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Me'n Polyamory
By PolyFido
I must have been around nine at the time. Or was it ten? I heard some boys talking behind my back about a girl who was described as "the local bike". I thought I knew what that meant, and I knew who "she" was; but I didn't know that "she" was having sex with anyone, let alone that "she" was having sex with more than one boy.
All that I knew about her up until then was that she was attractive and confident; a kind of goddess in the background of my life at school. She was often there but never close by. I liked her...at a distance. As a matter of fact, I liked her a lot; everything about her. Rather than thinking less of this girl for having sex with more than one boy, she went up in my estimation. I just wished that I could be one of the lucky ones. But I was too young. I was still a kid, and she was a voluptuous young woman already; mature beyond her years.
But this remark that I overheard by chance set me thinking about my values regarding sex and love. For a start, I thought that the kids who were talking about this girl in such a demeaning way were probably not among 'the lucky ones'. I was sure that none of them had had sex with her. They were younger, like me. If any of them had had her, I thought that they deserved a good kick up the arse for betraying this nubile goddess in such a vicious way.
Even if they were mad with jealousy, why weren't they grateful that such a person existed...in their small town; someone so courageous and generous of spirit that she was willing to take on the risks of having sex before it was even legal, let alone safe! Did she do it for money? The question never entered my head.
I made up my mind at that time that, if I should ever be lucky enough to find someone whom I truly loved, that I would let her have the boyfriends she wanted. From that time onward, I truly believed that a lover was somebody that deserved gifts and respect. And if the gift that a lover wanted was another lover...or maybe even more than one, well, if it made her happy, then what was the problem, besides the obvious?
I soon learned that having more than one lover was not a socially acceptable thing to do. Yes, you could change girlfriends and boyfriends, but the rule was that you only had one at a time. The done thing was to break up and move on. Anything else was taboo. It meant that someone was 'cheating'. The word hasn't changed, and neither have the attitudes.
So, reluctantly, I embarked on a course of serial monogamy, just like everyone else I knew. That lasted right through my twenties. I didn't like it. To me, the whole idea of a monogamous sexual relationship with just one person, for life, was crazy. I could not see how it would possibly work for either me or the vast majority of people. It seemed like monogamy was a kind of straight-jacket that everyone had to accept so that society would function. In the decades since then, the divorce statistics seem to back the views that I had as a boy.
There were glimpses of other things. I tried very hard to live a polyamorous lifestyle in my late twenties. At one time I was sleeping with two women in the same house. It was a nightmare. I still remember the shirt being ripped from my chest. The two women didn't talk to each other; they just communicated through me. It was always the same question: "What did she (i.e. that bitch) say about me?" It didn't work for me, or either of the women then, and I thought that it could never work with anybody. Polyamory seemed to be in the 'too hard' basket. In fact, I had never heard of the term 'polyamory', and it's only a recent addition to my vocabulary.
As an older man, I can look back on a life of...what? Well, I just can't say, exactly. My ex-wife, if she is still alive, would probably sue me. She was one of the two women in that torrid threesome. And, no, I wasn't married to her in the turbulent days when I was trying to have a relationship with two women in the one house.
I had several major live-in relationships after my divorce. Yet I can truly say that sex was not an essential factor in my deeper emotional connection with a certain woman. She was old enough to be my mother and we met later in life. We were together for twenty years. It was never romantic or sexual, but it was a deep soul connection nonetheless.
In my days as a soldier I saw that things could be different. Some of the army wives had other lovers when hubby was away in the bush playing war games. But it was all very hush hush. It was the kind of thing that you heard about as an item of gossip if you were not in 'the club'; not something that was an open and upfront lifestyle.
Why the secrecy? Here we had a profession that was actively engaged in taking the lives of strangers in a foreign land--and doing it on a massive scale--yet here they were, keeping quiet about what goes on in the married quarters when hubby was away. It was such a disappointment. Yet what was being reflected was the values of society at large. That was the problem. So maybe secrecy had a point?
I thought that social attitudes were going to improve when I left the army and went to university. It was the early seventies; Vietnam was pretty much over; and social experimentation of a kind was almost compulsory. For example, if you wouldn't partake of a joint when it was handed to you, it wasn't going to do much for your sex life; at least, not in the circles that I mixed in.
Did that mean that people were swapping partners and having orgies? Not in my neck of the woods. Yes, there was sex with different people, at times. But in the end, couples formed and that was, well...that! So much for sexual liberation. Coupledom became the norm along with the denim and the left wing politics.
All along I stuck to my guns: If I had a girlfriend that wanted other lovers, she could have them, and I said so. I wasn't going to complain if I missed out and she was out there having fun. Sexual freedom wasn't a trade so far as I was concerned; it was a right. As a matter of fact, I thought that it was just inevitable that two people in a relationship were going to get bored with one another; couple sex gets boring, no question. So it's only fair that each person should be free to have a good time.
Eventually I got married, and, yes, my wife did have other lovers. Did I complain? No. But my wife refused to discuss her other relationships with me. I didn't pressure her. Was I disappointed? You betcha. We had spent the seventies doing therapy and she still couldn't even mention her sexual stuff, let alone talk about it freely. The worst thing was that she blamed me for the breakdown in our relationship.
Our marriage was built on deceit and secrecy when it came to sex; just like everything else. The final straw was that she thought that I didn't love her because I didn't get jealous about her secret love life; a love life that I wasn't supposed to even know about in the first place!
She had it all wrong... I just didn't love her any more, because of the other things. The relationship was over. It had been for a while. Sharing the same bed with someone when it's all over can be torture and a very lonely experience. Many of you have probably been there.
The idea that polyamory was a reality that could actually happen (with little effort from me) was just not on my mind. Yet that's just what's happened recently. And I'm so deeply grateful. So I'm going to make an effort to understand polyamory better, and to help people find out whatever they need to know about it. It's a learning curve; but one that I happily embrace
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Tax planning and preparation form a winning combination for our successful individual and business clients. Whether you are an individual or a multi-tiered partnership, our experienced staff can develop tax strategies that take advantage of new tax laws and legislation. We provide tax support strategies for all states and handle multi-state taxation as well. We also provide fast refunds through the use of electronic filing. Ask about our Accuracy Guarantee.


text:
The first constitution of the independent state was adopted in Halifax on December 18, 1776.
Delegates to the provincial congress meeting in Halifax on November 13, 1776, appointed a committee to prepare a constitution. Delegate Samuel Ashe took a leading role in the proceedings. The committee reviewed sets of instructions from Mecklenburg and Orange counties. Of particular value to their deliberations was a letter from William Hooper of Hillsborough, a signer of the Declaration of Independence. With his letter, the elder statesman enclosed copies of other recently adopted state constitutions.
The committee set to work and, a month later, presented proposals to the full body. The provincial congress made minor adjustments. On December 17, 1776, the delegates approved the declarations of rights, and, on December 18, 1776, the new state constitution. The full body debated the various provisions over the course of six meetings. Although approved separately, the two documents are usually treated as one whole. Over the years there has been disagreement as to the precise location in Halifax where the first state constitution was drafted and adopted.
John V. Orth, North Carolina Constitution: A Reference Guide (1993)Jerry L. Cross, "Historical Research Report for the Constitution House, Halifax, North Carolina," (unpublished report prepared for the Division of Archives and History, 1973)
Hugh T. Lefler and Albert Ray Newsome, History of a Southern State: North Carolina (1954)
R. D. W. Connor, History of North Carolina, I (1919)
W. C. Allen, History of Halifax County (1918)
US 301 Bypass at US 301 Business north of Halifax

redpajama_set_name: RedPajamaCommonCrawl


Given the fast pace of today's society, it's understandable if you feel overwhelmed at times with the challenge of keeping your children safe.
May is National Youth Traffic Safety Month. Our car accident attorneys believe that awareness and education are the keys to protecting your family.
With warm weather coming and the school year ending, I'd like to give you some information you can use to avoid auto accidents from occurring when your children get behind the wheel. Unfortunately, the summer months are when car accidents skyrocket for young adults.
Motor vehicle crashes were the leading cause of death in 2008 for children ages 3 to 14 and for 15- to 20-year-olds nationwide. In Michigan, car accidents are also the leading cause of death for children ages 0 to 15. Because of these statistics, National Youth Traffic Safety Month is now an annual campaign focusing on saving young people's lives and promoting traffic safety during prom, graduation and throughout the summer.
2. Put children younger than 7 years old or under 4 foot 9 inches in a car or booster seat. Research by the National Highway Traffic Safety Administration on the effectiveness of child safety seats has found them to reduce fatal personal injury by 71 percent for infants and by 54 percent for toddlers in passenger cars. Use the National Highway Traffic Safety Administration's Child Safety Seat Inspection Station Locator to find a certified technician who will inspect your car seat, and show you how to correctly install it.
3. Slow Down! In 2008, speeding was a contributing factor in 31 percent (11,674) of all fatal car crashes (37,261) nationwide. Today, we all seem to be in a hurry wherever we go. Remind yourself every time you start the car to drive at posted speeds – risking injury is not worth saving five minutes of drive time.
4. Children are also at risk as pedestrians and while riding bicycles. The most effective way to prevent bicycle injuries, especially traumatic brain injury, is by equipping your children with properly fitted bicycle helmets every time they ride. Children walking to school or in their neighborhood should exercise caution crossing roads and obey all traffic signals.
My next blog will address the top five hazardous actions by teen drivers. Meanwhile, I wish you all safe driving.
– Steven M. Gursten is recognized as one of the nation's top experts in serious car and truck accident injury cases and automobile insurance No-Fault litigation.
Michigan Auto Law is the largest law firm exclusively handling car accident, truck accident and motorcycle accident cases throughout the state. We have law firm locations in Sterling Heights, Ann Arbor, Detroit, Grand Rapids and Farmington Hills. Call (800) 777-0028 if you've been injured in an auto accident, and would like to speak to an attorney.


