text:
Publishing Technology and CNPIEC launch CNP eReading platform
New channel unveiled for international academic publishers to access growing Chinese market
Beijing, China - 28 August 2013. Publishing Technology and CNPIEC, the China National Publications Import and Export (Group) Corporation, are proud to announce the unveiling of the CNP eReading platform at the Beijing International Book Fair this week.
At launch, the site carries over 200,000 individual book titles from more than 300 international publishers, extending their reach into the rapidly growing Chinese market and attracting new readers to digest their academic content for the very first time.
Following a deal signed at the London Book Fair in 2012, the digital hub has been developed by Publishing Technology, a world-leading provider of content solutions for publishers, on a custom-built online publishing platform. CNPIEC, the largest and most highly-developed government sanctioned publication import and export group in China, will manage the new site which is available in both English and Mandarin.
The CNP eReading platform presents international publishers with an exclusive opportunity to showcase their foreign language journals and ebooks to previously untapped Chinese audiences. The site is set to increase its presence dramatically in the next year with the continued assistance of Publishing Technology, who will invite its 250 ingentaconnect customers to host over 12,000 journal publications on the platform.
In order to assist this outreach programme, Publishing Technology's extensive ingentaconnect client list of international publishers have been invited to the First International Journals Conference in China – held at the Wuhan International Expo Centre (September 14-16).
Mr Bogen Liu, Vice President of China Publishing Group and President of CNPIEC, commented: "We are thrilled with the CNP eReading platform. We have worked closely with Publishing Technology to deliver a product which has the potential to greatly widen the amount and diversity of foreign language content available in China. Publishing Technology's affiliation with publishers across the globe has significantly extended the volume of international content we can offer our customers."
George Lossius, CEO at Publishing Technology, stated: "We are extremely proud to play a continued role in the advancement of Chinese library digitisation and increasing the opportunities for international publishers to gain exposure in China. Our partnership with CNPIEC is helping the platform grow in strength and depth, whilst our connection with local digital publishing experts is benefiting our international clients. Our strong presence within China allows us to provide appreciable value and assistance to publishers who are understandably eager to meet the growing appetite for digital content in the world's largest growing market."
- ENDS –
Publishing Technology plc:
Publishing Technology is the world-leading provider of content solutions that transform business. We cover the publishing process from end to end with content systems, audience development and content delivery software and services. Combining our unmatched publishing knowledge, global operations and perpetual support model with our advance operations system, ingentaconnect scholarly portal, pub2web custom hosting platform and PCG (Publishers Communication Group) sales and marketing consultancy, we offer the industry's only full spectrum of solutions to help publishers move their content forward. Listed on the AIM market of the London Stock Exchange, the company operates jointly from Europe (Oxford) and North America (Boston and New Jersey), with local offices in Brazil, India, China and Australia. Assisting 400 trade and scholarly publishers for over thirty years, Publishing Technology solves the fundamental issues content providers face.
Visit publishingtechnology.com, follow on Twitter @publishingtech, or connect on LinkedIn.
CNPIEC:
China National Publications Import & Export (Group) Corporation (CNPIEC) is the largest and most highly-developed cultural products import and export group in China. Its product structure incorporates core businesses of import of publications, export of publications, conferences and exhibitions in China and abroad, overseas publishing, global distribution and other international trade businesses.
As the largest importer of overseas publications into China, CNPIEC has over 10,000 domestic institutional clients and over 60% of the domestic market for imported publications. Based on its excellent brand reputation, CNPIEC has become the sole partner in the newsstand projects in 2008 Olympics Games, the 2010 Shanghai World Expo and Guangzhou 2010 Asian Games, and it's also an "appointed supplier" for the United Nations' book procurement. CNPIEC's Publications Bonded Warehouse is the first bonded warehouse exclusively for stocking imported publications in China.
CNPIEC is striving to create China's largest service platform for media information and materials on education, science, culture, health, economy, national defence and military services, and to turn CNPIEC Online into an internationally-leading, domestically-unrivalled platform for information services.
Emily Taylor Gregory, Publishing Technology plc
Email: emily.taylor.gregory[at]publishingtechnology[dot]com


Monika joined Burns + Beyerl Architects as a Project Architect in December 2012. She received a Bachelor of Architecture in 1988 from the University of Illinois at Chicago, with concentrated studies in the History of Art and Architecture. During her third year at UIC, Monika attended the E'cole de Architecture in Versailles, France.
Monika was born and raised on the north side of Chicago. She began her career in her first year of college, working for several small Chicago firms specializing primarily in custom single-family home renovation. She has also worked on several school, commercial office, restaurant and religious renovation projects throughout the Chicago and neighboring suburban area, often serving as the onsite architectural representative. Monika excels at coordinating the architectural design with the various consultant designs into a built environment, which meets the Client's desired program. When it comes to design renovation, Monika believes in enhancing the heart and soul of the existing structure, bringing it to its' full potential.


Currently WHMCS shows a warning for failure to use TLS or SSL for SMTP even when the MTX is on the local machine. In this scenario the use of TLS or SSL is not only useless but adds additional server load with no added benefits. Please do not show this warning when the SMTP server is set to "localhost" or "127.0.0.1"


text:
Los bodos, también llamados boros, son una etnia del noreste de la India, en el estado de Assam. Se clasifican dentro de un grupo étnico mayor denominado Bodo-Kachari. La población más importante en la que habitan los bodos y el centro de su cultura es la ciudad de Kokrajhar.

Durante mucho tiempo ha existido una lucha para promover la cultura e identidad de los bodos. En la actualidad, los bodos disponen del Consejo Territorial Bodo (the Bodo Territorial Council), una institución administrativa autónoma que ejerce su jurisdicción sobre Kokrajhar y sus alrededores.

El Frente Nacional Democrático de Bodoland (FNDB) reclama además la secesión de Bodoland y es una guerrilla activa en ciertas áreas.

Referencias
 The Bodos: Children of the Bhullumbutter, editado por Thomas Pulloppillil y Jacob Aluckal, (primera edición 1997). (en inglés)
 Bodo-English Dictionary, de Moniram Mochari, 1981. (en inglés)

Enlaces externos
 bodoland.org (en inglés)
 National Democratic Front of Bodoland (en inglés)

Etnias de India
Assam

redpajama_set_name: RedPajamaWikipedia


The 70th birthday is a perfect opportunity to celebrate the hard work and commitment of NHS staff, profiling their skills, experience and successes, whilst recognising the challenges they face.
NHS England and NHS Improvement are encouraging everyone to get involved – from NHS trusts, GPs, clinical commissioning groups and local authorities to charities, patient groups, unions and professional bodies.
It's also a time to look back over the last 70 years, celebrating key developments and breakthroughs, and to look forward and build confidence about the NHS's long term future.
In addition, it's a great opportunity to let people know about the many ways in which they can support the NHS – from volunteering and using services wisely through to giving blood and raising money for NHS charities.
Celebrations will start in January 2018 and will undoubtedly peak in July.


text:
Jamal A. Badawi () is an Egyptian-Canadian author, preacher and speaker on Islam.

Life
Badawi completed his undergraduate studies at Ain Shams University in Cairo. He left for the United States in the 1960s and completed his Masters and doctorate, both in the department of Business Administration, at Indiana University Bloomington. He has been serving as a volunteer imam of the local Muslim community in the Halifax Regional Municipality since 1970. He cites Hassan al-Banna as an influence.

Badawi was formerly a professor in the Sobey School of Business, Saint Mary's University in Halifax, Nova Scotia.

Badawi is married and the father of five children, and grandfather of 23.

Career
In addition to his participation in lectures, seminars and interfaith dialogues in North America, Badawi was invited as a guest speaker in various functions throughout the world. He is also active in several Islamic organizations, including the Canadian Council on American-Islamic Relations, the Fiqh Council of North America (FCNA), the Muslim American Society (MAS), and the European Council for Fatwa and Research (ECFR). He is also a member of the Islamic Society of North America (ISNA) and the founder and chairman of the Islamic Information Foundation, a non-profit foundation seeking to promote a better understanding of Islam and Muslims towards non-Muslims. He has lectured extensively in North America and abroad, and speaks on a variety of topics including Islam and Christianity and is a guest scholar at The American Learning Institute for Muslims. In 1997, he debated Christian apologist William Lane Craig over the concept of God in Christianity and Islam.

According to a 26 September 2005 diplomatic cable from the American Embassy in Ottawa, Canada's capital, Jamal Badawi is someone who has "broad influence among Muslim youth" and is "involved in countering extremism" and "promoting tolerance."  According to Gofran Sawalha of Middle East Eye, he is among "the best contributors to Islamic knowledge in America".

Badawi has authored several books and articles on Islam. He also researched, designed and presented a 352-segment television series on Islam, aired in Canada, the US and other countries.

In 2017, Badawi stated that "[t]errorism has no religion" in response to the Quebec City mosque shooting. In 2018, he expressed concern regarding the presence of the National Citizens Alliance at the Annapolis Valley Apple Blossom Festival in Kentville, Nova Scotia.

