text:
Annual General Meeting
Please join us for our Annual General Meeting. In addition to regular business, members will review the Executive reports from key areas, club financials and vote on next year's Executive Committee. Members interested in joining the Executive are encouraged to speak with club President Keith Robinson in advance of the meeting.
Reports, financials and club by-laws will all be posted online in the Members Only section, under Executive Cmte, then 2021 Annual General Meeting
ALL Members are encouraged to attend to ensure we have quorum for all votes taken.
Zoom Meeting coordinates are as follows:
Meeting ID: 822 2453 0033
Passcode: 079743
PLEASE NOTE: Visitors are welcome but only current membership may vote. Guests are invited to join an introductory meeting to see how they like the group, then encouraged to sign up online and become a regular active member of the Guild. Please let the host know (verbally or via Chat) that you are a guest.


text:
Fort Australia
Based in Byron Bay, on the east coast of Australia, Fort has been around since early 2000. With five releases under their studded belt and a history of rocking venues throughout the country, Fort has toured with the likes of Clutch, FuManchu, Nebula, Brant Bjork and the Bro's, the Datsuns and the Black Keys to name a few. Band "taking hiatus" as of Jan.17 2010...may never be back. 3 of them have formed new band with new singer, it is called OX, in the studio now doing 2 demo songs for their myspace page.

subset: commoncrawl


text:
–photo courtesy of White Marsh Volunteer Fire Company-
A multi-vehicle crash was reported by the White Marsh Volunteer Fire Company on Saturday afternoon in White Marsh.
Four vehicles were involved in the crash on July 30 at 4 p.m. at the intersection of Ebenezer Road and Pulaski Highway.
It was reported that eight were injured and transported for medical care.


text:
MANILA – The low pressure area (LPA) that the weather bureau has been monitoring is less likely to develop into a tropical cyclone, a forecaster said Thursday.
The LPA was last tracked 200 kilometers south southeast of General Santos City.
"This is also outside the Philippine Area of Responsibility (PAR) and is embedded along the Intertropical Convergence Zone (ITCZ). It has a slim chance to develop into a tropical cyclone within 24 hours," said Philippine Atmospheric, Geophysical and Astronomical Services Administration (PAGASA) forecaster Daniel James Villamil.
He, however, said that the LPA is near the landmass of Mindanao, so it will cause scattered rain showers and thunderstorms over the island.
The LPA and ITCZ will also cause the same weather condition over Eastern and Central Visayas, Bicol Region and Palawan.
Meanwhile, scattered rain showers and thunderstorms will also prevail over Quezon, Aurora, Isabela and mainland Cagayan due to the shear line.
The rest of the country will have isolated rain showers caused by localized thunderstorms.
Moderate to strong winds and moderate to rough seas will also prevail over extreme Northern Luzon and the eastern section of the country.
Elsewhere, winds will be light to moderate with slight to moderate seas. (PNA)


text:
Overview of Andrew Hildebrand
On 5/12/2019, Andrew was reported to the police as missing and a welfare check was initiated. Andrew's vehicle was located in Fond du Lac County by the boat launch for Auburn Lake on 05/14/2019. The Fond du Lac Sheriff's Department conducted an extensive ground and aerial search in that location but did not locate him. Andrew is familiar with the Kettle Moraine State Forest area and is believed to be in possession of a .22 rifle. Andrew was last seen in Oshkosh, WI. (Information retrieved from NamUs.gov)


text:
All-Ordinaries Stock Index
What Is the All-Ordinaries Stock Index?
The All-Ordinaries ( or "Kangaroos" for shoptalk) is a stock index contained common shares from the Australian Stock Exchange (ASX). The All-Ordinaries Index is the most quoted benchmark for Australian equities. The ASX is responsible for ascertaining and distributing the index and its returns.
Understanding the All-Ordinaries Stock Index
The All-Ordinaries Index is market weighted and incorporates around 500 companies. The All-Ordinaries Index began in January 1980, and is otherwise called the "All Ords." It is the most established index of shares in Australia. The market capitalization of the companies remembered for the All Ords index adds up to north of 95% of the value of all shares listed on the ASX. The 3-letter exchange ticker in Australia for the All Ordinaries is "XAO."
For a company to be remembered for the index, they must meet the requirements laid out by the ASX. The All Ords does exclude the value of any dividends paid to shareholders and subsequently doesn't mirror the total returns produced using share market investments during that period. An index that considers both price developments and dividends is called an accumulation index. There is an All Ordinaries Accumulation Index, but this isn't consistently quoted in the finance media.
At the point when laid out, the All Ords had a base index of 500; this means that assuming the index is at present at 5000 points, the value of stocks in the All Ords has increased ten times since January, 1980. On 3 April 2000, the All Ords was rebuilt to cover the 500 biggest companies by market capitalization. This concurred with the presentation of new benchmark indices like the S&P/ASX 200. The significance of the All Ords has been altogether diminished by these new indices.
All-Ordinaries Index Inclusion Requirements
To be remembered for the All-Ordinaries index, a company must have a market value of no less than 0.2 percent of all domestic equities quoted on the ASX and must keep an average turnover on the ASX of no less than 0.5 percent of its quoted shares each month.
There's a different scope of companies that fulfill these criteria, and their market values shift widely. This means that share price developments for companies with bigger capitalizations largerly affect the All Ordinaries Index than do smaller companies.
The All Ordinaries Index portfolio is refreshed toward the finish of each and every month to guarantee that the companies included keep on gathering the criteria for inclusion. It is additionally refreshed during the time when there are changes in the portfolio companies, including delistings, options and capital recreations.
- The All-Ordinaries (XAO) is the benchmark stock market index for Australian equities.
- The index is contained the 500 most important publicly traded Australian corporations, and has been in presence starting around 1980
- Otherwise called the 'Kangaroos', the index is built and kept up with by the Australian Stock Exchange (ASX).


text:
Dr. Stephanie Anderson
Monday, January 14, 2019 | 9:30 – 11:00 am | Scarfe Room 1107
Presentation Title
Toward Tomorrow: Expanding Social Studies Education for a New Age in Canada
Short Bio
Dr. Stephanie Anderson is a post-doctoral scholar with the University of Pennsylvania. She holds a Ph.D. & M.Ed. in Curriculum and Pedagogy (The University of British Columbia) and a B. Ed. and B.A (Honours) in History and French (Queen's University) and has over 20 years of experience teaching social studies throughout Canada. Dr. Anderson's research interests center on the intersection of history education, museum studies, decolonization, and historical consciousness. She is published in national and international journals and is an accomplished presenter at both academic and teaching conferences. Dr. Anderson's recent article for the Canadian Journal of Education received the R.W.B. Jackson Award for most outstanding 2017 English-language article. In September 2018, she was awarded the Emerging Scholar Award at the 11th International Conference on the Inclusive Museum, in Granada, Spain.


text:
Because of the consistently developing events, ordinarily, through the start of inventive degrees of progress seen over the earlier many years, engineers have arranged PC programs that can make: game plans of numbers which can build up the victorious possibilities of lottery players. These ventures figure for every one of the possible number mixes that a particular get-together of numbered game set contains. A couple of individuals find that such records become especially obliging to them since they are the ones who should be certain that they would not lose for their lottery bets. These individuals will enter the game set numbers, hold on for the IT program to deliver likely mixes; they will just put down bets for these, after the program has completed its endeavor. The most basic brand name, of what has been as of late communicated is: these players will conceivably put their best if they are sure that the program they are relying upon contains a best lottery programming stage.
While looking for PC programs which pass best lottery programming purchasing a genuine thing is ideal on. Buy the thing clearly from its specialist or straightforwardly from a retail store retail shop. The reason for why this is being prescribed to you is truly clear. Online togel macau dealers will endeavor to trade things which are divided which mean they have not been totally attempted. A portion of the time, other than not having been totally attempted, the gaming programming that a buyer has purchased turns out to be an online freeware version or a primer transformation. These versions will overall give you sham results, your number records would not be done and they might stop to work commendably, after some time. In addition to these, one ought to guarantee that the program they are buying is commonsense. You can check its overall quality through up close and personal investigation about it, in case you have the opportunity.
Scrutinize what others need to state about the program. Purchase a magazine that gives genuine reviews and updates concerning these gaming programming. Consistently be wary. Have some familiarity with your PC's functioning structure and do not tolerating something that would not be great with it. These undertakings do not come humble, but the awards are abundance. In case you endeavor to ask the players who have won by and large, why they approach the lottery in a legitimate way, they are well headed to respond with answers that do not refute the past explanation. They will probably incorporate that victorious with their best lottery writing computer programs was a consequence of the immediate and straight framework for the said game that their program gave. It moreover helps that these winners were adequately sharp to practice sagacious industrialism while they were searching for their favored things.


text:
This Week in USA
Orphan 55 (Season 12: Ep 3)
8:00pm-9:07pm EST (01:00 GMT) < > « » < > « »
BBC America (adverts during broadcast)
Featuring: Jodie Whittaker, Tosin Cole, Mandip Gill, Bradley Walsh, James Buckley, Laura Fraser
The Doctor and her friends discover that the luxury resort where they are holidaying is hiding a number of deadly secrets. What are the ferocious monsters attacking Tranquillity Spa?
Synopsis Source: BBC America schedule
Ratings Information:
Consolidated figures: 0.468m viewers (evening), 0.10% audience share, 60th place - Nielsen - original cable shows by day, P18-49 %
Rankings are recorded by Nielson's Adult 18-49 score for original cable shows, indicating the percentage of the participating audience in that age category watching the programme. Orphan 55 achieved a rating of 0.10, placing it 60th overall in the chart; it was the only programme on BBC America to make the Top 150. 468,000 viewers watched in total, making it the 73rd most watched programme of the day.
Highest rated shows in the timeslot and of the evening were TLC's 90 Day Fiance with an 0.84 audience share (2.7m viewers) and Real Housewives on Bravo with 0.73 (2.0m); other channel offerings included Obsession: Dark Desires on Investigation Discovery (29/0.17=8/767K), a Special Presentation on Hallmark (32/0.17/1.813m), Holmes and Holmes on Home and Garden TV (35/0.15/989K), Naked and Afraid: Alone on Discovery(38/0.15/620K), Ray Donovan on Showtime Prime (56/0.11/923K), Life Liberty and Levin on Fox News (67/0.09/1.369m), A Beautiful Place to Die on Hallmark Movies (74/0.08/1.295m), and Dr Pol Compilation on Nat Geo Wild (129/0.04/355K).
Taking into account the full week (Mon-Sun), the episode was ranked 507th overall in the P18-49 rankings, and 585th in terms of total viewers. It remained the top programme for BBC America, with the only other programme to feature during the week being the film The Hunger Games; Mockingjay on Thursday at 8:00pm - it was watched by 127K viewers, and had a P18-39 share of 0.04, making it 983rd overall.
[source: Showbuzz Daily]