He has 1 child, a son.
He has disavowed both - Thats my issue with him.
It's just sick and wrong I tell ya, sick and wrong.
I'm new on this Forum and all I know is what I read. For me Julie was Julie, that meaning nothing but an Avatar posting its opinions and knowledge here. The reputation is acquired with the reliability on what is said and as far as I know everybody learned a lot with Julies posts.
Just to clear up a thing or two. This thread did NOT get "Julie" Banned and things could have turned out in a more positive way had "Julie" chosen to play by FTE's rules. This thread was just used as an instrument to point out that someone needed to respond to an email.
I havn't been reading this trend till 2day. WOW hit me with a brick. All i know is JULIE/DAN/WHO EVER has helped me out from day one.Julie if you read this THANKS MY FRIEND. Guess what folks my name ain't FRED its DALE DEXTER KENT.
At least your name matches your gender!
My Road name is Grizz and I ride with BACA (Bikers Against Child Abuse) so that is where my username came from.
My real name is David and I can control my temper and post without belittling or degrading anyone. Although, sometimes my humor gets misinterpreted.
Bet ya all would never guess what my name is.
Like I said before, it's just sad that he/she had to paint herself/himself into a corner.
I've often noticed the threads where the arguementative belittling began that it was in the wee hours of the morning. Perhaps after a few cocktails? I dunno, I don't want to speculate that. But I've watched a few threads get edited before the morning crowd got on. I've known since day one, and never once bought into the story. On more than one occasion I've watched the correct answer be given to a question, and then dan comes on and posts the answer again. But the credit went to Dan because he was a "girl" and that chivalry always took first place. I was never mad that Dan answered the question, I was just upset that he got all the credit, and that his reputation points kept climbing. So I went and deleted a lot of my posts and wrote various things such as, "I know nothing" out of frustration. Petty on myseolf? Perhaps, but after awhile, I got to seeing things a little differently. The charade started to wear off, and that Dan was starting to actually "get it" with the being courteous to everyone else, and such. However, I never thought a question about PM's would get this far, and how the end result played out. That all said, I will regret how I reacted, and how Dan has acted. I never once asked him to get banned. I just hoped he would have come clean, but when you've gone as far as he did, it's hard to do that. And that is the corner he's painted himself into. All things being fair, I would say he knew his stuff, as well as a few others here. I would never say that anyone here didn't, unlike Dan in a few posts. Just ask Alanco.
I've seen this happen in several other forums and it always irks me. I believe in giving credit where credit is due. Whenever I would post something that I've learned from one of the guys in the Paint & Bodywork forum, I would always say where I got my info from.
I have dealt with Julie, and Dan before he was she, and they were both very easy to work with and helpful, Haven't dealt with Uncle Jim yet though.
The Stories didn't always quite fit together very well right around the time that Dan "Died" and up through to this ongoing drama, (as others have already pointed out ) and I always speculated something a bit odd, But Hey we are all here with mostly common interests in ford trucks and sharing info. and a lot of us are a bit odd ourselves in our own ways.
Like a couple advertisement Pm's a year, that is definitly the small stuff, Although I've been there before and it may very well be something else prior that set's you off and then the littlest thing happens and Kaboom a PM with an add show's up.
and don't worry about someone else beating you to an answer to a question, Like Carlene says, "give credit where credit is due"
Lifes to Short! Dont sweat the small stuff!
Julie really is my middle name.
I'm curious, how does one, he/she, join up in May 2008 and post 7,642 times to date? Just wondering.
Do I win a prize? An FTE shirt would be nice. TEE HEE.


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Touch Me (I Want Your Body) ist ein Lied von der britischen Popsängerin Samantha Fox aus dem Jahre 1986. Das Lied wurde von Jon Astrop, Pete Q. Harris und Mark Shreeve geschrieben. Es wurde im März 1986 als erstes Lied von ihrem Debütalbum Touch Me ausgekoppelt.

Die Single wurde ein weltweiter Erfolg und ist bis heute Fox' bekannteste und erfolgreichste Single. Unter anderem erreichte Touch Me (I Want Your Body) Platz 4 in den US-amerikanischen Billboard Hot 100, Platz 3 in ihrer Heimat Großbritannien und wurde ein Nummer-eins-Hit in Australien, Kanada, Norwegen, Schweden und der Schweiz. In Kanada stand ihr Titel für sieben aufeinanderfolgenden Wochen auf Platz 1, in Schweden sogar acht Wochen.

Inhalt und Musikvideo
Das Lied wurde 1986 in den Battery Studios in London aufgenommen. Touch Me (I Want Your Body) handelt von der Suche nach einem Mann und dem Wunsch nach Sex.

Das Musikvideo wurde im Frühjahr 1986 gedreht. Es zeigt, wie Samantha Fox und ihre Band das Stück auf einer Bühne vor ihrem Publikum spielen. Im Video trägt sie eine offene Jeansjacke und sehr enge Jeans, die ihre Kurven und ihren Hintern betonen sollen. Während die Band im Hintergrund der Bühne spielt, tanzt Samantha Fox im Vordergrund mit zwei Gitarristen. Einige Male geht Fox direkt ins Publikum und singt das Lied neben ihren Fans. In einer Szene des Musikvideos kommt ein Fan auf die Bühne und Fox hält ihn fest und zieht ihn dann zu sich, damit beide Körperkontakt haben, dabei tanzen beide und stellen intimen Sex nach. Danach macht Fox das gleiche bei einem der Gitarristen. In einigen Szenen des Videos schießen Paparazzi Fotos von Samantha Fox, auf denen man ihr Gesicht sieht und sie einen Orgasmus vortäuscht. In einer weiteren Szene schaukelt Fox mit einem Seil durch das Publikum. Darauffolgend macht Samantha ihren Schlagzeuger mit Wasser nass.

Der Liedtext selbst beinhaltet viele sexuelle Anspielungen: "Like a tramp in the night, I was begging for you, to treat my body like you wanted to" und im Refrain "Touch me, touch me, I wanna feel your body, your heartbeat next to mine, touch me, touch me now!" (deutsche Übersetzung: "Berühr' mich, berühr' mich, ich möchte deinen Körper spüren, deinen Herzschlag neben mir, berühr' mich, berühr' mich jetzt"). Außerdem sind zu Beginn des Liedes, sowie bei 2:33 einige Stöhn-Sounds von Fox zuhören.

Versionen und Veröffentlichungen
7" Single:

 a side: Touch me (I want youre Body) (3:46)
 b side: Never Gonna Fall In Love Again (5.10)

12" Single:

 a side: Touch Me (I Want Your Body) (Extended Version) (5:19)
 b side: Never Gonna Fall In Love Again (5:07)

Auszeichnungen

Weblinks

 Günther feat. Samantha Fox – Touch Me

Einzelnachweise

Lied 1986
Popsong
Dancesong
Nummer-eins-Hit
Samantha Fox

redpajama_set_name: RedPajamaWikipedia


Hi Gals I have miscarried 3 times. I went through all the testing and found out I have genital herpes. I also found out the one of my anti-platelet antibody came out positive. My doctor also told me that I had an active herpes infection when I was pregnant. After some research I found several scientific papers concluding that if there is active herpes infection during pregnancy it produces antibodies towards platelets and this can cause blood flow problems to the fetus and ultimately cause fetal death. Anyway, my question is does anyone on this forum with genital herpes have or had a successful pregnancy?


The Courthouse in Grange, Co. Sligo opens as the Spanish Armada Interpretive and Visitor Centre every Saturday and Sunday throughout the Summer. Find out about Sligo's Spanish Armada past. It's free!
Closed off season. Can be opened by appointment.
Sign up to receive news and updates on our event and other news from the Spanish Armada.


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Daniiar Martovich Duldaev (, born 27 October 1992) is a Kyrgyzstani tennis player.

Duldaev has a career high ATP singles ranking of 607 achieved on 14 July 2014. He also has a career high ATP doubles ranking of 484 achieved on 12 May 2014.

Duldaev represents Kyrgyzstan at the Davis Cup where he has a W/L record of 8–0.

Future and Challenger finals

Singles: 3 (1–2)

Doubles 7 (2–5)

Davis Cup

Participations: (8–0)

   indicates the outcome of the Davis Cup match followed by the score, date, place of event, the zonal classification and its phase, and the court surface.

External links

1992 births
Living people
Kyrgyzstani tennis players
Male tennis players
Sportspeople from Bishkek
Tennis players at the 2010 Asian Games
Tennis players at the 2014 Asian Games
Asian Games competitors for Kyrgyzstan


Here's a list of new and continuing Walmart deals you will find in stores now. Grab Kellogg's breakfast shakes for $1.24 each, Quaker Select Start oatmeal for 88¢ and more!
If you find any awesome deals of coupons that match up well at Walmart, please let me know in the comments below.