Publications
 1,000 Questions on Islam Islamic Book Services,
 Selected prayers Oscar Publications, Dehi 2007;
 Gender Equity in Islam: Basic Principles American Trust Publications, 1995;
 Leadership: An Islamic Perspective (jointly with Beekun Rafiq Issa) Amana Publications, 1999;
 Muhammad in the Bible Small pamphlet, 1982
 Status of Women in Islam Small pamphlet, 1976
 Muslim Woman's Dress According to the Qur'an and the Sunnah and Islamic Ethics Small pamphlet, 1980
 Polygamy in Islamic Law Small pamphlet
 Islam: A Brief Look Small pamphlet

References

Year of birth missing (living people)
Living people
Canadian anthropologists
Canadian Muslims
Ain Shams University alumni
Indiana University Bloomington alumni
Egyptian emigrants to Canada
Academic staff of the Saint Mary's University (Halifax)


text:
Q: Правильное создание потока Этот код сильно нагружает процессор. Как понимаю, простаевается поток все время.
Пробовал несколько вариантов, но все равно нагрузка не меняется

for (;;)
{
    Process[] processes = Process.GetProcesses();
    for (int i = 0; i < processes.Length; i++)
        if (processes[i].ProcessName.ToUpperInvariant().Contains("DNSPY"))
            Environment.Exit(0);
    Task.Delay(100);
}

A: Task.Delay работает только в асинхронной функции, при условии, что его дожидаются через await. Если у Вас отдельный Thread для этой задачи, то можно использовать Thread.Sleep(100);
пример:
private async Task DoSomeWorkAsync()
{
   for (;;)
   {
       Process[] processes = Process.GetProcesses();
       for (int i = 0; i < processes.Length; i++)
          if (processes[i].ProcessName.ToUpperInvariant().Contains("DNSPY"))
              Environment.Exit(0);
       await Task.Delay(100);
    }
}

Либо:
private void DoSomeWork()
{
    new Thread(() => {
      for (;;)
      {
         Process[] processes = Process.GetProcesses();
         for (int i = 0; i < processes.Length; i++)
            if (processes[i].ProcessName.ToUpperInvariant().Contains("DNSPY"))
                Environment.Exit(0);
         Thread.Sleep(100);
      }
    }).Start();
}

Вариант с Task предпочтительней.

redpajama_set_name: RedPajamaStackExchange


text:
Curious Theatre's The Secretary Is Right on Target
DCPA Announces Hamilton On-Sale Dates, With Scalper Warnings
Punk Rock Karaoke invites you to sing along to Bikini Kill and Jawbreaker for a good cause
Bree Davies
Bree Davies | June 20, 2013 | 9:15am
Rebel girls and rude boys, this one's for you: tonight's Punk Rock Karaoke invites everyone to sing along with his or her favorite songs for a good cause, as the Denver Health Collective hosts a traveling repertoire of punk hits. The karaoke project makes its first stop in Denver -- which also acts as a benefit for the Collective -- bringing with it a 500-plus song catalog of songs by Fugazi, Bratmobile, The Clash and more.
In advance of the gathering tonight, June 20, co-organizer Patrick Tyrrell spoke with Westword about the Punk Rock Karaoke Collective's formation, the importance of collaborating with grassroots organizations and why he geeks out over seeing Bikini Kill and Jawbreaker duke it out for the most requested karaoke spot.
See also: - The ten best places to sing karaoke in Denver - Derailer bicycle collective celebrates its tenth anniversary - Fat Mike of NOFX on pornography, punk-rock parenting and how America is sinking
Westword: How did Punk Rock Karaoke start?
Patrick Tyrrell: So, there was a pretty small group of us in Chicago -- and each of us was organizing or involved in various campaigns and community groups. A lot of us were musicians, too, or at least musically inclined. We had been batting around this idea of doing a more fun fundraising project that involved karaoke. One of our organizers had the idea for a punk rock flavor, because it was the kind of music we were all into.
Because a lot of us were musicians, we thought, hey, whatever songs we can't find that are already out there, we can make ourselves. That initial idea in 2011 is what spurred forth this project, two years later. What started in Chicago has now expanded to the Northeast and also become this nationwide tour.
When we first started (making karaoke tracks) we were recording in a recording studio and it was a fun project. To make so many of the songs ourselves and then see people perform them -- it was just a good way to combine our love of music and performing with making sure a good cause gets supported, and to let people know about what is going on in their community.
It is a great concept, because when you look at the majority of karaoke catalogs, it is very Top 40-heavy -- but there is definitely a nationwide audience out there for punk-inspired performances.
I've seen lots of themed karaokes, and even punk rock karaoke itself -- we didn't invent that specific concept at all. I don't think anyone else has done it as a fundraiser or as a not-for-profit venture, but there is definitely similar versions of this karaoke. A lot of times it is a live band karaoke. But to my knowledge and from what I've seen, we have the largest custom catalog of punk rock songs that are prerecorded and more like regular karaoke. I think we also have the largest catalog in general -- we have over 500 songs that are all tailored to a punk rock flavor, with classic punk songs and modern stuff, too.
How do you pick the songs that will be made into tracks and used for Punk Rock Karaoke?
A lot of it is from our audience -- and we have a sense from being fans of the genre what the kind of standards are. We know we need certain bands, like The Clash or Ramones. People love Jawbreaker and all of those bands. But we get a lot of feedback from people so, one thing we tried to do was have a good representation of female-fronted bands. Aside from what we knew, we heavily solicited people that we know that were in Riot Grrrl bands or that performed in punk bands and got recommendations like L7, Bratmobile and Sleater-Kinney.
We collect them in a giant list and go with how much time we have to record, how complicated the songs are and then also, just a general sense of what people are requesting -- we let our audience guide us.
As a musician, I see it being a fun challenge to sort of become a catalog of songs that you love.
Oh yeah. I know so many stupid punk songs. (Laughs) I know them by heart. We've been doing this for a long time, and I really don't remember recording a lot of the first tracks -- but it's funny because sometimes we'll be doing an event and I'll realize, oh, that's me playing the guitar solo. I don't remember even doing it, but I know at one point I had committed it to memory.
The fact that you go in and record the tracks seems so integral to this working -- live music is one thing, but knowing every guitar tone or hearing a pedal being pushed in an actual recording is so key to knowing the whole song.
Our producer, his name is Joe Tessone -- who runs Mystery Street Recording Company, the studio we worked out of originally -- would spend hours just tweaking the tone of everything. He'd line up the original recording and our track and he's such a great producer, he would get it so it (sounded) indistinguishable. He would play a song side by side, and I couldn't tell if it was the original or ours. For the touring edition of a Punk Rock Karaoke event, what does the night look like?
It really varies per town; some places are way more high energy than others. Normally, you show up and the first hour is a little more laid back -- we take requests and people mill around. No one wants to sing first, so there's usually more shyness happening than anything. Once we get through the first ten songs or so, the ball gets rolling. After that point, it is really high energy as the night goes on.
People do massive sing-alongs when a popular song comes up -- people rush the stage. We've had crowd surfing, we've had mosh pits, we've had rowdiness. I think in a lot of ways, most nights just feel like a show, you know? But it's a show where you get to see your friends performing. We try to foster a really participatory environment; we want to break down the barriers between the audience and the performer. We have three mics on stage and we always encourage people, if they like the song, to get up and sing with strangers. Great duets can happen spontaneously with two people that have never met each other before, but just love the band so much that they know what to do.
Because the venue we're doing it for in Denver is a little more D.I.Y., I think it will definitely feel more like a show. It's never like regular karaoke where you have one person singing awkwardly and everyone's just sitting there tapping their feet. It's never been like that.
What songs do you see performed or requested most often?
We actually keep data on everything -- we keep track of everything that's been requested. "Rebel Girl" by Bikini Kill and "Boxcar" by Jawbreaker are both tied at the top. For us, it is really geeky, but it is really exciting because Jawbreaker has beat Bikini Kill by one or two requests for the entire project. We've all been wondering, is Bikini Kill going to unseat Jawbreaker?
Then, the rest of the top five are: "Baby, I'm an Anarchist" by Against Me, "Linoleum" by NOFX, "Deceptacon" by Le Tigre and "Last Caress" by Misfits. So, that's the top requests, but there are certainly lots more -- like Fugazi, Descendents and Black Flag.
That's interesting data, and what sticks out to me is that two of Kathleen Hanna's projects -- Bikini Kill and Le Tigre -- are in the top five.
That's a good point -- I actually hadn't noticed that until now. It's been great to watch "Rebel Girl" climb up and tie (with "Boxcar.") Whenever these songs come on, it is like the whole room explodes.
From the benefit standpoint, how do you select the organizations that Punk Rock Karaoke works with?
On a tour it's been a little bit different -- we've been working within the networks and contacts that we have in each city. But when we're doing events in Chicago and the Northeast, we tend to choose groups that are, first of all, based in the community. They have some sort of specific tie-in to a neighborhood or area of the city. We focus on organizations that are working by their own bootstraps -- they don't have a ton of funding coming in from grants or any other foundational support. Smaller projects that don't get traditional sources of funding.
We don't raise millions of dollars or anything -- but we keep in touch with all of our organizations and ask, like, hey, what did you use the thousand bucks or whatever we were able to raise for? We did a fundraiser for a music program and they bought instruments, or a group may use the money to buy food for their own events.
Sometimes groups pitch to us and send us a letter explaining what they need the money for and why they like the project. The last fundraiser we did in New York was for the New York Abortion Access Fund -- and they are an entirely volunteer program. They have lots of different types of fundraisers, but they are really grassroots. Basically they gave us a really compelling pitch: the money that Punk Rock Karaoke raises for us would go directly for abortion services. That is the kind of impact we're looking for.
Punk Rock Karaoke starts at 7 p.m. at the Denver Community Health Collective, 2727 West 27th Avenue; proceeds benefit the DCHC (suggested donation is $5 to $10). For more information, visit Punk Rock Karaoke's Facebook page for the Denver event.
Follow @WestwordCulture
Bree Davies is a multimedia journalist, artist advocate and community organizer born and raised in Denver. Rooted in the world of Do-It-Yourself arts and music, Davies co-founded Titwrench experimental music festival, is host of the local music and comedy show Sounds on 29th on CPT12 Colorado Public Television and is creator and host of the civic and social issue-focused podcast, Hello? Denver? Are You Still There? Her work is centered on a passionate advocacy for all ages, accessible, inclusive, non-commercial and autonomous DIY art spaces and music venues in Denver.
Twitter: @cocodavies
Colorado Creatives: Meghan Frank and Buba Basishvili
Art Attack: Fourteen Best Things for Denver Gallery Lovers to...
Book It: The Six Best Literary Events This Week
Museum of Outdoor Arts Hosts a Massive Rauschenberg Exhibition

