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Willireo
Everything posted by Willireo
Daily Dose #32: Over Now (Opening Scene from Criss Cross)
Willireo replied to Dr. Rich Edwards's topic in Summer of Darkness: Investigating Film Noir
- Discuss how the opening of this film exemplifies the noir style and substance. Driving, urgent music accompanies the appearance of the Universal International logo and continues as we see a documentary style aerial shot of Los Angeles City hall. It's the city, the big city (not the anonymous small midwestern city) it's night for night and the city hall dominates a maze of streetlights. Cut to a parking lot and passing cars weave in front of us revealing two people kissing. They talk as passionately as they kiss. The woman is fearful and the may tries to comfort her but he is equal
Daily Dose #31: No Escaping Noir (Scene from Brute Force)
Sam Levene is one of the great Noir stalwarts. He was in many films of the classic period (The Killers and Crossfire) and always turned in a great performance. As in "Desperate" the scene begins with the victim cowed in the foreground. The Captain's interrogation is cold and becomes increasingly violent. When we cut to the guards playing cards at the table we see their discomfort, even revulsion at what they can hear going on, but they don't do anything to interfere. They need those jobs. The overtones of Nazism had to be significantly felt by filmgoers in 1947. We have a prison, we have
Daily Dose of Darkness #28: Pure Score (Opening Scene of Elevators to the Gallows)
Here are a few discussion starters (though feel free to come up with your own): -- In what ways does Miles Davis' score (improvised while watching scenes from the movie) work with and contribute additional layers of meaning to Louis Malle's visual design? This clip was a pleasure to hear and watch. These questions are hard to answer because they're filtered through 5 decades of different understanding. Eddie Muller has pointed out elsewhere that strings are more prominent in classic Film Noir scores and we've come to associate smoky be-bop brass with Noir as a later accretion. Still th
Daily Dose of Darkness #30: Into the Darkness (Scene from Desperate)
I've never seen this movie. This clip has me eagerly anticipating Friday night. Describe how this scene uses cinematography to accentuate the brutal beating of Steve Randall (Steve Brodie). -- How do Mann and Diskant utilize different points of view to heighten the tension in this scene? As the scene opens Steve is foreground left and Walt is in hidden in darkness in the center. When one of the stooges tells Walt they got Steve for him Walt strides into the center of the frame growing in size and threat. He dominates the center of the frame, his bulk squeezing his henchmen on the left
Daily Dose of Darkness #29: Noir City (Opening Scene of The Asphalt Jungle)
Thanks for identifying Mr. Drucker.
-- Discuss how this film depicts and utilizes this "unnamed city." Additionally, why do you think the film is entitled "The Asphalt Jungle?" It's amazing to see a city so deserted. I have the feeling this is very early morning although I don't recall the time being established. The deserted city starts as the stage for two players. A patrol car and an unknown man in a suit. The patrol car passes the unknown man who, almost instinctively hides behind a column as the patrol car passes. We see the many continuing to walk down a rubble strewn alley. The patrol car gets it's closeup as
Daily Dose of Darkness #25: Time for a Heist (Opening Scene of Kansas City Confidential)
-- Discuss the role of time and timing in this scene. Time and timing are in counterpoint in this scene. The pace of the opening is leisurely. No quick cuts and no rapid movement on the part of people on the street, John Payne exiting his van, the armored car guards or Foster. Despite this there is an extreme tension built by Foster's concentration, the dramatic quiet music and the pressure of time. We see the bank clock, we see Foster start his stop watch, we see the bank clock we see the stop watch end at one minute we see Foster slowly cross the room and add what appears to be th
Daily Dose of Darkness #24: The Square Circle (Opening Scene from 99 River Street)
We open up as spectators at the fight. The world surrounding the boxers is complete darkness. We're below the action, it would be ringside seats but we're even closer to the action than that. Medium closeups as the fighters exchange blows. Ernie's eye gets cut? A dirty trick by sailor? That's never revealed. Having to protect the eye throws Ernie off and he's knocked to the ropes. We get an extreme closeup of our man on the ropes. That's where we find him as we transition from spectators at the fight to Ernies apartment where he's watching a replay of the fight on television.
Daily Dose of Darkness #23: Iverstown (Scene from The Strange Love of Martha Ivers)
It's strange seeing Kirk Douglas in a situation where he's not the alpha male. That said I heard John Wayne criticized him for his portrayal of the weak struggling VanGogh. We open on Sam and Walter getting reacquainted after over a decade has passed. The scene is smoothly lighted without shadowy expressiveness. Walter, on the right dominates the scene. He's larger and taller than Sam although not by much. We learn Sam has spent their time apart as a gambler and, by inference living an adventurous life. Walter has pursued a more conventional path and it's gotten him to be District Attor
Daily Dose of Darkness #22: It's in the Bag (Opening Scene of Too Late for Tears)
We open on a dark car driving on a winding road. It's shot day for night so we see more of the environment and that we're within sight of city lights. The dark car pulls off the road next to an up-thrusting obelisk telling us we're 3.5 miles from somewhere. Cut to a close up of the car. There's a man at the wheel and his face is obscured by a window mounted searchlight is in the foreground. Is this guy a cop? He checks his Gruen. It's 8:30. Cut to a light colored top-down convertible coming down the road. A medium closeup shows a man driving and a woman in the passenger seat. A
Daily Dose of Darkness #21: Criss Cross (Opening Scene from Strangers on a Train)
i should proof read these things. I meant "cut back and forth between spectators and brogues"
We open on a medium shot of the train station arched entry. I only knew it was a train station because of the film title. I assume this would have been more recognizable to filmgoers of the time. The arch is brightly illuminated while the interior is shadowy. Our POV is the shadowy interior so we're already in the darker world. A cab pulls into the station a porter enters frame from the left and opens the Diamond cab company door. Out comes a leather suitcase followed by a pair of lace up patent leather cap toed spectators that starting walking left. Quick cut to a cab at the mo
Daily Dose of Darkness #20: The Man in Charge (Opening Scene from D.O.A.)
Thanks for raising this point. I think there was an essay by Roger Ebert where he says that action screen left or moving screen left generally indicates something bad going on.
We start at the top of the Los Angeles City Hall. A building that's probably been in more films noir than Elisha Cook Jr. It's night so we know we're not there for a zoning appeal. Panning down we find ourselves sharing the point of view of a lone man, left of screen, momentarily stopped & staring at the building. We here purposeful dramatic music as the man moves to center screen and walks forward in a one point perspective shot of a shadowy hallway. He is the only person in the hallway. We see a sign for Police Department. Entering these doors the man stops and asks direct
Daily Dose of Darkness #19: Behind Bars (Opening Scene of Caged)
It's always great to see the Warner Brothers logo. It's the Superman of studios. We're disoriented during the initial run of titles and theme music although we can see from the crispness of the title "Caged" we're watching an A film. As we progress it becomes clear we're in a dark place, moving, looking through chicken wire, going somewhere by force. Cut to the wagon doors opening and a medium shot of an attractive frightened woman seated at the front of the van. "Pile out your tramps. End of the line" A powerful opening and the cruel description contrasts sharply with the well gr
Daily Dose of Darkness #18: Keep Driving (Scene from The Hitch-Hiker)
We open on a pair of legs in a wedge of light. A dry wind blows dust across dirty shoes and a sinister brass sounds three notes. Headlights move toward us out of the darkness and a black arm throws a thumb in the air. The arm and thumb cover nearly the entire frame commanding rather than requesting the car to stop. The stranger opens the back door. Light glints across his leather jacket. The sinister implications mount. The jacket looks sinister and could remind audiences of this time of nazi uniforms. The stranger sits in darkness again a wedge of light barely illuminates his
Daily Dose of Darkness #17: Deadly Kiss Me (Opening Scene of Kiss Me Deadly)
Kiss me deadly opens with disorientation. Christina is running on a divided highway. We don't know why but whatever it is must be bad. She is shoeless so, we infer, naked under the trench coat. She looks around not knowing what to do. In desperation she stands in front of an on-coming car. If she can't escape she's willing to die. Mike Hammer drives a sporty two door and listens to hip music. He has a meanness unlike other male protagonists in films noir we've previously seen. His first concern is his car. He looks angry and is threatening to Christina when he finally speaks to he
Daily Dose of Darkness #15: MGM Noir (Scene From The Postman Always Rings Twice)
-- How are the "entrances" of John Garfield and Lana Turner staged in this sequence? What do their "entrances" reveal about their character? Garfield's entrance begins with a voiceover and the establishing shot of a rural diner/gas station. His tone is matter of upbeat, matter of fact and carefree. He is a man lacking in responsibility or ambtion who has plenty of time to worry about his future. He's found a place where there's a "man wanted" and may his future starts there. I'd like to thank the fellow students who commented about the meaning of the D.A. saying "Maybe I'll see you
Daily Dose of Darkness #12: Calling on Four Million Dollars (Opening Scene of The Big Sleep)
I was struck by the description of Marlowe's suit in the novel. Having seen the movie first I had different expectations. Powder blue suit and blue shirt sounds so bright and cheery. Bogart's suit is clearly not powder-blue.
The scene opens with Marlowe's shadow on the door of the Sternwood mansion. He's already a dark presence. Marlowe enters an ornate hallway reeking of wealth. His loose stride and posture is in contrast to the formal hallway. He is polite enough to take off his hat. The flirtatious daughter descends the stairs. Her brightness and energy are also in contrast to the surroundings. She comes on strong to Marlowe but clothes the come-on in childishness. He's not thrown off and not uninterested. He handles the situation adroitly showing he's in command regardless of the situation. His
Daily Dose of Darkness #11: Crossing Over (Opening Scene from Border Incident)
The Naked City is considered a Film Noir, but I never thought it was. It always felt more like a police procedural and the documentary style enhanced that feeling.
Films Noir are very sympathetic with their characters. They may be criminals morally ambiguous, trapped in circumstances beyond their control, plagued by bad decisions or just trying to survive but, we always feel for them. Something inherited from the socially conscious gangster films. The documentary opening of Border Incident tells us to anticipate a story about the problems of real people in the real world. On the surface it's a world of plenty achieved by industriousness. In the middle there's opportunity for large numbers of people who crowd the border waiting to follow the proce
Daily Dose of Darkness #9: Showstopping (Scene from Gilda)
Sweet Smell of Success was the first film I thought of when music and Film Noir was discussed . The music itself was a strong element of the story as was jazz and jazz musicians as part of the underworld. Martin Milner was a straight-laced guy but no one would question a jazz musician being busted for possession. Made him an easy mark.
Gilda glows with energy and unbridled exuberance in this number. I hadn't previously noticed how she removed her glove like an experienced stripper. She's brassy and so is the music. All the men love it, even the bouncer, but Johnny's jealous and enraged. He pulls her away and she rebels at him stopping her from being honest with the world. She's not proud of what she is, but she doesn't want to hide it. If she was a hard-boiled detective she could be proud or at lease blasé about her experience with the low life.
June 8, 1949 FBI Reports Hollywood Figures as Communists
Willireo posted a topic in Summer of Darkness: Investigating Film Noir
It was June 8 1949 when the FBI named a number of Hollywood people as members of the Communist Party. The list included John Garfield and Edward G Robinson. Later Congress cited 10 writers and directors for contempt for refusing to disclose their political affiliations. They came to be known as the Hollywood 10 and were convicted and imprisoned. The atmosphere generated by these events is often cited as a contributing factor in the mood of Film Noir of the period.