text:
Women Talking by Miriam Toews is loosely based on real events that happened in a Mennonite colony in the early 2000s. In that way, I want to classify it as historical fiction, and yet 2009 seems to near to be considered "historical;" instead I've filed it under Feminist Fiction—which also feels weird, considering it's narrated by a man… Let's just say, I don't really know how to classify this book.
The whole of the novel is summarized in the title; it is quite literally a bunch of women talking. The women in question are part of a Mennonite colony and are the victims of heinous physical and sexual assaults that had been passed off as "demon attacks." Unable to read or write, they ask August Epp, a man who is an outsider in the colony, to transcribe their discourse as they decide the most correct course of action to take.
The Good
I'll start with the good. If you like ethics, the discussion will be very interesting. For each option, the women discus the religious, moral, and ethical ramifications. While I have seen reviews that complain the women are "too rational" in their thinkings, I think it is befitting a strict upbringing such as in a Mennonite society (disclaimer, I've never met a Mennonite). I think this would be a great book in an ethics class. It's definitely very heavy on the philosophy of right and wrong.
The Bad
There are three major issues with the book, and they're so big they overshadow everything!
A Man Narrating
When I first picked the book, I noticed reviews complaining that a man was narrating and thought those reviewers were taking feminism too far. Within the first few pages its made clear that the women can't speak the native tongue of the country they're in, but that they also can't read or write. August, our narrator, is there to translate their words to paper. For much of the book I thought this was to be presented to the men at some point or perhaps given to someone outside the colony, so it made sense.
However, most of the time August interjects his own thoughts into the narrative—in fact we learn quite a lot about his family and his backstory through his dictation. At one point, the women ask him a question and he writes at length about how he feels about being asked the question. He goes so far as to tell us he put the pen down to answer the question. By the time he tells us what his answer is, I've forgotten the question. It was around this time that I began to agree that using him as a narrator was a weird choice. Considering how un-meeting-minute-like the book reads, the author could have easily made one of the women the narrator and just mentioned August was taking notes.
The Book is Boring
The only reason I finished this book is because it's such a short read. A book of this size would normally take me an afternoon to finish, but this one took me 4 days! Because I gave up on a lot of books last year, I resolved to see more books through til the end this year. As such, I felt compelled to finish not because I actually cared what happened next but because I didn't want to add another book to the "did not finish" self.
Much of the conversation is repetitive and not much was done to differentiate the characters. I often forgot who was who and who was related to who. Aside from Salome being a bit feisty, there really wasn't much distinction. This makes it hard to really get in to the book.
Nothing Happens
Spoiler alert, but absolutely nothing happens. The women make up their minds what they will do and there is a brief scare that they have been found out, but aside from them carrying out their plan in the final pages, it is just them sitting in the barn talking. They spend a great deal of time discussing the unknowns if they leave the colony, as well as the knowns if they stay, but we never see either of these. We don't even see the reaction of the men to their decision (though that may be a subliminal message about their reactions not mattering).
Imagine I start a story telling you I have two pair of shoes: one pair is yellow and will take me to a mythical land where unicorns and mermaids exists, while the other pair is orange and will take me to any time on Earth I'd like visit. I spend 200 pages discussing which pair I wanted to wear and then on last page I reveal my decision, but never tell you what happens after…
This book wasn't for me; I think engaging with the questions the women ask (e.g., are we animals?) might be interesting in a group setting but not as a solo read. It did do two things for me though:
- Made me aware of a horrible tragedy that happened while I was in college
- Made me more curious about the link between fundamentalism, cults, and abuse of women—look for more posts on that soon.
- "Women Talking | Official Trailer". MGM, via YouTube; visited February 2023


text:
What Constitutes a Personal Injury Claim?
What Is a Personal Injury Claim?
A personal injury is a term used mainly in law. A personal injury claim is a right for a victim to claim damage due to accidental incidents or social wrongdoings. An injury can occur anywhere such as at a workplace, a friend's home, or inside a grocery store. A personal liability claim offers the victim a chance to sue for damages. Those damages may be from medical bills, legal fees, or mental illness sustained from the injury.
Anyone who has been a victim of a personal injury may make a claim. In Texas, victims have two years to file a claim of the date the injury occurred. A Houston personal injury lawyer can help you determine the right time to file your claim. Find a lawyer as soon as possible – do not wait longer than 30 days so you can begin the process and receive what you are owed.
How to File a Claim
Step 1: Medical Treatment
Begin with a medical evaluation and treatment. This should be done immediately after the accident has occurred. If you do not experience symptoms until hours later, you may still visit the doctor. If they find your injury is a direct cause from the accident you may be eligible for a claim. Continue to follow your doctor's orders during this time and continue to receive treatment. Whether it's through physical therapy, medication, or frequent visit, follow the advice of your doctor. Lastly, save all your medical records during this time.
Step 2: Contact A Houston Personal Injury Attorney
Personal injury claims give victims the right to get money from necessary medical expenses. You should not have to worry about paying hospital bills while recovering from a major injury. A personal injury attorney Houston will help you with the best legal advice to get the money you deserve. Even if you do not believe your injury is big enough to file a claim – legal advice will help you understand what rights you have as a victim.
Do not pass on the chance to have your medical bills paid for by the person who caused your injury. Getting in touch with a lawyer will put you at ease with the entire situation. Continue to research what you are entitled to and contact a lawyer today who specializes in personal injury claims.
Step 3: Settlement or Lawsuit
Most cases do not reach court proceedings. Everyone involved would rather settle a case outside of court through an attorney's advice than stand trial. Many people will find comfort in knowing they have settled on a deal that both parties agreed upon. However, if chances are that a claim turns into a lawsuit then a personal injury attorney Texas will help with the case.
Your lawyer will prepare you for the best and worst outcomes of filing a personal injury claim. It's best to be prepared from the start. So, remember to document everything about your medical history from the injury. It will be a lengthy process but worth the time and effort to have your medical debt wash away so you can finally recover from your sustained injuries.
Finding A Trusting Lawyer
McLemore Law Firm has dedicated and educated attorneys that will guide you through this stressful process. Filing a personal injury claim will be made easy when following the advice of counsel. Give us a call today to see what can be done about your injury: 713-529-3678.


text:
Sworn law enforcement officers, firefighters and emergency medical technicians (EMTs) would get a one-time $1,000 for working during the COVID-19 pandemic under a plan proposed by Gov. Ron DeSantis. Revenue for the bonuses would be derived from federal funds awarded to Florida under the COVID-19 Relief Bill.
"We don't know exactly how much we're going to get because it is tied to unemployment, but we expect it to be between $9 and 10 billion," DeSantis said during a press conference in Tallahassee on March 16.
The bonuses, which will be made one-time cash payouts, recognize the performance of Florida's first responders during the pandemic.
"We know that the pandemic put an awful lot of strain on our first responders – sworn law enforcement, EMTs, firefighters," DeSantis said. "We believe we should recognize their sacrifice over the last year."
All told, the bonus payments will total about $208.4 million and be administered by the Florida Department of Economic Opportunity (DEO) and "other agencies," he said.
Exactly how much of that bonus revenue will fall to first responders in Hernando County is still uncertain.


text:
Alabama Sheriff Legally Took $750,000 Meant To Feed Inmates… | NPR
American Correctional Association Code of Ethics
The past and future of U.S. prison policy: Twenty-five years after the Stanford Prison Experiment. | Haney, C., & Zimbardo, P. (1998). American Psychologist, 53(7), 709–727. https://doi.org/10.1037/0003-066X.53.7.709
Attica's Ghost: A savage beating, a culture "beyond repair." | The Marshall Project
Torture at Abu Ghraib | The New Yorker
The Stanford Prison Experiment
The Lifespan of a Lie: The most famous psychology study of all time… | Medium
Professionalism in Corrections | W. Kohnke (2001) American Jails, 15(1), 75-78
5 Steps to Better Ethics in Corrections | Corrections One
Federal Prisons Official Used Prison Labor for Work on His Church | The Appeal


text:
Another review had posted that she is real, nice, but somewhat layed-back, almost to the point of lethargic. For the most part I agree. Would I see her again? Yes. Would I recommend her? Yes, provided she's your thing.
Contact/screening process very straight forward. She was prompt and reliable.
The simple fact that, based on a previous review, she wasn't a rip-off, combined with my hard-on for Maria Carrie, which Camille kind of got that look goin, was enough for me. She is attractive, with an awesome, tight little slender body. You can tell she works out regularly. The photos on Eros do not show how tight her abs are. Late 20's, early 30's. She's intelligent and real. Greeted with great hug, nice LFK. From there she was a bit layed-back. Perhaps she was role-playing the shy, timid girlfriend. If that's your thing, she does it well. French was covered briefly, then onto CG, mish, doggie. Great view all-around. I would have liked her to be a little more into it (she wasn't screaming out my name), but she was good, though layed-back.
Good rate, great value, I believe I paid $200 for the hour.


text:
There are no products to list in this category.
Limestone is a common type of carbonate sedimentary rock which is the main source of the material lime.
Limestone Tiles are renowned for their insulation properties. Out of many natural stones, limestone is the least likely to get cold. Although, when used outdoors the light color of limestone reflects the sun rays keeping the tile cool under foot. Limestone Tiles are the finest solution for many purposes.
Some of the most stunning buildings in the world have been created using limestone. For instance, The Pentagon and Empire State Building are both made from limestone. Some structures were built hundreds of years ago although the light coloring of the limestone makes them appear to have been built yesterday.
There are no products to list in this category.


text:
Recreation and Sport Studies
RSS3911 | Practicum 1 | 1 ch (1C/L) (EL) |
Relates theory to practice through professional career and field experiences. Students should expect to participate in at least 40 hours of learning. Course deliverables and learning outcomes will be determined with the Faculty. Faculty approval is required prior to any service commitment or registration procedures. Prerequisites: Must have completed 48 ch and have an AGPA of at least 2.5. |


text:
For the last 15 years, Jackson has been without a public bowling alley. Jessica MacGregor, the owner of the new boutique bowling alley called "Hole Bowl", changed that when they opened their doors 2 months ago. We talked about the challenges in doing so, how the sport creates a valuable social hub, and what's in store in the future.
High-altitude buzz
A 20-mile hike in the Tetons rewarded bumble bee researcher Ellen Keaveny with a special Bombus occidentalis sighting.


text:
Home › Forums › General Issues › Removing Paragraph Tags › Reply To: Removing Paragraph Tags
If anyone else is looking for a solution to removing <p> tags from around images, in the ACF WYSIWYG editor, just use the following code in your functions:
function filter_ptags_on_acf_images($acfcontent){ return preg_replace('/<p>\s*(<a .*>)?\s*(<img .* \/>)\s*(\/a>)?\s*<\/p>/iU', '\1\2\3', $acfcontent); } add_filter('acf_the_content', 'filter_ptags_on_images');
Welcome to the Advanced Custom Fields community forum.
Browse through ideas, snippets of code, questions and answers between fellow ACF users
Open a new forum thread
Helping others is a great way to earn karma, gain badges and help ACF development!
We use cookies to offer you a better browsing experience, analyze site traffic and personalize content. Read about how we use cookies and how you can control them in our Cookie Policy. If you continue to use this site, you consent to our use of cookies.