redpajama_set_name: RedPajamaCommonCrawl


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Urban Fantasy: Fiction, Art, Culture, Entertainment is a new Facebook group dedicated to all things Urban Fantasy: culture, literature (fiction, nonfiction & poetry) music, pictures, film, television, technology, games, fashion, lifestyle, events, cosplay, comics, manga etc.
We're also at the grimmer, darker, grittier occult detection, technomancy, rather than paranormal romance, end of the Urban Fantasy spectrum.
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text:
Q: Openssl_decrypt returns empty output Hello everyone i was trying to decrypt some encrypted strings i encrypted with python .
the problem is that sometimes it decrypts correctly and sometimes it gives empty output for no obvious reason and i couldn't find any solution for it.
here's the code i'm using to decrypt on PHP .
knowing that online AES decryption tools decrypts it correctly.
$rtk=base64_decode('zgdHfETipvp1E5m3ix5NFOLuX8N0+zAIBzg+GOq0cTQ=');
$method = "aes-128-ecb";
$key =  'aaaaaaaaaaaaaaaa';

$email=openssl_decrypt($rtk, $method, $key,OPENSSL_RAW_DATA);

i would apreciate your help  !
EDIT :
The python code i used to encript the string :
import pandas as pd
from Crypto.Cipher import AES
import names
import urllib.parse
import base64
from Crypto.Util.Padding import pad
from Crypto.Util.Padding import unpad

email="zqeafzeqaf23@example1.com"
key = b'aaaaaaaaaaaaaaaa'
data = email.encode('ascii', 'ignore')
cipher = AES.new(key, AES.MODE_ECB)
b64string = base64.urlsafe_b64encode(cipher.encrypt(pad(data,16)))
print(b64string)

A: Your Python code uses urlsafe_b64encode, but your PHP code uses the normal base64 variant (in fact your example data in the PHP code contains a + character so couldn't have been produced by that Python code).
This could explain why the decryption is failing. If the url safe base 64 output from Python contains a - or _ character, PHP will simply strip that character from the string before decoding the rest of it. This will leave a string that is not a multiple of the AES block length and the decryption will fail.
You should ensure you are using the same base 64 variant to encode and decode. It doesn't look like PHP provides a URL safe variant, you might need to use something like strtr before decoding to convert the base 64 into a normal variant:
$data = strtr($data, '-_', '+/');
$decoded = base64_decode($data);


text:
Nila Nilova
[[STREAMING##]] Blue Jackets vs Jets live free 3 December 2022
Columbus Blue Jackets at Winnipeg Jets - 12/2/22 NHL Picks and PredictionWinnipeg Jets (14-6-1) vs Columbus Blue Jackets (7-12-2) Game Info: Friday, December 2, 2022 at 8:00 pm (Canada Life Centre) Betting Odds: Winnipeg Jets -245 / Columbus Blue Jackets +195 --- Over/Under: 6. 5 Click Here for the Latest Odds Where to Watch: ESPN+ Stream live sports and exclusive originals on ESPN+. Sign up now! The Columbus Blue Jackets will play the Winnipeg Jets on Friday night at Canada Life Centre in Winnipeg, Manitoba.
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Winnipeg is 12-2 in their last 14 home games and 7-19 in their last 26 games against the Metropolitan. The under is 15-5-2 in Winnipeg's last 22 home games. The over is 5-1 in these two team's last 6 head-to-head meetings. Columbus comes into this game on a three game losing streak, but has played better defense in their last four games. Winnipeg has won three games in a row and is coming off of a 5-0 win against Colorado. The Jets are playing great right now and have one of the best goalies in the league.
Outside of Columbus the Jackets surrender a whopping 4. 5 goals per contest — the most in the league. Other than Johnny Gaudreau and Boone Jenner they simply don't have the firepower to make up for such poor goaltending. Jets 3-way ML at -150 available at time of publishing. Playable at that number. SportsbookSet WagerPayoutSelect Odds-SportsbookPayoutPlace BetSignUp$16.
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75 goals against average and a. 899 save percentage. The Winnipeg Jets are 14-6-1 this season after they defeated the Colorado Avalanche by a score of 5-0 in their last game. Winnipeg was out shot by a total of 40-23 and won 63% of the faceoffs in the game. The Winnipeg Jets are averaging 3. 24 goals per game, 2. 52 goals against per game, 20. 9% on the power play, and 82.
The Columbus Blue Jackets are 7-12-2 this season after they lost to the Vegas Golden Knights in a shootout by a score of 3-2 in their last game. Columbus was out shot by a total of 36-28 and went 0-2 on the power play in the game. The Columbus Blue Jackets are averaging 2. 86 goals per game, 4. 10 goals against per game, 15. 4% on the power play, and 79% on the penalty kill. Johnny Gaudreau has led the Blue Jackets with eight goals, 14 assists, and 65 shots on goal this season. The projected starting goalie for Columbus is Joonas Korpisalo, who is 3-4-1 this season with a 3.
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1% on the penalty kill. Josh Morrissey has led the Jets with five goals, 20 assists, and 38 shots on goal this season. The projected starting goalie for Winnipeg is Connor Hellebuyck, who is 11-5-1 this season with a 2. 29 goals against average and a. 931 save percentage. Columbus is 17-35 in their last 52 games and 1-6 in their last 7 games against the Central. The over is 14-6 in Columbus' last 20 games.
Blue Jackets vs. Jets Tickets 2022 - SeatGeek
Goaltender Connor Hellebuyck and the Jets are allowing just 1. 6 goals per game at home. That's incredibly low — the best mark in the league, in fact. Only 1 other team is below 2 goals allowed per game at home, and that's the 19-3-0 Boston Bruins. The Jackets have been the exact opposite in terms of goaltending. They are allowing 4. 1 goals per game, which is the 2nd most in the NHL, and that gets even worse on the road.
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Winnipeg Jets vs. Columbus Blue Jackets odds, tips and
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They took down Nathan MacKinnon and company 5-0, which made them the first team in the Central Division to reach the 14-win mark. Winnipeg has been off since that game, which bodes well when facing the worst team in the Eastern Conference. The Blue Jackets haven't been on the ice since Monday night, when they actually forced the Golden Knights to a shootout. While that was surprising, they have still lost each of their last 3 games — all of which were at home. Now they have to head up to Canada to take on a Jets team that has been great on their own ice. Their shutout over the Avs perfectly illustrated why they are so dominant in Winnipeg.
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Did you go and see Lum?
Polly and Martin are moving back home.
Caroline is a heavy reader.
Everything Barrio did, Juliane criticised.
I'm pretty sure Sergiu's rich.
Liza and Daniel got married a few years ago.
Takeuchi told me not to worry.
Geoff doesn't like washing windows.
How long have you been Leung's supervisor?
Torsten didn't go to work all last week.
I saw Claude and could barely recognize him.
Chet is wearing a biohazard suit.
I think Takao looked better before he shaved off his beard.
Cindie agreed to meet with Carolyn.
Pratap didn't get what he wanted.
Sean wants to borrow some money.
Where's your sense of humor, Mohammad?
Sarah had a good time.
Lenora wanted to know why Boyd didn't like him.
Izzy was rushed to a local hospital.
Do you really think Scot should resign?
Randolph kissed Darren and then she slapped him.
What does Sanity intend to do about it?
Come on, Matti. Let's get out of here.
Sanand and Nicolette were both alcoholics.
Kris hid behind her large sunglasses.
Shaw and Shamim are having a good time.
I should've given Knapper my old trombone.
I barely ever talk to Mohammad.
Bret has been reading a book all afternoon.
Randell, I want you to listen to me.
Kyung snatched defeat from the jaws of victory.