Jarrett Jack is not shy in crunch time. Quite the opposite, actually.
The veteran point guard seems to play with more confidence down the stretch of close games, as was the case Wednesday in the Knicks' 122-199 double overtime loss to the Bulls at Madison Square Garden.
At 34, Jack became the oldest Knick to register a triple-double since Mark Jackson in 2002 and the first to achieve the feat since Carmelo Anthony in 2012, finishing with 16 points, 10 assists and 10 rebounds in 40 minutes.
As the moment got bigger and the pressure got greater, Jack became more heavily involved in the action. He buried a 17-footer with 1:21 remaining in the fourth quarter to knot the score at 105-105. In the first overtime, he hit a key floater on the baseline after throwing an inbounds pass off the back of Chicago's Kris Dunn, and assisted on Porzingis' buzzer-beating dunk that kept the game going.
Yet Jack also made some costly mistakes, including a turnover in the waning seconds of the second overtime with the Knicks trailing 118-116. Moments later, he attempted the potential game-tying 3-pointer at the horn but could not convert.
Hornacek praised Jack's overall effort, which was vital for the Knicks to even make it to that point.
"Yeah, he had a couple turnovers, but great all-around game for him," he said.
Jack's night was spoiled by the final outcome, which had the veteran wishing he had done even more out on the floor.
"The triple-double is cool, but we lost," said Jack, who scored 10 of his points in the fourth quarter and the two overtimes. "That's just the way I look at things. I look at things in terms of wins and losses. I got to go out there and do more, whatever it is. Maybe I have to push the ball more. Maybe there was a loose ball that I didn't get, or a rebound that I didn't get.


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Chiranjeevi completes dubbing for Sye Raa Narasimha Reddy in just 10 days!
According to a report Chiranjeevi has taken just 20 hours within a period of 10 days to finish dubbing for his role in Sye Raa Narasimha Reddy.
In Com Staff June 28 2019, 5.17 pm
Megastar Chiranjeevi's upcoming film, Sye Raa Narasimha Reddy, is one of the most talked-about films currently. By now everyone knows that the film has a huge cast which includes Chiranjeevi, Amitabh Bachchan, Kichcha Sudeep, Nayanthara, Vijay Sethupathi, Tamannaah, Jagapathi Babu and Brahmaji among others. Updates about the film have been coming in regularly. Just a few days back, it was reported that the shooting for this film has been completed and Chiranjeevi has already started dubbing for his parts. Now, the latest updates suggest that the actor only took 20 hours to finish off the complete dubbing part. Quite impressive, isn't it?
A report in a leading media suggests that Chiru has taken just 20 hours within a period of 10 days to finish dubbing for his role in Sye Raa Narasimha Reddy. This means that the post-production work is going on at a very fast pace and we might get to see the film right when it is expected. Chiranjeevi is, reportedly, playing the role of freedom fighter Uyyalawada Narasimhareddy in this movie. This film is tentatively releasing on October 2nd. Apparently, the makers have employed several top studios to execute the VFX work. According to reports, once the studios deliver the output by August end, a release date will be announced for the film. It is being reported that the teaser of the film will be out on Independence Day, which is on August 15th.
Bollywood star Amitabh Bachchan plays Gosayi Venkanna, the guru of the freedom fighter, while Vijay Sethupathi is playing Raaja Pandi, a Tamil speaking person on whom Uyyalawada had a lot of faith in. Few stills have also been disclosed from the sets, which took the internet by storm. Now, our wait may finally come to an end, if the teaser is released in August. Stay tuned.
Amitabh BachchanBrahmajiChiranjeeviGosayi VenkannaJagapathi BabuKichcha SudeepnayantharaRaaja PandiSye Raa Narasimha ReddyTamannaahtamilTrending In SouthUyyalawada NarasimhareddyVijay Sethupathi
nextKadaisi Vivasayi: Vijay Sethupathi to play the role of a mentally challenged person in this one!

redpajama_set_name: RedPajamaCommonCrawl


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tofu and tempe : all the same?? » Beeleden,beelden en Ban Ki Moon die "alles schijnt te begrijpen"............
Rocket hits central Israeli town; 2 hurt
Two lightly injured when rocket hits building in Rishon Lezion; another projectile seriously wounds one person in Ashkelon. IAF warns northern Gaza residents to evacuate
communities persisted on Tuesday, the IDF continued to gear towards a ground invasion of the Gaza Strip.
Two people were lightly injured when a rocket hit a six-storey building in Rishon Lezion. Four others suffered panic attacks. An air raid siren sounded in the central Israeli city around 6 pm.
German FM en route to Cairo; Morsi: Conflict to end Tuesday
Saturday: Iron Dome missile nearly hits Ashdod homes
Direct hits on Ashdod building, Eshkol house
"We heard a blast," Naor Eini, a resident of the nearby Holon, told Ynet. "We look out of the window and saw smoke. We haven't heard the siren here."
Direct hit in Rishon Lezion (Photo: Avi Moalem)
The rocket attack (Photo: Asi Amar)
Rocket damage in Rishon (Photo: Naor Eini)
A short while earlier, one Israeli was seriously wounded when a rocket hit a house in Ashkelon, and seven were lightly injured by a rocket barrage that was fired on the southern city of Ashdodaround 4 pm. Three of the victims in Ashdod were hurt when a projectile hit a store in a commercial center, while the others were injured when a rocket exploded in an open area in the city. Another salvo was fired on the Eshkol Regional Council, hitting open areas and failing to cause injuries or damage.
The owner of the store that was hit in Ashdod said that his customers and employees ran for cover when they heard the air raid siren.
"My entire store was destroyed and my car was totaled," he said.
Store damaged in Ashdod (Photo: Noam Dvir)
Meanwhile, the army airdropped leaflets on the Gaza Strip's northern neighborhoods in the afternoon, warning residents to evacuate the region, an IDF official told Ynet. The Al-Arabiya news network reported that residents of the Strip's southern and eastern neighborhoods received similar instructions.
IAF aircraft struck a Hamas operative trying to fire an antitank missile towards Israel from Gaza's Dhaniya Airport. A direct hit was identified. According to the IDF, it was the third time in two hours that terrorists attempted fire antitank missiles on Israeli civilians and troops. The IAF has also bombed a Hamas operative while he was trying to launch a rocket.
Leaflets scattered in Gaza
Earlier Tuesday, a rocket was fired from Gaza toward Jerusalem for the second time since Operation Pillar of Defensebegan. The projectile hit an open area in Gush Etzion. Minutes before the siren sounded in the capital, six people were injured in two rocket attacks on the Eshkol Regional Council.
The siren went off in Jerusalem just as Foreign Minister Avigdor Lieberman was on the phone with his Japanese counterpart, Koichiro Gemba. Lieberman was forced to cut the conversation short and seek shelter.
Returning the call after the rocked had hit, Lieberman asked Gemba to try and imagine a situation in which he was sitting in the foreign ministry in Tokyo and had to leave a meeting or disrupt a phone call due to a rocket attack. He said that the operation in Gaza was imperative.
The foreign minister later met with UN Secretary-General Ban Ki-moon, who was at the Ben Gurion Airport at the time of the siren.
Lieberman thanked the secretary-general for his efforts to bring about peace to the region. However, the foreign minister said that the public outcry against an Israeli ground offensive in the Strip are empowering Hamas and prolonging the hostilities.
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Home > News and Events > Recent News > News 2019
Dr. Greg Simon wins national suicide prevention award
Kaiser Permanente physician-scientist will receive American Foundation for Suicide Prevention's Lifesavers Research Award.
Greg Simon, MD, MPH, will receive the American Foundation for Suicide Prevention (AFSP)'s Research Award at its Lifesavers Gala 2019 on May 7 in New York. The AFSP will also honor four others, including Anderson Cooper of CNN and CBS News.
Dr. Simon is a psychiatrist at Kaiser Permanente in Washington and a senior investigator at Kaiser Permanente Washington Health Research Institute. A University of Washington research professor of psychiatry and behavioral sciences, he leads the national Mental Health Research Network.
"We want to prevent suicide, so we must keep getting better at predicting it—although unfortunately we may never be able to predict every suicide," Dr. Simon says. In 2017, 14 in 100,000 Americans died by suicide, with rates rising in recent years, especially in rural areas, according to the National Center for Health Statistics.
"There's a conspiracy of silence around suicidal thoughts, because it's awkward to discuss," Dr. Simon says. "So we've found that we have to incorporate talking about it into our standard care. Our suicide prevention work is a great example of how research and care keep influencing each other to improve our patients' health. When research springs from clinicians' and patients' questions, 'learning health systems' can put results into practice much faster than the oft-cited 17-year lag."
It all started when Kaiser Permanente in Washington began routinely using a mental health questionnaire—a recognized hallmark of good care—in mental health and primary care. Now standard practice calls for all Kaiser Permanente Washington patients to be asked routinely to answer this questionnaire at least yearly at primary care visits—and at every visit for mental health concerns.
One question asks whether patients have thought, in the last two weeks, about wanting to harm themselves. First, Dr. Simon and his colleagues proved in 2013 that patients who answer "yes" all too often attempt suicide. Then the next year they implemented a standard process for follow-up to help prevent these patients from harming themselves, including personal safety plans, safe firearm storage, and disposing of extra medications. This work has spread to some other Kaiser Permanente regions—and beyond, including some other integrated health systems around the country.
Recently, Dr. Simon and his colleagues have used machine-learning models to develop and test a more sophisticated—and accurate—way to predict risk of suicide, by combining mental health questionnaire responses with information from electronic health records, including prior suicide attempts and mental health and substance use diagnoses.
"We're excited that Kaiser Permanente in Washington plans to put this new research into practice soon," Dr. Simon says. "And now we're conducting a large trial here to test two different outreach programs to prevent suicide attempt in people who are at high risk."
This work complements Kaiser Permanente's ongoing efforts to emphasize the importance of mental health and wellness and its impact on total health: mind, body, and spirit in unison. A research brief sums up Kaiser Permanente's research on mental health.
Gregory E. Simon, MD, MPH
Senior Investigator
Access researcher directory
What inspires Dr. Greg Simon to study mental health?
Kaiser Permanente Share profiles longtime psychiatrist, KPWHRI investigator, and Mental Health Research Network leader.
Read about it in Healthy Findings.
Predicting suicide attempts and suicide deaths using electronic health records
National Institute of Mental Health (NIMH) News, July 12, 2018
Suicide prevention: Research network finds new way to predict risk
Led by KPWHRI scientists, study aims to target those who need interventions most.
Read it in News and Events.
KPWHRI News
Rebecca Hughes
rebecca.f.hughes@kp.org