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Home > University Subject Rankings > Top Universities for Psychology in 2015
Interested in studying abroad?
Check out our comprehensive guides
Top Universities for Psychology in 2015
11:45AM May 5, 2015
**This article covers the 2015 psychology ranking. Click here to discover the top universities for psychology in 2019.**
This fascinating and fast-evolving field of enquiry is a hugely popular field of study and research worldwide. The QS World University Rankings by Subject 2015 features a ranking of the world's 200 top universities for psychology, based on a methodology which draws on employer and academic reputation, and data on research citations.
For a general overview of the top psychology schools in each world region, plus the overall top 10, see below. Or, to search and compare using the interactive ranking table, click here.
Top 10 Universities for Psychology in the World
Based on the QS World University Rankings by Subject 2015
University of California, Los Angeles (UCLA)
University of California, Berkeley (UCB)
UCL (University College London)
See the full psychology ranking >
Top universities for psychology in the US and Canada
The US claims a mammoth share of the top universities for psychology, including six in the top 10 alone. Outside of the top 10 there are another 57 US universities featured, including 33 in the world's top 100. Within the top 50 are New York University (NYU, 41st in the QS World University Rankings® 2014/15), University of Pennsylvania (13th in the world rankings), Columbia University (14th in the world rankings), the University of Chicago (11th in the world rankings), Massachusetts Institute of Technology (MIT, 1st in the world rankings), University of Illinois at Urbana-Champaign (63rd in the world rankings), University of Minnesota (119th in the world rankings), University of Texas at Austin (79th in the world rankings), University of Wisconsin-Madison (41st in the world rankings), Duke University (25th in the world rankings), Northwestern University (34th in the world rankings), Brown University (52nd in the world rankings), Ohio State University (109th in the world rankings) and the University of California, San Diego (UCSD, 59th in the world rankings).
Meanwhile neighboring Canada has a total of 16 top psychology schools featured, including seven within the world's top 100. In the world's top 50 are the University of Toronto (20th in the world rankings), the University of British Colombia (43rd in the world rankings) and McGill University (21st in the world rankings).
Learn more about studying in the US and Canada with our regional guide.
Top universities for psychology in Europe
After the US, the UK claims the next highest number of top universities for psychology, occupying four of the top 10 positions. As well as those listed in the table above, also in the top 50 are King's College London (KCL, 16th in the world rankings), Cardiff University (123rd in the world rankings), the University of Edinburgh (17th in the world rankings), London School of Economics and Political Science (LSE, 71st in the world rankings) and the University of Manchester (30th in the world rankings).
Meanwhile Germany is home to 13 of the world's top psychology schools, with two ranked among the global top 100: Humboldt-Universitat zu Berlin (134th in the world rankings) and Ludwig-Maximilians-Universität München (52nd in the world rankings).
Following on is the Netherlands, with nine schools featured in the psychology ranking, all among the top 100. In the top 50 are the University of Amsterdam (50th in the world rankings), Vrije Universiteit Amsterdam (171st in the world rankings), Erasmus University Rotterdam (90th in the world rankings) and Radboud University Nijmegen (156th in the world rankings).
Six more top psychology schools are claimed by Spain and five by Belgium, while Switzerland, Sweden and Italy each have four. Among these countries, universities ranked within the top 100 for psychology include Belgium's University of Ghent (129th in the world rankings) and KU Leuven (82nd in the world rankings); Switzerland's University of Geneva (85th in the world rankings) and the University of Zurich (57th in the world rankings); Sweden's Karolinska Institute; and Italy's Università di Bologna (UNIBO, 182nd in the world rankings).
Beyond Sweden, the other Nordic countries offer an additional six entrants in the ranking, with two each for Denmark, Finland and Norway. These are Denmark's Aarhus University (96th in the world rankings) and the University of Copenhagen (45th in the world rankings); Finland's University of Helsinki (67th in the world rankings) and the University of Turku (208th in the world rankings); and Norway's University of Bergen (155th in the world rankings) and the University of Oslo (101st in the world rankings).
Lastly in Europe, there are two entries in the psychology ranking from France within the world's top 200, including the Université Aix-Marseille (341st in the world rankings), and one each for both Austria and Ireland: Universität Wien (University of Vienna, 156th in the world rankings) and Trinity College Dublin (TCD, 71st in the world rankings) respectively.
Learn more studying throughout Europe with our country guides.
Top universities for psychology in Asia
An additional 12 of the world's 200 top psychology universities are claimed by Asia. Hong Kong and Singapore each have three representatives in the ranking, including the University of Hong Kong (HKU, 28th in the world rankings), the Chinese University of Hong Kong (CUHK, 46th in the world rankings), the National University of Singapore (NUS, 22nd in the world rankings) and Nangyang Technological University (NTU, 39th in the world rankings) – all ranked among the world's top 50 for psychology.
China and Japan each have two schools featured, including China's Peking University (57th in the world rankings) and Japan's University of Tokyo (31st in the world rankings), both within the top 100 worldwide. South Korea and Taiwan each make one appearance, with Seoul National University (SNU, 31st in the world rankings) and National Taiwan University (NTU, 76th in the world rankings) respectively.
See the latest QS University Rankings: Asia to discover more top universities in Asia, or learn more about studying in the region with our country guides.
Top universities for psychology in Australia and New Zealand
Australia boasts 17 of the world's top universities for psychology, while New Zealand has four. Australia's representatives within the top 50 are the University of Melbourne (33rd in the world rankings), the University of New South Wales (48th in the world rankings), the University of Queensland (UQ, 43rd in the world rankings), the University of Sydney (37th in the world rankings), Australian National University (ANU, 25th in the world), the University of Western Australia (UWA, 89th in the world rankings) and Monash University (70th in the world rankings).
Two of New Zealand's offerings are also among the world's top 50 psychology schools: the University of Auckland (92nd in the world rankings) and the University of Otago (159th in the world rankings)
Find out more about studying in Australia and New Zealand with our country guides.
Top universities for psychology in Latin America
There are four Latin American universities featured in the psychology ranking, with one entrant each for Argentina, Brazil, Chile and Mexico. In order, these are the Universidad de Buenos Aires (UBA, 198th in the world rankings), the Universidade de São Paulo (USP, 132nd in the world rankings), Pontificia Universidad Católica de Chile (167th in the world rankings) and the Universidad Nacional Autónoma de México (UNAM, 175th in the world rankings).
Top universities for psychology in the Middle East & Africa
Finally, Israel and South Africa each claim one entrant each – these are Tel Aviv University (195th in the world rankings) and the University of Cape Town (141st in the world rankings).
Want more content like this? Register for free site membership to get regular updates and your own personal content feed.
See the full ranking of top psychology schools worldwide
Petra T, Atiqa F & 4 others saved this
Written by Laura Tucker
Laura is a former staff writer for TopUniversities.com, providing advice and guidance for students on a range of topics helping them to choose where to study, get admitted and find funding and scholarships. A graduate of Queen Mary University of London, Laura also blogs about student life.
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text:
Season & Episode Discussion
[Spoilers] Season 12 Discussion Thread
By Tensor
July 6, 2018 in Season 12
12 minutes ago, genecollie said:
First all scenes were in swing sets, hard to see. Mayim was not there, maybe because of holiday. Still fun.
Pre-taped, Shamy in apt, Sheldon is photoshopping people in their wedding pictures, the Wright brothers, and he get a email from Tam. He is coming to CalTech with his son who will be attending school there and wants to see Sheldon. Amy asks who Tam is, Sheldon is very angry at Tam and won't tell Amy why.
In Bernadette's office, Penny and Bernadette want to check out Anu, Bernie calls her dad to do a police check on her, he says no.
Pre-taped, Amy asks the gang in Lenny's apt, what they know about Tam, they haven't heard about him, but check Sheldon's enemy list. They find Tam and it only says he knows why. They decide to find him on campus.
Penny and Bernadette go to the hotel where Anu works to check her out, they make plans with her to go out to dinner. She gets reservation for a hard to get into restaurant and Bernadette says if she can get in anywhere why would she want to marry Raj.
Pre-taped, at CalTech Quad, the boys check out every Asian man until they find Tam. They ask about Sheldon and he is unaware Sheldon is angry at him. Sheldon comes over and calls the boys traitors and says Tams knows why I am mad and Tam says no I don't.
Penny, Bernadette, and Anu is nice restaurant. See LaBron James. They talk and Anu says she tried falling in love and it didn't work and thought my parents are happy, I should do it their way.
Sheldon's office, Leonard and Sheldon talking about how it stupid for Sheldon to angry at Tam and Leonard for taking Tam's side.
Pre-taped, Shamy in apt taking about why he is angry. He seems when he came to California, Tam was supposed to come too, but he stayed in Texas with his girlfriend. Amy says it worked out okay, you have new friends and a good life.
Back in the restaurant, the girls are drunk and Anu is questioning them about why is Raj still single. They say many embarrassing things, like bathing with his dog, crying while listening to music, and others.
Sheldon's office, Sheldon forgives Tam and Tam says I didn't know you where angry. When I didn't come to California I knew you would do well and I was in love with my wife wanted to stay in Texas. I have had a good life. Sheldon responses you are so brave.
Cafe, Tam is leaving and Sheldon makes Leonard drive him to the airport.
In Bernadette's office, Bernadette is showing Penny a blur in LaBron's intsagram photo and says it's her. The phone rings and Raj yells why did you tell Anu I use Secret deodorant, it was secret. Don't talk to her anymore. The girls decide to see Anu again to fix it, maybe she can get them in another fancy restaurant.
Sorry I left out most of the jokes, it was funny. At least, you the plot.
Thank you so much for the information! Hope you had an amazing time!
chucky 2336 posts
joyceraye 719 posts
Chrismo 714 posts
bfm 637 posts
Thanks so much for the spoilers @genecollie I hope you had a wonderful time at the taping! 😊
@genecollie Thank you so much! I really appreciate your detailed report, hope you had a great time.
hokie3457
@genecollie : Thank you so much for sharing! It sounds like a lot of fun! I hope you laughed a lot and really enjoyed yourself!!!! Also, your heading says "New Member", so welcome and thank you, thank you, thank you again!
Thank you so very much @genecollie! You're AMAZING!!!
But I have no words to describe how truly upset I am now for what the writers are doing with Lenny. My terrible prediction was true. I can't believe it. I so so hope they change that later. This is just unbelievable. They had Penny say various time that she wanted kids, and there wasn't doubt there. So suddenly she doesn't?!?! And Leonard so quickly accepts this?!?! I just... I just can't, I really don't know what to write. Right now my desire to watch the season sank down.
Chrismo
2 minutes ago, bfm said:
I disagree. This episode makes it IMO so obvious they will have kids.
That won't necessarily calm you down now, but there are 20 more episodes to come...
4 minutes ago, veejay said:
And you made my point there are 20 more episodes. There not going to have a Lenny pregnancy storyline. Another clue for me is Penny rented the batmobile.
My only hope is that it turns out this whole ..something.. is out of fear they/she won't be good enough, maybe because of this test. Something like that did happen before ("The First Pitch Insufficiency"). Her not wanting kids did not happen.
1 minute ago, Chrismo said:
What does the batmobile have to do with that?
Strawberry PopTart Fan
I feel your pain. But, she may change her mind or have an unplanned oops and then be happy after all. Plus, it's very cute that she rented the Batmobile for Leonard. She loves him. And bless Leonard, he loves Penny so much, even if she doesn't want to have children, he's ok.
One way or another, I think they will have a happy ending.
1 minute ago, Strawberry PopTart Fan said:
Of course they will be happy, and of course they love each other.
But this is totally out of the blue and against what they previously wrote and performed. And Leonard just being okay with it so quickly seems unrealistic to me. It is a big change. He's not gonna leave her over it but I would expect him needing more time to process it.
4 minutes ago, Strawberry PopTart Fan said:
There is still 20 episodes to go. She already got the batmobile for him
MsGreentea
It really stinks because who else but Leonard really is supposed to be a parent?
We know the two man-children are parents or will be. But one thought this was episode three, clearly written before everyone knew this was the last season (at least for sure). They would have had the table read for this maybe a week after the announcement. But the idea you can give Leonard a ride in the bat mobile instead of a kid?
10 minutes ago, bfm said:
Let me invite you to a virtual beer among friends...
...they don't always look so skeptical.
Just now, bfm said:
But this is totally out of the blue and against what they previously wrote and performed. And Leonard just being okay with it so quickly seems unrealistic to me. It is a big change. He's not gonna leave her over it but I would expect him needing more time to process it.
Quick reminder: How many times have they talked about kids and what exactly did Penny say? Cause most I remember was something along the lines of "sure but maybe later". The thing is, women putting off having kids over and over again like that can be a sign of being uncertain or not even wanting them deep down and being unable to admit it. So I don't think it's completely unbelievable that she comes to this conclusion (maybe also she has a weak moment because of the quiz or something?).
I do agree on Leonard though. He already picked names and everything so that's weird. But then again there's the suggestion he's not completely okay after the talk with Penny's dad? But I guess we'd need to see how that plays out.
I'm pretty certain that Lenny may end up with a sort of mishap and get pregnant anyway and the writers may think that this is more in character for Lenny because of course they do. I just hope it's not too messy in the end. (Penny really really not wanting a baby and remaining more or less against it would be awful in terms of storytelling. ugh)
For the Marines it's Semper Fidelis; the Coast Guard Semper Paratus. My motto for Penny & Leonard going forward is "Always Faithful; Always Prepared".
3 minutes ago, April said:
The thing is, she did not say this in a wishy-washy way. There was no hesitation there. And she had at least three chances to show such hesitation.
And from what I understand the one who is said to not be okay with it is Wyatt.
And I sure do not want them to have kids if Penny doesn't want it, that would be terrible.
genecollie
Thank you for the nice comments, I have lurking for years and felt a owed you all a report. Sorry I left out so many words, but got home very late after the taping. I had a good time, won candy for being married the longest. (which also make me feel very old). Raj really looked happy after the proposal, not like when he tried to break up with the girl friend in the graveyard. A funny thing when Leonard got in the Batmobile he drove without Penny and then came back and said get in Robin.
1 minute ago, genecollie said:
So happy you had a good time!!! Also "get in Robin"! A great addition to your prior reports! Thans again @genecollie !!!!
I had fears they might do a story like that when it was announced he was turning up, not a fan of that kind of story and putting pressure on his daughter. But it's very cliche, happens a lot in real life 'When are you going to give me some grandkids?'
9 minutes ago, hokie3457 said:
Very well said!
2 minutes ago, Jonny said:
Yeah but at least it sounds like a very short, unimportant bit.
At least they had the mercy to not rub it in with Sheldon saying something about their children being imaginary.
That is up for interpretation, I guess. I don't think it was super hesitant and unsure (that also wouldn't really fit Penny in these kinds of situations). But it also wasn't a super strong confirmation. It's easy to say "sure, one day" or something - even without hesitation. In the end it simply might be you interpreting the writing/acting differently than it was intended by the writers.
Honestly, I wouldn't be surprised if at some point we got an interview about how and why Lenny are "just not that kind of couple" similarly to what we got for the wedding. yikes
Maybe. It wasn't really clear to me what's happening. Pronouns can cause misunderstandings.
Most importantly, Sheldon wasn't in the Batmobile!


