Zaratan - Arte Contemporânea is pleased to announce the opening of The Advantages of Poisonous Plants, a flash exhibition by Roi Carmeli, which presents a series of new works produced during his residency in Lisbon.
BIO: Roi Carmeli is a visual artist and musician based in Glasgow. He graduated with a Bachelor of Fine Arts from the Bezalel Academy of Art and Design, Jerusalem (2010) and obtained a master degree in Fine Arts from the Glasgow School of Art, Glasgow (2015).
During his education, Roi spent a year as exchange student at the California College of Art, San Francisco, as well as residencies at Corning museum for glass, NY (2012), Scottish sculpture workshop, UK (2016) and Tenjiamyama art studio, JP (2018), among other places. His work has been exhibited in Israel, USA, Scotland, England, Japan, Czech Republic, and the Netherlands.
narratives. He distinctively uses sound not as background, but as a tangible material in and of itself.


text: Taylor Frankie Paul, a 28 year old Tiktoker has revealed the details of her custody agreement with her soon-to-be ex-husband, Tate Paul, with a literal song and dance. her triplet alphas chapter 9 Andrew Tate is currently in jail and cannot agree to or sign a contract for a fight until his legal disputes are cleared. However, the Romanian resident squared off against Jake Paul last year ...Paul Tate Married Relationship facts. Doris Tate · His Other Children · Children Together · Her Other Children · Who's In The News?...Tate's custom manufacturing capabilities and innovative line of products provide data centers with an optimized floor-to-ceiling solution. Our Products Access Floors Finishes Airflow Panels and Controls Structural Ceilings Containment Accessories Contact Tate [email protected] 1-800-231-7788 Featured Case Studies Kroon Hall etsy chunky knit blanket The Squaw: Directed by Gerald Mayer. With James Arness, Dennis Weaver, Milburn Stone, Amanda Blake. A young man is very unwelcoming to the young Indian bride his father brings home. euromillions hotpicks predictions Sharon Tate Murders Crime Scene331,587 views Sep 13, ... Police guard the $200,000 rented estate of movie director Roman Polanski after his wife, actress Sharon Tate, and four other persons were found slain August 9th. ... Sharon was eight and half months pregnant with her and Polanski's first child, ...Ruby Landry-Dumas (later Ruby Andreas) is the main character in The Landry Series by V.C. Andrews. She is the daughter of Pierre Dumas and Gabrielle Landry. She is also the twin sister of Giselle Dumas, the half-sister and wife of Paul Tate, wife of Beau Andreas, and mother of Pearl Andreas, Pierre Andreas, and Jean Andreas Gabrielle Landry, Ruby's mother, died shortly after Ruby was born. As ...Jan 2, 2023 · Photo by Clive Brunskill/Getty Images. The tennis player is playing his first Wimbledon this week so we find out more about Tommy Paul's ex-girlfriend, Kiki Passo, … reverie + oak presets freeSharon Tate won the first of many beauty contests before her first birthday. Her looks, charm, and grace would be her passport into the acting world, initially as an extra in movie shot on location in Italy, where her army officer father was posted in the early 1960s. ... Her parents were Paul James Tate (1922-2005) and Doris Gwendolyn Willett ...What happened between Taylor Paul and her former husband, Tate? Paul announced her split from her husband in a viral TikTok, ending their marriage after first tying the knot in 2016. Tate...15 de jul. de 2022 ... Tate and I are divorcing, however, I always love him deeply, ... Paul's divorce comes in the aftermath of the swinging scandal that rocked ... no mercy in mexico link Tate Paul's wife is Taylor Frankie Paul who is also a social media star and a popular TikToker in the USA. They got married in September 2016. They first met the same year in the month of January. and has married after 8 months of dating. Taylor Frankie Paul and her husband Tate PaulBy Stacy Carey / March 24, 2021 9:41 pm EST. Paul McCartney lost his first wife, Linda McCartney, in 1998 and he's been quite open in the years since then regarding how heartbreaking the loss was ...Larenz Tate And Wife Tomasina's Sweet Love Through The Years ... The Tates welcomed the birth of their first born, son Miles Xavier Tate, 10, on July 29, 2006. getty images26 de abr. de 2022 ... Learn about Taylor Frankie Paul's Age, Height, Weight, Net Worth, TikTok, Instagram, Family, Husband, Tate Paul, Children, Son, Daughter.Taylor Frankie Paul was married to Tate Paul after meeting him in January 2016. On August 7, 2016, the couple got to know each other after dating for seven to eight months. AdvancedMD currently employs Tate as a district manager. On August 31, 2017, Taylor Frankie Paul gave birth to their first child, a daughter Indy Paul.Paul Tate. 1,037 likes. Musician/band timing chain replacement ford f150 My First Wife is an excruciatingly emotional film about the break-up of a marriage in contemporary Australia. There's not much new in it, and the dialogue isn't especially brilliant, but it's exceeding well done and sharply observant. Director and co-author Paul Cox based the story partly on events in his own life, and it shows.His mother has been dead almost a year, and Cully is troubled by his pa's carousing. When they discuss his getting married again, it seems like a good idea until he returns from a business trip with a new wife, an Arapaho squaw, who he has conflicted emotions about. — DrDOS north america Plot summary Plot synopsis Genre Western Certificate TV-PGSharon Marie Tate was born on January 24, 1943, in Dallas, Texas, the eldest of three daughters to Colonel Paul James Tate, a United States Army officer, and his wife, Doris Gwendolyn (née Willett). The family are of English, Scottish, Swiss-French, and Swiss-German descent. [8] taurus horoscope for january 2023 26 de mai. de 2022 ... The internet was abuzz with reports that the TikToker and her husband Tate Paul were separating. The influencer has since revealed that the two ...Tate is an American Western television series starring David McLean that aired on NBC from June 8 until September 14, 1960. [1] [2] It was created by Harry Julian Fink (the creator of Dirty Harry and T.H.E. Cat ), who wrote most of the scripts, and produced by Perry Como 's Roncom Video Films, Inc., as a summer replacement for The Perry Como Show . slap battles secrets Sep 24, 2021 · Steve McQueen's Serial Infidelity Saved His Life in 1969 When He Did Not Go to the Dinner Where Sharon Tate Was Killed. By Camila Santiago Sep 24, 2021. 02:00 A ... Judge Judy Caused Husband of 45 Years to Leave His First Wife and Regretted Divorcing Him 13 Years Later. February 09, 2022.Sharon tate was married to acclaimed director, Roman Polanski, and had just earned her first Golden Globe nomination for her role in Valley Of The Dolls. But then, on August 9, 1969, she was ...The prosecution also showed crime scene photos during the testimony of its first witness, Bobby Jo. You are interested in: White house farm murders crime scene photos. With plenty of ad-libs, Cid leads his (fake) organization against the foul (yet surprisingly real) Diabolos Cult. See for yourself - below are rare crime scene photos showing ...4 hours ago · The beautiful Sharon Tate would have celebrated her 80th birthday on Jan. 24, 2023. ... Smokey Robinson reveals he cheated on his first wife with Diana Ross, more … quant internships reddit Jul 15, 2022 · Taylor Frankie Paul, a 28 year old Tiktoker has revealed the details of her custody agreement with her soon-to-be ex-husband, Tate Paul, with a literal song and dance. Weeks after announcing they had split over 'soft swinging' drama, Taylor Frankie Paul, who shares two young children with Tate Paul, answered a fan's question about …Taylor Frankie Paul was married to Tate Paul after meeting him in January 2016. On August 7, 2016, the couple got to know each other after dating for seven to eight months. AdvancedMD currently employs Tate as a district manager. On August 31, 2017, Taylor Frankie Paul gave birth to their first child, a daughter Indy Paul. powerapps patch table Tate Britain Free Guided Tours and Talks Tours run most days at 12.00, 13.00 and 14.00 Learn about the art in Tate Britain with a talk on a number of artworks or a guided tour around … Free Tate Britain Tour Tate Britain Members Welcome Tour 10 Feb 2023, 11 Feb 2023, 10 Mar 2023, 11 Mar 2023 Take a free welcome tour of Tate Britain Now booking There is no one currently known as Andrew Tate's wife. Although there is no definitive answer to the question of whether or not Andrew Tate is married, it seems likely that he is not. There are no public records of him being married, and he has not been seen with a wife or partner in any of the many public appearances he has made over the years.As enviable as their relationship would become, when he first met Woodward in 1953, Newman was still married to his first wife, Jackie Witte, with whom he had three children. The handsome 28-year-old was making his Broadway debut in the play Picnic. It was a hot August day, and Woodward, then 22 and an understudy in the play, was sweating in ...Tate Paul (@mrtatepaul) on TikTok | 166.8K Likes. 94.2K Followers. Booking: [email protected] Check out my Soundcloud.Watch the latest video from Tate Paul (@mrtatepaul).Witch or Wife: Directed by William Asher. With Elizabeth Montgomery, Dick York, Agnes Moorehead, Irene Vernon. When Larry and Louise Tate stumble upon Samantha and Endora having lunch in Paris, she can't return since they think that Darrin sent her over for a mortal vacation. farm for sale bridgnorth Ben Affleck's wife Jenner Lopez and his ex-wife Jennifer Garner came together for the former lovers' daughter's musical recital.The Argo star was seen with his newly blended family...Tom Brady and Gisele Bündchen to file for divorce today. When Elise Tate and husband Golden Tate decided to teach their then-9-month-old daughter sign language, they didn't realize how ... gumtree housekeeping jobs london Tate Britain Free Guided Tours and Talks Tours run most days at 12.00, 13.00 and 14.00 Learn about the art in Tate Britain with a talk on a number of artworks or a guided tour around … Free Tate Britain Tour Tate Britain Members Welcome Tour 10 Feb 2023, 11 Feb 2023, 10 Mar 2023, 11 Mar 2023 Take a free welcome tour of Tate Britain Now booking Jan 13, 2023 · Tate – who has had a long-standing beef with Paul, which involved name-calling on both sides – re-ignited the feud from a Romanian jail, where he is being held on sex-trafficking and rape charges. Tate on Wednesday tweeted: "Lol. Even in Romanian Prison they talk about Logan Paul being a scammer." Tate may have been referring to …Nov 10, 2022 · Dominic Smithers. Jake Paul and Andrew Tate have finally met face-to-face. The pair of internet stars appear to be lining up a fight, with them seen standing up to one another bare-chested in a ...The presenter and his now-wife Charlotte Godfrey first met on the set of Flog It!, which Paul has been presenting on since the show first aired in 2002. At the time Charlotte worked as a ... can unvaccinated children travel to usa Jan 20, 2023 · Andrew Tate and his brother Tristan have had their detentions in Romania extended for a second time. The siblings are to be held in custody until 27 February …26 de abr. de 2022 ... Learn about Taylor Frankie Paul's Age, Height, Weight, Net Worth, TikTok, Instagram, Family, Husband, Tate Paul, Children, Son, Daughter.Both became good friends with Shelley Winters, who played a key role in introducing Jones to his future wife, Susan Strasberg. They married in 1965, after living together, when Strasberg became pregnant. Served six months in a military stockade on Governor's Island in New York, NY, for going AWOL from the army. farms for sale gloucestershire Aug 6, 2022 · Shanti Das. Andrew Tate says women belong in the home, can't drive, and are a man's property. He also thinks rape victims must "bear responsibility" for their …Just because you're married doesn't mean you have to sleep together every night. Influencer Taylor Paul shared a video revealing the reasons why she and her husband sleep in separate bedrooms....Aug 7, 2022 · Tate first called out Logan's brother Jake making a $3m fight offer back in 2020, when the YouTuber was setting out on the early stages of his boxing career. He reiterated that offer again more ... trauma healing retreat ontario By The Editors of Encyclopaedia Britannica. Paul McCartney's first wife was Linda McCartney (formerly Linda Eastman), an American photographer, musician, and entrepreneur, whom he married in 1969. They formed the band Wings, which toured the world and became the best-selling pop act of the 1970s, with 27 U.S. Top 40 hits.Jan 2, 2023 · Tristan Tate currently doesn't have a wife after he broke up his relationship with Bianca Dragusanu. Tristan Tate is a former athlete and businessman from Luton. In 2011, he appeared on the British reality television program Shipwrecked: The Island, where he gained notoriety for his outspoken views. how to access gstt email from home By The Editors of Encyclopaedia Britannica. Paul McCartney's first wife was Linda McCartney (formerly Linda Eastman), an American photographer, musician, and entrepreneur, whom he married in 1969. They formed the band Wings, which toured the world and became the best-selling pop act of the 1970s, with 27 U.S. Top 40 hits. weight gain simulator game 27 de abr. de 2022 ... Taylor Paul said she sleeps in a different bedroom to her husband, Tate. (TikTok). READ MORE: 'My family is putting too much pressure on me ...Paul Tate, 82, a former Army intelligence officer who went undercover seeking clues to the murder of daughter Sharon Tate by the followers of Charles Manson, ... Tate and his wife, Doris, wrote ...Side By Side Books Showing 1-28 of 28 Side by Side: Student Book 2 (Paperback) by Steven J. Side by Side Book 1 (2nd Edition) 2nd Edition by Steven J. It's also owned and operated by Holocene. pdf - Free ebook download as PDF File (. Anger at Tate for not listening, and being so stubborn. The riddle appeared in an 1847 edition of The.As excited as he is to fight Tate, Paul doesn't believe the 35-year-old will ever agree to share the ring with him. ... LeBron James' first NBA coach Paul Silas dead at 79. ... US journalist's cause of death remains a mystery as wife pays tribute. Latest. TRAGIC DEATH. Soccer journalist Grant Wahl dead aged 48 in Qatar while covering World Cup ...23 de mai. de 2022 ... Taylor Frankie Paul is married to Tate Paul, whom she adores. The couple first met in January 2016 and began dating shortly after. On August 7, ... lima mora jevremovic Tate Paul's wife is Taylor Frankie Paul who is also a social media star and a popular TikToker in the USA. They got married in September 2016. They first met the same year in the month of January. and has married after 8 months of dating. Taylor Frankie Paul and her husband Tate PaulView the profiles of people named Tate Paul. Join Facebook to connect with Tate Paul and others you may know. Facebook gives people the power to share...Apr 19, 2007 · Tate. @Tate. ·. Jan 15. Gerhard Richter (born 1932) is one of the most important German artists in history. Enjoy the rich surfaces of the artist's abstract works, painted to the music of avant-garde composer John Cage, on free display at Tate Modern. bit.ly/3QH3o1W. 19 de set. de 2022 ... Tate has been relatively quiet on his social media accounts following his divorce after his ex-wife Taylor Frankie Paul admitted to "soft ...In reference to the apostle Paul, he was not married during his years of travel and ministry, but many believe he was a widower. Paul offers marital advice that is very romantic and positive, suggesting that he once was married. lucky mobile Here's a look at Pollard and their relationship. 1. Pollard Works at Harvey Nash in Los Angeles. According to her LinkedIn bio, Pollard is a director of business development at Harvey Nash. She ...2023-01-23T01:00:00 | Joshua Boren was a police officer in Utah. In 2014, after failing to show up to work, officers went to his house and saw cause for concern through the window of his house. When they got inside, they found that Joshua Boren had murdered his wife, two children, and his mother-in-law and then took his own life. Join Mike and Gibby as … lifa app template notion View the profiles of people named Tate Paul. Join Facebook to connect with Tate Paul and others you may know. Facebook gives people the power to share...September 20, 2022 Taylor Frankie Paul's lucky charm, Brayden Rowley, has been pursued by a number of suitors. She may be moving in with Rowley despite the rumors of their breakup. In addition to being an American artist, Taylor Frankie Paul is a well-known web-based entertainment creator and novelist. hampole dog rescue doncaster Sharon Tate, in full Sharon Marie Tate, (born January 24, 1943, Dallas, Texas—killed August 9, 1969, Los Angeles, California), American actress and model. The second wife of film director Roman Polanski, Tate was brutally murdered (along with four others) by followers of Charles Manson at the couple's rented home in Los Angeles in 1969 (see Tate murders). Tate was born to Paul James Tate ...Aug 7, 2022 · Tate first called out Logan's brother Jake making a $3m fight offer back in 2020, when the YouTuber was setting out on the early stages of his boxing career. He reiterated that offer again more ... minecraft altsMisogynist influencer Andrew Tate is feuding with fellow influencer Logan Paul again: this time, from a Romanian jail. Tate — who has had a long-standing beef with Paul, which involved name-calling on both sides — re-ignited the feud from a Romanian jail, where he is being held on sex-trafficking and rape charges. Tate on Wednesday tweeted ...Golden and Elise Tate Got Married in Cabo! After a somewhat lengthy engagement (not to mention helping plan other people's romantic proposals through the Golden Future Foundation), Golden and Elise Tate finally tied the knot on March 11, 2017.Fifty years after her tragic murder at the hands of Charles Manson's cult followers, actress Sharon Tate is in the limelight again - her innocent and playful persona embodied by Margot Robbie in... indeed jobs burlington iowa By Stacy Carey / March 24, 2021 9:41 pm EST. Paul McCartney lost his first wife, Linda McCartney, in 1998 and he's been quite open in the years since then regarding how heartbreaking the loss was ...Tate is an American Western television series starring David McLean that aired on NBC from June 8 until September 14, 1960. [1] [2] It was created by Harry Julian Fink (the creator of Dirty Harry and T.H.E. Cat ), who wrote most of the scripts, and produced by Perry Como 's Roncom Video Films, Inc., as a summer replacement for The Perry Como Show . hvt smart contract address Paul Tate Paul and Ruby were once high school sweethearts. They truly loved each other and even talked about marriage. Octavious and Gladys Tate opposed their relationship and it wasn't until Grandmere Catherine told Ruby that Paul was her half brother that Ruby broke up with him.By Paul . The part of Louise, Larry Tate's wife, was played by two different actors during the eight year run of Bewitched. The first Louise was Irene Vernon who appeared in 11 episodes during the first two years of Bewitched. At the end of the second season she retired from the show to, as the story goes, pursue real estate opportunities in ...Geoff Tate (born Jeffrey Wayne Tate, January 14, 1959; he later changed his first name to Geoffery or Geoffrey) is an American singer and songwriter. He rose to fame with the progressive metal band Queensrÿche, who had commercial success with their 1988 album Operation: Mindcrime and 1990 album Empire.Tate is ranked fourteenth on Hit Parader ' s list of the 100 Greatest Metal Vocalists of All ...No. 2 Marriage 4 Children. Contents: He is a singer and musician, best known as the vocalist of the metal band Queensr che. His birth name is Jeffrey Wayne Tate. In the Hit Parader's list of the 100 Greatest Metal Vocalists of All Time, he was ranked 14 and was placed 9th on 2015 OC Weekly's list of the 10 Best High Pitched Metal Singers. border collie puppies for sale ontario The most famous victim was actress Sharon Tate, wife of film director Roman Polanski, who was killed in her Los Angeles home along with three guests. The True Story of Charles Manson and Sharon Tate Is Devastating. late in the night between the 8th and 9th of august, 1969, tate, who was eight months pregnant at the time, was murdered in her los angeles …Aug 6, 2022 · Shanti Das. Andrew Tate says women belong in the home, can't drive, and are a man's property. He also thinks rape victims must "bear responsibility" for their …4 de ago. de 2022 ... ... Taylor Frankie Paul introduced TikTok followers to her new boyfriend, Dakota Mortensen, amid her nearly-final divorce with ex Tate Paul. ue5 landscape material not showing Logan Paul is hoping to face Andrew Tate next (Image: @jakepaul/JEFF FM) Paul has been a vocal critic of Tate's rhetoric and was a staunch supporter of his ban from social media earlier this year.Colonel Paul Tate spent the night before the funeral, beside his daughter's casket. He had had them open the casket so he could have one last look at Sharon, who lay peacefully in her favourite Pucini dress Roman had picked out, and his grandson who was swaddled and tucked under his mother's arm. Then he closed the lid and said good-bye.30 de mai. de 2022 ... She's married to Tate Paul, and they often create content together. ... Taylor Frankie Paul Announces Her Divorce. mobile homes for sale in caravan parks Sep 23, 2022 · Tylor is married to Tate Paul. Tate Paul and she met for the first time in January 2016 and began dating. Tate proposed to Tylor on August 7, 2016, after 8 … autotrader campervan for sale Logan Paul is hoping to face Andrew Tate next (Image: @jakepaul/JEFF FM) Paul has been a vocal critic of Tate's rhetoric and was a staunch supporter of his ban from social media earlier this year.Taylor Frankie Paul was married to Tate Paul after meeting him in January 2016. On August 7, 2016, the couple got to know each other after dating for seven to eight months. AdvancedMD currently employs Tate as a district manager. On August 31, 2017, Taylor Frankie Paul gave birth to their first child, a daughter Indy Paul.Paul Tate Paul and Ruby were once high school sweethearts. They truly loved each other and even talked about marriage. Octavious and Gladys Tate opposed their relationship and it wasn't until Grandmere Catherine told Ruby that Paul was her half brother that Ruby broke up with him.16 de ago. de 2022 ... Paul announced the end of her marriage during a May 25 TikTok live stream in which she told viewers she and her husband Tate Paul were in an ... ocr health and social care level 3 unit 3 mark scheme Paul Tate, 82, a former Army intelligence officer who went undercover seeking clues to the murder of daughter Sharon Tate by the followers of Charles Manson, ... Tate and his wife, Doris, wrote ...Dec 30, 2022 · Born Emory Andrew Tate lll at Walter Reed Army Hospital in Washington, DC, on Dec. 1, 1986, Tate has said he came out of the womb a fighter. He was named after his dad, a black military veteran ... free shemale facial galleries Aug 6, 2022 · Shanti Das. Andrew Tate says women belong in the home, can't drive, and are a man's property. He also thinks rape victims must "bear responsibility" for their …Aniston's first serious relationship was with the late Daniel McDonald. The duo dated for five years in the early 2000s, with Aniston later citing the actor as the one who got away. "He was my...You heard it here first: Tate McRae is taking over the music scene. There's a 90% chance Tate's viral song "She's All I Wanna Be" has landed on your TikTok For You Page and if you haven't...Paul Tate Married Relationship facts. Doris Tate · His Other Children · Children Together · Her Other Children · Who's In The News?...Taylor Frankie Paul, 28, has blown up #MomTok after announcing on TikTok that she and her husband, Tate Paul, are getting a divorce In a bombshell TikTok Live, she opened up about their... casino royal club no deposit bonus 2022 house. She is first seen as Samantha greets her after dinner and offers to get her a drink. Samantha defers to Louise, as the Boss' wife, by referring to her as "Mrs. Tate," though Louise refers to Samantha by her given name. Louise isNov 11, 2022 · KSI is also active in boxing and recently created history when he beat two opponents on the same night.He has fought Logan before and defeated him, but recently stated that he has returned to the boxing realm with the final goal of beating Jake. Jake Paul wants to fight Andrew Tate in 2023. Jake Paul and Andrew Tate have been going back …Jun 28, 2020 · Peter Paul's first wife files a complaint against his marriage with Vanitha Vijayakumar; It's a baby boy for Daniel Annie Pope; Fast-track your journey to being a …The beautiful Sharon Tate would have celebrated her 80th birthday on Jan. 24, 2023. ... Smokey Robinson reveals he cheated on his first wife with Diana Ross, more news. Edgy and Elegant! 3 bedroom house to rent in oldbury Paul McCartney Remembers Late First Wife Linda With Instagram Post: 'Beautiful Memories' On April 17, the 75-year-old legend took to the platform to share a picture of his first wife Linda on ...The Tate family name was found in the USA, the UK, Canada, and Scotland between 1840 and 1920. The most Tate families were found in USA in 1880. In 1840 there were 95 Tate families living in Virginia. This was about 15% of all the recorded Tate's in USA. Virginia had the highest population of Tate families in 1840. Mae Brussell was a case of the latter, admitting that the government was up to no good, but then preventing you from seeing what they were really up to. As alternative as Brussell seemed to be, she never got to the truth. She always fingered the CIA, which turns out to be true enough, but beyond that she was always in sea of molasses.Tate, who rose to fame in the middle of 2021 through videos making misogynistic claims about women and deployed an army of diehard fans to propagate his content, was banned from nearly every... craigslist vancouver b c Ben Affleck's wife Jenner Lopez and his ex-wife Jennifer Garner came together for the former lovers' daughter's musical recital.The Argo star was seen with his newly blended family...May 27, 2022 · Tate Paul Age. Tate Paul was reportedly born in 1996. That made him 26 years of age in 2022. He is 2 years younger than his wife Taylor who was born on May … vnjshr 10 de nov. de 2022 ... He beat Deji Olatunji, a YouTuber and brother of KSI, by knock out. His boxing nickname is The Problem Child. His first professional fight came ...By The Editors of Encyclopaedia Britannica. Paul McCartney's first wife was Linda McCartney (formerly Linda Eastman), an American photographer, musician, and … ground floor flats to rent Taylor Frankie Paul, 28, has blown up #MomTok after announcing on TikTok that she and her husband, Tate Paul, are getting a divorce In a bombshell TikTok Live, she opened up about their...May 26, 2022 · Some users have shared photos of a woman they claim is Rowley's wife. Taylor has been married to Tate Paul since 2016. The couple are parents to son Ocean and a daughter Indy.Misogynist influencer Andrew Tate is feuding with fellow influencer Logan Paul again: this time, from a Romanian jail. Tate — who has had a long-standing beef with Paul, which involved name-calling on both sides — re-ignited the feud from a Romanian jail, where he is being held on sex-trafficking and rape charges. Tate on Wednesday tweeted ...May 26, 2022 · Some users have shared photos of a woman they claim is Rowley's wife. Taylor has been married to Tate Paul since 2016. The couple are parents to son Ocean and a daughter Indy. why does fetterman cover his neck

