative report by Fox Undercover's Mike Beaudet below.
View our main website at LubinandMeyer.com
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Q: Magento ProductController append categoryId to products I am adding a mass action to add a category. I am most of the way there I only have one function left to figure out.
Clr\Categorymassaction\controllers\Adminhtml\Catalog\ProductController.php

    class Clr_Categorymassaction_Adminhtml_Catalog_ProductController extends Mage_Adminhtml_Controller_Action
    {
        public function massCategoryAction()
        {
            $productIds = $this->getRequest()->getParam('product');
            $cat = $this->getRequest()->getParam('Category');
            if (!is_array($productIds)) {
                $this->_getSession()->addError($this->__('Please select product(s).'));
                $this->_redirect('*/*/index');
            }
            else {
                    $cat = $category['label']->getCategoryId();
                    foreach($productIds as $product) {
                        //Process $cat into categoryId append categoryId to $productId
                       $cat->setPostedProducts($product);

                    }
              //Save product
              $cat->save();
            }

}
}

Clr\Categorymassaction\Model\Observer

class Clr_Categorymassaction_Model_Observer {
    public function addCategoryMassAction(Varien_Event_Observer $observer)
    {
        $block = $observer ->getBlock();
        if ($block instanceof Mage_Adminhtml_Block_Catalog_Product_Grid) {
            $block->getMassactionBlock()->addItem('Clr_Categorymassaction', array(
                'label'     =>  Mage::helper('catalog')->__('Add to Category'),
                'url'       =>  $block->getUrl('*/*/massCategory', array('_current' => true)),
                'additional'=> array(
                    'visibility'    => array(
                        'name'          =>'Category',
                        'class'         =>'required-entry',
                        'label'         =>Mage::helper('catalog')->__('Categories'),
                        'type'          => 'select',
                        'values'        => Mage::getModel('Categorymassaction/system_config_source_category')->toOptionArray(),
                        'renderer'      => 'Categorymassaction/catalog_product_grid_render_category',

                        )
                    )
                ));
        };
    }

}

One last thing
 class Clr_Categorymassaction_Model_System_Config_Source_Category
    {
        public function toOptionArray($addEmpty = true)
        {
            $options = array();
            foreach ($this->load_tree() as $category) {
                $options[$category['value']] =  $category['label'];
            }

            return $options;
        }

I am mostly in trouble here because I am refactoring, Flagbit_changeattributeset and Vuleticd_AdminGridCategoryFilter. I know what I need to do (at least I think I do) I just don't know how to finish this off. Thanks for your eyes and ears if you read it all.
UPDATE: The observer from Vuleticd_AdminGridCategoryFilter had this additional code
                    'filter_condition_callback' => array($this, 'filterCallback'),
                    )
                )
            ));
    };
}
public function filterCallback($collection, $column)
{
    $value = $column->getFilter()->getValue();
    $_category = Mage::getModel('catalog/category')->load($value);
    $collection->addCategoryFilter($_category);

    return $collection;
}

This was used to apply the filter to the grid. What I am trying to do is instead of using the dropdown to filter column fields; use the dropdown to trigger the ProductController to pass the selected items a new categoryid.

A: https://magento.stackexchange.com/questions/67234/productcontroller-for-mass-action  Asked this question over at magento's stackexchange figured I would post the link here for posterity.

redpajama_set_name: RedPajamaStackExchange