text:
DEFANGING BIG TECH
Mayor Jeff Lehman: In these noisy times, we need local, trusted sources of information more than ever
'False information and the resulting mistrust of all information is probably the greatest threat to the vaccination rollout ahead of us. The local lens has length and depth — and that creates trust with its audience,' Barrie Mayor Jeff Lehman writes.
By Jeff LehmanContributor
Thu., Feb. 4, 2021timer3 min. read
When I was nine years old, a tornado ripped through my neighbourhood in Barrie, Ont. In moments, a verdant, muggy spring afternoon had become a postapocalyptic landscape of ruined homes and lives. Eight lives were lost, and 600 homes were damaged or destroyed.
In the days to follow, news helicopters and camera crews descended; my hometown was suddenly national news … for a single cycle.
The most powerful (and lasting) story of the Barrie tornado wasn't the devastating storm or the rubble-strewn front yards. That story was the response to the tornado — the way we reacted, assisted and rebuilt. People were pulled from cars and ruined homes by anonymous heroes. Teams of residents, organized by police and firefighters, worked through the night in search of missing people, then clearing rubble and salvaging pets and treasured possessions. Homeowners painted messages on their ruined garage doors ("Gone with the Wind" was a favourite). Issues like the fragility of some home insurers were exposed, leading ultimately to changed regulations. And the stories of generosity and compassion were endless.
These stories would never have been told if not for local media. The national networks left in a day or two. But in 1985, Barrie had two daily newspapers, a network TV station and four radio stations. And their journalists, cameras and tape recorders told the stories of the humour, heartache and humanity of the response to the tornado. It was only because of local journalism that the people of Barrie came to know themselves in that time of adversity.
The story of our response to the tornado is remembered (and occasionally romanticized) whenever our community faces challenges, including right now amid a global pandemic. It's invoked by leaders, including myself, when we need to reach a collective high bar. In this way, local journalism built a lasting, undefinable-yet-essential aspect of my community's identity.
The local lens has length and depth — and that creates trust with its audience. That trust is more important now than it has ever been before. Today, Barrie is a city of 150,000 — three-quarters or more of whom were not here for the tornado — and the media landscape is much different. We consume "news" on a cluttered digital commons which, while open to all, has enabled a cacophony of conspiracy theories, look-alike websites and deceptively patriotic propaganda machines. All the more important, then, that we have journalism motivated by reporting objective truths. COVID has raised the stakes. False information and the resulting mistrust of all information is probably the greatest threat to the vaccination rollout ahead of us; never has the need for trusted sources been greater.
The nuances of local context are essential, and local journalists can create a contract of trust by being embedded in their communities in a way no remote reporter can achieve. Local journalists live the impacts of local choices and leadership, can analyze and criticize the longitudinal impacts of decisions and issues, and see the dimensions of a story that require immersion.
The local lens has length and depth — and in doing so has impact. While network coverage may reach more eyeballs, the reach of local stories is evident in the reaction from the community. How do I know this to be true? As mayor, I've been doing town hall meetings in rec centres and library meeting rooms for more than 10 years — and now do them online. I get questions from residents that begin with "I read in the paper how … " or "I saw on the 6 p.m. TV news ... "
The newspaper or TV referenced was usually the local outlet — but what's also interesting is this: what they read, heard or saw made them ask a mayor a question. What they read, heard or saw had impact — it cut though the noise.
It is said that all politics is local. In this noisiest of times, when journalists share an equal status on platforms with bots, we need to double down on local, trusted sources of information. Our choice of information sources is in our control, and we need local choices now more than ever.
Jeff Lehman is the mayor of Barrie, Ont.
'You're going to be left with the junk': What happens when Google hides the news?
As COVID shrank your world, it made big tech even bigger. Now, things may never be the same again
What would life be like without Google's search engine? Australia might be about to give Canada a preview


text:
from ronin.cli import cli
from ronin.contexts import new_context
from ronin.java import JavaCompile, Jar, JavaClasses
from ronin.phases import Phase
from ronin.projects import Project
from ronin.utils.paths import glob, input_path, join_path
from ronin.utils.platform import which

with new_context() as ctx:

    project = Project('Java Swing Hello World')

    # Compile
    Phase(project=project,
          name='compile',
          executor=JavaCompile(),
          input_path=join_path(ctx.paths.root, 'src'),
          inputs=glob('**/*.java'))

    # Jar
    Phase(project=project,
          name='jar',
          executor=Jar(manifest=input_path('MANIFEST.MF')),
          extensions=[JavaClasses(project, 'compile')],
          output='hello',
          run_output=1 if ctx.build.run else 0,
          run_command=[which('java'), '-jar', '{output}'])

    cli(project)

redpajama_set_name: RedPajamaGithub


The SIRIN LABS R&D team has created a protocol to seamlessly exchange between tokens, we call it the Token Conversion Service (TCS). We've recently published a video of the TCS in action. If you haven yet watched the video, here's the gist: to make a purchase that requires SRN on your FINNEY™ with only ETH in your wallet, currently, you would need to go to an exchange, buy SRN for ETH, and then go back to the website to finalize your purchase. What we have done is simplified and streamlined this process.
The TCS can, in real-time, perform all the necessary transfers and transactions needed to make a purchase using a coin you do not currently hold in your wallet. This service works in the background of your phone in order to facilitate a purchase using cryptocurrency in a mobile environment. The significance of this development is monumental and worthy of attention.
It has always been the position of SIRIN LABS that one of the largest hurdles to creating mainstream adoption of blockchain technology and cryptocurrency is the user experience. Similar to Linux, cryptocurrency and blockchain, in its current form, is entirely manual. You need to understand exactly what you're doing, and how to navigate the appropriate sites (Etherscan, MEW, etc.).
A novice couldn't sit down at a computer, easily purchase cryptocurrency, and then use it, regardless of the platform. For those of us who see crypto as the future, this poses a huge problem, namely, in order for cryptocurrency to succeed in the long-term, it requires mass-adoption, but the average person has no incentive to learn how to operate on the blockchain in its current form, and has no interest in committing the requisite time needed to do so.
Our Founder and Co-CEO, Moshe Hogeg, has been preaching for quite some time that users shouldn't need to understand the technology to enjoy it. When a teenager gets their first smartphone and uses the GPS navigation, they don't know how the phone is able to provide them turn-by-turn navigation to get them to their destination. Nor do they care that the amazingly advanced piece of technology in the palm of their hand is able to seamlessly communicate with satellites in geosynchronous orbit around the Earth. All they care about is that it tells them the correct directions from A to B. The average person doesn't care how the technology works; they care only that it does work.
This is the vision behind both the SIRIN OS and the FINNEY smartphone. To give each and every person simple, streamlined, and intuitive access to cryptocurrency and the blockchain, while maintaining the necessary security needed to keep stored currencies and transactions safe.
The goal for the SIRIN LABS FINNEY™ phone is to allow any person to pick up a device, use cryptocurrency, and operate on the blockchain completely intuitively, without ever needing to know details such as which blockchain is being used, or how many places after the decimal are needed is a — which is why we're so excited about this new development. The ability to simplify and provide an incredibly streamlined user experience is a major step towards creating mass appeal for both cryptocurrency and blockchain.