redpajama_set_name: RedPajamaCommonCrawl


Private, docent-led tours are a wonderful way for groups of 10 or more to enjoy the museum and share a fun learning experience. Let us help create a memorable visit tailored to your group's needs and interests. Group visits must be scheduled at least four weeks in advance. Contact us today to get started!
Private, docent-led tours are available to groups of 10 or more. We can accommodate up to 50 people at a time.
Our docents will lead a 45-minute highlights tour, providing a broad overview of the museum with a focus on key works of art. There is much more to see, so you may want to plan for additional time to explore on your own after the tour.
Use the "Request a Tour" button to access the online reservation request form. Tours must be scheduled at least four weeks in advance. We will contact you by email to confirm your tour date and time. If we are unable to accommodate your group on the date(s) you requested, we will work with you to schedule your visit for an alternate date.
A $50 deposit is required to secure a tour date, and must be paid within two days after you receive confirmation that your date has been scheduled. Call 727-892-4200 ext. 1021 to pay by Visa or Mastercard.
The balance of your admission is due two business days prior to your visit. Be sure to have an accurate headcount when you make your final payment, as tickets are nonrefundable. Call 727-892-4200 ext. 1021 to pay by credit card. If you are paying by check, please ensure that it arrives at least two days prior to your visit.
Located inside the museum, The Canyon Cafe welcomes groups for lunch, dinner and drinks. Reservations are required for groups, as seating is limited. To make a reservation, please contact the Cafe's General Manager at jess@datztampa.com or 727-892-4200 ext. 1202.
Individual vehicles (height limit of 6'10") may park in the city garage located above the museum for an hourly rate of $2. The garage entrance is on the back of our building, on 1st Ave South. Metered street parking is also available in the vicinity.
Buses and large vans may pull up to the museum's front entrance at 150 Central Ave. to allow passengers to disembark. We will provide directions to an offsite parking lot upon arrival.
If you need to cancel your tour reservation, please do so in writing. Your deposit is fully refundable if you cancel no less than 30 days prior to your tour date. Or you may reschedule your visit based on museum availability.
For more information, please call 727-892-4200 ext. 1021 or email education@thejamesmuseum.org.


text:
Cantatrix sopranica L. et autres récits scientifiques es un libro póstumo del escritor francés Georges Perec (1936-1982), publicado en 1991 en la colección «Librairie du XXe siècle» de la editorial Seuil y que no ha sido traducido al castellano.

El libro recopila cuatro artículos pseudocientíficos, escritos entre 1974 y 1980 como parodias y aperturas del concepto de literatura.

Contenido

Referencias

Procedencias

Bibliografía

Libros de Georges Perec
Libros de 1991
Libros póstumos

redpajama_set_name: RedPajamaWikipedia


text:
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Earthquakes Devastate Italy
Italians try to pick up the pieces after two earthquakes in the northeastern region of the country strike
Italians have been wracked with sadness and fear as two earthquakes left a path of destruction in northeastern Italy. They are glued to the TV checking in on the residents of the region, most of whom are now living in tents, getting updates on aftershocks, listening to stories of tragedy and survival, and wondering how all this property damage is going to weigh into an already difficult and growing economic crisis.
Two earthquakes hit the provinces of Mantova, Modena, and Ferrara on May 20 and 29, according to the the Italian news agency AGI. As I write this AGI is reporting that the magnitude and number of aftershocks is finally falling with only six yesterday and the highest having a magnitude of only 2.6. To put the number in perspective, the agency reported 30 aftershocks on May 29 and the quake had a magnitude of 5.3. It killed a total of 19 people.
The images we here in Italy have been seeing on the news are devastating. There are buildings that were hundreds of years old laying in bits and pieces on the street. People – tired, hungry, and now perhaps poor – in tents not far from the debris. And the stories people keep discussing over coffee and in passing on the street are enough to make you cry.
One man had just been telling his colleague that he wanted out of a factory job that was going nowhere. He was determined to find something else because he was so unhappy with his work life. The next day, he would die in the factory as the earthquake shook and the building fell upon him. Other stories were tragic but uplifting. There were the firemen who saved a woman who had been trapped for a day under debris. What saved her? A piece of furniture in her kitchen, which protected her from life-threatening injuries.
Reports about the lost Parmigiano-Reggiano cheese, which had been aging for years and now is stuck under piles of garbage and debris and the possibility of having lost balsamic vinegar of Modena have been flying about. Some shoppers are stockpiling these goods because they're concerned the prices will go up even further and they won't be able to afford them. Indeed, after the loss of life and property, people are wondering about how this will factor into the economic crisis hitting all of Europe. As Greece and Spain aim to stay alive, Italy's barely safe itself. Before the earthquake, the news was full of stories about those who were committing suicide because they are unemployed, poverty stricken, and hopeless. Repairing these damaged cities, helping these interrupted businesses, and getting people back to work and school and back into normal homes as opposed to tents costs money, which is something Italy doesn't really have at the moment. What is the country to do?
Thinking about the loss of life and having to rebuild entire cities, many of which have a history that goes back thousands of years and now has been somewhat lost is too much to bear. It has gotten the Italians to be even more introspective than usual. Many of them are considering what would happen if a quake struck them, especially here where I am in Ischia, a province of Naples that has a history of seismic activity. Ischia was itself a volcano before becoming an island.
Today's Ischitani – at least those of a certain age – remember the 1980 earthquake that struck Napoli. They all know exactly where they were when the earth began to shake. Many of them say that in the seconds just before the quake, they felt ill and thought they were experiencing some sort of imbalance and then the earth literally moved. Most, rather than taking cover in a doorway or under a table, ran to the nearest exit to get outside. Now, they realize it would have been better to do what they were told and protect their heads from falling debris. They vividly recall sleeping outside for days afterward as they felt aftershocks. Word of any possibility of nearby Vesuvius awaking from its slumber or these other quakes in Italy shifting the earth are among their greatest nightmares. They're not the only ones worrying. An article in Time this week describes the writer's concerns about tremors in Rome, where earthquakes hardly ever a hit and aren't a real threat. While Italians are known to be passionate people who can make the earth move figuratively, they don't want it to move literally.
Di Meglio is the Guide to Newlyweds for About.com, and you can follow her life and work at the Two Worlds Web site.
Article Published 6/4/2012
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Hello, I'd like more information about the listing I found on Homes.com at 18 Alfred Buck Lane.
This home is located at 18 Alfred Buck Lane Pittsford, VT 05736 US and has been listed on Homes.com since 24 March 2019 and is currently priced at $209,000, approximately $138 per square foot. This property was built in 1997. 18 Alfred Buck Lane is within the school district(s) Pittsford School District and Otter Valley Union High School District 8 with nearby schools including Lothrop School and Otter Valley Uhsd 8. Similar properties nearby are 587 Fire Hill Rd, 350 Oxbow Rd, 4152 Us Route 7, and 129 Elm St.