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Home News Record turnout for Beaulieu's bumper Simply Land Rover
Record turnout for Beaulieu's bumper Simply Land Rover
Posted	24.06.2019
Beaulieu's Simply Land Rover was the biggest and best yet, with record numbers of Land Rovers and rally participants turning out for the busy event which saw the return of the popular Simply Rummage and a fully-booked Forest Drive.
542 Land Rovers of all ages and varieties rumbled into the grounds of the National Motor Museum for the rally on Sunday June 23rd, an increase of 8% from 501 vehicles in 2018, while the total of 1,285 rally participants was 18% up on 2018's 1,091 show-goers.
The much-loved Discovery was well-represented in its 30th anniversary year within the varied Simply Land Rover line-up, from Spartan early three-door versions up to the modern-day Discovery Sport SUV. Also on show were examples of the rugged Land Rover Series I dating as far back as 1949, the latter-day Defender, luxury Range Rover and even the military Land Rover 101 Forward Control.
Beaulieu's Forest Drive was fully-booked with enthusiasts eager to put their off-road driving skills to the test. Held in association with Land Rover Experience West Country, this action-packed feature took in a newly-extended course as Land Rovers of all ages were put through their paces.
Returning due to popular demand was Simply Rummage, a Land Rover-themed autojumble that made its successful debut at last year's show. Buyers snapped up bargains amongst the spares and accessories on offer, which ranged from seats, wheels and body panels to a rolling chassis. There were even complete vehicles for sale at the show, including an ex-Manchester Police 1973 Range Rover restoration project.
Winning the People's Choice Award with the most votes from show-goers were Michael Jones and his son Louie for their 1993 Land Rover Defender 90. Michael from Eastleigh had carried out a lot of work over the past three years to equip the 4X4 for off-road adventures, including fitting an impressive roof tent. "Really it's just a big Tonka Toy," said Michael. They were presented with a Beaulieu trophy and Autoglym car care pack by Beaulieu's Commercial Director Stephen Munn.
Beaulieu's Stephen Munn presents People's Choice winners Michael and Louie Jones with their trophy
Also scoring highly were runners-up Adrian, Kirsty and Tom Paget from Malmesbury, Wiltshire, and Fred Barclay from Devon. The Paget family's immaculate 1966 Series IIA had a fascinating tale to tell, having been bought new by Kirsty's mother and since restored to its original specification. "I have many childhood memories of this Land Rover," said Kirsty. "It will certainly stay in the family."
A People's Choice runner up, this Land Rover Series IIA had been owned by the Paget family since new
Fitted with a huge crane and laden with a tree trunk, Fred Barclay's hard-working 1999 Defender 110 was also a real crowd-pleaser. "I put the crane on with a friend, strengthened it and changed the colour," said Fred. "It's an everyday work vehicle and very practical." Both Fred and the Paget family were presented with Autoglym prizes and tickets for a behind-the-scenes tour of the National Motor Museum Trust.
Belonging to People's Choice runner up Fred Barclay, this Defender is fitted with a fully-functioning crane
Show regulars the Solent & District Land Rover Club showcased a selection of their members' vehicles, including a Defender tipper truck and a pair of Discoverys that had been modified for extreme off-road terrain. Also putting together popular club stands were The Discovery 3 & 4 Owners Club, Range Rover P38 Owners Club, LR Ladies Club, The Land Rover Series One Club, Range Rover Owners and Hants & Berks 4X4 Response.
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Q: Android WebView opens uTorrent When utorrent is installed on phone, clicking on a link in app webview opens utorrent instead of loading link!
even using javascript doesn't solve the problem:
window.location.href='index.html'

it still brings up utorrent.
There's no problem if utorrent is not installed.
I couldn't find anything about this in web.
What is the problem?

A: When the user clicks on an ordinary link in a WebView, Android, by default, will start an ACTION_VIEW Intent on the URL in the link. Typically, this will bring up the user's default Web browser, though in some cases it may bring up another app, as apparently is the case here.
If you do not want the default behavior, you need to create an instance of a WebViewClient subclass, where you implement shouldOverrideUrlLoading() to do something else instead of the default behavior. You then attach the WebViewClient instance to the WebView via setWebViewClient().

A: You'll have to create a WebViewClient to prevent that behavior:
public class webViewClient extends WebViewClient {
    @Override
    public boolean shouldOverrideUrlLoading(WebView view, String url) {
        view.loadUrl(url);
        return true;
    }
}

And then set it to your WebView like this:
webview.setWebViewClient(new webViewClient());

A: uTorrent app may specify an activity that can be opened by an intent containing an html link. If you don't have anything else that can open links (ie. some kind of browser), or if you've set uTorrent as default app to launch for links, it will open automatically.
Try going to Settings > Apps and click on uTorrent. If you can, click the "clear defaults" button, then try opening your link again.


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Statements are not accurate. Teams can begin setting a 10 man practice squad on Sunday. Rounding off the night is Super Bowl hopefuls Oakland Raiders' trip to Washington to face the Redskins in Sunday Night Football, from 1.15am.. "So Reggie was obviously a great player at Alabama," Reid said.
Who Receives Super Bowl Rings? By Jennifer Mueller Made of white or yellow gold and studded with diamonds and other gems, the coveted Super Bowl ring represents persistence and sacrifice in pursuit of professional football superiority. But an intriguing prospect is http://villa-micka.de/buy-methotrexate-2-5mg/ online Order Purchase wholesale jerseys Cheap Order http://www.consulpri.com.br/?p=866 Malcolm Carter, a six foot six, 192 pound junior player with the Ottawa Sooners who ran a 4.6 second 40 but laid out to make an outstanding catch in the one on one drills..
An example of a widely known pay for play site is also a company that is well represented in airing and promoting games in professional sports: Fox Fantasy Sports. "This is a huge opportunity for us," she adds, noting how NFL games offer the kind of reach that is hard to come by these days..
Some of the combatants wore jerseys of the opposing teams that day: the Rams and the Dallas Cowboys.. NFL teams may not have signed him to play quarterback this season, but they could not keep Kaepernick off the field.. To Benjamin Franklin Parkway and 24th St.
In a statement Sunday, Kraft said he was disappointed by the tone of the comments made by the president. Smith has a homefield advantage and better weapons at his disposal, but this Broncos defense is legit and brings heat from everywhere. Best friend qualifies for a number of mitigating factors that would support his early parole and has been discipline free during imprisonment, Bisbee said Thursday.


Camping is one of the best ways you can bond with your family and friends, it is a way for everyone to get together away from normal daily distractions and get back to life's simple pleasures. It's an activity everyone can participate in and lets you reconnect with nature. If If you live in the city and you want some time off the busy city streets, you can consider camping as a getaway watching the stars before you go to sleep.
Not only for families, camping is also really popular with schools. Maybe you are a teacher, planning to suggest this activity to your school and looking to gather abit of info. Below are some of the best outdoor camp providers in South Australian that might be able to point you in your quest.
Beyond the Classroom – is a part of Surf & Sun that promotes tourism and awareness. Camps are run in a very distinctive and exciting way with emphasis on ecological consciousness without taking the fun out of the table. Our events will be most likely adored by your family members, a group of friends and students wanting to enjoy and experience an out-of-school adventurous activity. Surf & Sun also offer other activities like surf lessons Middleton, bike rentals and kayaking.
Wilderness Escape Outdoor adventures – also a school camp in Adelaide South Australia, aside from camping they also offer various activities that you can participate with your family.
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If you are still looking for other recreational outdoor activities to keep either you, your family or friends busy, you can contact Surf & Sun for more activities you can choose from.