The Wnt/β-catenin signaling pathway plays an important role in the development of second heart field (SHF Isl1+) that gives rise to the anterior heart field (AHF) cardiac progenitor cells (CPCs) for the formation of the right ventricle, outflow tract (OFT), and a portion of the inflow tract (IFT). During early cardiogenesis, these AHF CPCs reside within the pharyngeal mesoderm (PM) that provides a microenvironment for them to receive signals that direct their cell fates. Here, N-cadherin, which is weakly expressed by CPCs, plays a significant role by promoting the adhesion of CPCs within the AHF, regulating β-catenin levels in the cytoplasm to maintain high Wnt signaling and cardioproliferation while also preventing the premature differentiation of CPCs. On the contrary, strong expression of N-cadherin observed throughout matured myocardium is associated with downregulation of Wnt signaling due to β-catenin sequestration at the cell membrane, inhibiting cardioproliferation. As such, upregulation of Wnt signaling pathway to enhance cardiac tissue proliferation in mature cardiomyocytes can be explored as an interesting avenue for regenerative treatment to patients who have suffered from myocardial infarction.
To investigate if Wnt signaling is able to enhance cellular proliferation of matured cardiomyocytes, we treated cardiomyocytes isolated from adult mouse heart and both murine and human ES cell-derived matured cardiomyocytes with N-cadherin antibody or CHIR99021 GSK inhibitor in an attempt to increase levels of cytoplasmic β-catenin. Immunostaining, western blot, and quantitative PCR for cell proliferation markers, cell cycling markers, and Wnt signaling pathway markers were used to quantitate re-activation of cardioproliferation and Wnt signaling.
N-cadherin antibody treatment releases sequestered β-catenin at N-cadherin-based adherens junction, resulting in an increased pool of cytoplasmic β-catenin, similar in effect to CHIR99021 GSK inhibitor treatment. Both treatments therefore upregulate Wnt signaling successfully and result in significant increases in matured cardiomyocyte proliferation.
Although both N-cadherin antibody and CHIR99021 treatment resulted in increased Wnt signaling and cardioproliferation, CHIR99021 was found to be the more effective treatment method for human ES cell-derived cardiomyocytes. Therefore, we propose that CHIR99021 could be a potential therapeutic option for myocardial infarction patients in need of regeneration of cardiac tissue.
Cardiovascular disease is currently the leading cause of death globally and is expected to rise in future as the incidence and prevalence of chronic diseases associated to cardiovascular risk factors increases . In conjunction with preventive medicine, cardiac regeneration therapy may therefore be the next best solution to overcome this problem. Efforts are now being focused on improving the efficiency of cardiomyocyte proliferation to preserve cardiac function after an injury [2–4]. Hence, understanding the mechanism of cardiovascular development that leads to cardiovascular proliferation is pivotal in the advancement of therapeutic approach aimed at cardiac pathology.
The Wnt/β-catenin signaling pathway in particular plays an important role in the development of second heart field (SHF Isl1+) that gives rise to the anterior heart field (AHF) cardiac progenitor cells (CPCs) for the formation of right ventricle (RV), outflow tract (OFT) and portion of inflow tract (IFT) [5–9]. During early cardiogenesis, these AHF CPCs reside within the pharyngeal mesoderm (PM) that provides a microenvironment for them to receive inputs and direct their fate. N-cadherins are Ca2+ dependent transmembrane proteins that associate with cytoplasmic catenins, such as the Wnt signal transducer β-catenin, to form adherens junction protein complexes which enable intercellular adhesion while simultaneously sequestering β-catenin to the plasma membrane. Unbound β-catenin are able to undergo nuclear translocation, forming complexes with the TCF/LEF family of DNA binding proteins to activate downstream Wnt effector genes, consequently promoting cardioproliferation and preventing premature differentiation of CPCs . As such, cadherins are generally treated as negative regulators of the Wnt signaling pathway. However, N-cadherin, although weakly expressed in AHF CPCs, play a significant role in their cell fate and development by promoting intercellular adhesion between AHF CPCs while also being crucial to maintain high Wnt signaling activity . On the contrary, adult myocardium express N-cadherin strongly, and yet have low levels of Wnt signaling activity and regenerate at a very low frequency, with an estimated rate of 0.5–1% per year [10–12]. We therefore attempted to revert matured cardiomyocytes into a more proliferative state by activating high levels of Wnt signaling. We posit that N-cadherin expression is necessary to protect a cytoplasmic store of β-catenin from proteolytic degradation via the GSK ubiquitination complex. These stores result in high Wnt signaling in CPCs due to the presence of Wnt signaling ligands binding and releasing β-catenin, while in matured cardiomycytes, β-catenin is highly sequestered and do not contribute to Wnt signal transduction. These stores of β-catenin, if released and made available in matured cardiomyocytes, should be able to undergo nuclear translocation and activate Wnt signaling to promote cardioproliferation.
In this study, we demonstrated that re-activation of Wnt/β-catenin signaling pathway can be achieved in matured mouse and human ES cell-derived cardiomyocytes by using N-cadherin antibody or GSK inhibitor, CHIR99021, releasing sequestered β-catenin in the N-cadherin-based adherens junctions or by preventing the ubiquitination-degradation of β-catenin, respectively [13–15]. We further validated the mechanism, demonstrating the translocation of cytoplasmic β-catenin into the nuclei to re-activate Wnt signaling leading to cardioproliferation. This study has therefore provided an avenue for the expansion of matured cardiomyocytes in vitro and perhaps regenerative medicine through enhanced cardiomyocyte proliferation to preserve cardiac function after myocardial injury.
The human ES cell line (H7) was cultured in feeder-free condition on Matrigel (BD, Franklin Lakes, NJ). The cells were maintained in mTeSR™1 medium (Stemcell Technologies) and passaged with collagenase IV (1 mg/ml) enzymatic treatment. To differentiate human ES cells towards cardiomyocytes, we adopted the protocol established by Lian et al. [16, 17]. The Cdh2 mouse knockout ES cells were cultured and differentiated towards cardiomyocytes as described by Soh et al. . In this study, matured ES cell-derived cardiomyocytes were cultured for more than 2 months from the initial contraction to ensure sufficient cardiomyocyte maturation .
Single-cell suspension was obtained from cardiomyocytes derived from both murine and human ES cells. The cells were stained using vascular cell adhesion molecule (VCAM-1) and SIRPα/β antibodies, respectively. Briefly, staining of mouse cardiomyocytes was achieved with rabbit anti-VCAM1 monoclonal antibody (1:50) (Cell Signaling Technologies) in the presence of blocking buffer consisting of 5% FBS and 2% BSA in PBS for 90 min at 37 °C, followed by donkey anti-rabbit IgG Alexa Fluor 594 at 1:1000 dilution (Invitrogen) for 1 h. Human ES cell-derived cardiomyocytes, on the other hand, were stained with PE/Cy7-conjugated anti-human CD172a/b (SIRPα/β) antibody at 1:300 dilution (Biolegend). Cardiomyocytes were subsequently purified via fluorescence-activated cell sorting (FACS). Matured human ES cell-derived cardiomyocytes were treated with either 100 μM of TBP or 100 nmol/L of EDN1 to induce cardiac hypertrophy.
Matured cardiomyocytes were isolated from mice that are at least 2 months old according to published protocol . The isolated cardiomyocytes were maintained in medium comprising of RPMI and B27 supplement .
For cultured cell samples, 2 × 106 cells were harvested and lysed in 800 μl of TRIzol reagent (Invitrogen). The samples were allowed to stand for 5 min at room temperature, after which 160 μl of chlorofoam was added to allow for phase separation by centrifugation at 12,000×g for 15 min at 4 °C. Following that, the aqueous phase was transferred to a fresh tube, and equal volume of isopropanol was added and mixed. RNA samples were allowed to precipitate at room temperature for another 10 min. The precipitated RNA samples were pelleted by centrifugation at 12,000×g for 15 min at 4 °C. For cDNA synthesis, RNA samples (500 ng) were reverse transcribed to obtain cDNA using the iScript cDNA Synthesis kit (BioRad). Primer sequences are provided in Additional file 1: Table S1. Quantitative PCR analyses were performed using SYBR Green Master Mix Reagent (Applied Biosystems) on an ABI Viia7 Real-Time PCR System. The threshold cycle (Ct) was determined to be ≥ 35. Each experiment was repeated at least twice. Standard deviations (s.d.) of the means in qPCR experiments were obtained from three independent experiments.
Immunocytochemical analysis was performed using the respective antibodies: mouse anti-PY654-β-catenin (membrane bound) and anti-PY489-β-catenin (nuclear) (both from Hybridoma bank) and mouse anti-cardiac troponin T (1:400; Abcam ab8295). Briefly, cells were first harvested and washed once with PBS. Fixation of cells was achieved with 4% paraformaldehyde for 30 min, followed by treatment with 0.3% Triton X to permeabilize cells, if necessary. The cells were then blocked with PBS containing 5% fetal bovine serum and 1% BSA for 30 min at room temperature. Primary antibodies were added at respective dilutions and incubated for 1 h at room temperature. After washing, the cells were incubated for another 45 min at room temperature without light exposure with either 1:1000 diluted Alexa Fluor 488 or Alexa Fluor 594 secondary antibodies. Nuclei were counterstained with 4,6-diamindino-2-phenylindole (DAPI). The cells were observed under a fluorescent microscope (Nikon TS-100).
Fluorescent images were acquired by an automated microscope (PerkinElmer Operetta) at ×20 magnification. Image analysis was performed using the Columbus software (PerkinElmer), by first recognizing and outlining cellular nuclei based on DAPI staining. For each experiment, equal number of SIRPA-positive cardiomyocytes (1 × 105 cells) was seeded to compare proliferation of treated cells (N-cadherin antibody or CHIR99021) vs control (untreated). Quantification of cardiomyocytes was performed by summing all cardiac troponin T (cTnT)-positive cells (relative cutoff intensity > 150) in the treated wells vs the control. For the analysis on cardiomyocyte proliferation, the double-positive cells (Ki67+/cTnT+ or BrdU+/cTnT+) in each of the treatment conditions have been calculated as a percentage of all the single-positive cells (cTnT+).
Whole-cell lysates were generated by scraping cultures of matured cardiomyocytes treated with CHIR99021, N-cadherin antibody, or combination for 1 day and resuspended in RIPA lysis buffer (1% Nonidet P-40, 0.5% deoxycholate, 5 M NaCl, and 1 M Tris pH 7.4) for 10 min at 4 °C and then centrifuged at 11,400g for 20 min at 4 °C for the isolation of the cytosolic fractions. Quantification of protein concentration was determined using Pierce BCA Protein Assay kit (Thermo-scientific). Protein lysate of 60 μg per sample was loaded onto 4–20% Mini-PROTEAN TGX Stain-Free Precast Gels (Bio-Rad), resolved by electrophoresis, and transferred to nitrocellulose membrane (Bio-Rad). Membranes were blocked with 5% skimmed milk in TBS + 1% Tween-20 and probed overnight with primary antibodies. The antibodies were rabbit anti-β-catenin antibody (1:2500; Abcam ab32572) and mouse anti-β-actin antibody (1:1000; Cell Signaling). Membranes were washed and incubated for 1 h with their respective IgG-HRP-conjugated secondary antibody (Santa Cruz) in 5% skimmed milk in TBS + 1% Tween-20 and developed using ECL substrate (Bio-Rad).
Results involving two experimental groups expressed as mean ± s.d. were tested for statistical significance using Student's t test, two-sided based on assumed normal distributions. P values < 0.05 were considered statistically significant. The error bars in graphs with three or more experimental groups refer to standard error of means of stated replicates. A non-parametric test (Kruskal-Wallis one-way analysis of variance) was used to assess whether the mean values of the experimental groups could be considered to be significantly different compared to the control group at 95% confidence level.
Earlier data demonstrated that the loss of N-cadherin seen in AHF-Cre; Cdh2fl/fl mutant mice embryo causes lower levels of both membrane and nuclear β-catenin due to cytoplasmic degradation of β-catenin. Resultant downregulation of Wnt signaling led to the decrease in proliferation of CPCs, premature differentiation of AHF CPC to cardiomyocytes in the pharyngeal mesoderm, and disrupted growth of the OFT and RV . As N-cadherin was found to be weakly expressed in proliferative CPCs and highly expressed in less proliferative cardiomyocytes, we therefore hypothesized that N-cadherin expression plays an important role in regulating the Wnt/β-catenin signaling pathway for proliferation and regeneration in mature cardiomyocytes. To test this hypothesis, we sought to re-activate Wnt signaling in matured mouse cardiomyocytes and assess their proliferation capabilities. Immunostaining for BrdU, a proliferation marker, was performed to assess proliferation of matured mouse cTnT+ cardiomyocytes. To affect availability of cytoplasmic β-catenin for nuclear translocation, we treated mouse cardiomyocytes with either N-cadherin antibody, which could potentially mimic Wnt signaling ligand binding or destabilize the adherens junction for the release of β-catenin, or CHIR99021, a GSK inhibitor. The results showed that more BrdU and cTnT double-positive cells were found in cardiomyocytes of either treatment, suggesting that upregulation of Wnt signaling via cytoplasmic β-catenin availability is able to enhance proliferation in matured mouse cardiomyocytes (Fig. 1a, b). These results were validated by quantitative PCR, whereby the cyclin genes known for their roles in cell cycle such as CDK1, CCNA2, CCNB1, and CCNB2 were observed to be significantly upregulated in both N-cadherin antibody- and CHIR99021-treated cardiomyocytes, suggesting that both treatments successfully increased cytoplasmic β-catenin levels, reactivate Wnt signaling, and result in the matured mouse cardiomyocytes entering a more proliferative state (Fig. 1c).
While previous studies have reported that cadherins generally function as a negative regulator of Wnt/β-catenin signaling due to their role in sequestering cytoplasmic β-catenin at the cell membrane, they also play an important role in Wnt/β-catenin signaling during development [5, 20]. We investigated the changes of cytoplasmic and nuclear β-catenin levels after treatment with N-cadherin antibody to uncover the interaction between N-cadherin-based adherens junctions and the canonical Wnt signaling pathway. Immunostaining results and quantification of N-cadherin antibody-treated matured cardiomyocytes showed lower levels of membrane β-catenin but higher levels of nuclear β-catenin as compared to control (Fig. 2a–d). These results confirmed that there was translocation of β-catenin from the membrane to the nucleus upon treatment with N-cadherin antibody. We reasoned that N-cadherin antibody neutralizes N-cadherin signaling, preventing their ability to sequester β-catenin at the cell membrane which consequently leads to the release of membrane-bound β-catenin, resulting in larger reservoir of cytoplasmic β-catenin that undergoes nuclear translocation to activate Wnt signaling. Quantitative PCR results of these N-cadherin antibody-treated cardiomyocytes showed that many Wnt signaling-related genes were upregulated (Fig. 2e). We figured that treatment with GSK inhibitor, CHIR99021, should achieve Wnt activation through the same pathway due to the increased availability of cytoplasmic β-catenin levels. Therefore, we performed quantitative PCR on matured cardiomyocytes treated with N-cadherin antibody and the GSK inhibitor, CHIR99021, and observed upregulation of the main Wnt signaling effectors, Axin2 and Lef1 (Fig. 2f). Taken together, these results prove that neutralizing N-cadherin signaling results in high levels of cytoplasmic β-catenin for nuclear translocation to promote the re-activation of Wnt signaling in matured cardiomyocytes.
To further support our hypothesis that neutralizing N-cadherin signaling results in increased proliferative capacity of matured mouse cardiomyocyte through the re-activation of Wnt signaling, we treated N-cadherin knockout mouse ES cell-derived cardiomyocytes with N-cadherin antibody or CHIR99021. The mouse ES cell-derived cardiomyocytes were first purified using fluorescence-activated cell sorting (FACS) based on their expression of vascular cell adhesion molecule-1 (V-CAM1)  (Fig. 3a); thereafter, the cardiomyocyte numbers between treatments and the control were compared (Fig. 3b). Significant increase in numbers of cardiomyocytes was observed only for CHIR99021 treatment, compared to the control, and as expected, N-cadherin antibody failed to have significant effects on cardiomyocytes derived from Cdh2 knockout mouse ES cells. This result supports our earlier hypothesis that cardiomyocyte proliferation through N-cadherin antibody treatment was due to the release of sequestered β-catenin at the cell membrane, which undergoes nuclear translocation to activate Wnt signaling. Consistent with the above data regarding increased cell numbers and proliferation, we observed upregulation of genes involved in cell cycling (Ccnb1, Ccnb2, and Ccnd2) and Wnt signaling (Wnt1, Wnt2, Wnt6, Axin2, and Lef1) in CHIR99021-treated cardiomyocytes (Fig. 3c, d). While Ccnd2 activity is required for cell cycle G1/S transition, the family of cyclin B genes are essential mediators of cell cycle progression in the G2-M phase transition and promotes cell proliferation [22, 23]. In contrast, cell cycling genes were not significantly upregulated in N-cadherin antibody-treated cardiomyocytes. As cardiomyocytes derived from Cdh2 knockout mouse ES cells do not form N-cadherin-based adherens junctions, our results confirm that N-cadherin antibody is only able to activate Wnt signaling and proliferation through interactions with N-cadherin.