It's the end of the season so it wasn't hard to be honest.
15 win streak, undefeated from legend 1 to KOG!
Since X sabers made it to the top in the last meta weekly at Duel Links Meta, I'd say the game has gotten way more fun and playable thanks to the last banlist. I hope they add all the synchro support soon.
Oh man, Im glad they added 5d's I think they are by far the best series!
Glad onslaught is pretty much dead and buried.
Does this still count as lucky as someone who win a lottery?
Jeez ! congratulations bud !
The new white dragon has so much in it man. That card is not okay it is both normal monster and effect monster. Moreover it doesnt need to be double summoned to have its effect used. Just normal summon it and you have a free cosmic cyclone. It is not okay to have a deck that can summon 3 7+ level monsters in one turn. Archfiends needed a trap card to be effective and all but blue eyes meta is cancer. When i play against a yubel deck or something i have fun because i usually use SSA or Aromage but when i play against that blue eyes deck it is cancer. They are pretty consistent and you have to have hard backrow based deck for that deck.
Blue eyes need a trap card?
Any deck where you can summon 2 monsters with 3000+ attack is basically broken. It's a tier 0/1 deck and the tiers aren't necessarily that close to each other.


From the Departments of Neurology (B.Z., J.L., X.L., X.G., D.Z., Z.H., H.S.) and Neurobiological Laboratory (Y.Z., C.J.), West China Hospital, Sichuan University, Chengdu; and Department of Neurology (W.G.), Leshan People's Hospital, People's Republic of China.
Isaacs syndrome is an acquired peripheral nerve hyperexcitability syndrome characterized by twitching, painful cramps, stiffness, delayed muscle relaxation, and sweating.1 Needle EMG findings include continuous single motor unit discharges occurring as doublets, triplets, and multiplet single unit discharges firing at higher rates (150–300 Hz; neuromyotonic discharges) and at lower rates (less than 60 Hz; myokymic discharges).2 Isaacs syndrome has been reported in patients with thymoma, small-cell lung carcinoma (SCLC), or lymphoma.2 Antibodies specific for voltage-gated potassium channels (VGKC) have been reported as a serologic marker of Isaacs syndrome. Antibodies specific for the neuronal ganglionic acetylcholine receptor (AChR) are sometimes encountered in some patients.1 It has not been previously associated with sarcomatoid carcinoma (SC) or antibodies against γ-aminobutyric acid B (GABAB) receptors, and the latter often present in limbic encephalitis and other uncommon clinical syndromes (such as cerebellar ataxia).3 Here we describe a case of Isaacs syndrome with a positive test for anti-GABAB and anti-AChR antibodies and broad tumor screening for this case of SC.
Accepted in final form July 10, 2018.


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Regular Colleges or Educational Institutions Sports Education and Training Master of Physical Education (M.P.Ed)
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Bhupatinagar, Purba Medinipur , Midnapore (West Midnapore (Paschim) Dist.) - 721425
Mugberia Gangadhar Mahavidyalaya was established on 2nd of July, 1964 as a co-education college by ...
University of Kalyani, Nadia
PO Kalyani , Nadia (Nadia Dist.) - 741235
The University of Kalyani was established in 1960
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We are thrilled to welcome Jessica Mailhot as the Director of Forever Lancaster!
This newly created position will spearhead a strategic initiative from the Community Foundation focused on the education and building of our community's endowment. Forever Lancaster will focus on legacy giving that supports causes from families, youth, education, and health, to the environment, arts, culture, and economic development. As Director, Jessica will serve as an expert and resource to individuals, professional advisors, and nonprofits across our community seeking to make a long-term impact on Lancaster County.
Jessica brings a depth of experience from the both the public and private sector. A graduate of the University of Pennsylvania, she most recently served as Executive Director and Market Manager of Lancaster Central Market. Her board participation includes Lancaster Historical Commission, Lancaster Creative Factory, and Penn Alumni Club of Lancaster.


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Vladimír Borovička (born 25 March 1954 in Děčín) is a Czech football coach.

Playing career
Borovička played as a goalkeeper. He played for Bohemians Prague and Dukla Banská Bystrica in the Czechoslovak league.

Coaching career
Borovička worked as a Zenit St. Petersburg's caretaking manager in May 2006 after the departure of the fellow Czech Vlastimil Petržela. He since returned to his coaching duties.

References

External links
  Profile at the official FC Zenit St. Petersburg website

1954 births
Living people
People from Děčín
Czechoslovak footballers
Czechoslovakia international footballers
Czechoslovak expatriate footballers
Bohemians 1905 players
FK Dukla Banská Bystrica players
Expatriate footballers in Austria
Czech football managers
Czechoslovak football managers
FC Zenit Saint Petersburg managers
Russian Premier League managers
AC Sparta Prague managers
Expatriate football managers in Russia
Czechoslovak expatriate sportspeople in Austria
Czech expatriate sportspeople in Russia
Bohemians 1905 managers
Association football goalkeepers
Sportspeople from the Ústí nad Labem Region

redpajama_set_name: RedPajamaWikipedia


A long time ago I inquired about SkyDSL's internet service and, to my surprise, they required cable connection to have the internet connected. I heard of their service being pretty good and I wanted to try it but I already had Destiny cable installed so I didn't go with it and chose BayanDSL. Now that I've moved to another place, I decided to give their free trial a go.
My choices were PLDT or SkyDSL, PLDT takes 2 weeks to process application (not to mention that I applied for their service before and they actually lost my application) while SkyDSL offered to connect us within 2 days (I moved their visit as I wasn't available otherwise it would've been a day). I was lucky enough to be covered by their service, and I chose the 6 mbps trial service (double of what I had before of 3 mbps). The free trial guys came very early in the morning and were able to connect me within the next few minutes.
I was actually amazed by the consistency of their speed, I tested it throughout the day and everything was loading fast. I was surfing in fast speeds and webpages seemed to load really fast and I was even able to watch Youtube videos (some full movies) without having to stop to buffer. I was also amazed with how SkyDSL downloads Torrents, I had some torrents download at 600+ kb/s and took half the time to download than what I had with BayanDSL. Overall, I would certainly recommend their service based on my Free Trial.
Their prices are pretty good as it the P3,999 comes with a gold Skycable subscription, it also comes with a IWantV service which allows you to watch local shows online on demand.
Skydsl modem's adaptor was not functioning after 5 days, it might be that the test unit was old or a factory defect, It was later replaced by another adaptor. Customer Service also needs improving, I remember calling at 5pm in the afternoon to have the adaptor replaced I had to wait more than 15 minutes to talk to a representative to which I got cut off due to bad signal and had to wait another 15 minutes more. The good thing was that the representative was very helpful and was able to get me scheduled to replace the defected adaptor.
Overall, I love SkyDSL's service! The speeds are really consistent and torrent speeds are at the upper limits. I just hope that it would be the same service once I do get the actual paid service. I would certainly recommend this service to others, you can go for their 15 days free trial (deposit of P1000 required for modem). You can visit their website at MySky or call (632) 381-0000.