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Despite recent news, Waterloo community and RIM employees remain confident
Digging around the internet today there are plenty of headlines proclaiming the death of RIM. Microsoft rumors, Android rumors, network sale rumors. You name it, someone probably wrote about it by now. However, despite all of those headlines and bad outlook there is still plenty of faith in RIM to be found. RIM employees are still proud of where they work, RIM's hometown of Waterloo still has faith in the company and I'm positive it doesn't stop there. As noted in The Record, many folks spoke about the recent job cuts, RIM's future and what it all means to Waterloo:
"We'd certainly prefer not to see the recent job cuts. However, we are still very confident in RIM's ability to remain a major player in the global hand-held market place," Accelerator Centre CEO Tim Ellis said in an email.
"My heart goes out to the employees that are going to be affected,We don't know how many are going to be affected in our community, but it's significant for people. It's our friends and our families and our neighbours . . . I think it's important that we're supportive." said Waterloo Mayor Brenda Halloran.
"I think there's a lot of resiliency in the community," Regional Chair Ken Seiling said. "I'm optimistic that (RIM) will continue in one form or another."
"All I can say is, I'm going to keep doing this until they tell me to stop." - Alex Kinsella, Senior Public Relations and Social Media Manager, RIM
At this point, there is no denying that RIM has some major hurdles to overcome. That's stating the obvious. I won't even bother pretending I have the solution for them, I'll leave that to the armchair critics. However; I will say that for a company proclaimed dead by the masses they sure seem to have a lot of fight left in them.
Via: The Record


We went from frozen tundra to tropical rainforest in about 3 days!
Here is the scene at the start.
Normally I would never set up with this situation, but I found this great scene with a flowering apple tree, and my spot was shaded so I knew I'd be cool.
There were some large clouds that moved over the scene putting various parts into light/shadow.
Began with the panel in shadow ended with it in light.
"I'm very new to plein air and I'm curious how you are getting your lines on your canvas to match up perfectly with your background in your photos? You're obviously getting perfect scale drawing as well."
I learned how to draw, (or render) when I was a teenager, in classes and informally, still lives, figures, etc. I had a lot of teachers and studied great art and art history and still do, or try to. I believe drawing is the base skill for just about everything, and feel like I should be drawing now. When I'm using paint I'm not thinking about lines, I'm thinking about shapes. About copying the shapes I see in nature onto my panel. I think about their relative value in comparison to other relative values. If you really look at your subject, you may be surprised that things don't have outlines or clearly defined edges, that things don't even look like things. In daily practice, I don't want to think too much. I want to respond to the day and the scene in front of me, much of the "drawing" aspect of my work is automatic, I measure, line things up, and take a lot of what I've learned for granted. So now you're thinking I'm a pompous jerk, but all the practice that's gone into crafting a scene in paint is just that. Great art can be anything, but it must also convey the intent of the artist, it must be seen as only you can see it. Have fun, don't think too much and go on to the next.


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No one expected CNN to backstab this Biden official live on air
People have come to expect that CNN will protect Joe Biden like a praetorian guard. But all of that could be changing.
And No one expected CNN to backstab this Biden official live on air.
Even Joe Biden can't ignore the flood of illegal immigrants coming across the southern border anymore.
With a new Republican majority in the House of Representatives, Biden knows he will have to face a series of politically embarrassing investigations and hearings about his open borders policies that let more than 5 million illegal immigrants try to enter the country.
Biden gave a bunch of fake executive orders to make it look like he was doing something at the border.
Homeland Security Secretary Alejandro Mayorkas also talked to the media to spread the false story that the Biden administration cared about securing the border.
"We have accomplished more than 7,000 arrests. We have dedicated really untold resources, personnel, technology, investigative capabilities to break up these smuggling organizations, to disrupt them. You and I have both seen too much tragedy on the border. It's precisely why we're trying to build safe and lawful pathways," Mayorkas said on CNN.
Anchor Poppy Harlow pushed back and asked Mayorkas if the fact that more than 2.4 million illegal immigrants would cross the border in 2022 was a "crisis."
Mayorkas lied and said that the invasion at the border was "managed in an orderly way." He then asked Congress to give illegal immigrants amnesty.
"We have seen the situation at the border managed in an orderly way, we have seen it in extraordinarily challenging circumstances as well," Mayorkas added. "You can rest assured, Poppy, that we're doing everything that we possibly can to build a system that provides humanitarian relief in a safe and orderly way while trying to persuade Congress to fix what is a broken system."
Harlow tried to find out if what was going on at the border was a crisis.
"I understand that, but just what you're seeing the 20 times you've been there, the record number of migrants at the southern border, and last year it was nearly 2.4 million," Harlow continued. "If that's not a crisis, Secretary, what is?"
Mayorkas again wouldn't admit that the situation at the border was real. Instead, he tried to explain it in terms of how people move around the world.
"You know, Poppy, we have seen 2.4 million encounters at our southern border and it is reflective of the greatest level of displacement of people in the world since World War II," Mayorkas declared. "It is reflective of a migration challenge that is gripping the entire hemisphere."
Mayorkas said that Biden knew what was going on at the border, even though he had never been there.
Mayorkas also tried to convince Americans that the government cared about the border by saying that he had been there a dozen times.
"The President knows the border very well," Mayorkas stated. "He has had his Secretary of Homeland Security visit multiple times since the very initiation of the administration."
This is a strong criticism of Biden's policies of open borders.
Mayorkas has been to the border many times, but illegal immigrants are still getting into the country. This shows that he is ineffective and incompetent.
The administration also doesn't want to admit that there is a crisis at the border because that would make the new GOP House majority more likely to try to impeach Biden and Mayorkas.
But even CNN has to admit that what is happening at the border is clear to all Americans.
Previous Joe Biden's FBI and DOJ have been smacked with NEW investigations
Next Joe Biden will be terrified by Donald Trump's inquiry regarding confidential papers
Tucker Carlson just exposed Biden's connections to Antifa
A former Clinton advisor just ended Joe Biden's career in one breath


text:
IATC young critics' seminar in Bucharest, October 30 to November 4, 2013 – Report
Home IATC young critics' seminar in Bucharest, October 30 to November 4, 2013 – Report
It was with great pleasure that IATC held a very successful international young critics' seminar in Bucharest, as part of the programme of the National Theatre Festival of Romania, between October 30 and November 4, 2013. Young critics from countries as diverse as South Korea, New Zealand, Canada and, of course, Romania itself worked in two language groups; one in French (monitored by experienced critic Jean-Pierre Han from France), the other in English (monitored by established critic Mark Brown from Scotland/UK). As often in the recent past, the overwhelming majority of applicants to the seminar (and therefore of participants) were women, suggesting that the future of theatre criticism internationally will be increasingly female
The work seen by the seminar participants included an acclaimed staging of Molière's The School for Wives, presented by the Liviu Rebreanu Company of Târgu-Mures, and a production of Chekhov's The Seagull, by the German State Theatre of Timisoara, in which the audience joined the actors on the stage of the National Opera Theatre.
IATC thanks the festival for the superb organisation and hospitality it offered to the seminar.
Mark Brown and the English-speaking group in Bucharest.