text:
A stoppage time goal from Shane Long was enough to see Saints through in the FA Cup this evening.
In a drab game befitting the half empty stands and dire atmosphere at St Marys, Long's goal was one of few moments of action in the whole game.
Claude Puel named a young side, with six academy graduates starting, and while they had the better of the match, they offered very little.
Jordy Clasie had a couple of early efforts, as did Jay Rodriguez, but Norwich sat back in a 5-4-1 formation and allowed the hosts to have the ball in non-threatening areas.
There was a lack of width and very little penetration in a performance that is typical of Southampton this term. The team's lack of confidence was epitomised when Shane Long and Sam McQueen both passed up the opportunity to hit an inviting Norwich clearance, leaving it to each other with the visitors breaking up the pitch and earning a free kick.
There was very little improvement in the early stages of the second half, with Saints pedestrian and unimaginative. James Ward-Prowse shot straight down McGovern's throat and Rodriguez failed to get Cuco Martina's cross under control, before the first genuine chance came nearly an hour in. Josh Sims brilliantly beat Sebastien Bassong in the box, and slid a ball across towards Rodriguez for an easy tap in, unfortunately Ivo Pinto slid in to clear the ball behind.
The game pressed on with poor quality on display from both sides, Long firing a shot just wide just after the hour mark. With the substitutes warming up on the sidelines, the hosts looked to be improving - Rodriguez hooking an effort close to goal.
Then, as if to halt his side's brightening display, Puel chose to put defensive midfielder Oriol Romeu on in a game his side were dominating. Former Norwich man Nathan Redmond also came on with Harrison Reed and Jay Rodriguez making way.
Moments before the subs, Josh Murphy picked out Kyle Lafferty in the box, who stretched but couldn't get on the end of it in what was a reminder of how the game was poised.
Ward-Prowse looked to have finally broke the deadlock with 15 minutes remaining, his curling free kick rippling the side netting. Saints were getting closer. Minutes later, the Englishman had another free kick further out, which saints duly wasted with one of their favoured short set plays.
Clasie's effort was spilled by the 'keeper with Long lurking, before the Irishman nicked the ball from Pinto, feeding Sims whose chip drifted well over the bar. Substitute Sergi Canos had a chance to score with his first touch, failed to make it count before Martina hit a shot way over the bar.
Saints upped the pressure late on, with Redmond snatching at three separate chances before Long finally settled the replay in injury time. Sam McQueen's ball was seemingly wasted by the Irishman, who was fortunate to bury the rebound.
Saints will now face Arsenal in the fourth round, whom they dumped out of the League Cup earlier this season. Next in the league are Champions Leicester, with Liverpool after that and a chance at making it to a Wembley final.
Saints: Lewis; Martina, Yoshida (c), Stephens, McQueen; Clasie, Reed (Romeu 70), Ward-Prowse; Sims, Long, Rodriguez (Redmond 70).
Subs: Taylor, Cédric, Gardos, Tadić, Romeu, Redmond, Højbjerg.
Norwich: McGovern; Pinto, Bennett, Klose, Bassong, Martin (c); Jo. Murphy (Ja Murphy 72), Godfrey (Grant 85), Brady, Pritchard; Lafferty (Canos 79).
Subs: Jones, Naismith, Canos, Ja. Murphy, Grant, Ramsay, Middleton.


text:
Colorful bursts themed desk name tags for your students desks.
These colorful bursts-themed name tags can be used as name tags on your students' desks or for equipment labels to use around the classroom.
You can print out the desk name tags and write your students' names on them, then cut and laminate to protect them from general wear and tear. Stick them on your students' desks with scotch tape or contact to create a vibrant classroom environment.


text:
ABC's NASHVILLE has reached the midpoint of its season as one of the better dramas on network television, and also one of the more frustrating. Particularly galling is that it seems to falter worst at the things other, less interesting shows do with much less effort.
Plotting, for example. Within the show's mostly parallel structure following three women at their various points in the Nashville music hierarchy (they intermix occasionally, as in last week's benefit concert set-piece), much of the story concerning veteran Rayna Jaymes (Connie Britton) has been consumed by the sadly tiresome saga of her husband Teddy (Eric Close) and his run for Mayor. This kind of tawdry political dirty tricks material is the meat and potatoes of any number of serialized dramas, from The Good Wife to Scandal–hell, even Parenthood had a political storyline last season. The Nashville version has been mostly dull. That's partly because Teddy, along with Rayna's evil political kingpin father Lamar (Powers Boothe), are the flimsiest, most cardboard characters on the show, but also because series creator Callie Khouri, showrunner Dee Johnson and the other writers seem to be going through the motions, with no interest in the race or the warmed-over scandals it uncovers. (There's a particular irony in the fact that Johnson was, for the past season, the showrunner on Boss, which was nothing but political manipulation and personal trauma.) If there's a way to make Teddy's embezzlement of $2 million several years ago (which he subsequently repaid) the stuff of high–or even low–drama, the show hasn't found it. Tonight's addition of a failed suicide attempt by Teddy's co-conspirator Peggy (Kimberly Williams-Paisley), with whom he did not, repeat not, have an affair, didn't help. The fact that Connie Britton, one of the great television actresses of her generation, has no better substance to work with than this–plus pining over old boyfriend Deacon (Charles Esten)–is downright heartbreaking.
Similarly, for several successive episodes newly-crowned country music crossover superstar Juliette Barnes (Hayden Panettiere) was mostly concerned with her addict mother and a ridiculous scandal involving Juliette shoplifting something from a drugstore that in real life wouldn't have lasted beyond a single news cycle. (The recent storyline that had Juliette getting involved with a Tim Tebow-like religious athlete, though, has been more entertaining.) And the day angelic songbird Scarlett O'Connor (Claire Bowen) gets a storyline that goes beyond her tremulously uncertain choice between bad boyfriend Avery (Jonathan Jackson) and sweet co-songwriter Gunnar (Sam Palladio) will be a milestone for the show.
And yet, there's more than a little that makes Nashville worth watching. Khouri's portrait of the music industry feels informed and much smarter than other shows in this field have been. (It probably doesn't hurt that Khouri's husband is the great T-Bone Burnett, who produces the excellent original music for the series and is also a producer of the show.) There's a real feel for Nashville as a music community, a genuine interest in the lines of communication that run from the top of the business to the outskirts, and the ways that the pressures of the business prey on its people. Every so often, when Britton gets her teeth into a real piece of material, she imparts half a dozen conflicting emotions where most actors would be lucky to find two. Panettiere and Bowen, in their wildly conflicting tiger vs mouse ways, give strong, enjoyable performances as well.
Things may be looking up. Tonight's episode, written by Jason George and directed by Wendey Stanzler, suggests that the show may be ready to move out of its ruts, as Rayna left Teddy, Juliette proposed to her Tebow clone, and there seemed to be the real possibility of a Rayna/Juliette tour that would certainly liven things up, as the interactions between those two characters have been show high points. Perhaps Nashville can find its way to a comfort zone that mixes compelling storylines with the examinations of character and setting that seem to interest its creators.
Nashville has received its back order for a full season, although its ratings–while OK for a 10PM drama–have been nowhere near the down-the-middle mainstream hit ABC was hoping for. Conventional wisdom has it that it's very difficult for a serialized show to acquire new viewers once its characters and storylines have been established, but that rule doesn't seem to apply to cable, where DVDs and streaming in the off-season have sometimes built audiences dramatically in a second season. Perhaps, if Nashville can find itself, and if the network gives it a renewal, the same could happen here. First, though, the series needs to figure out the notes to its particular song.


text:
Logan is a writer who enjoys all things anime and TTRPG, and any form of geek media. He gets very nervous when it comes to deadlines and socializing, but knows he is a great writer. Despite not having the courage to submit his work to agents, Logan has booklets and binders of his work piled around his house. He one day knows he'll publish a book, but his insecurity gets in the way.
Logan can sometimes be a know-it-all but loves his wife Nadine and his friends. He also loves his anime waifu, and has a huge collection/secret stash of adult anime games, figurines and dakimakura. He is 100% an otaku! He once worked with Reginald, Nate, Alliard and ??? in an experimental program, but…more on that later. No spoilers! With his commitment to all things geek, he's also stumbled into life's oddities causing him to have issues getting along with others. For years before Nadine came into his life, Logan was committed to being a shut-in out of fear for the world. With baby steps he leaves the house from time to time but still enjoys his life online playing games and creating visual novels. |