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<title>Uses of Interface net.sf.jasperreports.engine.JRImage (JasperReports 5.6.0 API)</title>
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net.sf.jasperreports.engine.JRImage

Packages that use JRImage

Package
Description

net.sf.jasperreports.engine

Provides access to the library's main functionality through facade classes for compiling, filling
 and exporting reports and also publishes the library's main interfaces and classes.

net.sf.jasperreports.engine.base

Contains base implementations for the library's main interfaces.

net.sf.jasperreports.engine.convert

net.sf.jasperreports.engine.design

Contains design time implementations for the library's main interfaces as well as report compiling tools.

net.sf.jasperreports.engine.fill

Contains fill time implementations for the library's main interfaces and the entire
 engine used in the filling process (the actual core of JasperReports).

net.sf.jasperreports.engine.util

Contains utility classes for the core library.

net.sf.jasperreports.engine.xml

Contains classes for loading report templates from XML files.

Uses of JRImage in net.sf.jasperreports.engine

Methods in net.sf.jasperreports.engine with parameters of type JRImage

Modifier and Type
Method and Description

void
JRExpressionCollector.collect(JRImage image)

void
JRVisitor.visitImage(JRImage image)

Uses of JRImage in net.sf.jasperreports.engine.base

Classes in net.sf.jasperreports.engine.base that implement JRImage

Modifier and Type
Class and Description

class
JRBaseImage

The actual implementation of a graphic element representing an image.

Methods in net.sf.jasperreports.engine.base with parameters of type JRImage

Modifier and Type
Method and Description

void
JRBaseObjectFactory.visitImage(JRImage image)

Constructors in net.sf.jasperreports.engine.base with parameters of type JRImage

Constructor and Description

JRBaseImage(JRImage image,
           JRBaseObjectFactory factory)

protected JRBaseImage()
        {
                super();
        }

        /**
 Initializes properties that are specific to images.

Uses of JRImage in net.sf.jasperreports.engine.convert

Methods in net.sf.jasperreports.engine.convert with parameters of type JRImage

Modifier and Type
Method and Description

void
ConvertVisitor.visitImage(JRImage image)

Uses of JRImage in net.sf.jasperreports.engine.design

Classes in net.sf.jasperreports.engine.design that implement JRImage

Modifier and Type
Class and Description

class
JRDesignImage

The actual implementation of a graphic element representing an image, used at design time.

Uses of JRImage in net.sf.jasperreports.engine.fill

Classes in net.sf.jasperreports.engine.fill that implement JRImage

Modifier and Type
Class and Description

class
JRFillImage

Methods in net.sf.jasperreports.engine.fill with parameters of type JRImage

Modifier and Type
Method and Description

protected void
JRTemplateImage.setImage(JRImage image)

void
JRFillObjectFactory.visitImage(JRImage image)

Constructors in net.sf.jasperreports.engine.fill with parameters of type JRImage

Constructor and Description

JRFillImage(JRBaseFiller filler,
           JRImage image,
           JRFillObjectFactory factory)

JRTemplateImage(JROrigin origin,
               JRDefaultStyleProvider defaultStyleProvider,
               JRImage image)

Uses of JRImage in net.sf.jasperreports.engine.util

Methods in net.sf.jasperreports.engine.util with parameters of type JRImage

Modifier and Type
Method and Description

void
JRVisitorSupport.visitImage(JRImage image)

void
JRDelegationVisitor.visitImage(JRImage image)

void
UniformElementVisitor.visitImage(JRImage image)

void
JRApiWriter.writeImage(JRImage image,
          java.lang.String imageName)

Uses of JRImage in net.sf.jasperreports.engine.xml

Methods in net.sf.jasperreports.engine.xml with parameters of type JRImage

Modifier and Type
Method and Description

void
XmlWriterVisitor.visitImage(JRImage image)

void
JRXmlWriter.writeImage(JRImage image)

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text:
Arctic Portal
AP Community
Arctic Data Committee (ADC)
ADC meetings
ADC members
ADC web site
Committee on Observations and Networks (CON)
CON Meetings
CON Members
CON Inventory work
CON Work Plan
CON Terms of Reference
Road Map Task Force (RMTF)
Goal 3 Task Force
H2020 Task Force
SAON Initiating Group
Stockholm workshop
Aim & format
Material presented
Edmonton workshop
Helsinki workshop
St Petersburg workshop
Agency Officials workshop
Data Management workshop
SAON News
SAON Committee on Observations and Networks (CON)
The Committee will meet 9th December in Ottawa, Canada.
The Chair of the Committee is Lisa Loseto (Lisa.Loseto(at)dfo-mpo.gc.ca).
1st International Arctic Products Validation & Evolution (APVE) Workshop
http://calvalportal.ceos.org/events/apve-workshop
Permafrost Research Priorities Survey
The International Permafrost Association (IPA) and the Climate and Cryosphere project (CliC) are seeking your input. We invite you to participate in a survey on 'Permafrost Research Priorities', which aims at identifying the top priorities in permafrost research. The process will span much of 2014 engaging the research community and will result in a short publication listing and putting into context research priorities. The document shall become the benchmark against which permafrost research should be gauged starting in 2015. The final document of priorities will be sent to national and international funding agencies, international organizations, policy makers, and others with interests in supporting permafrost research. It will form one of the outputs of the International Conference on Arctic Research Planning (ICARP III).
We are interested in collecting forward-looking research questions (max. 3) from individuals with professional interests in understanding physical, biogeochemical, ecological, and social processes that affect permafrost areas in the Arctic, the Antarctic and in mountain regions.
Read more: Permafrost Research Priorities Survey
Arctic Observing Assessment: Phase II
Thank you to all that contributed to the first phase of the Arctic Observing Assessment. In recent weeks, a first round of anonymous input about societal priorities in the Arctic was received by phone, e-mail, and the web. These responses were wide-ranging. A draft attempt has been made to group these responses into categories that would serve as the basis for the next steps of the Assessment.
At this time, 11 categories have been identified and made public in an online survey. Your input is again welcomed for the next step of the Arctic Observing Assessment for the Interagency Arctic Research Policy Committee (IARPC) and the Sustaining Arctic Observing Networks (SAON). Please visit http://bit.ly/ssipcomment to review these categories and their descriptions and provide feedback about missing categories or edits to category descriptions.
Input is requested by August 8, 2014.
Read more: Arctic Observing Assessment: Phase II
Arctic Observing Assessment
Your input is needed to help identify Societally Significant Information and Products (SSIPs) to assess the capacity of observational networks in the Arctic.
The Arctic Observing Assessment (AOA) is being conducted to support the U.S. Interagency Arctic Research Policy Committee (IARPC) and the international Sustaining Arctic Observing Networks (SAON) objectives for observing coordination and data interoperability. Details about the assessment, its relationship to IARPC and SAON, including FAQ's, are available on the ArcticHub (http://www.arctichub.net) under the Arctic Observing Assessment header. In this first step of the assessment, we are building a list of Arctic societal priorities that will help scope the effort.
There are two methods for submitting Arctic societal priorities to this effort – via an anonymous online web form (http://bit.ly/ssip2014) or via e-mail . Online and e-mail submissions will be compiled into one list. Additional priorities will be sourced from available Arctic vision, strategy, and priority documents and reports. Examples of Arctic priorities that have been drawn from documents and could scope this effort include food security, freshwater security, and coastal vulnerability.
A full list will be circulated via the ArcticHub. The deadline for input via the form and e-mail is July 11, 2014. We encourage you to distribute and share this message widely to make this opportunity for input known to all.
There will be future opportunities to provide input into next stages of this assessment. Please visit the ArcticHub (http://www.arctichub.net) for updates.
SciDataCon 2014
The World Data System (WDS) and Committee on Data for Science and Technology (CODATA), interdisciplinary bodies of the International Council of Science (ICSU), will be convening the International Conference on Data Sharing and Integration for Global Sustainability (SciDataCon 2014) in New Delhi from 2-5 November 2014 (www.SciDataCon2014.org/).


LONGLAC MAN ACTING STRANGE IN GERALDTON DISTRICT HOSPITAL ON CRACK AND METH PLEADS GUILTY AND GETS A SUSPENDED SENTENCE AND PROBATION.
Appearing in courtroom 102 before His Honour Dino DiGiuseppe is a 54 year old GUY FORTIER of Longlac to plead guilty today to possession of crack cocaine.
He's about 5'5″ and a bit pudgy. He has a very noticeable walk related to a disability he claims he got operating heavy equipment at the Greenstone Landfill sight.
Federal Crown Attorney Alec Hardiejowski is present along with defence attorney Gil Labine.
Admitted facts before the court were that March 2,2018, GUY FORTIER was admitted to Geraldton District Hospital for medical treatment. On March 9,2018 the hospital staff noticed FORTIER behaving very "strange". The Doctor requested a urine test from Fortier.
The urine test came back positive for drugs which were not prescribed to him.
Out of concern for other patients in the immediate vicinity, hospital staff searched the duffel bag that Fortier brought to the hospital with him.
They found a pill container with pills of methamphetamine in it, along with .1 gram of crack cocaine. Greenstone OPP were called and charges were laid.
Fortier did not want to leave his illegal drugs at home, so he brought them to hospital. He mistakenly took a methamphetamine pill instead of a sleeping pill as an explanation for his strange behavior.
The court hears that Fortier receives long term disability and CPP.
He also has previous drug conviction from 2011. His Honour wonders how someone on limited income can afford expensive drugs but agrees to the joint submission of 12 month suspended sentence and probation. The possession of methamphetamine charge is withdrawn by the crown. He's also ordered to pay a 100$ victim surcharge fine.


Allow us to introduce you to two of our own corporate hot air balloons, G-OOEY 'Exclusive One' and G-UNKY 'Exclusive Two'.
One of the key criteria of any marketing company is their confidence in their own products. This is another factor setting us apart from our competition as we invest heavily in our own branded hot air balloons and marketing programs.
'Exclusive One' is a Kubicek balloon built in the Czech Republic using only the highest grade of polyester fabrics ensuring that the balloon has a life expectancy of 1,000 hours. At 77,000 cubic feet, we use it as a camera platform for filming our client's balloons and also as marketing for Exclusive Ballooning. In fact, One's first public outing at the Exclusive Cup resulted in victory of the entire event.
'Exclusive Two' is One's baby brother at just 50,000 cubic feet. Built in Spain by Ultramagic balloons, it has a fully branded matching collapsible basket meaning the balloon is easy to transport. Two's first outing was at the European hot air balloon fiesta in Spain where it won the fly in to the factory.
It is now most famous for being the only balloon in the world to fly in every U.S. state, demonstrating how flexible and portable our product is.
If you'd like to find out more about what our balloons can do for your brand, connect with us today.