To assess possible applications for regenerative medicine in humans, we generated human ES cell-derived cardiomyocytes to investigate if induced proliferative effects observed in the mouse model could be replicated in human cell models. To ensure that the human ES cell-derived cardiomyocytes used expressed a matured phenotype sufficient to model adult cardiomyocytes, the cardiomyocytes used in this study were cultured for at least 2 months post-initial contraction . We performed gene expression analysis on cardiomyocytes, which were cultured for up to 1 month post-initial contraction. The results revealed downregulation of early key transcription factors governing cardiogenesis such as NKX2.5 and MEF2C and upregulation of cardiac maturation and ion channel markers such as MYH7, KCNQ1, SCN5A, HCN4, and CACNA1G at the end of the time-course experiment (Fig. 4a). Furthermore, consistent with other studies [18, 24], a concomitant shift in metabolism from glycolysis to β-oxidation was observed as the differentiated cardiomyocyte matures (Fig. 4b). Therefore, we concluded that the cultured cardiomyocytes used, 2 months post-initial contraction, are sufficiently mature to model adult cardiomyocytes.
To investigate if the enhanced proliferative effect induced by upregulation of Wnt signaling could be replicated in adult human cardiomyocytes, we treated human ES cell-derived cardiomyocytes with N-cadherin antibody and CHIR99021 and immunostained for Ki67 as the marker for cellular proliferation. Initial testing with N-cadherin antibody (1:1000) showed promising results, with quantification of immunostaining results showing ~ 1.0-fold increase in Ki67+ population of cells in the cultured human ES cell-derived cardiomyocytes compared to a non-treated control (Fig. 4c–e). Thereafter, we varied concentrations of N-cadherin antibody and CHIR99021 to determine the optimal concentrations of both treatments that would result in the highest increase in Ki67+ population within the cardiomyocytes (Additional file 1: Figure S1A-B). The optimal concentrations were determined to be 1:1000 N-cadherin antibody and 1 μM CHIR99021, with further increased concentrations of either resulting in decreased cellular proliferation or increased cell death respectively (data not shown). Consistent with the effects of CHIR99021, quantitative PCR results showed that different concentrations of the GSK inhibitor do not significantly affect transcription of CDH2 or CTNNB1, genes that are upstream of β-catenin signaling. On the other hand, dose-dependent upregulation of both downstream effectors AXIN2 and LEF1 was observed (Additional file 1: Figure S1C).
We then investigated if there are synergistic effects between N-cadherin antibody and CHIR99021 treatments. Immunostaining for Ki67+ and quantitative analysis on day 7 after treatment revealed that a combinatory treatment of N-cadherin antibody (1:1000) and CHIR99021 (1 μM) resulted in cardiomyocyte proliferation rates at an average between the more effective CHIR99021 treatment and the less effective N-cadherin antibody treatment (Fig. 5a–e). N-cadherin antibody treatment and CHIR99021 treatment resulted in a ~ 40% and ~ 100% increase in cardiomyocytes expressing the proliferation marker respectively, while the combinatory treatment resulted in a ~ 50% increase (Fig. 5b, c). Furthermore, flow cytometric analysis revealed that N-cadherin antibody and CHIR99021 both resulted in an increase in total cardiomyocyte number in culture compared to the control treatment, with the combinatory treatment also resulting in an average between the two treatments (Fig. 5d, e).
In order to demonstrate a direct correlation between unbound β-catenin in the cytosolic fraction with cardiomyocyte proliferation, we performed western blot on cytosolic fractions of control cardiomyocytes and cells treated with CHIR99021, N-cadherin antibody, or combinatory treatment for 1 day. Longer treatment duration was not preferred in this case as the release of membrane-bound β-catenin after treatment with N-cadherin antibody would not be observed if the lysates were collected at a later time point. Consistent with the results thus far, increased levels of unbound β-catenin compared to the control were observed after 1 day of treatment (Fig. 5f). As illustrated by the western blot results, stabilization of cytoplasmic β-catenin via CHIR99021 treatment results in its slow build-up for nuclear translocation, but the majority will still be sequestered at the plasma membrane due to high N-cadherin levels in matured cardiomyocytes. As expected, N-cadherin antibody treatment which acts as a ligand releases the larger pool of β-catenin observed (Fig. 5f), which is still susceptible to GSK-tagged degradation, possibly explaining the more significant upregulation of Wnt signaling effects from CHIR99021 treatment due to inactivated GSK. Finally, a dual treatment of CHIR99021 and N-cadherin antibody would therefore result in an even larger increase in cytoplasmic β-catenin, as the protein is both being released from the plasma membrane and being stabilized in the cytoplasm.
In addition to the upregulation of cytoplasmic β-catenin, more cardiomyocytes were stained positive for PHH3 and Survivin, a marker specific for G2-M phase that marks entry into the mitotic process and eventual division, when the cells were treated with CHIR99021 (Fig. 5g). All these results taken together suggest that both N-cadherin antibody and CHIR99021 induce dose-dependent cardiac proliferation through Wnt signaling and can possibly be used to revert mature human cardiomyocytes into a more proliferative state, up to a maximal dosage.
Despite that, we reasoned that excessive amounts of N-cadherin antibody hindered the formation of adherens junctions, inhibiting cell adhesion between cardiomyocytes leading to the loss of cardiomyocytes and inefficient proliferation. Sufficient amounts of free N-cadherin are required for proper cardiomyocyte proliferation, adhesion, and overall cardiac function. Similarly, we reasoned that high concentrations of CHIR99021 are cytotoxic due to the pleiotropic effects of GSK inhibitor which affects several other pathways. Overall, CHIR99021 at its optimal concentration exhibits the best potential as a therapeutic agent for cardiac regeneration as it promotes the maximal re-activation of Wnt/β-catenin signaling for cardiomyocyte proliferation without disrupting the necessary role of N-cadherin in cellular adhesion or inducing cytotoxicity.
Many known cardiopathies and injuries affect mainly the ventricles, causing inflammatory responses leading to scarring, fibrosis, and hypertrophy of the mammalian heart, referred to as ventricular remodeling. Wnt/β-catenin signaling has been implicated in cardiac remodeling regulation as an injury response. In the mouse model, numerous works have suggested Wnt/β-catenin signaling to play a part in regulating cardiac hypertrophy and preventing apoptosis. However, there has been little evidence uncovering the mechanisms and pathways involved. Wnt ligands have been shown to be upregulated alongside their feedback regulators Dkk1 and Dkk2 as a response in mouse models with induced cardiac injuries such as myocardial infarction [25–27].
In the current revision, we utilized human matured cardiomyocytes modeling cardiac injury to uncover how Wnt signaling is affected by cardiac injury. Cardiac hypertrophy has been induced as a form of heart injury in both animal and human models using oxidative stress treatments [28, 29]. Tert-butyl peroxide (TBP) is an oxidizing agent, commonly used as an alternative to hydrogen peroxide in stimulating oxidative stress in cells and tissues, while Endothelin-1 (EDN1) can result in cardiovascular fibrosis and hypertrophy due to increased collagen synthesis and reduced collagenase activity in cardiac fibroblasts [30, 31]. We treated human matured cardiomyocytes with either Endothelin-1 (EDN1) or Tert-butyl peroxide (TBP) to model cardiac injury and looked at changes in the transcript levels of CDH2, CTNNB1, AXIN2, and LEF1, effectors within the Wnt pathway (Additional file 1: Figure S2).
From the results, we observed that cardiomyocytes treated with EDN1 or TBP expressed reduced levels of CDH2 transcription by ~ 0.5-fold while CTNNB1 expression increased by ~ 2-fold. We hypothesize that diseased cells are undergoing a phenomena of dedifferentiation, thereby reducing surface N-cadherin and cell-cell contact and proliferation in an attempt to repopulate after cardiac injury. Dedifferentiation of cardiomyocytes for proliferation was first shown to occur in the naturally regenerating Zebrafish heart . Several other researchers have also observed dedifferentiation when regeneration is induced in other models [33, 34]. We further investigated AXIN2 and LEF1 expression levels to understand this phenomena. AXIN2, being an immediate downstream target of β-catenin, shows a similar upregulation pattern to CTNNB1 and serves as a negative feedback regulator of β-catenin. Interestingly, LEF1, the effector of proliferation signaling, is significantly downregulated upon TBP or EDN1 treatment. These results indicate that injured cardiomyocytes do not have a proliferative response despite the Wnt signaling received, suggesting that the upregulation of Wnt signaling via CHIR99021 treatment immediately after injury induced by ROS or EDN-1 may not result in cardiac regeneration in vivo. Nonetheless, CHIR99021 can be used in combination with simvastatin that has recently demonstrated to inhibit neural cell apoptosis and promote locomotor recovery via activation of Wnt signaling pathway by enhancing Lef1 expression after spinal cord injury .
Previous studies have shown that sustaining high Wnt signaling activity in AHF CPCs during early cardiogenesis promotes proliferation and prevents premature differentiation of CPC . In this study, we further document the mechanism for the re-activation of quiescent Wnt signaling in matured mouse and human cardiomyocytes that leads to cardioproliferation by maintaining high levels of cytoplasmic β-catenin for nuclear translocation using N-cadherin antibody and GSK inhibitor, CHIR99021 (Fig. 6). Earlier studies have reported that N-cadherin sequesters cytoplasmic β-catenin at the cell membrane, preventing its proteolytic degradation by the proteasome complex while simultaneously preventing the β-catenin from exerting its Wnt signaling effect through nuclear translocation . We demonstrated that neutralizing N-cadherin signaling results in an effect similar to Wnt ligand binding and release of membrane-bound β-catenin, causing an increase in Wnt signaling. Although cadherins generally function as a negative regulator of Wnt signaling, they play a crucial role in protecting a cytoplasmic store of β-catenin from proteolytic degradation which can be released upon Wnt ligand binding to activate the Wnt pathway. Confirmatory treatment of cardiomyocytes derived from Cdh2 knockout mouse ES cells with N-cadherin antibody validated that the Wnt signaling upregulation in wild-type mouse treatments was solely due to the release of β-catenin sequestered and protected by proteolytic degradation by N-cadherin. However, it is important to note that N-cadherin also plays a role in cell adhesion which is essential in determining the cardioproliferation efficiency. We observed that concentrations of N-cadherin antibody beyond 1:1000 were less effective in inducing cardioproliferation as they cause cells to detach from each other leading to the loss of cardiomyocytes and inhibition of cell-cell adhesion necessary for proliferation.
While β-catenin/Wnt signaling is one such pathway that the cadherins regulate, there are several other proliferative and survival pathways that require N-cadherin/adherens junctions-mediated cell-cell adhesion. For instance, cadherin-mediated cell-cell contact is necessary for mechanical coupling between cells to activate mechanical signaling through both the actomyosin cytoskeleton and microtubule networks, shown to be crucial for cell division and transcriptional regulation [36, 37]. The closely related Notch and Hedgehog signaling pathways have also been shown to be regulated by N-cadherins in neural progenitor cells (NPCs). Notch1 and its ligand Dll1, distributed around adherens junctions, determine the apical polarity and guide asymmetric neurogenic division for NPCs and their differentiated progenies . Alpha-catenin, which is also sequestered at the adherens junction, highly upregulates Hedgehog signaling leading to excessive NPC proliferation when deleted . In skin tissue, α-catenin tightly regulates the activity of YAP1, the transcriptional effector of the Hippo pathway, which controls epidermal stem cell proliferation and tissue expansion . Additionally, endothelial cells maintain cell-cell contact through molecular players such as the cadherins and gap junctions and have been shown to increase cell proliferation via PI3K-dependent signaling pathway . In matured myocardium, continuous expression of N-cadherin is essential for normal cardiac electrophysiological activity due to their functions in gap junctions .
In our study, we demonstrated that CHIR99021 treatment exhibited the best potential for cardiac regeneration as it is not only more efficient in inducing cardioproliferation, it also does not interfere with the function of N-cadherin required for cell adhesion and cardiac electrophysiological activity . Consistent with this, recent studies have demonstrated that CHIR99021 alone can promote cardiomyocyte proliferation in neonatal atrial human cardiomyocytes , emphasizing again the feasibility of using CHIR99021 as a potential drug that can endogenously repair injured cardiac tissue and promote cardiac regeneration. We found that CHIR99021 exhibits cytotoxic effects at a concentration of more than 1 μM, plausibly due to several reasons which were not uncovered in this study. GSK functions in several intracellular signaling pathways which result in CHIR99021, a GSK inhibitor to have several reported pleiotropic effects. One such pathway affected would be the nuclear translocation of transcription factor EB which leads to the activation of lysosomal/autophagy pathway, stimulating lysosomal biogenesis resulting in GSK inhibitor-induced apoptosis [44–46]. High concentrations of GSK inhibitor were also reported to cause centrosome aberrations, deformation in mitotic spindles, chromosome instability, and subsequently cell death from mitotic catastrophe .
While re-activation of Wnt signaling by CHIR99021 induces cardioproliferation required for cardiac regeneration, unregulated activation of Wnt signaling has been associated with many cardiac diseases such as cardiac hypertrophy and even cancer [48, 49]. While activated Wnt signaling can be useful in promoting cell proliferation, studies have also shown that constitutively activated β-catenin can result in the hyperactivation of Wnt signaling. Conditional inactivation of adenomatous polyposis coli (APC) which forms part of the β-catenin destruction complex and increased expression of β-catenin/Lef-1 fusion protein can lead to cell death by apoptosis in mammalian intestinal epithelia [50–52]. Collectively, the results from the usage of different concentrations of N-cadherin antibody and CHIR99021 observed in our experiment support the evidence that the cellular responses to Wnt/β-catenin signaling is dosage dependent [53, 54]. Apart from that, several other studies have also reported that the Wnt/β-catenin signaling has directional and graded activity across the ventricular wall to regulate cardiomyocyte proliferation, with decreasing gradient from the outer to the inner myocardium [9, 55–57]. Therefore, it is important to consider the spatiotemporal pattern of Wnt/β-catenin signaling when testing potential therapeutic agents such as CHIR99021 to improve cardiomyocyte proliferation in the future.
The current study focuses on laying the groundwork for the use of matured cardiomyocytes that were expanded in vitro for cell-based therapy. While the data showcases evidences that suggest plausible application for CHIR99021 to be utilized directly as an in vivo therapy for heart regeneration by stimulating endogenous cardiomyocytes to proliferate, we found it unwise due to reports of uncontrolled activation of the Wnt pathway causing potential deleterious and cancerous effects [48–51]. Until we can sustainably release CHIR99021 in a controlled and targeted manner in an injured heart, it would be challenging to demonstrate application of the findings in vivo.
We have demonstrated in this study that activating Wnt signaling pathway by availing cytoplasmic β-catenin for nuclear translocation can enhance proliferation of matured cardiomyocytes. More importantly, we revealed that CHIR99021 is a promising candidate for the in vitro expansion of matured cardiomyocytes. The current study also provided evidence of dosage-dependent cellular responses to Wnt/β-catenin signaling using N-cadherin antibody and CHIR99021, providing a plausible avenue in the near future to activate Wnt signaling for endogenous cardiac regeneration using activators of canonical Wnt signaling pathway.
Yong Fan and Beatrice Xuan Ho contributed equally to this work.
Publication fee was covered by the Institute of Molecular and Cell Biology, A*STAR.
This work is supported by the Agency for Science, Technology and Research (Singapore) and by a grant from National Medical Research Council to B.S.S (NMRC/OFYIRG/0017/2016). This work is also partially supported by grants from National Natural Science Foundation of China to Y.F. (grant number 81871162 and 81570101), Guangdong Province Science and Technology Project (2017A020214005) and the Guangdong Science and Technology Project (201803010048). Beatrice Xuan Ho and Jeremy Kah Sheng Pang are supported by the National University of Singapore graduate scholarships.
Please contact the corresponding authors for data requests.
NSY and SBS conceived and designed the experiments. FY and HBX performed most of the experiments and analyzed the data. PMQN, PJKS, and HJH assisted in the experiments. SBS and PJKS wrote the manuscript. All authors read and approved the final manuscript.
The experiments regarding animal research were approved by the Institutional Review Board at The Third Affiliated Hospital of Guangzhou Medical University. The experiments using human cells and mice were approved by the ethics committee of The Third Affiliated Hospital of Guangzhou Medical University, and all animal care and experiments were carried out in accordance with the institutional ethical guidelines for animal experiments.
Robertson C, Tran DD, George SC. Concise review: maturation phases of human pluripotent stem cell-derived cardiomyocytes. Stem cells (Dayton, Ohio). 2013;31(5). https://doi.org/10.1002/stem.1331.
Li D, Wu J, Bai Y, Zhao X, Liu L. Isolation and culture of adult mouse cardiomyocytes for cell signaling and in vitro cardiac hypertrophy. J Visual Exp. 2014;(87). https://doi.org/10.3791/51357.
Ozhan G, Weidinger G. Wnt/β-catenin signaling in heart regeneration. Cell Regeneration. 2015;4(1). https://doi.org/10.1186/s13619-015-0017-8.