text:
- Research
- Open access
- Published:
"Evaluation of ROS1 expression and rearrangements in a large cohort of early-stage lung cancer"
Diagnostic Pathology volume 18, Article number: 70 (2023)
ROS1 fusion is an infrequent, but attractive target for therapy in patients with metastatic non- small-cell lung cancer. In studies on mainly late-stage disease, the prevalence of ROS1 fusions is about 1–3%. In early-stage lung cancer ROS1 might also provide a fruitful target for neoadjuvant or adjuvant therapy. In the present study, we investigated the prevalence of ROS1 fusion in a Norwegian cohort of early-stage lung cancer. We also explored whether positive ROS1 immunohistochemical (IHC) stain was associated with certain mutations, clinical characteristics and outcomes.
The study was performed using biobank material from 921 lung cancer patients including 542 patients with adenocarcinoma surgically resected during 2006–2018. Initially, we screened the samples with two different IHC clones (D4D6 and SP384) targeting ROS1. All samples that showed more than weak or focal staining, as well as a subgroup of negative samples, were analyzed with ROS1 fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS) with a comprehensive NGS DNA and RNA panel. Positive ROS1-fusion was defined as those samples positive in at least two of the three methods (IHC, FISH, NGS).
Fifty cases were IHC positive. Of these, three samples were both NGS and FISH-positive and considered positive for ROS1 fusion. Two more samples were FISH positive only, and whilst IHC and NGS were negative. These were also negative with Reverse Transcription quantitative real time Polymerase Chain Reaction (RT-qPCR). The prevalence of ROS1 fusion in adenocarcinomas was 0.6%. All cases with ROS1 fusion had TP53 mutations. IHC-positivity was associated with adenocarcinoma. Among SP384-IHC positive cases we also found an association with never smoking status. There was no association between positive IHC and overall survival, time to relapse, age, stage, sex or pack-year of smoking.
ROS1 seems to be less frequent in early-stage disease than in advanced stages. IHC is a sensitive, but less specific method and the results need to be confirmed with another method like FISH or NGS.
Targeted therapy has been a game-changer in the treatment of metastatic lung cancer, providing effective treatment opportunities and improving overall survival in late-stage disease [1, 2]. However, for early-stage disease, the treatment opportunities have developed more slowly. There is now a focus on the potential for targeted therapy in early-stage disease, and more research is needed on the prevalence and characteristics of relevant targets in this setting.
The ROS1 fusion protein is an attractive therapeutic target for patients with metastatic non-small cell lung cancer (NSCLC). Several therapies are available and recommended by the European Society for Medical Oncology (ESMO) [3] and National Comprehensive Cancer Network (NCCN) [4]. In studies done mostly on advanced stage disease, the prevalence of ROS1 rearrangements in NSCLC is about 1–3% [5,6,7,8]. But there is now a growing interest for the prevalence of ROS1 fusions in early-stage lung cancer. For EGFR mutated resected early-stage NSCLC, adjuvant therapy with tyrosine kinase inhibitor have showed promising results [9]. And perioperative targeted therapy can also be feasible for other oncogene addicted NSCLC. Recently, case reports have showed effect of crizotinib as neoadjuvant or adjuvant therapy in a ROS1 translocated setting [10, 11], and there is also ongoing trials investigating this further [12]. This implies that ROS1 fusions in early stage lung cancer can be a target for neoadjuvant or adjuvant therapy in the future.
Two meta-analyses indicate that ROS1 fusion is more common in late-stage disease [6, 8]. It is also shown that the prevalence is higher in women, never-smokers, adenocarcinomas and in patients of Asian ethnicity. Two previous studies on early-stage lung cancer found a prevalence of ROS1 fusion of 0.4–1.2% in adenocarcinomas [13, 14]. These studies were based on one immunohistochemical (IHC) clone and fluorescence in situ hybridization (FISH).
The validity of fusion detection methods, i.e. detection of clinically relevant fusions, remains challenging. Until recently, ROS1 FISH analysis was regarded as a gold standard. However, new methods like next-generation sequencing (NGS) have revealed conflicting results [15]. An emerging strategy in fusion detection diagnostics is the reliance on a combination of several methods, including IHC, FISH, NGS and Reverse Transcription quantitative real time Polymerase Chain Reaction (RT-qPCR). NGS is now more widely available and ESMO recommends the use of NGS in patients with non-squamous NSCLC [16]. For detection of ROS1 fusion, RNA-NGS is preferred over DNA-NGS [15, 17, 18].
IHC-based methods are generally feasible, cost-effective and widely available in pathology labs. These methods are based on immunologic principles, where labeled antibodies bind to specific antigens such as cell proteins. When a part of the ROS1 gene is fused with another gene and the kinase domain is included in that fusion, the subsequent gene expression can lead to an increased ROS1 expression. IHC can therefore detect increased protein expression, but it cannot distinguish whether this is the normal/wild type ROS1 protein or a chimeric protein as a result of gene fusion [19]. For ROS1 IHC there is still no consensus on cut-off levels, though several studies have focused on different IHC clones for ROS1-detection [14, 15, 20,21,22,23]. Only a few studies have used NGS as part of the test algorithm [15, 20, 22]. To our knowledge, there are no published studies based on a diagnostic algorithm combining different IHC-clones, FISH and comprehensive NGS-panels.
This retrospective study aimed to determine the prevalence of ROS1 fusion in a cohort of Norwegian, early-stage resectable lung cancer. We used ROS1 IHC screening and performed FISH and NGS on IHC positive samples, and described relevant challenges in the interpretation of the test results from all three methods. In cases with positive FISH and negative IHC/NGS cases, we have also used RT-qPCR. In addition, we explored associations between ROS1 fusion or IHC positivity, and clinical, histopathological and comprehensive genetic characteristics.
We used biobank specimens from a cohort of surgically resected lung cancer patients at Oslo University Hospital. This biobank has a connected database with histopathological-, biological- and clinical information. The surgery and sampling was done during the period 2006–2018, with a median follow-up exceeding 5 years. Mortality data were imported from the Norwegian Population Registry, which is updated monthly. Two patients were lost to follow up (emigration), and was considered negligible in the analysis. We also excluded patients with carcinoid tumour and thymoma. Written informed consent was obtained from all patients and the project was approved by the regional ethics committee (1904/2009).
The staging has been done according to the latest TNM classifications of malignant tumours at the time of surgery, but for the FISH positive cases we restaged the samples to the current edition [24]. For adenocarcinomas with ROS1 fusion, we have also used the proposed new grading system for invasive pulmonary adenocarcinoma [25].
Specimen characteristics
In this study we used full size slides from formalin-fixed paraffin embedded (FFPE) blocks, tissue micro arrays (TMA), and DNA and RNA extracted from fresh frozen material or FFPE blocks.
The full size slides were taken from FFPE blocks from resections in the diagnostic biobank at Oslo University Hospital. These blocks were sliced at the Department of pathology at Vestfold hospital trust.
Tissue microarray blocks were made from specimens from all the resections, each block consisting of specimens from 25–30 patients. All specimens were represented in triplicates, each core being one mm in diameter, and all tissue cylinders were harvested from the original FFPE blocks after careful selection by a trained pathologist. The TMA blocks were made and sliced at the Department of pathology at Oslo University Hospital. The majority of the TMA blocks have been used in previous projects [26,27,28].
DNA and RNA were extracted from fresh-frozen tissue obtained at surgery. In the few cases of an insufficient number of tumour cells in the frozen tissue samples, the extraction was done from FFPE-blocks.
Positive external control in ROS1-IHC can be difficult to find [29], because of the lack of reliable naturally ROS1 positivity in normal tissue. We used known positive FFPE tissue from a patient with a CD74-ROS1 fusion (confirmed with FISH and NGS) as a positive control. This tumour tissue was also strong and diffuse positive with the two different ROS1 IHC clones (D4D6 and SP384).
Assay methods. Immunohistochemistry
We performed ROS1 IHC analyses by use of two different ROS1-directed antibody clones: ROS1 (D4D6) Rabbit mAb (Cell Signaling, 3287) and the VENTANA ROS1 (SP384) Rabbit Monoclonal Primary Antibody (Roche Diagnostics, 790–6087). The slides were stained at the Department of Pathology at Vestfold Hospital Trust on a VENTANA BenchMark ULTRA system. This is a fully-automated IHC staining platform. For details about the protocols, see Additional file 1: Table S1. The microscope was an Axio Imager.A2 (Zeiss, item no. 490022–0009-000).
We used both a qualitative and a semiquantitative scoring system. All samples were initially grouped into one of seven predefined groups; Negative, weak and focal, weak and diffuse, moderate/strong and focal, moderate/strong and diffuse, ambiguous and too few viable tumour cells (<10 viable tumour cells). Negative samples were defined by lack of tumour cell positivity under 400x (40x objective) magnification. Moderate/strong positivity was defined by dark brown staining with 25 - 200x magnification (2,5-20x objective). Weak positivity was defined by light brown staining, visible at 100-200x (10-20x objective), but requiring of 400x (40x objective) to see clearly. Diffusely positivity was defined by more than 50% of tumour cells with positive staining, including weak positivity, while focal positivity was defined by less than 50% of tumour cells with positive staining. Ambiguous samples were samples that could not be easily classified due to unspecific staining. Within the TMAs, the staining intensity and distribution were scored by assessing the cores from each case as one unit.
All ambiguous samples were reexamined by an additional pathologist and were classified in a consensus meeting. To assess the inter-observer variability and ensure standardized categorization, a random sample of five of the TMA-blocks (114 cases) were examined by two pathologists individually. Any discrepancies in the interpretation were noted and discussed in a consensus meeting in order to optimize reliability.
We also used a combinative semiquantitative scoring system on both the TMA slides and full size slides. In order to compare results, we used the H-score [30] which has been used in several recent papers on ROS1 expression [14, 15, 20, 21, 23]. We used the same formula as Huang et al. 2019 [22]: (1x (percentage of relevant cells with 1+ staining) + (2x (percentage of relevant cells with 2+staining) + 3x (percentage of relevant cells with 3+ staining). Staining intensity was defined as absence of staining (0), weak staining (1+), moderate staining (2+) and strong staining (3+). We used the definition of 0, 1+, 2+ and 3+ from Conde et al. 2019 [15]: Negative staining (0), which was defined as an absence of expression; weak staining (1+), which involved the use of a 40x objective; moderate staining (2+), which required the use of a 10x or 20x objective and strong cytoplasmic staining (3+), which was clearly visible with the use of a 2x or 4x objective. See Additional file 1: Table S2 for details about the scoring system and definitions.
To assess the heterogeneity of ROS1 expression, any IHC-positive sample with more than weak and patchy staining was also reexamined with IHC on full size sections from the original FFPE blocks. Heterogeneity in staining can be due to technical issues like fixation, edge artifacts or biological heterogeneity in expression. Thereafter, the IHC-positive samples were grouped according to the percentage of tumour cells with positive IHC and staining quality (membranous/cytoplasmatic/nuclear, granular, diffuse). In addition, in order to reduce false negative IHC due to heterogenic expression, full size section IHC was also performed on a subgroup of TMA-IHC-negative cases where full size sections had already been prepared for a different study (cases with positive NTRK expression in IHC). Heterogeneity was defined as positive cases with areas of both 0 or 1 + in addition to 2 + or 3 + [15].
Assay methods. FISH
We tested IHC-positive cases (see definition of IHC-positive under variables) with FISH on full-sized slides. In addition, a sub-group of IHC-negative cases were also tested. FISH was performed at the Pathology department of Oslo University Hospital (Radiumhospitalet) with a dual break apart probe from Abbott (Vysis ROS1 Break Apart FISH Probe Kit, product number 08N29-021). The preparation was done according to the manufacturer's recommendations [31]. For the interpretation, we chose areas with the best preserved morphology and with clear signals. In the validation/verification of the test, the laboratory has found that a positive tumour is defined with a cut-off of at least 15% split signals, including isolated orange (5`) and green (3`) signals. Fused signals and break apart, single green /orange signals were counted in at least 50 nuclei and also at least 200 signals.
Break apart split signals are green and orange signals separated by at least 1 signal diameter. An isolated 5'or 3'pattern means that an orange or green signal is present alone or together with fused or break apart signals. A fused signal is either break apart signals separated with less than 1 signal diameter or a completely fused signal that appears yellow.
Assays methods. NGS
NGS was performed on the same samples as FISH. Isolated DNA and RNA were extracted from fresh-frozen tissue (stored at -80 °C) with AllPrep DNA/RNA/miRNA Universal Kit (Qiagen, 80,224) on the QIAcube (automated, spin-column-based nucleic acid extraction from Qiagen). The concentration was measured with Nanodrop (Thermo Fisher Scientific) and Qubit Fluorometer (Thermo Fisher Scientific). We used Agilent 2100 Bioanalyzer (Agilent Technologies) to analyze the RNA-quality. The tumour percentage was evaluated on frozen sections. If the percentage was below 10%, and there were no relevant findings (EGFR-, KRAS- or BRAF-mutation, or ROS1-, ALK- or RET-fusion), then NGS was repeated on slides from FFPE-blocks from the original resection. In these cases, the extraction was done from FFPE blocks using MagLead12gC (Biosystem, A1120) for DNA and Quick RNA FFPE kit (Zymo Research, R1008) for RNA.
For the sequencing, we used the Oncomine Comprehensive V3-panel (OCAv3)(Thermo Fisher Scientific, A35806). This panel covers 161 genes. The library preparation was done on an Ion Chef instrument (Thermo Fisher Scientific). The sequencing was done at both Oslo University Hospital and Vestfold Hospital Trust from July 2019 – September 2021. In Oslo, the sequencing was done on an Ion Torrent S5, and in Vestfold on an Ion Torrent S5XL. For the bioinformatical analysis, we used the Ion Reporter version 5.10–5.16 with a custom filter (Oncomine Variants, 5% CI CNV ploidy > = gain of 2 over normal). The sequencing of DNA and RNA based on tissue from FFPE blocks was done in Vestfold. In these analyses, the material was more fragmented, and therefore there was more "noise" in the sequencing results. To improve interpretation, we therefore heightened the quality requirements of the original filter in these cases (coverage > 1000 reads, Phredscore > 20, allele frequency above 5%). The same quality requirements were also used in sequencing performed on fresh material, but in these cases the heightened quality requirements were not included in the filter. See Additional file 1: Table S3 for details about the quality requirements. Fusion transcripts with unknown gene partner, can be reported as a non-targeted fusion by a combination of primers used for different targeted isoforms [32].
Assays method. RT-qPCR
In cases with positive FISH and negative IHC/NGS, RT-qPCR was performed with Idylla GeneFusion Assay RUO/1.1 (Biocartis AO121/6). Eluat of RNA extracted for NGS was used in the procedure. The most common fusion partners (CD74, SDC4, SLC34A2, EZR, TPM3, GOPC and LRIG3) are included in the kit. In addition the method can detect if there is an expression imbalance between the 5´and 3´end of the ROS1 gene. An expression imbalance indicates that there can be a fusion with a partner other than those detected by the kit [33].