Complete characterization of the biological properties of hepatitis B virus (HBV) variants requires the generation of full-length genomes. The aim of this study was to develop new tools for the efficient full-length genome amplification of virus from samples with low viral loads. Rolling circle amplification (RCA) was used to amplify full-length HBV genomes from both sera and liver biopsy samples from chronic HBV carriers. Serum-derived relaxed circular HBV DNA could be amplified only after completion and ligation of plus-strand DNA. Covalently closed circular DNA (cccDNA) from liver biopsies could be amplified directly from as few as 13 copies, using RCA, followed by a full-length HBV PCR. Three serial liver biopsy samples were obtained from a lamivudine-resistant patient who cleared detectable serum HBV after adefovir dipivoxil was added to the lamivudine therapy and then seroconverted to anti-HBs. Only the genomes from the last biopsy specimen obtained after the emergence of lamivudine resistance contained the lamivudine resistance-associated mutations rtL180M and rtM204V ("rt" indicates reverse transcriptase domain). Defective genomes were also found in this biopsy sample. Genomes cloned from the liver biopsy specimens were transfected into HuH7 cells to study their replication competence and their susceptibility to lamivudine. RCA is a powerful tool for amplifying full-length HBV genomes and will be especially useful for the study of occult or inactive HBV infections and patients undergoing antiviral treatment. It can also be used to probe HBV cccDNA, the crucial intermediate in viral persistence and the archive of resistance mutations.
Chronic hepatitis B virus (HBV) infection is widespread, with an estimated 370 million carriers worldwide (1). Despite constraints imposed by a complex genetic architecture involving overlapping genes, the virus is highly variable, with eight known genotypes designated A to H (20).
The complete molecular and biological characterization of HBV variants requires the isolation of full-length genomes. This has been done classically by amplifying overlapping fragments from serum-derived relaxed circular genomes (RC-DNA) (4, 21). However, the discontinuous structure of RC-DNA means that the amplification of at least one of the fragments can be problematical. Increasing the number of PCRs ineluctably augments the probability of incorporating PCR errors, and further steps are necessary to construct a full-length HBV genome. Günther et al. have developed an elegant method for amplifying full-length HBV genomes in a one-step PCR (7). However, this method requires serum with viral loads of >104 copies/ml of HBV DNA. In typical chronic HBV infections, viral loads usually range from 106 to 1010 copies/ml. However, for inactive carriers (individuals who are hepatitis B surface antigen positive [HBsAg+]) with minimal virus replication and for occult HBV-infected patients (HBsAg−), serum HBV DNA levels are usually very much lower. Consequently, few occult HBV infections have been studied at the genomic level and then sometimes only after the occult infection has been reactivated. We therefore wanted to develop a method for the efficient and high-fidelity amplification of full-length HBV genomes from biological samples with low levels of HBV DNA. The method chosen was rolling circle amplification (RCA), also called ramification amplification (26). This method relies on the properties of the phi29 phage DNA polymerase that possesses a strong 3′ exonuclease (proofreading) activity, that is very processive, i.e., is able to polymerize more than 70,000 nucleotides without detaching from the template, and that can displace previously elongated strands (2). On a circular template there is linear amplification (70,000 nucleotides represents more than 20 HBV genomes), and more importantly, there is branching amplification, as displaced strands reveal more binding sites for the amplification primers. The end product is a high-molecular-weight double-stranded DNA containing multiple copies of the initial target. RCA exhibits less amplification bias and greater yield, product length, and fidelity than PCR (5, 13). We have adapted this method to amplify full-length HBV genomes from the covalently closed circular DNA (cccDNA) form of the genome found in the nuclei of infected hepatocytes and from RC-DNA found in circulating virions. This method has been used here to study, as an extreme example, the residual cccDNA genomes from a patient who has seroconverted to anti-HBs following successful add-on antiviral therapy.
HBV cccDNA extraction from liver biopsy specimens.Liver biopsy specimens were collected from a chronic hepatitis B carrier and an HBV-negative patient and stored at −80°C. The chronic carrier has been followed for several years in our laboratory (17). cccDNA was extracted as described previously (24), and the copy number was quantified using quantitative PCR (23). Ten-fold serial dilutions of the cccDNA stock were carried out using material extracted from the HBV-negative liver biopsy the same way.
Isolation of DNA from serum.Nucleic acids were extracted from 150 μl of serum from a chronic HBV carrier, using a QIAamp UltraSens virus kit (Qiagen, Courtaboeuf, France). This patient has also been previously studied in our laboratory (16), and the complete sequence (of genotype C) is known.
Completion and ligation of the HBV RC-DNA plus strand.Plus-strand DNA was completed using the endogenous polymerase activity of HBV nucleocapsids before extraction. Seventy-five microliters of 3× reaction buffer was made fresh before use (21.6 μl of Tris-HCl [pH 7.6], 0.8 M; 10.8 μl of MgCl2, 0.8 M;10.8 μl of NH4Cl, 1.2 M; 10.8 μl of 2-mercaptoethanol, 3%; 10.05 μl of NP-40, 20%; 2,2 μl of each deoxynucleoside triphosphate [dNTP], 10 mM; quantity sufficient for 75 μl with sterile water), and then 150 μl of serum was added. The reaction was carried out for 12 h at 37°C in a humid chamber. The sample was then treated with 10 U of RNase-free DNase I for 10 min at 37°C, followed by extraction as described above for serum samples. For ligation, 8.5 μl of the extraction product was added to 1× ligation buffer and 1.5 U of T4 DNA ligase (Promega, Charbonnières, France) in a final volume of 10 μl. The ligation reaction was carried out for 16 h at 16°C, followed by heating at 65°C for 10 min.
PCR amplification of whole HBV genomes.Full-length HBV genomes were amplified using a one-step PCR method (7). Briefly, the method was originally based on the unusual structure of HBV RC-DNA, which has a short and A/T-rich terminal redundancy on minus-strand DNA but can also be used to amplify unit-length HBV genomes from RCA products. The sense and antisense primers are situated at the terminal redundancy and partially overlap (Table 1). In both primers, the recognition site for LguI (Sap I) is immediately adjacent to the overlapping sequences. LguI cuts outside its recognition site, and cleavage of PCR products occurs within the overlap sequences, liberating a full-length HBV genome that can circularize without the gain or loss of nucleotides.
Rolling circle amplification.Amplification primers for both plus and minus strands were chosen from well-conserved regions of the HBV genome (Table 1). To protect primers from the strong proofreading activity of the Phi29 polymerase, the two 3′ bases were phosphorothioate modified. Appropriate amounts of DNA (shown in legends to Fig. 1 to 4) were mixed with the eight primers at a concentration of 0.5 μM each and a 1× Phi29 buffer in a final volume of 10 μl. The DNA mixture was denatured at 95°C for 3 min and then cooled to room temperature in stages: 50°C for 15 s, 30°C for 15 s, and 20°C for 10 min before it was placed on ice. Sample mixtures were combined with 10 μl of reaction mixture containing: 1× Phi29 buffer, the eight primers at a concentration of 0.5 μM each, 0.4 mg/ml of bovine serum albumin, 4 mM of dithiothreitol, 2 mM of each dNTP, and 10 U of the Phi29 DNA polymerase (New England Biolabs, Worcester, MA). Reactions were carried out at 30°C for 18 h and terminated at 65°C for 10 min.
RCA using different HBV DNA templates. Southern blot hybridization of RCA products. H2O, negative control; RC, RC-DNA; CP, RC-DNA with plus-strand completed using the endogenous polymerase activity of HBV virions. CPL, CP ligated after DNA extraction. CCC, cccDNA.
RCA product analysis.First, 2 μl of RCA products was run on an 0.8% agarose gel and stained with ethidium bromide. Second, 4 μl of RCA product was digested in a final volume of 10 μl with 3 U of SpeI (New England Biolabs) that cuts only once in most HBV genomes. Digestion products were analyzed on a 1% agarose gel. Full-length genomes amplified by RCA and then genomic PCR were cloned into pCR2.1-TOPO (Invitrogen, Cergy Pontoise, France) and fully sequenced.
Transfection of HuH7 cells.HuH7 cells were grown in Dulbecco's modified Eagle's medium-F12 supplemented with 10% fetal calf serum and antibiotics. Cells were seeded at a density of 5 × 105 cells per well in six-well plates 24 h before transfection. Full-length HBV genomes were excised from cloning vectors using SpeI or LguI (an isoschizomer of Sap I; Fermentas, Vilnius, Lithuania), depending on how they had been cloned. Total DNAs (HBV genome plus vector) were treated with phenol-chloroform, ethanol precipitated, and resuspended in sterile water at an approximate concentration of 1 μg/μl. Exact concentrations were measured by A260. The DNAs were transfected (2 μg per well for single transfections, 1 μg per well of each DNA for cotransfections) using 2 μl per μg of DNA of TransIT-LT1 (Mirus, Madison, WI) according to the manufacturer's instructions. Medium was replaced on the following day (day 1) with medium containing 0, 10, or 100 μM lamivudine (LAM). Two wells were used for each DNA and LAM concentration. Medium contents were changed each day until day 5, and the spent medium was clarified and stored at 4°C until analysis was carried out. HBsAg levels in medium samples pooled from the different days were quantified using a commercial enzyme-linked immunosorbent analysis, MonoLisa HBsAg Ultra (Bio-Rad, Marnes-la-Coquette, France), and a standard curve generated by using purified HBsAg. To be within the linear range of the assay (A450, <1), samples were diluted with normal human serum. On day 5, core-associated intracellular HBV DNA was isolated as described previously (11), electrophoresed, blotted onto a nylon membrane, and hybridized using a 32P-labeled full-length HBV genomic probe. Hybridization signals were quantified by using a PhosphorImager.
Nucleotide sequence accession numbers.The sequences of the HBV genomes have been deposited in GenBank under accession numbers AM295796 to AM295800 and AM410963.
Adaptation of RCA for the amplification of RC-DNA and cccDNA HBV genomes.After the primer and phi29 polymerase concentrations were optimized, RCA was performed with different HBV DNA templates, as follows: (i) the partially double-stranded RC-DNA (6); (ii) the RC-DNA after the plus-strand DNA was completed using the endogenous HBV polymerase activity; (iii) the treated RC-DNA, ligated after extraction; and (iv) the liver-derived cccDNA (6). Southern blot analysis (Fig. 1) shows that RCA could efficiently amplify only completely circular DNA templates such as HBV cccDNA and the completed/ligated serum HBV DNA. Amplification of serum HBV RC-DNA or HBV RC-DNA that had only been completed but not ligated did not amplify. Also, ligation alone did not permit amplification (not shown). Completion of plus-strand DNA is therefore essential, and it is probably this strand that is ligated. Consequently, in the absence of pretreatment, RCA can discriminate highly between HBV cccDNA and RC-DNA. The pre-S/S region of the genome amplified from serum-derived DNA was directly sequenced, and the sequence was identical to that of the genome originally amplified by conventional techniques (16).
Serial 10-fold dilutions of HBV cccDNA, extracted from the liver tissue of a chronic hepatitis B carrier and quantified by real-time PCR, were used to determine the assay sensitivity. Electrophoretic analysis and ethidium bromide staining show that DNA amplification occurred for all samples tested, including the control reaction mixture without the template added and the sample from an HBV-negative patient (Fig. 2A). Consequently, even though HBV-specific primers were used, nonspecific amplification can occur. This can be explained by self-priming and extension of the primers and by nonspecific amplification that occurs because RCA is carried out at 30°C. However, after Southern blotting and hybridization of the same gel, using a specific HBV oligonucleotide probe, a hybridization signal was observed only with HBV DNA samples and down to the level of 10 copies/reaction (Fig. 2B). Two methods were used to recover unit-length HBV genomes. First, RCA products were digested with SpeI, which cuts only once in the HBV genome. A clear and specific band of linear full-length HBV DNA (verified by hybridization [not shown]) can be seen (Fig. 2C), and exploitable amounts are obtained with the reaction initiated with 102 copies of HBV cccDNA. Second, an HBV genomic PCR (7) was performed with the RCA products. A clear ethidium bromide-stained band of the expected size can be obtained from the RCA reaction product originally amplified from 10 copies (Fig. 2D). Thus, the sensitivity of this combination is 10-fold higher than that of the restriction method and is comparable to the sensitivity of real-time PCR. Finally, we have tested the HBV genomic PCR directly with the same serial dilutions of HBV cccDNA without prior amplification by RCA. Analysis shows only a weak band with the sample initially containing 103 copies/reaction (Fig. 2E). Consequently, the association between RCA and specific full-length HBV PCR enables a significant increase in sensitivity.
Amplification of HBV genomes from cccDNA. The number of copies of cccDNA initially present in each RCA reaction is shown at the top. M, molecular weight marker (the sizes of selected bands are shown on the left). H2O, negative control. 0, negative control (the normal human serum used for the cccDNA dilutions). (A) Ethidium bromide-stained agarose gel of the raw RCA products. (B) Southern blot hybridization of the same gel. The position of the wells that retain some of the RCA products and the position of the high-molecular-weight products (>10 kb) that migrate in the gel are shown. (C) Ethidium bromide-stained agarose gel of the RCA products digested with SpeI. (D) Ethidium bromide-stained agarose gel of the products of a PCR performed with the RCA products. (E) Ethidium bromide-stained agarose gel of the products of a PCR performed directly with cccDNA. In panels C, D, and E, arrows on the right point out full-length (3.2-kb) HBV genomes.