text:
Bushey Cemetery
Bushey Cemetery is a 16-acre site of outstanding natural beauty situated within the meadows of London's green belt. Established in 1947 it is one of the most significant Jewish burial sites in the UK. The extension will allow for a further 8,000 burials over the next 60 years.
Laid out along a north-south axis the buildings facilitate the movement of mourners
© Lewis Kahn
Rammed earth prayer halls taking shape on site
Large corten steel doors complement the raw material palette of rammed earth and english oak
© James Stephenson
Designed around the existing landscape the buildings are very much part of their setting
Section Prayer Hall
Section Cohanim Room
Detail - Cohanim room-prayer hall junction
Designed around the landscape the buildings are part of their setting and when the cemetery is fully occupied will be returned to the earth and the site to the green belt.
A linear arrangement of buildings laid out along a north-south axis facilitates the movement of mourners. A timber colonnade forms the processional route to the prayer halls which are entered from the west and exited to the east.
Discretely embedded into a corner of the sloping site are two monolithic prayer halls. Built of rammed earth this organic material defines the overall design and was chosen for its symbolic and practical sensitivity to the Jewish faith, echoing the sentiment of the deceased being laid to rest in plain wooden caskets, returning to the earth.
A 10-year journey taken alongside the Jewish community required Waugh Thistleton to develop an understanding of the processional nature of the Jewish faiths' practice of burial. With no established typology the form and layout of the buildings was not determined by the religion per se, but grew from an understanding of the process of Jewish burial and how this might relate to the site.
Flood risk was intrinsic to the site and therefore design and arrangement of the buildings. The new landscape uses a series of interconnecting swales and ponds to collect surface water from the highly impermeable clay soil.
Positioned in the lowest corner of the site the cemetery buildings maintain open views to the fields beyond the site. The prayer halls face east to Jerusalem and are embedded within the landscape. Sitting amongst the planting and oak trees they are cradled by the larch colonnade which masks their scale with one of human interaction. The varied colour and texture across the expansive walls of the prayer halls allows these imposing, tomb like structures to recede into the landscape.
Bushey Cemetery is built of the earth, a reference to the simplicity of the Jewish traditions. The austerity of the rammed earth reflects the simple burial traditions.
Rammed earth has been used for centuries but is relatively unknown in the UK. A moistened mixture of soil is compacted into a formwork which, when dried forms a dense, monolithic wall. Adding a small amount of lime/cement improves strength and durability over time. This is known as stabilised rammed earth (SRE).
There is no design standard in the UK for SRE. The team therefore had to justify the proposed design with repeated testing. A rammed earth specialist was engaged to ensure potential issues with the material and construction process could be addressed. Strength, durability, colour and texture were explored to provide a bespoke solution for the project.
SRE has strong sustainability credentials. Precise proportions of sand, limestone and earth mixed with a little cement and water, result in low embodied energy. The reception and mortuary buildings are constructed from well insulated pre-fabricated timber cassettes. PV panels provide energy while the high thermal mass will retain heat gains.
Bushey, United Kingdom
Waugh Thistleton Architects
Andrew Waugh (United Kingdom); Anthony Thistleton (United Kingdom)
Landscape architect: J&L Gibbons Structural engineering: Elliot Wood Construction: Earth Structures Planning Consultant: Dalton Warner Davis Quantity surveyor: Deacon & Jones Construction company: Buxton Mechanical: PR3
£3,235 €/m2
United Synagogue
LatLng: (51.655490, -0.350520)


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This concise Highlights Report has been prepared to provide the City of Toronto, TTC and Metrolinx with a snapshot of the feedback captured at the public meeting held on June 17, 2015. A more detailed report of the feedback captured during this phase of consultations will be prepared in the coming days.
On Wednesday, June 17, 2015, the City of Toronto, City Planning Division (Transportation Planning), the TTC and Metrolinx, hosted a public meeting on four key transit projects currently being planned. The meeting was held at the Spring Garden Baptist Church, Toronto.
Following an introductory presentation on Coordinated Network Transit Planning given by Mike Logan (Senior Transportation Planner) and David Cooper (Senior Transportation Planner) at 7:00 PM, participants had the opportunity to ask questions of clarification as well as provide feedback.
Approximately 25 people attended the public meeting, including York Region Councillor Jim Jones (City of Markham).
The discussion captured during the question and answer period following the overview presentation is summarized below. Questions are noted with a "Q", comments with "C" and answers with "A". Answers were provided by Mike Logan, David Cooper and Paul Millett (Chief Project Engineer, Engineering, Construction and Expansion Section, TTC).
C. The Scarborough Subway route should go up Midland to Sheppard, and the Sheppard line should be linked to form a loop.
A. Thank you for your input. We are considering the Scarborough Subway interface with the Sheppard East LRT.
Q. Regarding the Relief Line, when is construction anticipated to begin and finish?
A. After the Project Assessment is complete and the recommendations approved by Council, a Transit Project Assessment Process (TPAP) takes a further 6 months which is the final approval under the Environmental Assessment Act. The detailed design phase would likely be another 2-3 years and construction could take 8-10 years. The timeline depends on overall political support and funding. If approval and funding is received the project could be operational in 12-15 years.
Q. I am aware that GO lines are mostly on CN tracks. What are the options for signaling in the future to get the headways down as you expand in order to get more people riding on those trains?
A. Representatives from Metrolinx are here tonight and can speak with you about this in more detail, but generally, there are certain levels of rail operation rules based on the type of the train (diesel or electric). There are complexities when passenger and freight services share a corridor. Metrolinx is looking at which corridors can be electrified through their EA process and other assessments.
Q. Why does the Eglinton Crosstown LRT not continue to the Mississauga Airport Corporate Centre?
A. The base case assumption is that the LRT will be continued to the airport employment area – this extension has already been approved but is not funded. City Council has asked the project team to look at the feasibility of a heavy rail corridor instead of the LRT. The study will determine whether or not those are feasible options.
C. SmartTrack should have a spur to Scarborough Centre so that one train could go to Scarborough Centre and the next could go to Markham. The same could be done at the Mississauga Airport Corporate Centre (one train going there and the next train going on to Brampton). SmartTrack should be using as much of the existing infrastructure and going many different places rather than being asynchronous.
A. Thank you for your input. That may be an option. The direction from Council was that the Scarborough Subway be an extension from Kennedy Station though Scarborough Centre to Sheppard. RER and SmartTrack currently do not contemplate going into Scarborough Centre. Operating a spur with alternating destination would have implications on the timing of the service schedule and might make it impossible to deliver the 15 minute frequency of service. That is one of the issues that will be carefully considered in the Eglinton West feasibility study. We welcome the opportunity to have a further discussion on this.
Q. Who has the final say on the decision in terms of the final designs?
A. The project team will be completing the studies for the Relief Line and Scarborough Subway Extension. We will then submit a recommended alignment and station concepts to City Council. At that point we will be asking for authority to undertake the final review using the Transit Project Assessment Process (TPAP) which results in the authority to construct. SmartTrack is at a different stage in the process. Council has asked us to look at feasibility of the various options and it will be up to Council to direct us to do further work based on the results of the feasibility study.
Q. Do you have a cost estimate of routing to the Mississauga Airport Corporate Centre?
A. The project team will be looking at the cost of the various alternatives as part of the feasibility study.
C. Markham has to grow by 200,000 people over the next 25 years and we have one stop at Markham Centre.
A. In the coming months, the project team will be consulting with the Town of Markham, City of Mississauga, York and Peel Regions.
Q. When I'm on the GO train at Oriole station I have to walk through the snow to get to the Sheppard subway. The stations should be connected so you don't have to walk outside. The GO train stop needs to be moved.
A. The work being done near the Leslie/401 intersection will result in Metrolinx moving Oriole station north to Sheppard and therefore much closer to the TTC's Leslie Station. Metrolinx will be consulting on that in the coming months.
Q. Will the Yonge subway be extended to Highway 7?
A. Through the Yonge Relief Network Study there is a discussion happening on the next steps. The Relief Line study is also underway and will help us understand more about relieving some of the pressures on the existing Yonge subway, and we will better understand the capacity of the Yonge Subway and the potential to extend the line through our modeling that will be wrapping up over the summer.
Q. Is anyone looking at affordability of fares?
A. Part of the work that Council has directed staff to do on SmartTrack is to review fare structures and integration of fares. We will be modeling various fare structure scenarios with a consideration of a hybrid of a GO and TTC fare and the current fare structure. We will be reporting back to Council this Fall. Metrolinx is also undertaking a more detailed fare integration study.
Through a dotmocracy exercise at the meeting, most of the meeting participants generally agreed with the results of the evaluation for the Connections to the Danforth Subway, the results of the evaluation for Key Activity Areas West and East of the Don River and the evaluation of the Downtown connections.
Through a dotmocracy exercise at the meeting a majority of the meeting participants indicated a preference for Corridor D by a factor of 2:1 over Corridor B.
Five more public meetings are scheduled during this phase of consultations, after which a more detailed report of all consultation activities will be made available. Comments must be submitted by July 3, 2015 to ensure inclusion in this report.