Dependent and independent variables
The main outcome of this study was ROS1 fusion positivity and ROS1 IHC-positivity. Positive ROS1-fusion was defined as samples positive in at least two of the three methods (one of the IHC-clones, FISH, NGS). Positive ROS1 IHC-staining was defined as focal moderate/strong, diffuse weak or diffuse moderate/strong staining, while negative ROS1 IHC-staining was defined as clearly negative or focal and weak staining.
The independent variables included the clinical parameters like smoking status (former, current or never smoker), sex and age (continuous), stage, histopathological diagnosis and NGS-results.
Study design and statistical analysis methods
This was a retrospective, cohort study based on linked register data and biobank material. We examined the data by use of frequency tables. Associations between positive IHC-staining and epidemiological factors and specific mutations were tested by use of univariate logistic regression with positive IHC-staining as the dependent variable. To test for differences in heterogeneity between the two IHC-clones, we used a test for equality of proportions. We regarded two-sided P values < 0.05 as statistically significant. Differences in overall survival and time to relapse were assessed by use of Kaplan Meier plots and log rank test. STATA Release 16 [34] was used for statistical analysis.
The biobank contained tissue from 921 resection specimens. The median age was 67.5 years (range 39.2 to 87). The majority (93.1%) were current or former smokers with a median pack-year of 32.9. 83.7% were in stage I and II and 16.1% were in stage III and IV. The three most common histology subtypes were adenocarcinoma (58.8%), squamous cell carcinoma (32.6%) and large cell carcinoma (3.0%). See Table 1.
Distribution of tumour markers
We made 36 TMAs of the samples. Some of the cores in the TMA contained too few tumour cells to interpret the immunohistochemical analyses (32 for D4D6 and 21 for SP384). Twenty eight (D4D6) and forty (SP384) cases showed more than weak and focal staining and were defined as positive IHC. These cases went forward to further testing with FISH, NGS and full size section IHC. In addition, several negative and weak and focal cases were analyzed with this expanded testing (Fig. 1).
In total, 50 of the 921 cases were identified as positive by at least one of the IHC clones. Among these, 18 cases were identified as positive by both clones, whereas 10 of the D4D6-positive cases were negative with SP384 and 22 of the SP384-positive cases were negative with D4D6. SP384 had a higher number of IHC-positive cases than D4D6. In each scoring group except in the group with diffuse, moderate/strong staining, the mean H-score was higher with SP384 (Table 2).
The interobserver agreement was high in determining whether the cases were either negative/focal and weak/not enough tumour cells or strong/diffusely weak, with an observed agreement of 0.99 for D4D6 and 0.97 for SP384. The discrepant cases were mostly due to different interpretation of reactive pneumocytes and macrophages.
We performed 101 FISH analyses, with 27 and 37 of the D4D6 and SP384 positive cases respectively. FISH found five cases of ROS1-fusion. The percentage of positive FISH signals ranged from 16–64%. Surprisingly, two cases that were negative in IHC with both clones showed a split pattern (above 15%) with FISH. These two cases (case number 2 and 3, Table 3), were also completely IHC-negative (H-score 0) on the full slide sections.
One of the cases had a percentage of isolated/split signals of 16% (just above the threshold). This case was also negative with RNA-NGS. The other case had a percentage of isolated/split signals of 28%, which is low, but considered clearly positive. In this last case the NGS failed on FFPE material and fresh frozen tissue was not available. RT-qPCR was negative on these two discrepant cases, and there were no 3´-5´ ROS1 expression imbalance. The cases with positive FISH were in stage IIb-IIIa [24]. The adenocarcinomas are all grad 3/poorly differentiated tumours with predominantly (> 20%) high grade pattern (solid and trabecular/complex glandular patterns) [25].
With NGS we detected three cases with ROS1-fusion. These cases (case number 1, 4 and 5, Table 3) showed a strong, granular, cytoplasmatic staining without membranous attenuation and no heterogeneity with both clones and they were all FISH positive. All three cases had a TP53 mutation, the CD74 –gene was the fusion partner and they were all adenocarcinomas in stage IIb or IIIa. As shown in Fig. 1, 76 of the D4D6 and 64 of the SP384 negative samples were also analyzed with NGS, but we found no ROS1 fusion in this group.
Three cases were positive in at least two of the three methods (IHC, FISH, NGS). These were considered confirmed cases with ROS1-fusions according to our definition of ROS1-fusion positive (samples positive in at least two of the three methods). All of these cases were positive in all three methods. They showed diffuse and strong staining intensity, and all of them were adenocarcinomas (Fig. 2). Thus, the prevalence of ROS1 fusion was 0.6% in adenocarcinomas and 0.3% in the whole cohort.
There was no significant differences in heterogeneity between D4D6 and SP384 (p = 0.967). The proportion of positive cases with heterogenic staining was 48.3% and 48.8% for D4D6 and SP384 respectively. None of the three ROS1-fusion confirmed cases showed heterogenic staining.
Eight cases showed strong and diffuse staining with D4D6. Three of five cases where we could not confirm a ROS1 rearrangement showed a partly lepidic or acinar/tubular growth pattern in contrast to the confirmed cases that had solid or trabecular growth patterns. The non-confirmed cases also showed more heterogenic staining on the full size slides and H-score on these slides were from 100–240. We found the same pattern with SP384 where ten of twelve non-confirmed cases showed a lepidic or acinar/tubular growth pattern, and more heterogenic staining on full size slides.
Expression and ROS1 fusion related to standard prognostic variables, genetics, relapse and overall survival
Positive ROS1 IHC-staining was strongly associated with adenocarcinomas. The estimated ORs for positive IHC-staining was 20.1 (95% CI 2.7–148.4, p = 0.003) and 9.3 (95% CI 2.8–30.1, p < 0.001) for the D4D6 clone and the SP384 clone respectively. Positive IHC-staining with the SP384 clone was statistically less frequent among former and current smokers than never-smokers (OR 0.2, 95% CI 0.08–0.36, p < 0.001). The same association was not found for the D4D6 clone. For both IHC-clones, there were no associations between positive staining and pack-year of smoking, stage, sex or age.
We found TP53 mutation in all three cases with confirmed ROS1 fusions, and they had no other driver mutations like KRAS, BRAF or EGFR. Among the 104 NGS analyzed cases, the two most frequent mutations were TP53 (n = 49) and KRAS (n = 20). IHC-positivity was associated with KRAS mutation, both in D4D6 (OR 6.5, 95% CI 2.3–18.7, p = 0.001) and SP384 (OR 2.9, 95% CI 1.1–8.1, p = 0.04), but when we adjusted for adenocarcinoma histology there was no significant association. There were no statistically significant associations between positive IHC and TP53.
There was no statistically significant association between IHC expression and overall survival or time to relapse.
Two of the patients with ROS1 fusion (case number 4 and 5, Table 3) received adjuvant chemotherapy. Patient number 1 relapsed after 3 years, and died of lung cancer one month later. Patient number 5 relapsed after only 3 months, and died of lung cancer shortly after that. While patient number 4 has still not relapsed and is still alive eleven years after surgery.
One of the two patients with positive FISH and negative IHC/RT-qPCR/NGS (case number 2, Table 3) never relapsed and died after 14 years (other cause). Patient number 3 relapsed after almost 14 months and died 6 months later. None of these two were tested for ROS1 fusion.
In this study we aimed to map the prevalence of ROS1 rearrangement in a Norwegian cohort of early-stage resectable lung cancer, and see whether the tumours with ROS1 fusion or ROS1 protein expression were associated with specific epidemiological, histological or genetic characteristics.
The prevalence of ROS1 fusion in resected adenocarcinomas in this cohort was 0.6%. The SP384 was in general more often positive and therefore less specific, but both clones identified the three cases that were positive with both FISH and NGS. Positive staining was strongly associated with adenocarcinomas. Most of the cases with strong and diffuse ROS1 expression where we could not confirm a ROS1 fusion had an acinar or lepidic growth pattern, in contrast to the confirmed cases that showed solid or trabecular growth pattern. In SP384, positive ROS1 IHC-staining was more common among never-smokers than among former- and current smokers, but this association was not found when using the D4D6 clone. There were no associations between IHC based ROS1 expression and other prognostic markers like age, sex, stage or pack-year of smoking.
Strengths and limitations
This study was based on comprehensive cohort data from a large number of resected lung cancer patients, with the use of multimodal testing by use of two different IHC-clones, FISH, NGS and RT-qPCR. We believe these features are strengths of the study.
There are several limitations in this study, including the use of TMA and IHC as a screening method, the interpretation of IHC and FISH and detection of fusions with novel/uncommon partners. First, we used TMA in the IHC-based ROS1-screening process. TMA is a cost effective method for IHC-based mass screening. However, morphology assessment can be challenging on IHC-slides and even more challenging on TMAs. Morphology assessment is particularly important as macrophages and reactive pneumocytes can stain positive for ROS1. To reduce the risk of false positive staining, we included a slide from the TMA block with principal staining (hematoxylin and eosin stain), so that the morphology could be examined together with the IHC.
Second, since we only had a small area of the tumour in a TMA, tumour heterogeneity could give a wrong impression of the general protein expression in the entire tumour. The degree and direction of such heterogeneity related sampling bias in our data are unknown. However, previous studies have shown substantial correlation between TMA and full size slides [35]. In our study, the correlation between TMA and full size slides was also good (see Additional file 1: S7). Furthermore, we found that the NGS and FISH positive cases had a homogenous staining pattern, and this is consistent with other studies, especially with the SP384 [15, 21].
Third, we performed manual IHC scoring. Several conditions may affect the subjective interpretation of the intensity of staining. As highlighted by Giltnane et al., background staining, stromal staining and the order in which a slide is observed may be of importance [36]. In our experience, the calibration will also differ from day to day. The subjective assessment of IHC-staining represents a risk of misclassification bias. We attempted to reduce the impact of misclassification bias by using a precise definition in addition to having two pathologists independently calibrate the interpretation. One of the pathologists scored all the TMA twice to reduce the impact of day to day differences in calibrations.
In the FISH interpretation, we used the number of split apart signals/isolated signals and not the number of positive cells which has been more commonly used in previous studies [14, 15, 20, 21, 23]. For FFPE slides, tangentially cut nuclei can give both false negative (split signals not presented on the slide) and false positive results (fused signals are separated in the cutting giving an impression of a single signal). In the validation of the method, the laboratory found that this could be compensated for by counting signals instead of positive cells, an approach for which they have long experience. For cytological specimens, the other formula with a score based on the number of positive cells can be used, as whole nuclei can be evaluated.
Finally, as mentioned in the introduction, for sensitive fusion detection RNA-NGS is preferred over DNA-NGS [15, 17, 18]. However, for targeted RNA-panels like OCAv3, the sensitivity is best if the partner gene is known[37]. We can therefore not totally exclude false negative NGS in the cases with uncommon/novel fusion partners. Nevertheless, the OCAv3 can potentially detect novel/uncommon fusion partners by combinations of primers in the panel, and they are then reported as a non-targeted fusions. We did not find any non-targeted fusions. In addition, in cases with positive FISH and negative NGS/IHC, we performed RT-qPCR with the ability to find evidence of uncommon fusion partners with the 3´and 5´ expression imbalance. No cases with expression imbalance were detected.
Unexpected findings
Two cases were positive with FISH, but negative with IHC and RT-qPCR. One of them was also negative by NGS and the other one failed. It is known that IHC can give false positive results, and that ROS1 IHC always has to be confirmed with a supplementary test such as RT-qPCR, FISH or NGS. Even though FISH is considered to be the gold standard, other studies also found that positive FISH results are not always reproducible [15]. In cases with isolated 3`pattern there is a biological explanation for this, because this may represent a deletion of the segment of the gene containing the binding site for the 5`probe [15, 38, 39]. In cases with positive FISH and negative IHC, the fusion may have been inactivated after posttranslational modification [21] or it might be a non-functioning fusion where the kinase domain is not included in the fusion [29]. Since FISH break apart probes can detect fusions independent of the fusion partner, the discrepant cases might also be a result of false negative RT-qPCR and NGS. As mentioned earlier, novel/uncommon fusion partners can give rise to false negative RT-qPCR and NGS. However, both the Idylla GeneFusion assay (RT-qPCR) and OCAv3 (targeted RNA-NGS) may manage to detect non-targeted fusions were the fusions partner is not included in the kit [32, 33]. Thus, the negative NGS-finding and in addition negative RT-qPCR may imply that there are true false positive results from the FISH-analysis. None of these two patients have been treated with ROS1 inhibitors.
In total, 50 of the 921 cases were positive by IHC, but only three cases could be confirmed with NGS and FISH. Thus, positive IHC staining is much more common than the presumably clinically relevant ROS1 fusions in this study. In addition to the 47 FISH- and NGS negative cases with IHC-positive staining, we noted unspecific staining in macrophages, osteoclasts and reactive pneumocytes. As discussed, there might be several reasons for unspecific staining [19]. The different clones react to different epitopes on the antigen. The antigen, in this case, is the ROS1 protein, either wild type or chimeric/fused. Chromosomal rearrangement leads to activation of the ROS1-gene with sub-sequent overexpression of the chimeric ROS1 protein. Thus, IHC positive cases in wild type tissue/tumour cells may be due to either preanalytical conditions (too long or short fixation, crushed tissue, etc.), cross reactivity to other epitopes, or biological factors (necrosis, phagocytosis of antigens by macrophages, etc.) [19]. In addition, wild type ROS1 protein overexpression could also explain IHC-positivity, and ROS1 seems to be particularly enriched in lung tissue [40]. Lung cancer seems to be even more enriched, perhaps through genetic or epigenetic mechanisms [41]. Contrary to what is seen in ERBB2 amplification and Her-2 overexpression [42], ROS1 amplification does not seem to affect the ROS1 protein expression [43].
Based on our definition of fusion positivity (2 of 3 tests positive), we found no false negative FISH or NGS. False negative FISH might be because of complex fusions where signals appear normal. Biological reasons for false negative FISH are mostly due to intrachromosomal fusions like GOPC, also called FIG [15, 44].
False negative RNA-NGS is mainly seen in specimens with poor RNA quality, and it is important to evaluate the quality metrics in each sample [15]. The reasons for discrepancies in our two cases with FISH positivity are not clear. In both cases, we found a mix of isolated 3'signals and split signals, but in one of the cases, the percentage was just above the threshold.
Discussion on how the results from the study integrate with the existing evidence