Amplification and molecular characterization of HBV genomes from serial liver biopsy specimens.Three sequential liver biopsy specimens from a patient who had undergone several antiviral treatments were available (17). The first biopsy was performed before the patient entered a dose-finding entecavir trial. The patient was receiving the lowest dose (0.1 mg/kg of body weight/day), and after 6 months of treatment, HBV serum DNA levels had dropped by less than 1 log. A second biopsy was then performed, and the patient was switched to lamivudine but developed resistance to the drug. Line probe assays indicated that the HBV genome was genotype A and contained the mutations rtL180M/rtM204V ("rt" indicates reverse transcriptase domain), associated with lamivudine resistance. Adefovir dipivoxil was added to the regimen, and serum HBV DNA levels gradually declined to an undetectable level, accompanied by the loss of HB e Ag (HBeAg) and HBsAg, followed by seroconversion to anti-HBs 8 months later. The patient did not seroconvert to anti-HBe. The third biopsy was performed at 5 months after seroconversion, and over a year later, the serum HBV DNA level had become undetectable. The levels of cccDNA in the biopsy specimens, estimated by real-time PCR, are shown in Table 2. HBV genomes were amplified from all three biopsy specimens, using RCA, followed by the HBV genomic PCR (Fig. 3) and cloned. A genome (liver biopsy 1, clone 5 [LB1Cl5]) amplified from the first biopsy was fully sequenced. As expected, it was of genotype A and, consistent with the fact that the patient was HBeAg+, contained neither a precore mutation (22) nor core promoter mutations (12). Serum drawn at the same time the biopsy was performed was no longer available, and RCA could not be used to amplify circulating genomes. However, we did have DNA extracted from this serum, and full-length genomes were amplified using the genomic PCR. The PCR product was directly sequenced and was found to be identical to that of the clone amplified from the biopsy. One of the clones amplified from the biopsy (LB1Cl10) displayed a restriction pattern that was different from the other clones. It was fully sequenced and, apart from two single nucleotide substitutions, the only major difference from the first clone was a 129-bp deletion in the PreS1 region (Table 3). It eliminates residues 57 to 99 of the PreS1 protein and most of the S promoter. This genome is therefore probably not viable by itself but can potentially be trans complemented by a wild-type genome. One clone (LB2Cl6) amplified from the second biopsy was fully sequenced and was completely identical to LB1Clone5. Mutations that could explain the nonresponse to entecavir were therefore not seen with this clone. Two genomes isolated from the third biopsy (LB3Cl21 and LB3Cl24) were fully sequenced and were identical to that of LB1Clone5, with two important exceptions. These exceptions were that both clones contained the substitutions T667A and A739G that introduce the lamivudine resistance-associated mutations, rtL180M/rtM204V, into the viral polymerase. This shows that the genomes were amplified from the third biopsy, the only one performed after the emergence of lamivudine-resistant mutants. Therefore, more than a year after the disappearance of the serum HBV DNA, viable cccDNA genomes still persisted in the infected liver. However, two types of aberrant genomes were also identified. Some clones showed a restriction enzyme profile consistent with a large deletion. Two clones were fully sequenced, and both possessed an identical 388-bp deletion (positions 2025 to 2412), covering most of the core protein gene. The deletion creates a core-pol fusion that will produce a protein containing the spacer, reverse transcriptase, and RNase H domains of the viral polymerase. Both clones possess the two lamivudine resistance substitutions, and apart from these and the deletion, the sequence of one clone (LB3Cl1) is 100% identical to that of LB1Clone5, and the other clone (LB3Cl10) has a single nucleotide substitution. The other type of aberrant genome was found in a clone that contains not only the two lamivudine resistance substitutions plus one other single-nucleotide substitution but also a single-nucleotide insertion (LB3Cl11). The insertion, a G between positions 285 and 286, affects both the polymerase (P) and the surface antigen (S) genes. The insertion frame shifts codon 401 of the P gene into an unused open reading frame, where an additional 65 amino acids can be encoded. The 5′ end of the S gene is fused to the 3′ end of the P gene, encoding a chimera containing 43 residues of HBsAg fused to the last 445 residues of the polymerase.
RCA plus genomic HBV PCR with cccDNA extracted from serial liver biopsy specimens. Ethidium bromide-stained agarose gel of final products of combined RCA and PCR with material from serial liver biopsies. H2O, negative control; LB, liver biopsy; T+, positive control, using a plasmid containing a full-length HBV genome (of genotype C). Both the negative and the positive controls were also subjected to RCA, followed by the genomic HBV PCR.
Functional characterization of the amplified HBV genomes.To test the replication competence and the LAM sensitivity of the clones, LB1Cl5 and LB3Cl24 were transfected into HuH7 cells, and the cultures were treated with different concentrations of the drug. As a control, a wild-type genome of genotype D, 15803, that is known to be replication competent (11) was also transfected. To see whether the two types of aberrant clones can trans complement each other, LB3Cl1 and LB3Cl11 were transfected singly and cotransfected. With the exception of LB3Cl11, which has a frameshift mutation in the S gene, all the genomes were capable of synthesizing and secreting considerable quantities of HBsAg (Fig. 4A), including LB3Cl1, which has a deletion in the core/pol genes but whose S gene is intact. Intracellular HBsAg levels were also measured (not shown) but represented, at most, 4% of the total HBsAg. HBsAg expression is not affected by LAM treatment, at least on the time scale of this experiment. This is not surprising since LAM affects viral replication and not the synthesis of viral transcripts. In cells transfected with LB1Cl5 and LB3Cl24, in the absence of LAM, the levels of intracellular core-associated HBV DNA were comparable with that of 15803, showing that the genomes are replication competent (Fig. 4B). No core-associated HBV DNA could be detected in cells transfected with the defective clones LB3Cl1 and LB3Cl11, either transfected singly or cotransfected (not shown). In cells transfected with 15803 and LB1Cl5, HBV DNA synthesis is inhibited by LAM treatment in a dose-dependent manner, with 50% inhibition concentrations of 13.9 and 16.3 μM, respectively. At the doses tested, HBV DNA synthesis in cells transfected with LB3Cl24 is completely resistant to LAM treatment.
Transfection of HuH7 cells and lamivudine treatment. (A) A representative transfection experiment measuring the expression and secretion of HBsAg from cells transfected with cloned full-length HBV genomes and treated with lamivudine. (B) Southern blot of intracellular core-associated HBV DNA from cells transfected with cloned full-length HBV genomes and treated with lamivudine. 15803, a wild-type HBV genome of genotype D.
As shown here, RCA reactions can efficiently amplify full-length HBV genomes from circular templates, either cccDNA or RC-DNA that has been completed and ligated. That ligation alone is not sufficient for obtaining amplification suggests that it is completed plus-strand HBV DNA that is initially circularized, despite the fact that plus-strand synthesis is primed by a capped oligoribonucleotide (14). It is probable that this has been eliminated in some RC-DNA molecules. When coupled with a genomic PCR, RCA becomes as sensitive as real-time PCR but with the immense advantage of amplifying full-length genomes rather than small fragments. Using this coupled system, we have successfully amplified virus genomes from several patients whose occult HBV-infected sera could not be amplified using genomic PCR alone. In one case, we could show that the amplified genomes are replication competent (N. Martel and A. Kay, unpublished data). Another major advantage of RCA over PCR is its fidelity (5). Often, genomes amplified using the genomic PCR do not replicate in cell culture even if they are obtained from patients with high levels of HBV replication (8). The sequences of these genomes are often "normal," indicating that apparently anodyne mutations can have important effects on virus replication. If genomes can be amplified using only RCA, then this problem is minimized. If it is necessary to add on a genomic PCR, then this advantage is lost, but it should be possible to further optimize the RCA step and reduce the number of cycles in the genomic PCR. The reliance of RCA on a circular template for efficient amplification is also an advantage. In infected cells, only cccDNA will be efficiently amplified and not integrated HBV sequences. We have used RCA to specifically amplify cccDNA from cells transducted with an HBV/baculovirus vector (15). This ability to directly probe the cccDNA pool will be important for the study of HBV variants, especially the emergence of drug-resistant mutants. The variability of HBV is partly due to the replication strategy of the virus (6). Mutations originally arise in RC-DNA during error-prone reverse transcription and can be stably propagated only by their recruitment into the cccDNA pool. This can happen in two ways: the nucleocapsid containing the mutated RC-DNA can be recycled to the nucleus to maintain the cccDNA pool, or it can be secreted to infect a new hepatocyte, where RC-DNA is converted into cccDNA. In both cases, the mutated RC-DNA genome will be in competition with nonmutated genomes or other genomes newly mutated during the reverse transcription step. The emergence of HBV variants, including drug-resistant mutants, is therefore a long process that can take months or even years. The capacity to directly probe the cccDNA pool can therefore provide significant insights into this problem. In the case of drug-resistant genomes, these have to integrate the cccDNA pool before they can have an impact, and monitoring of this pool can predict future problems. Studies using RCA can investigate the kinetics of the integration of drug-resistant genomes into the cccDNA pool. This implies serial and closely spaced liver biopsies that cannot be done with human subjects but can be done using animal models, duck or woodchuck.
The cccDNA pool is indeed central to the problem of viral persistence. The half-lives of infected hepatocytes and cccDNA are long, and antiviral drug treatments can substantially reduce RC-DNA synthesis but have little effect on intracellular cccDNA levels (19, 23). Prolonged treatments are therefore necessary, and this increases the potential for the emergence of drug-resistant mutants. Even after apparent virus clearance, cccDNA can persist. This has been documented in the woodchuck model (18) and probably explains HBV reactivation (3, 9, 10). In this study, we have shown that a patient who has seroconverted to anti-HBs and has an apparently resolved HBV infection still retains cccDNA in the liver. The results also illustrate the crucial role of the cccDNA pool in the balance between maintaining genetic stability and reacting to selective pressure. Apart from the lamivudine resistance substitutions found in specimens from the third biopsy, the nondefective genomes isolated from the three biopsy specimens over a period of more than 4 years show, at most, a one-nucleotide difference. Conversely, all of the clones isolated from the third biopsy specimen contain the two lamivudine resistance mutations, showing that in the face of strong selective pressure, the plasticity of the HBV genome permits the acquisition and propagation of mutations. This shows that resistance mutations can be archived in cccDNA and persist even after seroconversion to anti-HBs. The host immune response is crucial for controlling such a latent infection, and if the patient subsequently becomes immunocompromised, then there is a risk of reactivation. Transfections of genomes amplified by RCA alone (LB1Cl5) or RCA followed by genomic PCR (LB3Cl24) are fully functional, showing that errors were probably not introduced during the amplification process. More importantly, this functional test also permits the confirmation of the phenotypes suggested by the sequences, LAM sensitivity for LB1Cl5 isolated from a biopsy obtained prior to LAM treatment of the patient and LAM resistance for LB3Cl24 isolated from a biopsy obtained not only after LAM-resistant variants had emerged in the patient but also after the patient had seroconverted to anti-HBs. In addition to functional genomes, the accumulation of defective genomes does seem to have occurred after seroconversion and instauration of a "silent" occult HBV infection. They must have been generated during or after the emergence of lamivudine resistance, since they contain the associated mutations. Such defective cccDNA molecules can persist because no viral promoters are affected. However, the retrotranscription of defective pregenomic RNAs into RC-DNA that can then be converted back into cccDNA to maintain persistence of the defective genomes requires trans complementation, probably by nondefective genomes. It was possible that the two types of defective genomes could trans complement each other (25). However, cotransfection of the two types of defective genomes did not produce detectable levels of replicating HBV DNA. These genomes are therefore probably "dead end" products that will eventually be diluted out by hepatocyte turnover, unless reciprocal recombination between the two types of defective molecules generates a viable genome. It remains to be seen with other patients who have cleared HBV infection whether the persistence of cccDNA together with the accumulation of defective genomes is a general phenomenon.
In summary, RCA will be extremely useful for generating full-length HBV genomes, even from patients with low levels of virus replication, and will permit the direct probing of cccDNA, the central actor in viral persistence and the propagation of mutations. It will also be useful for the phenotypic testing of HBV variants, especially resistance mutations selected by antiviral therapy.
S.M. received a French government stipend.
Returned for modification 21 November 2007.
↵▿ Published ahead of print on 7 July 2008.
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Investigation as Earth tremor and rumble shake part of Cornwall
A local geothermal energy power plant stopped operations temporarily as a result
Charlotte Becquart
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People in the Redruth area were left in shock this morning after they felt the earth shake and heard a low rumble. The small-scale earthquake was felt in several villages including Frogpool, Lanner, Carharrack, Penhalvean and St Day.
Several residents took to the Lanner Life public Facebook group to report the "earth tremor" and several said it "scared the life out of them".
The small earthquake has not been recorded yet by the British Geological Survey, however a local geothermal energy power plant confirmed it was linked to its operations.
Following the incident, an investigation has been launched at United Downs Geothermal, located near Redruth, where drills are sent deep into the ground to extract heat for use in homes and businesses.
A spokesperson wrote online: "The testing at the United Downs site this morning has been stopped. There has been some micro-seismicity, which is being investigated."
A similar incident happened on September 30 of this year as locals said that their windows rattled and houses shook. Read that full story here.
Geothermal at United Downs confirmed a small earthquake was linked to testing at the site today. Operations temporarily ceased while teams attempted to understand the cause.