text:
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        public string RelationshipType { get; set; }
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        public string ReverseRelation { get; set; }
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redpajama_set_name: RedPajamaGithub


House prices are now rising at a gentle 2.5% a year, according to the latest figures from Halifax.
Consumer price index inflation, meanwhile, is at 2.7%. Wage inflation – excluding bonuses – is rising at 2.9% (we talk about wage inflation in the latest MoneyWeek podcast – check it out here).
In other words, house prices are now falling in "real" terms.
Are we on our way to our hoped-for "soft" landing?
To get an idea of how important house prices are to the UK economy, you merely need glance at the number of indices we have that claim to track prices in one way or another.
For inflation, we have quite a few indices, but they're mostly variations on a theme from the Office for National Statistics (a cynic might explain this as being the product of successive governments trying to gerrymander the figures lower – I won't comment).
Similarly, for employment, you'll get the odd private sector survey – recruitment agencies talking about ease of hiring and wages, for example – but largely we all focus on the official data again.
But when it comes to house prices – you're spoiled for choice. You could be reading about a new survey every other week.
We've got Rightmove, which looks at asking prices. We've got mortgage approvals, which looks at – well, mortgage approvals. We've got the official house price index from the ONS.
We've got the niche indices from upmarket estate agents, that look at prime central London property, or country houses. We've got Hometrack, which looks at trends across UK cities.
But probably the best known pair of indices – partly because they have some of the longest histories – are the ones compiled by Nationwide and Halifax. These two take their data from somewhere in the middle of the buying process.
So the latest Halifax house price index will probably grab a few headlines this morning. Prices apparently fell by 1.4% in the month of September, which is a big drop compared with expectations.
Looking at the month-on-month figure is fairly pointless. It's quite a "noisy" data set. However, the year-on-year data is equally striking. In the three months to September, prices rose by 2.5% compared to the same quarter last year. Analysts had expected a rise of 3.4%.
That means not only that house price growth is still slowing (last time, according to Halifax, the annual growth rate was 3.7%), but also that house price growth is now firmly below inflation.
In other words, house prices are falling in "real" terms.
This is good news. You want this to continue. Here's why.
From the homeowner's point of view, you shouldn't really care that much about what happens to the price of your house as long as you can continue to pay your mortgage.
But that's not the way things work. No one likes to see the value of their home fall, particularly if it then puts them into negative equity. So there's a psychological impact from falling house prices, which is not great for consumer confidence, and spending.
Moreover, widespread, sharp falls in house prices tend to be symptomatic of something else – usually surging interest rates and a struggling economy. Rising borrowing costs not only push up mortgage bills, they also squeeze over-indebted businesses. That drives up unemployment and results in more forced sellers.
So a big house price crash is a messy way to get back to some sort of sensible housing market.
Instead, if the price of your house stays broadly flat, but your wages go up, then you don't get the psychological hit from a falling house price, and hopefully your mortgage also becomes more affordable relative to your pay packet. That's ideal.
Meanwhile, if you're frustrated by the unaffordability of property, then you might hope for a big crash. And I sympathise. Trouble is, getting a home loan when prices are crashing tends to be difficult. That's why the inflationary option can be more attractive even for those who don't already own property.
Basically, you want your wages to rise more rapidly than prices. If prices are flat or gently falling but your wages are rising, then it gets easier to save for a deposit.
So having made the case – will this continue?
The good news is that a lot of heat has come out of the market. Buy-to-let investment is much less attractive these days. Not only are the tax changes still coming through, the fact that house prices are no longer soaring – and therefore no longer seen as a "sure thing" – will reduce the appeal of property as an investment.
Meanwhile, one thing mitigating against prices falling hard is that, despite mortgage rates ticking higher, as Samuel Tombs of Pantheon Economics points out, "households can offset the impact of rising mortgage rates on monthly repayments by taking out longer maturity loans, while payments as a share of disposable incomes still are low by past standards." He reckons that means that we're unlikely to see big falls in prices at a nationwide level.
Overall, I think a prolonged period of single-digit house price falls and above-inflation wage gains would be a nice way out of this affordability problem.
I think it's a definite possibility. That said, I'll admit that the experts at our recent property roundtable were less optimistic than me. But we can always hope.
(Subscribe to MoneyWeek now if you haven't already, to make sure you don't miss our future roundtables).