In this Norwegian cohort of early-stage lung cancer, the prevalence of ROS1-fusion in adenocarcinomas was 0.6%. This is consistent with other studies on early-stage disease. In the study by Selinger et al., 0.4% (1/278) of resected stage I-III adenocarcinomas had a ROS1 fusion and Warth et al. (both used ROS1 IHC and FISH) found a prevalence of 1.2% (5/405) in resected adenocarcinomas [13, 14]. In Warth`s study the majority of patients were in stage I-III, but 2.5% were in stage IV. Bergethon et al. (Reverse Transcription PCR /Sanger sequencing and FISH) found a prevalence of 0.6% in stages I and II and 2.8% in stage III and IV [5].
The three cases with confirmed ROS1 fusion had an H-score between 200 and 300 in both clones, and there was no heterogenic staining. Other studies that have used the two different ROS1 clones also found that the clones had good sensitivity, but the specificity was lower depending on the cut-off level [15, 23].
The average H-score is higher for SP384 in both studies, which is consistent with our findings. The two studies have used the same dilution, detection system and staining platform as we have. While Conde et al. also found that the D4D6 showed a more heterogenic staining pattern in ROS1-positive tumours, statistical difference in heterogenic staining was not found in our material. Both Conde et al. and Hofman et al. have reported a moderate or good interobserver agreement and that positive staining can be found in normal lung tissue as well, which is also consistent with our findings [15, 23]. In ROS1 FISH negative cases they also found some degree of staining in 13–32.1% and 3–20.3% cases analysed with SP384 and D4D6, respectively. Although mostly weak or focal, this emphasizes that ROS1 IHC can show of unspecific staining in non-rearranged cases.
We found a strong association between adenocarcinoma histology and IHC positivity, and for SP384 an association with never smoking. There was no association between positive IHC and overall survival, time to relaps, age, sex, stage or pack-year of smoking. Warth et al. also found an association with adenocarcinoma histology. They also found that ROS1 positivity was associated with longer overall survival and female sex [13]. One reason for this discrepancy may be that they defined positive IHC as cases showing any positivity, while we defined positive as more than focal weak staining.
All three cases with ROS1 fusion had a TP53 co-mutation and the CD74-gene as fusion partner. TP53 is known to be the most frequent co-mutation in ROS1 fusion and is associated with a shorter progression free survival with firstline crizotinib therapy [45]. CD74 is the most common ROS1 fusion partner [46].
The ROS1 prevalence in adenocarcinomas was 0.6% with our algorithm. Both IHC-clones showed strong and homogenous staining with an H-score above 200 in the three cases where both FISH and NGS confirmed the presence of ROS1 fusion. 50 of 921 cases were positive with IHC, but only three of these cases were confirmed with NGS/FISH. The cases with confirmed fusion showed a solid/trabecular growth pattern, in contrast to most of the non-confirmed cases that showed a lepidic or acinar/tubular growth pattern. We also found ROS1-FISH positive cases that could not be confirmed with IHC, NGS or RT-qPCR. This illustrates that we might need dual testing with two different modalities.
Currently, the prevalence of ROS1 fusion in late-stage disease in the Norwegian population is unknown, but as ROS1 testing has become mandatory in adenocarcinomas we will know more about this in the coming years. Relevant tests should be able to identify treatment responsive tumours. FISH ROS1 has been considered to be the "gold standard", however we need more data from treatment studies to determine which test and algorithm identifies the clinically relevant ROS1 positive tumours. Hopefully, more trials will lead to broader therapy options for both early and late-stage lung cancer .
Availability of data and materials
Some of the data generated or analyzed during this study are included in this published article and its additional information files. Further datasets are not publicly available. However, they can be made available from the corresponding author on reasonable request and after approval by the Regional Ethics Committee.
Fluorescence in situ hybridization
Next-generation sequencing
- RT-qPCR:
Reverse Transcription quantitative real time Polymerase Chain Reaction
Non-small cell lung cancer
European Society for Medical Oncology
National Comprehensive Cancer Network
Tissue microarray
Formalin-fixed paraffin embedded
Not applicable
Copy number variation
Hematoxylin azophloxine saffron
Odds ratio
Confidence interval
Howlader N, Forjaz G, Mooradian MJ, Meza R, Kong CY, Cronin KA, Mariotto AB, Lowy DR, Feuer EJ. The Effect of Advances in Lung-Cancer Treatment on Population Mortality. N Engl J Med. 2020;383(7):640–9.
Lu T, Yang X, Huang Y, Zhao M, Li M, Ma K, Yin J, Zhan C, Wang Q. Trends in the incidence, treatment, and survival of patients with lung cancer in the last four decades. Cancer Manag Res. 2019;11:943–53.
Clinical practice living guidelines metastatic non-small cell lung cancer [https://www.esmo.org/guidelines/guidelines-by-topic/lung-and-chest-tumours/clinical-practice-living-guidelines-metastatic-non-small-cell-lung-cancer] Accessed 26 Jun 2022.
NCCN Guidelines Version 6. 2+21 Non-Small Cell Lung Cancer [https://www.nccn.org/professionals/physician_gls/pdf/nscl.pdf] Accessed 13 Oct 2021.
Bergethon K, Shaw AT, Ou SH, Katayama R, Lovly CM, McDonald NT, Massion PP, Siwak-Tapp C, Gonzalez A, Fang R, et al. ROS1 rearrangements define a unique molecular class of lung cancers. J Clin Oncol. 2012;30(8):863–70.
Yang J, Pyo JS, Kang G: Clinicopathological significance and diagnostic approach of ROS1 rearrangement in non-small cell lung cancer: a meta-analysis: ROS1 in non-small cell lung cancer. Int J Biol Markers 2018:1724600818772194.
Bubendorf L, Buttner R, Al-Dayel F, Dietel M, Elmberger G, Kerr K, Lopez-Rios F, Marchetti A, Oz B, Pauwels P, et al. Testing for ROS1 in non-small cell lung cancer: a review with recommendations. Virchows Arch. 2016;469(5):489–503.
Zhu Q, Zhan P, Zhang X, Lv T, Song Y. Clinicopathologic characteristics of patients with ROS1 fusion gene in non-small cell lung cancer: a meta-analysis. Transl Lung Cancer Res. 2015;4(3):300–9.
Wu YL, Tsuboi M, He J, John T, Grohe C, Majem M, Goldman JW, Laktionov K, Kim SW, Kato T, et al. Osimertinib in Resected EGFR-Mutated Non-Small-Cell Lung Cancer. N Engl J Med. 2020;383(18):1711–23.
Zhao S, Zhu S, Lei X, Xu D, Shi T, Chen Q, Ren F, Chen G, Huang D, Xu S. Use of crizotinib as neoadjuvant therapy for non-small cell lung cancers patient with ROS1 rearrangement: A case report. Thoracic cancer. 2021;12(20):2815–8.
Chen AG, Chen DS, Li S, Zhao LL, Xiao MZ. Case Report: Adjuvant Crizotinib Therapy Exerted Favorable Survival Benefit in a Resectable Stage IIIA NSCLC Patient With Novel LDLR-ROS1 Fusion. Front Oncol. 2022;12: 837219.
Lee J, Wistuba I, Ngiam C, Yu W, Schulze K, Rocha M, Bara I, Carbone D, Johnson B, Kwiatkowski D, et al. P03.04 Phase II Study of TKIs as Neo(adjuvant) Therapy in Stage II–III Resectable NSCLC with ALK, ROS1, NTRK or BRAFV600 Alterations. J Thoracic Oncol. 2021; 16(3):S259-S260.
Warth A, Muley T, Dienemann H, Goeppert B, Stenzinger A, Schnabel PA, Schirmacher P, Penzel R, Weichert W. ROS1 expression and translocations in non-small-cell lung cancer: clinicopathological analysis of 1478 cases. Histopathology. 2014;65(2):187–94.
Selinger CI, Li BT, Pavlakis N, Links M, Gill AJ, Lee A, Clarke S, Tran TN, Lum T, Yip PY, et al. Screening for ROS1 gene rearrangements in non-small-cell lung cancers using immunohistochemistry with FISH confirmation is an effective method to identify this rare target. Histopathology. 2017;70(3):402–11.
Conde E, Hernandez S, Martinez R, Angulo B, De Castro J, Collazo-Lorduy A, Jimenez B, Muriel A, Mate JL, Moran T, et al. Assessment of a New ROS1 Immunohistochemistry Clone (SP384) for the Identification of ROS1 Rearrangements in Patients with Non-Small Cell Lung Carcinoma: the ROSING Study. J Thorac Oncol. 2019;14(12):2120–32.
Mosele F, Remon J, Mateo J, Westphalen CB, Barlesi F, Lolkema MP, Normanno N, Scarpa A, Robson M, Meric-Bernstam F, et al. Recommendations for the use of next-generation sequencing (NGS) for patients with metastatic cancers: a report from the ESMO Precision Medicine Working Group. Ann Oncol. 2020;31(11):1491–505.
Davies KD, Le AT, Sheren J, Nijmeh H, Gowan K, Jones KL, Varella-Garcia M, Aisner DL, Doebele RC. Comparison of molecular testing modalities for detection of ROS1 rearrangements in a cohort of positive patient samples. J Thorac Oncol. 2018;13(10):1474–82.
Benayed R, Offin M, Mullaney K, Sukhadia P, Rios K, Desmeules P, Ptashkin R, Won H, Chang J, Halpenny D, et al. High yield of RNA sequencing for targetable kinase fusions in lung adenocarcinomas with no mitogenic driver alteration detected by dna sequencing and low tumor mutation burden. Clin Cancer Res. 2019;25(15):4712–22.
Vyberg M. Anvendt immunhistokemi: Bioanalytikeruddannelsen København; 2007.
Wang W, Cheng G, Zhang G, Song Z. Evaluation of a new diagnostic immunohistochemistry approach for ROS1 rearrangement in non-small cell lung cancer. Lung Cancer. 2020;146:224–9.
Park E, Choi YL, Ahn MJ, Han J. Histopathologic characteristics of advanced-stage ROS1-rearranged non-small cell lung cancers. Pathol Res Practice. 2019;15: 152441.
Huang RSP, Smith D, Le CH, Liu WW, Ordinario E, Manohar C, Lee M, Rajamani J, Truong H, Li J, et al. Correlation of ROS1 Immunohistochemistry With ROS1 fusion status determined by Fluorescence In Situ Hybridization. Arch Pathol Lab Med. 2020;144(6):735–41.
Hofman V, Rouquette I, Long-Mira E, Piton N, Chamorey E, Heeke S, Vignaud JM, Yguel C, Mazières J, Lepage AL, et al. multicenter evaluation of a novel ROS1 Immunohistochemistry Assay (SP384) for detection of ROS1 rearrangements in a large cohort of lung adenocarcinoma patients. J Thorac Oncol. 2019;14(7):1204–12.
Brierley JD, Gospodarowicz MK, Wittekind C. TNM classification of malignant tumours, Eighth Edition. Hoboken: Wiley Blackwell; 2017.
Moreira AL, Ocampo PSS, Xia Y, Zhong H, Russell PA, Minami Y, Cooper WA, Yoshida A, Bubendorf L, Papotti M, et al. A grading system for invasive pulmonary adenocarcinoma: a proposal from the International Association for the Study of Lung Cancer Pathology Committee. J Thorac Oncol. 2020;15(10):1599–610.
Rud AK, Boye K, Oijordsbakken M, Lund-Iversen M, Halvorsen AR, Solberg SK, Berge G, Helland A, Brustugun OT, Maelandsmo GM. Osteopontin is a prognostic biomarker in non-small cell lung cancer. BMC Cancer. 2013;13:540.
McGowan M, Hoven AS, Lund-Iversen M, Solberg S, Helland A, Hirsch FR, Brustugun OT. PIK3CA mutations as prognostic factor in squamous cell lung carcinoma. Lung Cancer. 2017;103:52–7.
Lund-Iversen M, Grøholt KK, Helland Å, Borgen E, Brustugun OT. NUT expression in primary lung tumours. Diagn Pathol. 2015;10:156–156.
Tsao MS. IASLC atlas of ALK and ROS1 testing in lung cancer. International association for the Study of Lung Cancer. 2016.
McCarty KS, Jr., Miller LS, Cox EB, Konrath J, McCarty KS, Sr.: Estrogen receptor analyses. Correlation of biochemical and immunohistochemical methods using monoclonal antireceptor antibodies. Arch Pathol Lab Med. 1985; 109(8):716–721.
Vysis ROS1 Break apart FISH probe kit [https://www.molecular.abbott/int/en/products/oncology/vysis-ROS1-break-apart-fish-probe-kit] Accessed 26 Jun 2022.
Ion Reporter software 5.20 Help. Fusion calls in analysis results. [https://ionreporter.thermofisher.com/ionreporter/help/GUID-759AEE0F-3542-4774-AC5E-E630E2A4B1FB.html] Accessed 30 Apr 2023.
Technical sheet Idylla Genefusion Panel [https://www.biocartis.com/sites/default/files/2023-04/genefusion_ivd_-_technical_sheet_-_mar2023.pdf] Accessed 30 Apr 2023.
StataCorp.: Stata Stastitical Software: Release 16. In. College Station: StataCorp LLC.; 2019.
Camp RL, Charette LA, Rimm DL. Validation of tissue microarray technology in breast carcinoma. Lab Invest. 2000;80(12):1943–9.
Giltnane JM, Rimm DL. Technology insight: Identification of biomarkers with tissue microarray technology. Nat Clin Pract Oncol. 2004;1(2):104–11.
Qu X, Yeung C, Coleman I, Nelson PS, Fang M. Comparison of four next generation sequencing platforms for fusion detection: Oncomine by ThermoFisher, AmpliSeq by illumina, FusionPlex by ArcherDX, and QIAseq by QIAGEN. Cancer Genet. 2020;243:11–8.
Clavé S, Rodon N, Pijuan L, Díaz O, Lorenzo M, Rocha P, Taus Á, Blanco R, Bosch-Barrera J, Reguart N, et al. Next-generation sequencing for ALK and ROS1 rearrangement detection in patients with non-small-cell lung cancer: Implications of FISH-positive Patterns. Clin Lung Cancer. 2019;20(4):e421–9.
Gao X, Sholl LM, Nishino M, Heng JC, Jänne PA, Oxnard GR. Clinical implications of variant ALK FISH rearrangement patterns. J Thorac Oncol. 2015;10(11):1648–52.
Acquaviva J, Wong R, Charest A. The multifaceted roles of the receptor tyrosine kinase ROS in development and cancer. Biochim Biophys Acta. 2009;1795(1):37–52.
Lee HJ, Seol HS, Kim JY, Chun SM, Suh YA, Park YS, Kim SW, Choi CM, Park SI, Kim DK, et al. ROS1 receptor tyrosine kinase, a druggable target, is frequently overexpressed in non-small cell lung carcinomas via genetic and epigenetic mechanisms. Ann Surg Oncol. 2013;20(1):200–8.
Owens MA, Horten BC, Da Silva MM. HER2 amplification ratios by fluorescence in situ hybridization and correlation with immunohistochemistry in a cohort of 6556 breast cancer tissues. Clin Breast Cancer. 2004;5(1):63–9.
Huang RSP, Gottberg-Williams A, Vang P, Yang S, Britt N, Kaur J, Haberberger J, Danziger N, Owens C, Beckloff SE, et al. Correlating ROS1 protein expression with ROS1 fusions, amplifications, and mutations. JTO Clinical and Research Reports 2020:100100.
Yoshida A, Tsuta K, Wakai S, Arai Y, Asamura H, Shibata T, Furuta K, Kohno T, Kushima R. Immunohistochemical detection of ROS1 is useful for identifying ROS1 rearrangements in lung cancers. Mod Pathol. 2014;27(5):711–20.
Zhang Y, Zhang X, Zhang R, Xu Q, Yang H, Lizaso A, Xu C, Liu J, Wang W, Ou S-HI et al: Clinical and molecular factors that impact the efficacy of first-line crizotinib in ROS1-rearranged non-small-cell lung cancer: a large multicenter retrospective study. BMC Med 2021, 19(1):206.
He Y, Sheng W, Hu W, Lin J, Liu J, Yu B, Mao X, Zhang L, Huang J, Wang G. Different Types of ROS1 fusion partners yield comparable efficacy to Crizotinib. Oncol Res. 2019;27(8):901–10.
We gratefully thank Biomedical Laboratory Scientist Hege Elisabeth Jørgensen, research technologist Astrid Marie Dalsgaard, bioinformatics scientist Nermin Zecic and other Biomedical Laboratory Scientists at Vestfold Hospital Trust for their excellent technical assistance. We also thank Synneva Nebauer for valuable comments on the draft, and professor Åslaud Helland and professor Klaus Hermann Beiske for giving us insightful advices.
Open access funding provided by University of Oslo (incl Oslo University Hospital). South-Eastern Norway Regional Health Authority, 2022020.
Research fund Vestfold Hospital Trust, 198,223.
Research fund Department of Pathology, Vestfold Hospital Trust.
Roche Diagnostics and Pfizer: Research grant and immunohistochemical kits.
Author information
Authors and Affiliations
Study design: OTB, MLI, PS and APHD. Data interpretation, statistics and making figures: OTB, IJZE, JOM and APHD. FISH: AAW and MLI. RT-qPCR: MOL. Writing the manuscript.: OTB, IJZE, JOM, MLI, PS and APHD. Immunohistochemical analyses: PS and APHD. Making tissue micro arrays: MLI and OTB. Extraction nucleic acids from FFPE and sequencing: MOL. Interpretation of NGS results: MOL and APHD. All authors approved the final version of the manuscript.
Corresponding author
Ethics declarations
Ethics approval and consent to participate
Study participants submitted informed consent, and data were anonymized for the analyses. The study was approved by the Regional Ethics Committee of South Eastern Norway (REK), number 2009/1904, S-
Consent for publication
Not applicable.
Competing interests
Roche diagnostics: Contracted/Support Research Grant.
Pfizer: Contracted/Support Research Grant.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Additional file 1: Table S1.
Details on immunohistochemical protocols. Table S2. Details about the scoring system and definitions. Equation for a positive/rearranged ROS1 tumour with FISH. Table S3. Quality requirements NGS. Table S4. Comparing D4D6 and SP384. Table S5. Table including DNA NGS with CNV and hotspot mutation. Table S6. IHC and stage, sex and smoking. Table S7. TMA versus full size slides.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
About this article
Cite this article
Dyrbekk, A.P.H., Warsame, A.A., Suhrke, P. et al. "Evaluation of ROS1 expression and rearrangements in a large cohort of early-stage lung cancer". Diagn Pathol 18, 70 (2023). https://doi.org/10.1186/s13000-023-01357-1
DOI: https://doi.org/10.1186/s13000-023-01357-1