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The Museum has 2 entrances, the Cromwell Road entrance has steps. The Exhibition Road entrance is step-free, and best for entry with wheelchairs and pushchairs. Cromwell Road entrance: 10 steps via a ramped path at either side of the entrance. Exhibition Road entrance: there are no steps into the entrance lobby. Beyond that are 18 steps, or you can take the lift (this is a standard lift with doors). Wheelchairs are available on loan from the cloakrooms just inside the Exhibition Road entrance and behind the Central Hall by the main toilets. There is no charge for this service.
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text:
Паталеница е село в Южна България. То се намира в община Пазарджик, област Пазарджик.

География
Село Паталеница се намира на 15 км югозападно от град Пазарджик и след обединението си със село Баткун през 1955 г. се превръща в едно от най-големите села по северната страна на Родопския рид Къркария. Древните жители на региона са оценявали стратегическото значение на тези селища, разположени в началото на пътя между Тракия, речното корито на Чепинска река и долината на река Места. Край селото тече река Ерменска, за която се споменава и в книгата на географа проф. Иван Батаклиев "Пазарджик и Пазарджишко", София, 1969, с.72. Най-близките села наоколо са Црънча (2 км югоизточно), Дебращица (5 км югоизточно), Ветрен дол (6 км) и Варвара (6 км), а зад билото на Родопския рид Каркария – село Дорково.