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In the latest issue of Bioscience Reports (volume 39, issue 6), Khan et al. discuss the antimicrobial and antibiofilm properties of epsilon-poly-L-lysine(EPL)-catechol hydrogel against multidrug-resistant Acinetobacter baumannii in burn wounds. This cover features a scanning electron microscopy image of EPL-catechol hydrogel against multidrug-resistant Acinetobacter baumannii in a burn wound after 2 days of treatment.
Author Contribution
Research Article| June 14 2019
Knockdown of hsa_circ_0023028 inhibits cell proliferation, migration, and invasion in laryngeal cancer by sponging miR-194-5p
Xiaofeng Chen
Department of Otolaryngology, Ruian People's Hospital, No. 108 Wansong Road, Yuhai Street, Ruian City 325200, Zhejiang Province, China
Xiaoqing Su
Correspondence: Xiaoqing Su (suxqra@protonmail.com)
Chuansai Zhu
Jing Zhou
Biosci Rep (2019) 39 (6): BSR20190177.
https://doi.org/10.1042/BSR20190177
Revision Received:
Accepted Manuscript online:
Split-Screen
Xiaofeng Chen, Xiaoqing Su, Chuansai Zhu, Jing Zhou; Knockdown of hsa_circ_0023028 inhibits cell proliferation, migration, and invasion in laryngeal cancer by sponging miR-194-5p. Biosci Rep 28 June 2019; 39 (6): BSR20190177. doi: https://doi.org/10.1042/BSR20190177
Emerging evidences have proposed that circular RNAs (circRNAs) play a major role in carcinogenesis. Hsa_circ_0023028 has been reported to be aberrantly expressed in laryngeal cancer (LCa). However, the role and the mechanism of hsa_circ_0023028 in LCa have not been adequately studied. In the present study, we demonstrated that hsa_circ_0023028 expression was up-regulated in LCa tissues and cell lines. miR-194-5p was down-regulated in LCa cells. Functionally, knockdown of hsa_circ_0023028 inhibited the proliferation, migration, and invasion of LCa cells, as evidenced by the reduced number of 5-Ethynyl-2′-deoxyuridine (EdU)-positive cells and decreased number of migrated and invaded cells. Additionally, hsa_circ_0023028 was identified as an miR-194-5p sink. A negative correlation between miR-194-5p and hsa_circ_0023028 expression was observed in LCa tissues. Besides, down-regulation of miR-194-5p attenuated the inhibitory effects of hsa_circ_0023028 silencing on LCa cell proliferation, migration, and invasion. In summary, hsa_circ_0023028 functions as an miR-194-5p sponge to promote the proliferation, migration, and invasion of LCa cells.
competing endogenous RNA, hsa_circ_0023028, laryngeal cancer, miR-194-5p
Cancer, Cell Cycle, Growth & Proliferation, Cell Migration, Adhesion & Morphology, Molecular Interactions
Laryngeal cancer (LCa) is one of the most frequently encountered malignancy of the head and neck that affected more than 1.4 million people worldwide in 2015 [1]. Laryngeal squamous cell carcinoma (LSCC) is the most common type of LCa. LCa usually arises from the skin of the larynx. It is the eighth leading cause of cancer-related deaths worldwide, and caused over 105900 deaths in 2015 [2]. LCa clinically presents with hoarseness, breathing difficulties, persistent cough, and dysphagia. Family history, tobacco smoking, and alcohol consumption are considered to be important contributors to LCa. Stages I and II of LCa are often curable with surgery and radiotherapy, alone or in combination. While patients with advanced LCa (stages III and IV) are always treated with surgery and chemotherapy (alone or in combination), the prognosis is unsatisfactory [3]. Clinically, metastasis is regarded as the major obstacle in the treatment of patients with LCa, but the molecular mechanism of LCa progression is still unclear. Therefore, a detailed understanding of the mechanism involved in LCa progression is crucial for developing a novel and effective therapy for LCa.
Noncoding RNAs (ncRNAs) are a class of endogenous RNAs that are not translated into proteins. ncRNAs occupy more than 90% of the human genome and have become a major focus of research [4]. ncRNAs participate in the regulation of inflammatory signaling that underlie many diseases [5]. Abundant and functionally important types of ncRNAs include long ncRNAs, small ncRNAs, tRNAs, rRNAs, and circular RNAs (circRNAs). circRNAs are considered as a novel type of ncRNAs that are hallmarked by covalently closed continuous loops [6]. With the rapid development of next-generation sequencing, an increasing number of circRNAs has been discovered. circRNAs are widely represented in the eukaryotic transcriptome and are generated by back-splicing without the 3′ and 5′ ends [7,8]. Owing to their unique structure, circRNAs are able to counteract RNA exonucleolytic digestion, which makes them an ideal biomarker for disease diagnosis [9]. Recently, increasing evidence supports the notion that dysregulated circRNAs are tightly linked to the progression of human diseases [10]. circRNAs have been shown to function as key regulators of carcinogenesis. The expression of circRNAs in cancers has been found to be closely correlated with clinical features, such as tumor-node-metastasis stage, metastasis, and tumor size [11]. Furthermore, circRNAs have been reported to be implicated in the progression of human cancers by modulating the proliferation, invasion, apoptosis, and cell cycle of tumor cells [12]. Hsa_circ_0023028, a novel identified circRNA, has been reported to be located at chr11:66515849-66590145 in gene symbol C11orf80 and markedly overexpressed in LSCC tissues. Moreover, hsa_circ_0023028 level in LSCC tissues was shown to be closely related with the clinical characteristics of patients with LSCC, such as tumor grade, lymph node metastasis, primary location, and clinical stage [13]. However, very little information is available regarding the role of hsa_circ_0023028 in the occurrence and development of LCa.
microRNAs (miRNAs) are a type of small (∼21 nucleotides) ncRNAs that inhibit the translation of messengers RNAs (mRNAs), and have been implicated in a cohort of biological processes [14]. After nuclear processing, miRNA precursors are exported from the nucleus and form mature miRNAs in the cytoplasm. miRNAs assemble into the RNA-induced silencing complex and base-pair with the 3′-untranslated region (UTR) of their target mRNAs, leading to mRNA degradation and/or impairment of translation [15]. Alternatively, miRNAs can bind competitively to the AU-rich regions in the target mRNA 3′-UTR and inhibit the degradation of target mRNAs, thereby mediating mRNA up-regulation [16]. Emerging evidence suggests that miRNAs serve as key players in carcinogenesis [17]. Aberrant expression of miRNAs was reported in numerous malignancies, including LCa [18]. miR-194-5p, a vertebrate-specific miRNA, plays an important role in the progression of human cancers, including gastric cancer [19], hepatocellular carcinoma [20], LSCC [21], and gallbladder cancer [22]. Up-regulation of miR-194-5p was shown to cause a marked reduction in cell proliferation, suggesting its anti-tumor effect in gallbladder cancer [22]. However, little attention has been given to the biological function and underlying mechanism of miR-194-5p in LCa tumorigenesis.
Here, we aimed to explore the biological role of hsa_circ_0023028 in LCa, and identified hsa_circ_0023028 as an miR-194-5p sponge in carcinogenesis. Moreover, hsa_circ_0023028 promoted the proliferation and metastasis of LCa cells by sponging miR-194-5p. These findings suggest that hsa_circ_0023028 may possibly serve as a promising therapeutic target for LCa.
Patient samples
Twenty pairs of liquid nitrogen frozen LCa solid tumors and their corresponding adjacent noncancerous tissues were obtained from the Department of Otolaryngology, Ruian People's Hospital. Patients who participated in the present study were confirmed by pathological examination and did not receive any adjuvant therapy prior to operation. Protocols were approved by the Ethics Committee of Ruian People's Hospital and informed consent was obtained from all patients prior to participation in the present study.
Human LCa cells (Hep-2 and TU212) and normal nasopharyngeal epithelial cell line (NP69) were obtained from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). Cells were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; Invitrogen, Grand Island, NY, U.S.A.) and 1% penicillin/streptomycin (Solarbio, Beijing, China) in a humidified atmosphere of 95% air and 5% CO2 at 37°C.
Cell transfection
Small interfering RNA (siRNA) targeting hsa_circ_0023028 (si-circ), miR-194-5p mimic, miR-194-5p inhibitor, and matched controls were synthesized by GenePharma (Shanghai, China). Cell transfection was carried out using Lipofectamine 2000 reagent (Invitrogen) as per the manufacturer's instructions.
Detection of cell proliferation capacity
Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2′-deoxyuridine (EdU) assays. For CCK-8 assay, Hep-2 and TU212 cells were seeded into 96-well plates and incubated overnight in a humidified atmosphere of 95% air and 5% CO2 at 37°C. At the indicated time points of 0, 24, 48, and 72 h post-transfection, the transfected cells were incubated in RPMI-1640 medium containing CCK-8 reagent (10 μl; Solarbio). After 2 h of incubation in a 5% CO2 incubator at 37°C, the absorbance at 450 nm was measured using a NanoDrop spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, U.S.A.).
For EdU assays, Hep-2 and TU212 cells were transfected with siRNA specific for hsa_circ_0023028 (si-circ), miR-194-5p inhibitor, or siRNA negative control (si-NC) and incubated for 48 h in 24-well plates. Following this, cells were incubated with 10 µM EdU (Beyotime, Shanghai, China) for 2 h, followed by fixation in 4% paraformaldehyde for 15 min. After washing, cells were permeated with Triton X-100 for 10 min, and then incubated with the reaction mixture for 30 min. Thereafter, cells were stained with Hoechst 33342 for 5 min in darkness. The number of EdU-positive cells was counted under a fluorescence microscope.
Transwell migration and invasion assay
Transwell chambers (Corning, Steuben County, NY, U.S.A.) were used to assess the migration and invasion of Hep-2 and TU212 cells. Hep-2 and TU212 cells were transfected with si-circ, miR-194-5p inhibitor, or si-NC. At 48 h post-transfection, Hep-2 and TU212 cells were harvested and then seeded in the upper chambers pre-coated with fibronectin (for migration assay; Solarbio) or Matrigel (for invasion assay; BD Biosciences, Franklin Lakes, NJ, U.S.A.). The lower chambers were filled with RPMI-1640 medium containing 5% FBS. After 48 h of incubation at 37°C, the migrated and invaded cells were fixed in 4% paraformaldehyde and stained with 0.1% Crystal Violet (Solarbio) for 4 min. The number of cells that migrated or invaded to the lower side of the membrane was counted in five random fields using a light microscope.
Luciferase reporter assay