text:
Product may be fresh or frozen
All our meat products may be fresh or frozen, and this varies for each product on a daily basis. Whether a product is sent out fresh or frozen will depend on the stock situation on the date of shipment and on the shelf life of the individual product. Mixed boxes may consist of both fresh and frozen items. When we pack your order for collection or delivery fresh items are in a bag marked 'fresh items' and frozen items are in a bag marked 'frozen items' . In general our sausages and burgers will be frozen - we produce these in large batches and freeze on the day of manufacture so these items will normally be frozen unless we send them to you on the day we make them.
(A very few products in our range are routinely stocked in both fresh and frozen options e.g. Free range chicken supremes chilled and Free range chicken supremes frozen, or whole chickens chilled and whole chickens frozen.)
If you do not want an item as a frozen product please put it into your basket as usual but put a note in the 'NOTES TO VENDOR' box on the checkout - just say 'NO FROZEN PRODUCTS PLEASE'. When we prepare your order we will leave out any items which are only in stock as frozen items on the day your order is shipped or collected and we will refund you for the items which we cannot supply as fresh items on that day.


text:
WuXi Biologics, a global open-access biologics technology platform company, announced that the construction of new biologics drug substance manufacturing facility has commenced on the Industrial Development Authority's (IDA) greenfield site in Mullagharlin, Dundalk, Co. Louth in Ireland.
It's the company's first site outside of China and is supported by the Irish government through IDA Ireland.
The facility is designed to run both traditional fed-batch or perfusion process and continuous bioprocessing, a next generation manufacturing technology to be first implemented globally in the 64-acre campus. When completed, a total capacity of 48,000L fed-batch and 6,000L perfusion will be installed, representing one of the world's largest facilities using single-use bioreactors.
Company CEO Dr. Chris Chen said the facility "symbolizes the significant start of our global biomanufacturing network." WuXi Biologics is headquartered in Wuxi city, Jiangsu province, China.
(Source: WuXi Biologics)