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© Christian Pondella/Red Bull Content Pool
These 9 female climbers will inspire you to start scaling peaks today
From Lynn Hill's iconic ascent of The Nose on El Capitan to Angela Eiter's route up La Planta de Shiva 25 years later, these women and their legendary climbs are among the best ever.
By Will Gray
Published on 03/08/2021 · 2:33 PM UTC
Part of this story
Sasha DiGiulian
American climber Sasha DiGiulian has made a career of overcoming the odds, with more than 30 first female ascents to her name.
You won't find anyone more determined than serial winner Angela Eiter who became the first woman to conquer one of the world's toughest climbing routes.


Haus of Girls: Beach Vacay- Part 3.
As I sit and type this, the cool fall air is bellowing in through my bedroom windows, however, I can't help but long for the salty hot air that engulfed our family for an entire week back in July.
The sand, waves, and memories of our first family beach vacation will forever bring tears in my eye.
I hope that a beach vacay will become an annual tradition for Steve and I, along with our three blondies.
The girls hopped up in this lifeguard tower for a photo.
I never noticed Sophie's cute binocular pose until weeks after we got home. Seriously adorable.
Their first time seeing a Sea Turtle nest.
One of my favorite photos from our trip.
Our last night in Alabama called for a little impromptu photos session on the beach with mom.
Even Gracie's beloved "Ducky" was able to join us on the beach.
The girls of Haus of Girls.
One of the few family shots- thanks to a kind beach-walking stranger.
Waving goodbye to the Gulf Coast. See you next summer!
Great pictures! It looks like you had so much fun and made some fantastic memories!


Rollin' News - World Wide Roller Derby News | RDNation releases Windows Phone App Update!
RDNation is proud to announce its second release of its Windows Phone app. The app dives more into the league management of RDNation and it sets it self up to be more usable to the larger crowd of users using RDNation for league management.
With over 370 leagues currently using RDNation, its about time to release some mobile apps to help them manage derby better. So with that, the first app released is for the Windows Phone.
Viewing all the leagues signed up.
Viewing all the skaters signed up.
Viewing all the merchandise for sale on RDNation.
With internal features, we won't forget about the internal management features.
Forum functionality - Create, view and edit posts.
Members functionality - View members, set permissions, skater dates, insurance numbers.
Polls and Voting - Create, manage, vote and view results on the polls created by the league.
Dues and Payments - Pay dues, modify dues and create new dues items.
Calendar Functionality - Create events, modify, take attendance. Its all there inside the calendar.
Messaging - View messages, receive messages and create new messages.
Edit Profile - Edit your personal RDNation profile.
There were features in RDNation we didn't have time for, but we included links to those pieces of functionality.
Bout Challenges - Used for finding bouts to play at.
Official Requests - Used for finding officials to ref your games.
We hope you like the first release of the Windows Phone app which can be found by just typing RDNation on the windows market place. Android and iPhone are next up!


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Disability Discrimination Act 1992
The Disability Discrimination Act 1992 (DDA) is an Australian Federal Government Act.
The objects of the Act are:
(a) to eliminate, as far as possible, discrimination against persons on the ground of disability in the areas of:
(i) work, accommodation, education, access to premises, clubs and sport; and
(ii) the provision of goods, facilities, services and land; and
(iii) existing laws; and
(iv) the administration of Commonwealth laws and programs; and
(b) to ensure, as far as practicable, that persons with disabilities have the same rights to equality before the law as the rest of the community; and
(c) to promote recognition and acceptance within the community of the principle that persons with disabilities have the same fundamental rights as the rest of the community.
Australian Legislation
ComLaw Page
Disability Discrimination Act 1992 (PDF)
actsdfed