The hsa_circ_0023028 sequence containing wild-type or mutated miR-194-5p binding site (WT-circ; MUT-circ) was fused into pmirGLO vector (Promega, Madison, WI, U.S.A.). The luciferase reporter vector was co-transfected into Hep-2 and TU212 cells with miR-194-5p or miRNA negative control (miR-NC). The Dual-Luciferase Reporter Assay System (Promega) was used to detect the luciferase activities at 48 h post-transfection as per the manufacturer's specifications.
Quantitative real-time polymerase chain reaction
RNA extraction from the tissues and cells was performed using TRIzol reagent (Invitrogen) and the RNA concentration was assessed using a spectrophotometer. For miRNAs, TaqMan miRNA assay (Applied Biosystems, Foster City, CA, U.S.A.) was applied as per the manufacturer's instructions. miRNA and U6 primers were purchased from Applied Biosystems. For hsa_circ_0023028, SuperScript III Reverse Transcriptase (Life Technologies, Gaithersburg, MD, U.S.A.) and Power SYBR Green Master Mix (Life Technologies) were used for cDNA synthesis and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. qRT-PCR was performed using the CFX96 Real-time PCR system (Bio-Rad, Hercules, CA, U.S.A.). The primers' sequences used were as follows: hsa_circ_0023028: forward, 5′-GCC AAG GCT CAG CAG AAA CTA-3′ and reverse, 5′-TGT TGC TCC AAG ACC TTG TCC-3′; glyceraldehyde-3-phosphate dehydrogenase (GAPDH): forward, 5′-GGG AAA CTG TGG CGT GAT-3′ and reverse, 5′-GAG TGG GTG TCG CTG TTG A-3′; miR-194-5p: forward, 5′-GCG GCG GTG TAA CAG CAA CTC C-3′ and reverse, 5′-ATC CAG TGC AGG GTC CGA GG-3′; miR-145: forward, 5′-GTC CAG TTT TCC CAG GAA TCC CT-3′ and reverse, 5′-GCT GTC AAC ATA CGC TAC GTA ACG-3′; miR-1225-5p: forward, 5′-GTG GGT ACG GCC CAG T-3′ and reverse, 5′-CAC AGG GGC TCA GTC AGT C-3′; miR-1234: forward, 5′-GTG AGT GTG GGG TGG CTG-3′ and reverse, 5′-GGT GGT CAG GCC GAC TAG G-3′; miR-186: forward, 5′-AAG AAT TCT CCT TTT GGG CT-3′ and reverse, 5′-GTG CGT GTC GTG GAG TCG-3′; miR-433: forward, 5′-CTG GTA GGA TCA TGA TGG GAT-3′ and reverse, 5′-TCA ACT GGT GTC GTG GAG T-3′; U6: forward, 5′-GCT TCG GCA CAT ATA CTA AAA T-3′ and reverse, 5′-CGC TTC ACG AAT TTG CGT GTC AT-3′. Expression levels were normalized to GAPDH for hsa_circ_0023028 and U6 for miRNAs. The 2–ΔΔCt method was applied to determine the expression of hsa_circ_0023028 and miRNAs.
All experiments were performed in triplicate and statistical analysis was carried out using SPSS 20.0 software (IBM, Armonk, NY, U.S.A.). The results are expressed as mean ± standard error of the mean, and data were analyzed using Student's t test or one-way analysis of variance. A P-value less than 0.05 was defined as the level of significance.
Ectopic expression of hsa_circ_0023028 in LCa tissues and cell lines
Previously, microarray analysis showed that hsa_circ_0023028 was up-regulated in LSCC [12]. LCa tissues and paired adjacent noncancerous tissues were collected to evaluate the expression of hsa_circ_0023028 in LCa. qRT-PCR analysis revealed that hsa_circ_0023028 was highly expressed in LCa tissues as compared with their adjacent noncancerous tissues (Figure 1A). In line with this, the expression level of hsa_circ_0023028 was found to be higher, but, miR-194-5p was lower in Hep-2 and TU212 cells than that in NP69 cells (Figure 1B).
The expression of hsa_circ_0023028 characterizes LCa tissues and cell lines
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(A) A total of 20 LCa solid tumor samples and paired adjacent noncancerous tissue samples were obtained from patients with LCa. Hsa_circ_0023028 expression was detected to be up-regulated in LCa tissues using qRT-PCR. (B) Expression levels of hsa_circ_0023028 and miR-194-5p were validated in human LCa cell lines (Hep-2 and TU212) and normal nasopharyngeal epithelial cell line (NP69) using qRT-PCR. **P<0.01 and ***P<0.001.
Knockdown of hsa_circ_0023028 inhibits the proliferation of LCa cells
Since hsa_circ_0023028 is up-regulated in LCa, we knocked down hsa_circ_0023028 expression to assess its functional role in LCa. A specific siRNA targeting hsa_circ_0023028 was transfected into Hep-2 and TU212 cells, leading to down-regulation of hsa_circ_0023028 (Figure 2A,B). We found that down-regulation of hsa_circ_0023028 significantly reduced the viability of Hep-2 and TU212 cells compared with the control group (Figure 2C,D). In parallel, silencing of hsa_circ_0023028 in Hep-2 and TU212 cells resulted in a striking reduction in cell proliferation, as indicated by the CCK-8 (Figure 2E,F) and EdU assays (Figure 2G,H).
Hep-2 and TU212 cells were transfected with si-NC or si-circ. Cells were collected at 48 h after transfection. (A,B) qRT-PCR analysis of hsa_circ_0023028 expression in Hep-2 and TU212 cells. (C,D) CCK-8 assay showed the viability of Hep-2 and TU212 cells transfected with si-NC or si-circ. CCK-8 (E,F) and EdU (G,H) proliferation assay showed the cell proliferation ability of Hep-2 and TU212 cells transfected with si-NC or si-circ. **P<0.01 and ***P<0.001.
Knockdown of hsa_circ_0023028 inhibits the migration and invasion of LCa cells
To further elucidate the biological functions of hsa_circ_0023028 in LCa, a specific siRNA targeting hsa_circ_0023028 was used to down-regulate its expression in Hep-2 and TU212 cells, followed by transwell migration and invasion assays. As shown in Figure 3A,B, knockdown of hsa_circ_0023028 inhibited the cell migration ability of Hep-2 and TU212 cells compared with the control group. Meanwhile, compared with the control group, transfection of si-circ into Hep-2 and TU212 cells caused a marked reduction in cell invasion (Figure 3C,D).
A specific siRNA targeting hsa_circ_0023028 was transfected into Hep-2 and TU212 cells, and cells were collected at 48 h after transfection. (A,B) The migration ability of Hep-2 and TU212 cells transfected with si-circ or si-NC was determined using transwell migration assay. (C,D) Transwell invasion assay was performed to evaluate the invasion ability of Hep-2 and TU212 cells transfected with si-circ or si-NC. **P<0.01 and ***P<0.001.
Knockdown of hsa_circ_0023028 up-regulates the expression of miRNAs in LCa cells
Bioinformatics analysis that predicted the potential targets of hsa_circ_0023028 were miR-145, miR-1225-5p, miR-1234, miR-186, miR-433, and miR-194-5p, which share complementary binding sites with hsa_circ_0023028. To investigate the mechanisms that mediate the effects of hsa_circ_0023028 in LCa, we measured the expression level of miR-145, miR-1225-5p, miR-1234, miR-186, miR-433, and miR-194-5p in control and hsa_circ_0023028-silenced Hep-2 and TU212 cells. We found that the expression of miR-1234 and miR-194-5p was strikingly increased, but that of miR-145, miR-1225-5p, miR-1234, miR-186, and miR-433 did not show significant changes in hsa_circ_0023028-silenced Hep-2 cells. Of note, the expression levels of miR-194-5p and miR-1234 were obviously increased in the si-circ group, especially the expression of miR-194-5p (Figure 4A). Similar results were also seen in TU212 cells (Figure 4B). Therefore, in subsequent experiments, we focused on the role of miR-194-5p due to its marked up-regulation in hsa_circ_0023028-silenced Hep-2 and TU212 cells.
Knockdown of hsa_circ_0023028 up-regulates the expression of miR-194-5p in LCa cells
A specific siRNA targeting hsa_circ_0023028 was transfected into Hep-2 and TU212 cells, and cells were harvested at 48 h post-transfection. (A,B) qRT-PCR analysis of miR-194-5p expression after transfecting Hep-2 and TU212 cells with si-NC or si-circ. *P<0.05 and **P<0.01.
Hsa_circ_0023028 serves as a sponge for miR-194-5p
The expression of miR-194-5p was lower in LCa tissues than that in their adjacent noncancerous tissues (Figure 5A). Consistently, miR-194-5p expression was down-regulated in LCa cell lines (Hep-2 and TU212) compared with that in NP69 cells (Figure 5B). As indicated in Figure 5C, the sequence of hsa_circ_0023028 harbors a putative miR-194-5p binding site. Additionally, a negative correlation between hsa_circ_0023028 and miR-194-5p expression was observed in LCa tissues (Figure 5D). Notably, down-regulation of hsa_circ_0023028 by si-circ obviously increased the expression level of miR-194-5p in Hep-2 and TU212 cells (Figure 5E,F). To further validate the association between hsa_circ_0023028 and miR-194-5p, we performed luciferase reporter assay in Hep-2 and TU212 cells. Results revealed that the luciferase activity of WT-circ was reduced in the presence of miR-194-5p, but remained unaltered in the presence of miR-NC. Simultaneously, transfection with miR-194-5p or miR-NC did not affect the luciferase activity of MUT-circ in Hep-2 and TU212 cells (Figure 5G,H).
(A,B) The expression of miR-194-5p was evaluated in LCa tissues and cell lines using qRT-PCR. (C) The complimentary binding site for miR-194-5p in the hsa_circ_0023028 sequence is shown. (D) Correlation analysis of hsa_circ_0023028 and miR-194-5p expression in LCa tissues. (E,F) qRT-PCR analysis of miR-194-5p level after transfecting Hep-2 and TU212 cells with si-NC or si-circ. (G,H) Luciferase reporter assay showed the luciferase activity of Hep-2 and TU212 cells co-transfected with WT-circ or MUT-circ and miR-194-5p or miR-NC. **P<0.01 and ***P<0.001.
Down-regulation of miR-194-5p attenuates the inhibitory effect of hsa_circ_0023028 silencing on LCa cell proliferation, migration, and invasion
We further explored the potential involvement of miR-194-5p in mediating the effects of hsa_circ_0023028. Using an miR-194-5p inhibitor, we evaluated its ability to attenuate the effect of hsa_circ_0023028 silencing on the proliferation, migration, and invasion abilities of LCa cells. hsa_circ_0023028 silencing resulted in the upregulation of miR-194-5p (Figure 6A,B). CCK-8 assay showed that the viability of Hep-2 and TU212 cells was higher in the si-circ + miR-194-5p inhibitor group than in the si-circ group. Meanwhile, an increased number of EdU-positive cells was observed in the si-circ + inhibitor group as compared with the si-circ group, as indicated by the CCK-8 (Figure 6C,D) and EdU assays (Figure 6E,F). Moreover, transwell migration and invasion assays revealed that si-circ-mediated inhibition of cell migration and invasion in Hep-2 and TU212 cells was remarkably blocked by down-regulation of miR-194-5p (Figure 6G,H).
(A,B) Hep-2 and TU212 cells were transfected with si-NC, si-circ, or si-circ + miR-194-5p inhibitor. miR-194-5p level was detected. (C,D) CCK-8 assay showed the cell viability of Hep-2 and TU212 cells transfected with si-NC, si-circ, or si-circ + miR-194-5p inhibitor. (E,F) EdU assay showed the proliferation ability of Hep-2 and TU212 cells transfected with si-NC, si-circ, or si-circ + miR-194-5p inhibitor. (G,H) The migration and invasion of Hep-2 and TU212 cells transfected with si-NC, si-circ, or si-circ + miR-194-5p inhibitor were determined using transwell migration and invasion assays, respectively. *P<0.05 and **P<0.01.