text:
Lingoda Premier mois de l'abonnement mensuel à -20% - UNiDAYS réduction pour les étudiants mars 2024
En ligne
Premier mois de l'abonnement mensuel à -20%
En ligne
€20 Off on the sign-up deposit
Please verify your phone number. You will need it later on to unlock your offer.
We'll send a one-time verification code via SMS.
Enter the verification code we have sent to
Sorry, too many attempts! Please wait 24 hours before trying again.
You have entered the verification code incorrectly too many times. Make sure you have entered the correct phone number or change it below.
The code has expired. Please try again.


text:
HIGHLIGHTS: Day 4 – 52 SUPER SERIES Valencia Sailing Week
The day that Quantum Racing won the 2018 52 SUPER SERIES. Both the regatta, and the remaining season podium places still hang in the balance. Check back later today for racing.
The 52 SUPER SERIES Valencia Sailing Week takes place from 18-22 September. Here's how you can follow the 52 SUPER SERIES.
FULL RECAP: http://bit.ly/2NuVETd
Follow the 52 SUPER SERIES on:
Facebook: https://www.facebook.com/52SuperSeries/
Twitter: https://twitter.com/52superseries
Instagram: https://www.instagram.com/52superseries/
YouTube: https://www.youtube.com/user/52SuperSeries
Share this article:


text:
A1 Lithium Inc.
At A1 Lithium Inc., we believe that diversity and inclusion strategies are the catalysts for success and innovation in the workplace. We believe that differing opinions and lived experiences are valuable and serve to support our business overall. We are committed to engaging with our employees to provide a challenging, dynamic, inclusive, and diverse work environment that supports their professional development, as well as promoting a good work-life balance that prioritizes their overall health and wellness.
We will support initiatives that benefit the environment, human welfare, and education. This includes skills training that advance workforce talent pipelines for our customers and economic development in the communities in which we operate. We will preserve and promote the protection of human rights and welfare within our own business activities, as well as those of our supply chain, in accordance with our Human Rights Policy, Code of Ethics and Business Supplier Codes of Conduct.


text:
The Funky Biscuit Welcomes Guitarist Tab Benoit for Two Big Nights By SFL Music April 21, 2021 Blues Guitarist Tab Benoit Brings His 2021 Whiskey Bayou Revue To The Funky Biscuit in Boca Raton, 2 Big Nights ****Whiskey Bayou Records Artists Josh Garrett, Alastair Greene Joining Benoit on Tour For Selected Dates in April, May, June**** (BOCA RATON FL) – Renowned Blues guitarist Tab Benoit embarks on a year-long National tour, bringing his Whiskey Bayou Revue to South Florida's premier live music venue, The Funky Biscuit in Boca Raton, 303 E. Minzer, for two big nights, Monday, May 3 and Tuesday, May 4. Showtime each night is 7pm (Doors open at 5pm). Tickets are $60.-$75. Info: (561) 395-2929 or visit https://www.eventbrite.com/e/tab-benoit-w-special-guests-the-whiskey-bayou-revue-feat-josh-garrett-tickets-147870343177. Opening for Benoit is exciting new Whiskey Bayou Records recording artist, Josh Garrett. Benoit just concluded six big shows that The Houma, Louisiana-based co-headlined with another blues guitarist at the top of her game, Samantha Fish. Benoit and Fish also teamed last November at Nashville's famed Ryman Auditorium for a concert live streamed worldwide. "This is an artist whose playing speaks to you as much as his lyrics do," wrote Music Connection magazine, reviewing Benoit's Ryman show set. "What Tab lacked in theatrics he made up for with a warm and disarming stage presence and heartfelt playing." Benoit was selected a "30 Best Blues Guitarist In The World Today" by Guitar World in 2020 and also co-authored recently-released book, Blues Therapy, about the blues and mental illness. Benoit, who's always been known for his environmental activism, performed two nights in his hometown of Houma at the 16th Annual Voice of the Wetlands Festival. He also appears prominently in the IMax motion picture Hurricane on the Bayou, a documentary of Hurricane Katrina's effects and a call to protect and restore the Wetlands, and produced a CD to help restore that state's Coastal Wetlands. Benoit was inducted into the Louisiana Folklife Center Hall of Master Folk Artists in the Spring of 2020. Benoit has his own label, Whiskey Bayou Records, with partner and manager, Rueben Williams. The label has thusfar released albums by such established and up-and-coming artists as Eric McFadden, JP Soars, Alastair Greene, Damon Fowler, Eric Johanson, Jeff McCarty, and Dash Rip Rock. Tab Benoit – One Foot In The Bayou (Live) TAB BENOIT ON TOUR! SPRING/SUMMER 2021 ITINERARY Apr. 29 (Thur) GRAND OPERA HOUSE OF THE SOUTH Crowley LA Apr. 30 (Fri) SOUL KITCHEN MUSIC HALL Mobile AL May 1 (Sat) THE MOON Tallahassee FL May 3 (Mon) THE FUNKY BISCUIT Boca Raton FL May 4 (Tues) THE FUNKY BISCUIT Boca Raton FL May 5 (Wed) KING CENTER FOR PERFORMING ARTS Melbourne FL May 6 (Thur) THE WHARF Kissimmee FL May 8 (Sat) CAPITOL THEATER Clearwater FL May 27 (Thur) THE TRALF Buffalo NY May 28 (Fri.) ROBINS THEATER Warren OH May 29 (Sat.) TOKEN LOUNGE Westfield MI May 30 (Sun) WOODLANDS TAVERN Columbus OH June 2 (Wed) MOONTOWN BREWING COMPANY Whitestown IN June 3 (Thur) THE ACORN Three Oaks MI June 6 (Sun) CITY WINERY Chicago IL June 7 (Mon) CITY WINERY Chicago IL June 8 (Tues) CITY WINERY Chicago IL June 10 (Thur) THE SLOWDOWN Omaha NB June 11 (Fri) KNUCKLEHEADS SALOON Kansas City MO June 12 (Sat) COTILLION BALLROOM Wichita KS June 13 (Sun) TOWER THEATER Oklahoma City June 26 (Sat) FIRST FINANCIAL MUSIC HALL El Dorado, AR June 30 (Wed) STARGAZERS THEATER Colorado Springs July 3 (Sat) AVON PERFORMANCE PAVILION Avon CO July 4 (Sun) MESA THEATER Grand JunctionCO July 23 (Fri) THE 04 CENTER Austin TX July 24 (Sat) THE HEIGHTS THEATER Houston TX Aug 7 (Sat) THREE CAVES Huntsville AL Aug 12 (Thur) RHYTHM CITY CASINO RESORT Davenport, IA Aug 14 (Sat) BAYFRONT BLUES FESTIVAL Duluth, MN Aug 19 (Thur) RIVER CITY CASINO St. Louis, MO Aug 22 (Sun) THE KENT STAGE Kent, OH Aug 24 (Tues) BEACHLAND BALLROOM Cleveland OH Aug 25 (Wed) JERGELS RHYTHM GRILLE Warrendale, PA Aug 26 (Thur) LEESBURG THEATER Leesburg VA Sept 14 (Tues) RHYTHM ROOM Phoenix AZ Sept 15 (Wed) RHYTHM ROOM Phoenix AZ Sept 16 (Thur) BELLY UP TAVERN Solana Beach CA Oct 19 (Tues) RAMS HEAD ON STAGE Annapolis MD Oct 20 (Wed) RAMS HEAD ON STAGE Annapolis MD Oct 21 (Thur) THE BIRCHMERE Alexandria VA Oct 22 (Fri) ELEVATION 27 Virginia Bch VA Oct 24 (Sun) PARAMOUNT BRISTOL Bristol TN Oct 26 (Tues) SELLERSVILLE THEATER Sellersville PA Oct 28 (Thur) COHOES MUSIC HALL Cohoes PA Oct 29 (Fri) INFINITY MUSIC HALL Hartford CT Oct 30 (Sat) CENTER FOR THE ARTS Natick MA Nov 2 (Tues) CITY WINERY Boston MA Nov 3 (Wed) BULL RUN Shirley MA Nov 4 (Thur) SPIRE CENTER/PERFORMING ARTS Plymouth MA Nov 14 (Sun) TAFT THEATER Cincinnati OH *Direct Support for Tab Benoit: Josh Garrett (April 29-May 8); Alastair Greene (May 27-June 13). Both Garrett and Greene are Whiskey Bayou Records Recording Artists. Tab Benoit Interviews/Promotional Materials/Show Passes Available. Tab Benoit: Thunderbird Management/Rueben Williams (985) 688-5258 firstname.lastname@example.com tabbenoit.com facebook.com/TabBenoit twitter.com/TabBenoitLA Nashville Scene – TAB BENOIT INTERVIEW (November 2020) Rock And Blues Muse – TAB BENOIT/SAMANTHA FISH SHOW REVIEW Share It!
By SFL Music December 14, 2023 SOCIAL DISTORTION & BAD RELIGION AT REVOLUTION LIVE OUTDOORS Read More