В наше време село Паталеница продължава да привлича със своята магия и с кристално чистия си въздух и природа.

История
Паталеница е възникнала през средните векове, а Баткун е било селище с уредба като гръцките градове, в което е имало храм на Аполон. През Средновековието Баткунската крепост, позната и като Баткунион, е била най-големият град в Пазарджишко, наричан във византийските хроники "Блестящ и богат". Оттам са минали и войските на Филип и Александър Македонски. Днес до Баткун могат все още да се видят останките от светилището на Асклепий Земидренски датирано към I-IV век. В него са открити оброчни плочки на тракийския бог Херос, статуетки, жертвеник и много монети. Археологическите находки се съхраняват в Регионален исторически музей в Пазарджик.

От близкия манастир произхожда Баткунска хроника, свидетелстваща за насилията при налагането на исляма в Родопите.

През 1955 година към Паталеница е присъединено близкото село Баткун.

Културни и природни забележителности
 Баткунски манастир "Св. св. Петър и Павел" – основан през XII век. В него се намира скривалище на Васил Левски. Левски е основал тук местния революционен комитет.
 Баткунската крепост – "Баткунион"
 Църквата "Свети Димитър" – от XII век
 Народно читалище "Св. Пантелеймон-1909"
 Светилището на Асклепий Земидренски
 Църквата "Успение на Пресвета Богородица" в Паталеница
 Църквата "Света Богородица" в Баткун
 Параклис "Свети Георги"
 Паметникът с шадравана
 Късноантична крепост на връх Тепето
 Малката крепост на връх Калето

Редовни събития
 Патронът на селото е Свети Пантелеймон – 27 юли.
 От 1999 насам, в Паталеница всяко лято в края на юли Шекспирова театрална школа "Петровден" представя своите летни Шекспирови проекти. От 2017 година има и гостуващи представления на български театри и студенти от НАТФИЗ.

Други
 В Паталеница е запазен обичай в чест на свети Георги агнетата да се пекат на открито по улиците.

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Top Loop accommodates up to an 18g jump ring.
These Script Letters are punched out of metal sheet, just like our blanks, creating a nice and light charm. Great for a charm bracelet, necklace, or you can even cut the loop off, give it a quick filing and solder it onto a blank for a custom pendant!
Nickel silver is a metal alloy of copper, nickel and zinc. It is named for its silvery appearance, and contains no elemental silver. It can be stamped, polished and soldered just like sterling or copper.
Nickel silver, in its tarnished state, has an antiqued metal look. With a quick rub of aPro-Polish pad, they can be easily polished up to their grayish silver color.


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Dr. Sheldon K. Cho Joins Physician Team At Florida Pain Institute In Brevard County
By Space Coast Daily // December 22, 2015
seeing patients in titusville, merritt island
Dr. Sheldon K. Cho has joined Florida Pain Institute and is now seeing patients in Titusville and Merritt Island.
BREVARD COUNTY, FLORIDA – Dr. Sheldon K. Cho has joined Florida Pain Institute and is now seeing patients in Titusville and Merritt Island.
After spending more than 10 years practicing medicine on the West Coast, Dr. Cho decided to bring his medical expertise to Florida Pain Institute. He is a board certified Anesthesiologist with subspecialty qualifications in interventional pain medicine and substance abuse.
"I extremely happy to join the team of Physicians at Florida Pain Institute, and to call Florida home," said Dr. Cho.
Some of Dr. Cho's key areas of interest along with Interventional and Cancer Pain Management include various procedures under fluoroscopy including spinal cord stimulator trail and implant, spine Kyphoplasty, RFA, facet joint block, and epidural block.
He is also an Active member of the American Society of Addiction Medicine (ASAM), and an American Society of Interventional of Pain Physician (ASIPP).
Dr. Cho was born in Seoul Korea and received his Bachelors of Science degree from Seoul National University in 1988. He then continued his collegiate career at Seoul National University's College of Medicine, graduating in 1992.
After graduation, he interned in Long Island New York for several years at Nassau University Medical Center, Department of Surgery. He remained at Nassau University Medical Center, where he completed his residency with the Department of Anesthesiology in 1998. He then completed a Cancer Pain Medicine fellowship in New York, New York with Memorial Sloan-Kettering Cancer Center's Anesthesiology Department.
HOT OFF THE PRESS: Florida Pain Institute Magazine – Fall 2015
Dr. Cho continued his medical career as a Clinical Instructor at the University of Rochester Medical center, specializing in anesthesia regional block and pain medicine. In 2004 he moved to San Francisco California to take a position as a Clinical Professor at UCSF for the next four years.
Dr. Cho later went on to become the Medical Director at Anapa Pain Clinic in Los Angeles, specializing in Interventional Pain Management. While working at Anapa Pain Clinic, he served dual roles as the Medical Director at California Detox Clinic, where he specialized in addiction management.
Dr. Cho's certifications and qualifications include: American Board of Anesthesiology Certified; ASA, Pain Management Subspecialty; American Board of Pain Medicine Certified; American Board of Interventional Pain Physician; and American Board of Addiction Medicine Certified.
Patients can see Dr. Cho at the Florida Pain Institute Titusville location at 7455 South U.S. Highway 1; or at the Merritt Island office, located at 595 N Courtenay Parkway, Suite 101 on Merritt Island.
For more information or to make an appointment please call 321-784-8211 or visit FloridaPainInstitute.net