Stay tuned for further developments. People who care deeply about our community and have ideas about ways to improve it are encouraged to volunteer and/or run for the HRB. Please feel free to contact the HRB directly.


text:
@class PFImageView;
@interface PAPPhotoCell : PFTableViewCell

@property (nonatomic, strong) UIButton *photoButton;

@end


text:
Home > Celebrity > Directors
Real name Darren Aronofsky
Date of birth February 12, 1969
Occupation Filmmaker
Marrried
What is Darren Aronofsky relationship status? Single
What is his sexual orientation? Straight
Darren Aronofsky girlfriends list
3 Jennifer Lawrence Ex September 2016 October 2017 1 year 1 month
2 Brandon Milbradt Ex 2012 2015 3 years
1 Rachel Weisz Ex October 2001 September 2010 8 years 11 months
Couple story
Darren Aronofsky and Rachel Weisz
A super talented filmmaker and writer Darren Aronofsky was delivered to his parents Charlotte and Abraham, who worked as public school teachers. Also, the celebrity has a sister named Patti, who he is very close with.
His love for the moviemaking industry appeared when he saw Stars Wars. That's exactly when he started considering pursuing a career as a film director. In 1987 he entered Harvard University, where he learnt everything about filmmaking as well as got a degree in social anthropology in 1991. That same year he made his first short movie called Supermarket Sweep.
In order to boost career opportunities, Aronofsky moved to Los Angeles. In 1998 he released a TV project Pi that was presented at Sundance Film Festival, where he was awarded for the best direction. The success inspired Darren even more and in 2000 he presented Requiem for a Dream due to which he was nominated for an Oscar and Golden Globe.
Throughout his career, he introduced dozens of successful TV projects, including The Wrestler, Black Swan, Noah and many others. However, he never stops and keeps coming up with more and more interesting ideas for movies.
Are you eager to know more amazing information about the star's life? Have a look at these top 5 little-known facts:
He has a tradition not to shave his beard till the production is over.
He is known for his environmental activism.
He has a Humanitarian Award for PETA as well as Humane Society of the US.
He takes action against climate change.
He is a board member of The Sierra Club Foundation.
Tifany Landrob
Head Editor