Currently, there are several studies describing the roles of circRNAs in LCa. Fan et al. [23] identified 506 differentially expressed circRNAs in LSCC. They found that hsa_circ_0044520 and hsa_circ_0044529 expression levels were up-regulated in LSCC [23]. Wu et al. [24] found that 3715 circRNAs were differentially expressed in CD133+CD144+ cancer stem cells derived from human LSCC cells. Among these circRNAs, hg19_circ_0005033 was up-regulated, and down-regulation of hg19_circ_0005033 repressed the proliferation, migration, and invasion of these CD133+CD144+ cancer stem cells [24]. With regard to the normal tissues, hsa_circ:chr20:31876585-31,897,648 was validated to be down-regulated in LSCC tissues, indicating that it might serve as a novel tumor suppressor in LSCC [25]. In summary, previous literature shows that circRNAs play an important role in tumorigenesis. However, the specific circRNAs in LCa are just beginning to be characterized. In the present study, we first identified the function of hsa_circ_0023028 in LCa. We found that hsa_circ_0023028 could promote the proliferation, migration, and invasion abilities of LCa cells, indicating that hsa_circ_0023028 functions as a potential oncogene in LCa.
miRNAs have been extensively discussed in human cancers and serve as crucial regulators in numerous physiological processes. miR-194-5p has been shown to play an important role in tumorigenesis, although its role may vary in different cancers. Su et al. [26] identified the involvement of miR-194-5p in glioma. They demonstrated that miR-194-5p was down-regulated in glioma, wherein it functioned as a tumor suppressor to repress the malignant biological behaviors of glioblastoma stem cells [26]. Moreover, long noncoding RNA X-inactive specific transcript-induced down-regulation of miR-194-5p was shown to promote the proliferation, migration, and invasion of hepatocellular carcinoma cells by suppressing mitogen-activated protein kinase 1 expression [20]. Through targeted inhibition of lysosome-associated membrane protein-2, miR-194-5p repressed the cell proliferation ability of sunitinib-resistant ACHN cells in the presence of sunitinib [27]. Additionally, miR-194-5p was shown to be up-regulated in breast cancer tissues, and silencing of miR-194-5p repressed the cell proliferation, migration, and invasion of MCF-7 cells by promoting SRY-box 17 expression through the Wnt/β-catenin signaling pathway [28]. A recent study also showed that miR-194-5p was underexpressed in LSCC tissues and low expression of miR-194-5p was correlated with T stages, clinical stages, recurrence, and poor prognosis in LSCC. Furthermore, enhanced expression of miR-194-5p has been shown to repress the malignant phenotypes of LSCC by targeting Wee1 [21]. Li et al. [21] have demonstrated that miR-194-5p is a tumor suppressor in LSCC and they identified Wee1 as its downstream target gene. However, to date, the upstream regulatory mechanism of abnormal expression of miR-194-5p in LCa remains unclear. In the present study, down-regulation of miR-194-5p was validated in LCa tissues and cells, which was consistent with the previous study by Li et al. [21]. A negative correlation between miR-194-5p and hsa_circ_0023028 expression was observed in LCa tissues, suggesting the involvement of miR-194-5p in mediating the effects of hsa_circ_0023028 in LCa.
The interaction between ncRNA molecules is a novel and important regulatory mechanism in tumorigenesis [29]. Recently, the potential of circRNAs as gene regulators has been emphasized. It is noteworthy that circRNAs function as competitive endogenous RNAs to regulate gene expression through sponging miRNAs [30]. Due to the presence of multiple miRNA binding sites, circRNAs can serve as competitive inhibitors that inhibit the ability of miRNAs to bind their target mRNAs [31]. An example of this is the circRNA circMTO1, which harbors the miR-9 binding sites. circMTO1 serves as an miR-9 sink to increase the expression of the miR-9 target p21, thereby inhibiting the proliferation and invasion of hepatocellular carcinoma cells [32]. Li et al. [21] have shown the role of miR-194/Wee1 axis in regulation of the proliferation, migration, and invasion in LSCC cells. Based on [21], we delved more deeply into the regulatory mechanism of miR-194-5p in the proliferation, migration, and invasion of LCa cells. We reported, for the first time, the identification of hsa_circ_0023028 as an miR-194-5p sink, and demonstrated that hsa_circ_0023028 promoted the proliferation and metastasis of LCa cells by sponging miR-194-5p.
In conclusion, we first reported the up-regulation of hsa_circ_0023028 in LCa. Moreover, we showed that hsa_circ_0023028 promotes the proliferation and metastasis of LCa cells by sponging miR-194-5p. Hence, our research provides a better understanding of the relationship between circRNA deregulation and carcinogenesis.
The authors declare that there are no sources of funding to be acknowledged.
The authors declare that there are no competing interests associated with the manuscript.
Xiaofeng Chen, Chuansai Zhu, and Jing Zhou conducted literature search, carried out data extraction, and wrote the manuscript. Xiaoqing Su and Xiaofeng Chen designed the study. Chuansai Zhu and Jing Zhou performed statistical analysis. Xiaoqing Su revised the manuscript.
ACHN
renal adenocarcinoma cells
CCK-8
cell counting kit-8
circRNA
circular RNA
5-Ethynyl-2′-deoxyuridine
fetal bovine serum
GAPDH
glyceraldehyde-3-phosphate dehydrogenase
LSCC
laryngeal squamous cell carcinoma
miR-NC
miRNA negative control
ncRNA
si-NC
siRNA negative control
small interfering RNA
TNM
tumor-node-metastasis
untranslated region
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Do you want to know who Dizzy Gillespie idolized? Did you ever wonder who inspired Kenny Clarke and Art Blakey? Who was the pianist that mentored Monk, Bud Powell, Tad Dameron, Elmo Hope, Sarah Vaughan and Mel Torme? Who was Art Tatum's idol and Nat Cole's mentor? What musical quartet pioneered the concept adopted later by the Modern Jazz Quartet? Were you ever curious to know who taught saxophone to Stanley Turrentine or who taught piano to Ahmad Jamal? What community music school trained Robert McFerrin, Sr. for his history-making debut with the Metropolitan Opera? What virtually unknown pianist was a significant influence on young John Coltrane, Shirley Scott, McCoy Tyner, Bobby Timmons and Ray Bryant when he moved to Philadelphia from Pittsburgh in the 1940s? Would you be surprised to know that Erroll Garner attended classes at the Julliard School of Music in New York and was at the top of his class in writing and arranging proficiency?
Some answers can be gleaned from the postings on the Pittsburgh Jazz Network.
For almost 100 years the Pittsburgh region has been a metacenter of jazz originality that is second to no other in the history of jazz. One of the best kept secrets in jazz folklore, the Pittsburgh Jazz Legacy has heretofore remained mythical. We have dubbed it "the greatest story never told" since it has not been represented in writing before now in such a way as to be accessible to anyone seeking to know more about it. When it was happening, little did we know how priceless the memories would become when the times were gone.
Today jazz is still king in Pittsburgh, with events, performances and activities happening all the time. The Pittsburgh Jazz Network is dedicated to celebrating and showcasing the places, artists and fans that carry on the legacy of Pittsburgh's jazz heritage.
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ROGER HUMPHRIES BIG BAND @ RHYTHM HOUSE
Added by Dr. Nelson Harrison
Time: December 21, 2008 from 7pm to 10pm
Location: THE RHYTHM HOUSE
Street: 3029 Washington Pike
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Latest Activity: Dec 31, 2008
Come and celebrate Christmas, Hannukah, Kwanzaa, Eid and the Winter Solstice with the swingingest big band in the 'Burgh... tonight. Toast the season with great jazz. No cover, no minimum, WDUQ member discounts, dinner available, Pittsburgh Jazz Society member discounts. Be there or be square!
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Comment by Kenny Hawkins on December 23, 2008 at 8:37pm
Hey Nelson,
Sorry I couldn't be there to celebrate and participate in the great party. I am only 2,700 miles away and I was gonna come but my Leer Jet was down and I couldn't get a jitney from here to there. Later Brother
Comment by Rick Toliver on December 22, 2008 at 3:15pm
I'm sure all went well. I'm sorry I couldn't make it this time!!
Comment by Debi Arnett on December 22, 2008 at 1:53pm
Sorry Nelson, too far for a weeknight. Thanks for the invite. Deb
Comment by Amanda Sedgwick on December 22, 2008 at 8:58am
Oh, I am forced by circumstances to be square! Wish I was there to make it. Thank you for inviting me!
Comment by Jagsu on December 22, 2008 at 3:51am
That's a great photoof RH Factor at the Grill!
Comment by Gary Walker on December 22, 2008 at 3:05am
Play one for me guys and have a Merry Christmas Pittsburgh jazz friends
Comment by timm coxx on December 21, 2008 at 10:57pm
would love to atttend, but i'm only 700 miles from the jazzparty. drink a coupla cold ones for me, y'all. T.C.
Comment by Don Cerminara on December 21, 2008 at 10:47pm
THIS AN ESPECIALLY BIG "ENTERTAINING AT DON'S" TIME! WE'LL BE TIED UP, ALMOST EVERY NIGHT...'TAINING PEOPLE...BUT THANKS FOR THINKIN' OF ME
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A small congregation celebrates together each Sunday evening at 5:30pm in the Chapel, often followed by a cup of tea or coffee in the main house. Everyone is welcome.
Cottage (hallway shown here) are workspaces for lease by our Artists in Residence, who use the Old Friary as a reflective and sacred space for artistic creativity. Their artworks are often, among others, seen hanging in the main house.
Sea of Faith in Australia (SoFiA) meet at The Old Friary on the first Sunday of the month at 7:30pm in St Christopher's Lodge.
"SoFiA is a network of Australians who are interested in the non-dogmatic discussion of religion, faith and meaning. We want to explore for ourselves what we can believe and how we can find meaning in our lives.
SoFiA is not a church: it is a forum for discussing ideas, experiences and possibilities. Any who find themselves in sympathy with our aim - 'promoting the open exploration of religion, faith and meaning' - are most welcome to join us."
Heather and David Eckersley are members of the Northumbria Celtic Community in Australia. The Northumbria Community, founded in England, adheres to a celtic spirituality practice. The 3rd Sunday meditative service (see at left) is led by Heather as a member of the Northumbria Community.
for Monday on 3376 1830.