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JS001, a humanized IgG4 monoclonal antibody against the programmed death-1 (PD-1) receptor, blocks the interaction of PD-1 with its ligands and promotes T cell activation in preclinical studies. This phase I study is designed to evaluate the safety, tolerability, and clinical activity of JS001 in advanced melanoma or urologic cancer patients who are refractory to standard systemic therapy.
In the dose escalation cohorts, subjects initially received a single-dose, intravenous infusion of JS001, and were followed for 28 days followed by multi-dose infusions every 2 weeks. In the dose expansion cohorts, subjects received multi-dose infusions every 2 weeks. Clinical response was evaluated after each 8-week treatment cycle according to RECIST v1.1 criteria.
Thirty-six subjects diagnosed with advanced melanoma (n = 22), urothelial cancer (UC) (n = 8), or renal cell cancer (RCC) (n = 6) were enrolled. Melanoma subjects included 14 acral and 4 mucosal subtypes. JS001 was well tolerated, and no dose-limiting toxicity was observed. By the safety data cutoff date, 100% of subjects had treatment-related adverse events (TRAE) with most adverse events being grade 1 or 2, and ≥ grade 3 TRAEs occurred in 36%. Among all 36 subjects, 1 confirmed complete response (acral melanoma), 7 confirmed partial responses (2 acral melanoma, 1 mucosal melanoma, 2 UC, and 2 RCC), and 10 stable disease were observed, for an objective response rate of 22.2% (95% CI, 10.1 to 39.2), and a disease control rate of 50.0% (95% CI, 32.9 to 67.1). Clinical responses were correlated with PD-L1 expression on tumor cells, the presence of tumor infiltrating lymphocytes (TIL), baseline tumor volume, ECOG performance status, serum LDH levels, high percentage of activated CD8+ T cells and CD3− CD16+ CD54+ NK cells in the peripheral blood as well as tumor mutational burden (TMB).
JS001 was well tolerated and demonstrated promising anti-tumor activity in UC and RCC as well as in previously underexplored acral and mucosal melanoma subtypes. Subjects with an immune-active profile in the tumor microenvironment or in peripheral blood responded favorably to JS001 treatment. The completion of the current phase I study has led to the initiation of the first prospective anti-PD-1 registration trial in Asia focusing on acral and mucosal melanoma subtypes, representative of the regional disease epidemiology.
Clinical Trial ID: NCT02836795, registered July 19, 2016, retrospectively registered.
Immune evasion is one of the hallmarks of cancer. Tumor-specific T lymphocytes are often present in the tumor microenvironment, in the draining lymph nodes, and in the periphery, but are generally unable to control tumor progression due to a network of immunosuppressive mechanisms present in the tumor microenvironment [1–3]. CD8+ tumor infiltrating T lymphocytes (TILs) often express activation induced inhibitory receptors including CTLA-4 and PD-1 , while tumor cells frequently express immunosuppressive ligands, including PD-1 ligand 1 (PD-L1, also known as B7-H1 or CD274), that suppress T cell activation and effector function . Among the suppressive mechanisms, PD-1 and its ligands have emerged as a crucial pathway exploited by tumor cells to inhibit activated T cells in the tumor microenvironment .
PD-1 is an inhibitory member of the CD28 receptor family expressed on activated B and T lymphocytes and myeloid cells [6, 7]. Interaction of PD-1 and B7 family ligands PD-L1 and PD-L2 delivers inhibitory signals that contribute to the establishment and maintenance of peripheral immune tolerance [8, 9]. PD-L1, the primary PD-1 ligand in the peripheral tissue, is overexpressed in many solid tumors, and PD-L1 expression has been identified as a negative prognostic biomarker in various cancers associated with poor survival [5, 10].
In the past decade, PD-1/PD-L1 pathway blockade has been proven to be an effective approach in inducing durable anti-tumor responses in various cancer indications. Monoclonal antibodies (mAbs) that block the PD-1/PD-L1 pathway can enhance tumor specific T cell activation and effector function, reduce tumor burden, and improve survival [11, 12]. From year 2014 to 2017, FDA has approved two anti-PD-1 mAbs (nivolumab and pembrolizumab) and three anti-PD-L1 mAbs (atezolizumab, avelumab, and durvalumab) for human malignancies. Melanoma was the first indication approved for both nivolumab and pembrolizumab in 2014 [13, 14]. Melanoma has long been regarded as an immunogenic cancer type due to frequently observed lymphocyte infiltration into the tumor, as well as clinical response to immunotherapy of high dose IL-2 . Mechanistically, chronic sun exposure-associated ultraviolet (UV)-induced DNA damage is the leading cause for melanoma in the western population. Chronic sun damage (CSD) melanoma accounts for 95% of cutaneous melanoma in the USA and other western countries . In contrast, acral lentiginous melanoma (ALM) (~ 50%) and mucosal melanoma (MM) (~ 20%) are the two most common melanoma subgroups in Asia . Neither ALM nor MM are associated with chronic UV exposure and thus harbor much fewer DNA mutations . Retrospective studies showed immunotherapies were less effective treating ALM and MM when compared with CSD melanoma, presumably due to their low immunogenic properties and different molecular mechanisms for tumorigenesis .
JS001 is a humanized IgG4 mAb specific for human PD-1 . Here, we report the first-in-human dose-escalation study of JS001 conducted at Peking University Cancer Hospital in metastatic solid tumor subjects refractory to standard systemic treatment, including 22 melanoma, 8 urothelial carcinoma (UC), and 6 renal cell carcinoma (RCC) subjects. Among 22 melanoma subjects, 14 (64%) were acral and 4 (18%) were mucosal melanoma subtypes. JS001 had an acceptable overall safety profile up to 10 mg/kg Q2W with no novel safety signal identified when compared with marketed drugs in the same class. Objective clinical responses were observed at every dose level and in all three indications.
JS001, also known as TAB001 or toripalimab, is a humanized IgG4κ mAb specific for human PD-1 receptor and blocks interactions of PD-1 with its ligands, PD-L1 and PD-L2. JS001 contains the complementary-determining regions (CDR) of a murine antibody that binds to human PD-1 and human framework regions with limited back-mutations to the parental murine sequences. A serine to proline substitution was introduced at amino acid 228 (S228P) to minimize Fab arm exchange of the IgG4 molecule. JS001 is produced by recombinant DNA technology in a Chinese Hamster Ovary (CHO) mammalian cell expression system (LONZA).
Eligible subjects were between 18 to 70 years old with metastatic melanoma or urologic cancers who were refractory to standard systemic treatment. Subjects had measurable disease per RECIST v1.1, Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1, adequate organ and marrow function, willingness to provide consent for biopsy samples, and no history of autoimmune diseases or ongoing infections. Concurrent antineoplastic therapies, systemic steroid treatment, and prior immunotherapies with anti-CTLA4, anti-PD-1, and anti-PD-L1 were not allowed. Prior monoclonal antibody treatments were allowed but not within 3 months of the start of JS001 treatment. Other prior non-antibody immunotherapies were also allowed but not within 4 weeks of JS001 treatment.
This is a single-center, phase 1, open-label, 2-part (part A dose-escalation and part B dose-expansion) study to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics, immunogenicity, and antitumor activity of JS001, administered via intravenous (IV) infusion on an every 2-week (Q2W) dosing schedule in adult subjects with advanced melanoma or urologic cancers refractory to standard therapy (Clinical Trial ID: NCT02836795). This phase I study was approved by Peking University Cancer Hospital institutional review board.
In part A, the study could enroll up to 18 subjects with advanced melanoma or urologic cancers that were immunotherapy-naïve (no prior exposure to immunotherapy such as but not limited to anti-CTLA-4, anti-PD-1, or anti-PD-L1 antibodies). The planned cohorts in part A were 1 mg/kg, 3 mg/kg, and 10 mg/kg Q2W. A minimum of 3 subjects were initially enrolled at each dose level. Sequential subjects in a cohort received a single 60-min IV infusion of JS001 and were evaluated for toxicity for 28 days. DLT was defined as a treatment-related grade 3 or above AE or laboratory abnormality occurring within 28 days after the first dose. Provided there was no DLT in the initial 3 subjects, escalation to the next cohort could occur. If a DLT did occur, that cohort would be expanded to a total of 6 subjects. After 28 days following the first dose, subjects continued to receive JS001 at the intended dose level Q2W, followed by radiologic evaluation every 8 weeks (Q8W). Responses were evaluated by investigators using the Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 and Immune-related Response Evaluation Criteria In Solid Tumors (irRECIST). Subjects with progressive disease or an intolerant toxicity were taken off the study. Patients who initially developed progressive disease per RECISTv1.1 were allowed to continue therapy if investigator considered patients were benefiting from the treatment per irRECIST.
Any dose-escalation cohort that did not exceed the maximum-tolerated dose (MTD) could be expanded up to a maximum of 12 subjects in part B for further evaluation of safety, PK, pharmacodynamics, and clinical activity. In part B, subjects received JS001 through IV infusion at either 1, 3, or 10 mg/kg per dose Q2W, followed by radiologic evaluation Q8W until disease progression, development of intolerable toxicity, or voluntary withdrawal from the study. AEs are reported as of July 31, 2018. The latest survival follow-up was provided by July 3, 2018.
Archival or fresh tumor biopsies were obtained from subjects prior to JS001 treatment. Detection of PD-L1 and CD8+ T cells by immunohistochemistry (IHC) staining of formalin-fixed paraffin-embedded (FFPE) tumor tissue samples was performed at Peking University Cancer Hospital. Spring Bioscience (Roche) rabbit anti-human PD-L1 mAb (Clone SP142, Cat No.: M4422) and CD8 antibody (Clone 4B11, BioRad, Cat# MCA 1817 T) were used for the PD-L1 CD8 double staining assay. Citrate buffer pH 6.0 was used for antigen retrieval, and a TSA plus biotin kit (Perkin Elmer) was used for signal amplification, followed by development with Peroxidase Substrate Kit for PD-L1 (red color), and ImmPRESS AP reagent and Blue substrate kit (Vector lab) for CD8 (blue color) staining. Stained tumor tissue was evaluated for PD-L1 expression on tumor cells (TC) by a certified pathologist. PD-L1-positive status was defined as the presence of membrane staining of any intensity in ≥ 1% of tumor cells.
Human tonsil tissue was used as dual positive and negative tissue control including positive tissue elements (moderate to strong PD-L1 staining in lymphocytes and macrophages in germinal centers, with diffuse staining in reticulated crypt epithelial cells) and negative tissue elements (PD-L1-negative immune cells in the inter-follicular regions with negative superficial squamous epithelium).
Serum samples were collected 0.5 h prior to dosing and serially at the end of infusion (EOI), 0.5, 2, 6, 12, 24, 48, 96, 168, 336, 504, and 648 h after the first dose of JS001. An electrochemiluminescence (ECL) method was developed and validated for the detection and quantitation of JS001 in human serum, with lower limit of quantification (LLOQ) at 2.56 ng/mL. The capture reagent was biotinylated His-tag human PD-1 extracellular domain, and the detecting reagent was ruthenylated His-tag human PD-1. For all assays, the samples were incubated with labeled capture and detection reagents to allow for the formation of molecular complexes. The complexed samples were loaded into the wells of a streptavidin-coated carbon-electron plates (MSD). After unbound material was removed by washing, MSD read buffer was added and the bound complexes were detected by reading ECL signals using an MSD instrument. Standard curve was established with JS001 serially diluted in human serum. PK parameters for evaluation included area under the concentration-time curve (AUC), maximum observed concentration (Cmax), clearance (CL), volume of distribution (Vd), and elimination half-life (t1/2).
Serum sample was collected before the first dose of JS001 and every 28 days prior to dosing. Another ECL method was developed and validated for the detection and quantitation of anti-JS001 antibodies in human serum. In this assay, anti-drug antibodies (ADA) were complexed between biotinylated and ruthenylated JS001. The negative control was a human serum pool. Positive control samples were prepared by spiking the negative control serum pool with purified rabbit polyclonal anti-JS001 antibodies. For all assays, the samples were diluted at 1:10 in master mix and then incubated with labeled drug to allow for the formation of molecular complexes. The complexed samples were loaded into the wells of a streptavidin-coated carbon-electron plates (MSD). After unbound material was removed by washing, MSD read buffer was added and the bound complexes were detected by reading ECL signals using a MSD instrument. Samples that produced signals higher than the screening cut point (SCP) S/N = 1.05 were considered positive samples and were subject to the confirmation process in a competition assay in the presence of unlabeled JS001. The screening confirmation cut point (CCP) was determined to be 10.8%. If the response of the test sample was above CCP and confirmed, it was reported as an ADA-positive sample. The corresponding JS001 trough concentration was also evaluated in ADA-positive samples for neutralizing activity.
The pharmacodynamics study of JS001 was assessed by its ability to occupy its natural antigen, PD-1 receptor on peripheral T lymphocytes by flow cytometry, using subjects' whole blood samples. PD-1 receptor occupancy (RO) by JS001 on target cells was evaluated during treatment. For subjects enrolled in part A (dose escalation cohorts), whole blood samples were collected by phlebotomy from each subject prior to the first dose (day 0), 24 h (day 1), 96 h (day 4), 168 h (day 7), and every week until day 28, and thereafter, every 2 weeks prior to the subsequent dose until day 126, and then every 28 days prior to the next dose until the end of the study. For subjects in part B (expansion cohorts), whole blood samples were collected prior to the first dose (day 0) and every 2 weeks prior to the subsequent dose until day 98, and thereafter, every 28 days prior to the next dose until the end of the study. All samples were analyzed within 48 h of collection. For each sample, 2 aliquots of 100 μL of blood sample were washed twice in 1 mL of staining buffer (1% FBS in PBS) before being incubated at room temperature (RT) for 15 min with either 5 μg of JS001 for total PD-1 or 5 μg of human IgG4 isotype control antibody (anti-KLH antibody) for on-board JS001 (PD-1 RO). After an extensive washing, both tubes were stained at RT for 30 min with a cocktail of mouse anti-human IgG4 pFc'-PE (Southern Biotech), FITC mouse anti-CD3ε (BD Biosciences), PerCP/Cy5.5 anti-human CD8a (BioLegend), and APC mouse anti-CD45RA (BD Biosciences). Red blood cells were lysed by incubating stained samples at RT for 15 min with FACS lysing solution (BD Biosciences). Samples were acquired and analyzed using a BD FACSCanto cytometer.
Flow cytometry data was analyzed using FlowJo software. T cells were gated based on positive CD3 staining and low side scatter (SSC) level. Doublets and cell clumps were excluded by gating on FSC-H versus FSC-A channels. CD8+ T cell subsets were further gated from the CD3+ singlet population. T cell activation status was analyzed by CD45RA expression level. Activated T cells were defined as CD3+ CD45RA negative cells. The PD-1 staining was analyzed in both CD3+CD8+CD45RA− T cells and CD3+CD8−CD45RA− T cells. Consistent gating was used for all samples. PD-1 RO was calculated as percentage of on-board JS001 staining over total PD-1 staining. Parameters evaluated included the mean fluorescence intensity (MFI) of JS001 staining and RO (%) in activated CD8+ T cells (CD3+CD8+CD45RA−) and CD8− (CD4) T cells (CD3+CD8−CD45RA−). Occupancy above 80% was considered full RO. The results obtained were further analyzed using GraphPad PRISM 5.0 software.
Comprehensive genomic profiling was performed by next-generation sequencing with a 450 cancer-related gene panel (Yuansu Cancer Panel, OrigiMed) on both FFPE tumor and paired peripheral blood samples obtained from 23 subjects. Briefly, genomic DNA was extracted from unstained FFPE slides with tumor content no less than 20% and was fragmented to ~ 250 bp by sonication. A DNA library was constructed using KAPA Hyper Prep Kit (KAPA Biosystems). Hybridization capture was conducted with a custom panel covering a total of 2.6 Mb of the human genome and paired-end sequencing (2 × 150 bp) was performed by following manufacturer's protocols. Genomic alterations including single base substitution (SNV), short and long insertions/deletions (INDELs), copy number variants (CNV), and gene rearrangement and fusions were assessed. The tumor mutational burden (TMB) was estimated by analyzing somatic mutations including coding base substitution and INDELs per mega-base of the panel sequences examined. Driver gene mutations and known germline alterations in the single nucleotide polymorphism database (dbSNP) were excluded.
From March 30, 2016, through December 26, 2016, 36 subjects with advanced metastatic melanoma (n = 22), renal cell carcinoma (n = 6), or urothelial carcinoma (n = 8) refractory to standard systemic therapy were enrolled in three dose escalating cohorts at 1 mg/kg (n = 3), 3 mg/kg (n = 4), and 10 mg/kg (n = 3) and three expansion cohorts at 1 mg/kg (n = 12), 3 mg/kg (n = 11), and 10 mg/kg (n = 3) (Table 1, Table 2, and Additional file 1: Figure S1). The median and average ages were 51 years old, with 21 (58.3%) male and 15 (41.7%) female subjects. Twenty (55.6%) subjects had an ECOG performance score of 1, and 16 (44.4%) subjects had an ECOG score of 0. The enrolled subjects were heavily pre-treated with a median of 3 prior lines of systemic treatment (range 1–11). One hundred percent of subjects had prior systemic chemotherapy. Among 22 melanoma subjects, 14 acral (63.6%), 4 mucosal (18.2%), 1 CSD (4.5%), and 3 non-CSD subtypes (13.6%) (1 chest lesion, 1 hip lesion, and 1 unknown primary lesion) were enrolled.
JS001 was well tolerated, and no DLT or infusion reactions were observed in the study. By the cutoff date of July 31, 2018, 19 months after the last subject was enrolled, subjects had received 2 to 49 doses of JS001 (12.4 doses on average). Average safety follow-up time was 5.8 months (range 0.5–22.9 months). One hundred percent of subjects had treatment-related adverse events (TRAE), but mostly were grade 1 or 2 (Table 3). Treatment-related serious adverse events (SAE) occurred in 5 (14%) subjects (Table 3 and Additional file 1: Table S1). Treatment was permanently terminated in 5 (14%) subjects due to AEs. Treatment was temporarily paused in 6 (16.7%) subjects due to AEs, and all 6 subjects re-initiated treatment after AEs were resolved. Two TRAEs leading to death were attributed to disease progression and were determined to be possibly unrelated to JS001 by the investigator.
Grade 3 or above TRAEs occurred in 13 (36%) of subjects, including lipase increased (n = 4), anemia (n = 3), hypokalemia (n = 2), direct bilirubin (DBIL) increased (n = 2), and one occurrence each of ALT increased, amylase increased, proteinuria, creatine kinase increased, serum creatinine increased, hyperglycemia, low blood pressure, and kidney disease (Additional file 1: Table S2).
Common TRAEs included hyperglycemia (58%), proteinuria (50%), rash (44%), lipase increased (38%), anemia (33%), pruritus (25%), amylase increased (25%), leukopenia (25%), AST increased (25%), and ALT increased (22%) (Table 4). Immune-related AEs (irAEs) were of special interest as this class of drug promotes T cell activation and are known to cause autoimmune conditions. Observed irAEs included two hypothyroidisms (one grade 1 and one grade 2) and one hyperthyroidism (grade 1). The occurrence and severity of AEs/SAEs does not appear to be dose related. Overall, JS001 had an acceptable safety profile with no novel safety signal identified when compared with the two approved anti-PD-1 mAbs, nivolumab, and pembrolizumab.
Clinical response was evaluated using Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 by investigators every 8 weeks. As of July 3, 2018, 1 acral melanoma subject from the 1 mg/kg expansion cohort dropped out of the trial voluntarily on day 15 after receiving two doses of JS001 and no post-treatment radiographic imaging evaluation was available from this subject. This subject was still included in the intention-to-treat (ITT) population for efficacy evaluation. Among all 36 enrolled subjects, 1 confirmed complete response (acral melanoma), 7 confirmed partial responses (2 acral melanoma, 1 mucosal melanoma, 2 UC, and 2 RCC), and 10 stable disease (including 1 unconfirmed partial response (PR) of UC) were observed, for an objective response rate (ORR) of 22.2% (95% CI, 10.1 to 39.2) and a DCR of 50.0% (95% CI, 32.9 to 67.1) (Table 5). Clinical responses were observed in every dose level and in all three cancer types. The best objective response from the 1, 3, and 10 mg/kg dose cohorts was 21.4% (including 1 unconfirmed PR), 26.7%, and 33.3%, while the DCR was 64.3%, 46.7%, and 33.3%, respectively. No apparent dose-related clinical efficacy was observed. Seven out of 8 responding subjects had received at least two prior systemic treatments. The change of tumor size (sum of diameters of target lesions) over time and the best response from baseline were shown in spider plot (Fig. 1a) and waterfall plot (Fig. 1b).
The median time to response was 12 weeks (range 8 to 65.7 weeks), typical of immune checkpoint blockade therapy which requires time to elicit an anti-tumor effect. Notably, 1 UC subject maintained stable disease for 460 days until a PR was achieved. Responses were durable in most subjects as the median duration of response was 5.6 months (range from 1.8 to 17.7+ months).
Among 13 evaluable acral melanoma subjects, 1 confirmed complete response (CR), 2 confirmed PR, and 3 confirmed stable disease (SD) were achieved, for a 23.1% ORR and a 46.2% DCR. Meanwhile, 1 confirmed PR and 1 SD were observed in 4 mucosal melanoma subjects. The duration of response was 8.0, 8.4, 10.6, and 17.7+ months for 4 responding melanoma subjects who had received 2 to 5 lines of prior systemic treatment, indicating PD-1 blockade as a monotherapy could induce durable responses in heavily pretreated acral and mucosal melanoma subjects. One mucosal subject still had an ongoing response after receiving JS001 treatment for 19.5 months.
At the last survival follow-up on July 3, 2018, 21 subjects were deceased, 4 subjects were lost to follow-up, and 11 subjects were still alive. The median PFS was 2.8 months (range 0.5 to 21.9+ months). The median overall survival was 12.2 months (range 1.7 to 27.5+ months). There were no statistically significant differences by tumor histology in PFS and OS (Fig. 2), while RCC had the best median OS and PFS numerically. The 1-year survival rate was 50.0% for all subjects.
An ECL method was developed and validated for the detection and quantitation of JS001 concentration in human serum. JS001 serum concentration reached Cmax about 1 h after infusion and was slowly eliminated from the circulation. JS001 exhibited a dose-dependent linear PK profile. Accumulation of JS001 was observed when given as a multi-dose infusion. The measured serum half-life of JS001 was 6.2 ± 1.3, 7.7 ± 3.5, and 9.8 ± 4.1 days respectively after a single infusion, and 9.5 ± 2.0, 16.5 ± 7.8, and 13.9 ± 5.5 days respectively after multi-dose infusions of 1 mg/kg, 3 mg/kg, and 10 mg/kg given IV Q2W (Additional file 1: Figure S2A).
The serum JS001 trough concentration plateaued after approximately five consecutive doses. The accumulation index was 1.57 ± 0.19, 2.25 ± 0.77, and 2.00 ± 0.54 for the 1, 3, and 10 mg/kg cohorts, respectively. The serum concentration of JS001 over time is shown in Additional file 1: Figure S2B. An in vitro study had previously shown that the IC50 of JS001 blocking PD-1 receptor ligand binding was 3 nmol/L (~ 0.5 μg/mL) . The steady state trough concentration of JS001 in the trial was 8.9 ± 4.4, 37.8 ± 17.5, and 174.3 ± 95.6 μg/mL respectively for the 1 mg/kg, 3 mg/kg, and 10 mg/kg cohorts, indicating the PD-1 receptor will be fully occupied even at the lowest dose cohort of 1 mg/kg.
Serum samples were collected prior to the 1st dose of JS001 and every 28 days prior to dosing. The presence of JS001 ADA was determined using a validated Bridging-ECL method (Meso Scale Discovery, Inc.). No ADA-positive samples were detected in the 10 mg/kg cohort; 3 subjects in the 1 mg/kg and 4 subjects in 3 mg/kg cohort had ADA positivity detected. One subject in 3 mg/kg cohort had an ADA-positive sample before the first dose, reflecting a pre-existing condition. Accelerated JS001 serum clearance and reduced trough concentration coincided with the detection of ADA-positive signals in 1 subject in the 1 mg/kg cohort and 2 subjects in the 3 mg/kg cohort, indicating the presence of neutralizing ADA (8.3%, 3/36). These 3 subjects experienced progressive disease clinically, indicating neutralizing ADA might affect JS001 anti-tumor efficacy.
In this study, PD-1 RO was determined as the percentage of receptor bound JS001 to total PD-1 on human whole blood lymphocytes. Receptor bound JS001 was detected by an isotype-specific antibody that recognizes the IgG4 Fc portion of JS001. Total PD-1 was assessed by first incubating the cells with a saturating concentration of JS001 and then staining the cells with a PE-labeled anti-human IgG4Fc. JS001+ target cells were predominantly present in the CD45RA− T cell subset, which represents non-naïve or an activated T cell population. As shown in Additional file 1: Table S3, the majority of subjects in the 3 dose cohorts maintained full occupancy (> 80%) of the PD-1 receptor on the peripheral blood T lymphocytes over the course of treatment. As shown in Fig. 3a, the pharmacodynamic readout of the PD-1 RO had no correlation with clinical response, as responders (CR + PR + SD) and non-responders (PD) had similar RO during treatment.
The frequency and activation status (by CD45RA staining) of T lymphocytes in whole blood were also evaluated during the course of treatment. There was no major impact of JS001 on the frequency of activated CD8+ T lymphocytes in the peripheral blood in individual subjects (Additional file 1: Figure S3). Interestingly, when the percentage of activated CD8+ T cells of clinical responders (CR + PR + SD) and non-responders were compared, subjects with high percentage of activated CD8+ cells were enriched in the group with a PFS greater than 250 days (Fig. 3b). While the average percentage of activated CD8+ cells in all subjects was 65% before JS001 treatment, 4 out of 5 durable responders (PFS > 300 days) had activated a CD8+ percentage over 80% (of all CD8+ cells), indicating a potential correlation of favorable response to immunotherapy, with high percentage of activated CD8 cells in the peripheral blood prior to treatment. Similarly, subjects with a higher percentage of CD3− CD16+ CD54+ natural killer cells (≥ 26% of all lymphocytes) at baseline (n = 10) also responded better than subjects with less than 26% NK cells (n = 26) (40% ORR and 60% DCR versus 15.4% ORR and 46.2% DCR). The same 4 durable responders (PFS > 300 days) also had a starting NK cell percentage over 26% in the blood.
Previous studies have shown the PD-1 pathway blockade may be particularly effective in melanoma, NSCLC, and other solid tumors with abundant TILs and high PD-L1 expression , regardless of its histology origin. PD-L1 CD8 double IHC staining was performed on available subject tumor biopsy samples (n = 28) in this study. Spring Bioscience (Roche) rabbit anti-human PD-L1 mAb clone SP142  and anti-CD8 clone 4B11 were used. PD-L1-positive status was defined as the presence of membrane staining of any intensity in ≥ 1% of tumor cells in this study.
As shown in Fig. 4a, subjects with PD-L1-positive expression in their tumor biopsy benefited more from JS001 treatment than PD-L1-negative subjects (43.8% ORR versus 0% ORR, p = 0.0081) (Fig. 6b). The PD-1 expression high subgroup (> 50%) responded most favorably to JS001 treatment, with a 57.1% ORR and a 71.4% DCR. Furthermore, PD-L1+ subjects showed significant PFS benefit than PD-L1− subjects from JS001 treatment, HR = 0.36 (95% CI 0.14–0.93), p = 0.034 (Fig. 4b), and numerically better median OS, 613 days versus 448 days (Fig. 4c). But the OS difference was not statistically significant, p = 0.33. Notably, all CR/PR subjects were PD-L1 and TIL double positive. Co-localization of TILs and PD-L1+ cells were observed in tumor biopsies as 93.8% (15/16) PD-L1-positive tumors also harbored TILs. Consistently, TIL-positive subjects responded numerically better to JS001 treatment than TIL-negative subjects (31.8% ORR versus 0% ORR), but the difference was not statically significant (p = 0.11) presumably due to small sample size (Fig. 5). Nevertheless, PD-L1-negative and TIL-negative subjects still benefited from PD-1 blockade therapy as 50% of either group achieved stable disease after JS001 treatment.
Additional biomarkers or parameters analyzed for correlation with clinical efficacy included age, gender, baseline tumor burden, prior lines of treatment, ECOG status, and serum LDH levels (Fig. 5). Among the subgroups, ECOG performance status of 0, serum LDH level within normal range and low tumor volume (the sum of target lesion(s) diameter < 100 mm) at baseline had numerically better clinical response to JS001 treatment. However, only subjects with baseline low tumor burden (< 100 mm) had statistically better response than subjects with high tumor burden (≥ 100 mm), 36.8% ORR versus 5.9% ORR, p = 0.026 (Fig. 6 and Additional file 1: Table S4).
In summary, favorable clinical responses were correlated with PD-L1 expression on tumor cells, the presence of tumor infiltrating lymphocytes, ECOG status at 0, serum LDH in normal range, and low tumor volume prior to treatment.
Recent studies have found high TMB often correlated with a pre-existing adaptive immune response against the tumor and enhanced clinical efficacy in response to PD-1 blockade immunotherapy [24, 25]. Comprehensive genomic profiling was performed in this study by next-generation sequencing with a 450 cancer-related gene panel  (Yuansu Cancer Panel, OrigiMed) on both FFPE tumor and paired peripheral blood samples from 23 available subjects. The TMB was estimated by analyzing somatic mutations within the coding region of the cancer genome covering by the panel. As shown in Additional file 1: Table S5, the TMB was fewer than 10 mutations per Mb for all subjects expect one UC subject (152.8 Muts/Mb) and one acral melanoma subject (17.6 Muts/Mb), consistent with previous reports that acral and mucosal melanoma had limited DNA mutational burden  . Higher TMB levels appear to be correlated with better clinical response (Fig. 6). Multiple potential cutoff points were evaluated, 6 mutations per MB provided the most numeric difference in response rate between CR/PR and PD groups. Subjects with 6+ Muts/Mb (n = 11) achieved numerically better clinical response than subjects with fewer than 6 Muts/Mb (n = 12) (45.5% ORR and 54.5% DCR versus 16.7% ORR and 33.3% DCR) (Fig. 6a); however, the difference was not statically significant, p = 0.13 (Fig. 6b). TMBhigh and PD-L1+ populations were largely overlapping in this study. From 21 subjects with valid results in both TMB and PD-L1 expression, 9 out of 10 TMBhigh subjects were also PD-L1+, while 9 out of 12 PD-L1+ subjects were also TMBhigh.
Genomic profiling also identified frequent gene amplifications in available acral melanoma subjects (n = 7) in the study, including cell cycle regulators (Cyclin D1, CDK4, or CDK6) (5 out of 7), and genes locating in 11q13 such as fibroblast growth factors (FGF19, FGF3, and FGF4) and EMSY (a BRCA2 inactivating gene) (3 out of 7) (Fig. 7). Such clustered gene amplification was not observed in mucosal or UV-induced cutaneous melanoma, UC, or RCC subjects, suggesting a divergent tumorigenesis mechanism for the acral melanoma subtype. In comparison, RCC and UC tumors commonly harbor missense mutations in TP53 (5 out of 10) and SNVs at two hotspots in TERT promoter at nucleotide positions − 124 and − 146 upstream of ATG upregulating telomerase mRNA expression (6 out of 10) (Fig. 7), which occurred much less frequently in acral melanoma subjects in this study (0 out 7 for P53 mutation and 1 out of 7 for TERT promoter hotspots).
In recent years, the regulatory approvals and successful application of PD-1 pathway blockade treatment in a variety of solid tumors has highlighted the power of immunotherapy in controlling not only the classic immunogenic tumors, such as melanoma and RCC, but also malignancies historically considered resistant to immunotherapy such as NSCLC. The activity spectrum of PD-1 blockade was explored in this phase I dose escalation study with JS001, a new humanized IgG4 PD-1 mAb for treating acral (n = 14) and mucosal (n = 4) melanomas, together with RCC and UC subjects.
Unlike chronic UV exposure induced cutaneous melanoma, both acral and mucosal melanoma subtypes harbor much less DNA mutations  and were more refractory to standard chemotherapy as well as immunotherapy alone . In sharp contrast to the dominance of chronic-sun-damage (CSD) type melanoma in the western world (95% of all melanomas), acral and mucosal melanoma are the two most common melanoma types in Asia, accounting for over 70% of melanomas in that region. Previous melanoma immunotherapy research has not focused on these two distinctive melanoma subtypes with drastically different underlying disease mechanisms. In the current study, we prospectively enrolled acral and mucosal melanoma subjects consistent with the regional epidemiology and evaluated their response to PD-1 blockade immunotherapy.
In summary, JS001 had an acceptable overall safety profile up to 10 mg/kg Q2W with no novel safety signal identified. Confirmed objective clinical responses were observed in every dose level of 1, 3, and 10 mg/kg Q2W, and in both acral (1 CR and 2 PR) and mucosal (1 PR) melanomas. The response was durable in subjects who were heavily pretreated (2 to 5 lines of prior systemic treatment). JS001 was particularly effective in subjects with high PD-L1 expression and abundant TILs, regardless of tumor histology subtypes, indicating the pre-existing T cell response against tumors in the microenvironment was the predominant determinant for response to PD-1 blockade therapy. Consistently, higher TMB was also correlated with better clinical response (using 6 mutations/Mb as a cutoff). TMB was generally low for acral and mucosal melanoma subjects in the study (all except one had less than 10 mutations per Mb), consistent with previous reports. TMBhigh and PD-L1+ populations were largely overlapping in this study. From 21 subjects with valid results in both TMB and PD-L1 expression, 9 out of 10 TMBhigh subjects were also PD-L1+, while 9 out of 12 PD-L1+ subjects were also TMBhigh. For the double positive group, 1 CR, 4 PR, 1 SD (unconfirmed PR), and 3 PD were observed. The PD-L1+ and TMBhigh double positive group (n = 9) responded statistically better than the PD-L1− or TMBlow group (n = 12) to JS001 treatment, 55.6% ORR versus 8.3% ORR, p = 0.018 (Fig. 6b). However, PD-L1 expression was better than gene panel-derived TMB in predicting clinical response in this study, as only PD-L1-positive subjects had statically better survival benefits in PFS, HR = 0.36 (95% CI 0.14–0.93), p = 0.034 (Fig. 4 and Additional file 1: Figure S4).
Additional subgroup analysis found the general immune-competent status of a subject, including ECOG status, serum LDH levels, and baseline tumor volume which also had numerically better clinical response, whereas age, gender, and prior lines of treatment did not appear to play significant roles. Interestingly, 4 out of 5 durable responders (PFS > 300 days) had over 80% activated CD8+ T cells (of total CD8 T cells) and over 26% CD3− CD16+ CD54+ NK cells (of total lymphocytes) in the peripheral blood, whereas the average percentage of activated CD8+ T cells and NK cells in all subjects were 65% and 20% respectively, suggesting a potential favorable response to immunotherapy for subjects with a high percentage of activated CD8+ T cells and NK cells in circulation prior to treatment.
Genomic profiling of 23 available subjects also identified gene amplifications of cell cycle regulators (Cyclin D1, CDK4, or CDK6), fibroblast growth factors (FGF19, FGF3, and FGF4) and EMSY (a DNA repair inactivating gene) uniquely clustered in acral melanoma subjects in the study. Interestingly, 2 out of 7 acral melanoma subjects responded to JS001 treatment (1CR and 1PR), and both harbored EMSY gene amplification, which was suggested to be functionally equivalent to somatic BRCA2 loss-of-function mutations . These two subjects were also classified into the TMB high group in this study (higher than 6 mutations/Mb). Further correlation of EMSY gene amplification with response to PD-1 blockade needs to be investigated in a larger cohort of acral melanoma subjects. Genomic profiling in this study has thus confirmed the previous finding of frequent aberration in the CDK4 pathway of acral melanoma subjects  and suggested a combination potential of CDK4/6 inhibitor, FGF/FGFR inhibitor, or DNA repair targeting agent such as PARP inhibitor with PD-1 blockade in treating acral melanomas. In contrast, mucosal melanoma subjects had limited DNA mutations or gene amplification in this study. VEGF overexpression and the density of micro-vessel formation were associated with poor prognosis in mucosal melanoma . The combination of VEGF inhibition with PD-1 blockade might hold the key to enhance the response of mucosal melanoma to immunotherapy.
PD-1 blockade-based immunotherapy has drastically improved the survival of late stage metastatic cancer patients who had limited therapeutic options in recent years. It remains a challenge to further improve the response rate beyond the current ceiling of an average 20–30 percentile. Using melanoma as an example, biomarker-guided divergent combination strategies to treat different subtypes could be envisioned. We look forward to validating our findings in the ongoing registration trials of JS001 for 2nd line (NCT03013101) and 1st line (NCT03430297) treatment of metastatic melanoma subjects with emphasis on acral and mucosal subtypes.
JS001 was well tolerated in humans, with mainly grade 1 or grade 2 TEAEs. In this study, JS001 has demonstrated promising durable anti-tumor activity in UC, RCC as well as in previously underexplored acral (n = 13, 21.4% ORR, 42.9% DCR) and mucosal (n = 4, 25% ORR, 50% DCR) melanoma subtypes, which account for over 70% of melanoma patients in Asia and are not caused by chronic ultraviolet (UV) exposure. Subjects with an immune-active profile in the tumor microenvironment (PD-L1+, TIL+, and TMB high) or in peripheral blood (high percentage of activated CD8+ T cells and NK cells) responded favorably to JS001 treatment. The completion of the current phase I study has led to the initiation of the first prospective anti-PD-1 registration trial in Asia focusing on acral and mucosal melanoma subtypes, representative of the regional disease epidemiology.
Bixia Tang and Xieqiao Yan contributed equally to this work.
This study is sponsored by Shanghai Junshi Biosciences. The authors thank the patients who participated in this study and their families. We thank Margaret Fleming for editing the manuscript.
This work was supported by Beijing Municipal Administration of Hospitals Clinical medicine Development of special funding support (ZYLX201603) awarded to Dr. Jun Guo, Beijing Municipal Science & Technology Commission (Z161100000516062) awarded to Dr. Xinan Sheng and National Science and Technology Major Project "Key New Drug Creation and Manufacturing Program" awarded to Shanghai Junshi Biosciences.
BXT, XQY, HW, HF, SY, ZHC, and JG came up with the idea for, designed, and planned the study. BXT, XQY, XNS, LS, CLC, YK, LLM, BL, XB, XW, SML, LZ, JYY, DJ, LHD, and HFS acquired the data. BXT, XQY, KW, JWH, SY, ZHC, and JG analyzed the data. BXT, XQY, ZHC, and JG interpreted results. BXT, XQY, ZHC, and JG drafted the report. All authors read and approved the final manuscript.
The study was approved by regulatory and independent ethics committees at Beijing Cancer Hospital and done in accordance with the Declaration of Helsinki and the International Conference on Harmonization Good Clinical Practice guidelines. All patients provided written informed consent before study entry.
All patients provided written informed consent for publication.
Kai Wang and Jinwei Hu are employees of OrigiMed, Hai Wu, Hui Feng, and Sheng Yao are employees of Shanghai Junshi Biosciences Co. The other authors declare that they have no competing interests.


Here it is: my entrance onto the sport compact car scene. I bought it with some damage to the front, and with a salvaged title. I also sold the bags back to the guy, as I won't need them. I have all the parts to complete it. It has been undergoing a serious weight reduction program, including lightweight seats, removal of sunroof, (yep, I welded it shut) removal of decklid hinges, and many other non-essential items. The emblems are being shaved, and it will probably be lowered next year.
I recently got one seat mounted.
3/3/03 - Well, it's been a fun project, but I am moving on. The Saturn engine program has come to an end. I bought an Ecotec, and it makes much more sense to be using this engine for my racing program than the Saturn engine. So now I will assemble the engine, put the car back together, paint it, and sell it. Since I won't be turbocharging it, I am going to try to put the flat valves in the new style head to bring the compression to 10.2:1, throw the factory header on it, and call it good.
Pictures at Grattan Raceway, 2004.
12/21/03 - As I write this, I have been driving the car for a month or two. I decided to drive it in Winter to keep the pick-up away from salt. I don't have much in it, and used cars aren't worth much lately. The engine runs strong, and the car weighed in at 2180 with a half tank of gas! The engine sounds like it has a piston slap. It makes a bad noise as I pass through part throttle. If I press harder, it goes away. It makes the same sound as I begin to lift, but once I lift completely, it goes away. If I drive it full throttle, no noise. Sounds like a no brainer, huh?
8/20/04 - In May of this year, the knocking had gotten severe, and she went down in glory taunting a Mustang. I'm sure it was a V-6 Mustabg, but that's no excuse. It turned out to be two rod bearings. At 90,000 miles. Go Saturn. I rebuilt a SOHC to get it on the road quick. Had the head milled .027 to pep it up a bit. It runs good. I lowered it this year with Eibachs. It looks great now, but I really need new struts, it jumps around a lot over the bumps. The only thing left to do now is mild tint.