a small startup that turns text queries into data visualizations using generative AI announced a $5.75 million seed round Wednesday that was led by Eventbrite co-founder Kevin Hartz and includes investor Stanley Druckenmiller Vercel founder Guillermo Rauch and 49ers great Joe Montana Tako uses public and licensed data to instantly create slick-looking interactive charts and graphs it calls “knowledge cards.” Tako partnered with AI search engine Perplexity to offer the knowledge cards as part of some queries that centered around data Tako cofounder Alex Rosenberg said the goal is to create a kind of “Google Maps for knowledge,” allowing people to move through data on the internet graphically Rosenberg said the funding will allow the company to hire more people to expand partnerships attract talent and add capabilities to the product a software engineer who came from crypto-focused venture capital firm Paradigm “Their drive is unmatched,” said Hartz, who invested through his venture capital firm A*, short for A star capital Generative AI has upended the search engine for the first time in more than 20 years generative AI promises to deliver complete answers while offering links backing up its sources Companies like Perplexity have been on the leading edge of that shift which has forced even Google to rethink what it means to be a search engine Tako is another example of how the meaning of the term “search engine” is changing Researchers often use Google to find sources of trustworthy data which they then enter into a spreadsheet before turning it into a chart Tako compresses that process into a simple text query The company envisions partnerships like the one with Perplexity as a revenue driver but Rosenberg said the company is also planning to build a direct-to-consumer product line aimed at knowledge workers who use data Most publications no longer have graphics or research desks an area where Tako might be able to fill the void One unique thing about Tako is that customers won’t necessarily have to know what data they are looking for upload a written report and ask Tako to think of data that might help bolster it creating the appropriate charts and graphs The company said it plans to continue expanding its sources of data When Rosenberg compared Tako to a newspaper graphics department it brought back memories of my time at The Wall Street Journal when an entire team of graphics people would look at early drafts of articles and help conceive of charts and graphs to accompany it Most journalists don’t have anything close to that luxury these days so I instantly grasped the value of a tool like Tako The big question for any AI-enabled startup like Tako is how it’s going to defend its business from copycats — aka a “moat.” It takes time and effort to find and license sources of data that can be used for the charts The second is the honing of the text-to-chart AI model Tako’s technology requires proprietary code for searching and visualizing data directly Getting that right and connecting it to the natural language of AI prompts will take some time allowing Tako to keep gaining on would-be competitors Not everybody is convinced that AI search is going to be useful “Generative AI technology is supposed to transform the way we search the web. At least, that’s the line we’ve been fed since ChatGPT exploded on the scene near the end of 2022, and now every tech giant is pushing its own brand of AI technology,” Vox reported Sign up for Semafor .css-w8sqnb{text-transform:capitalize;}technology A Stuart woman drove her golf cart onto the patio dining area of Tako Tiki The golf cart was able to access the dining area via the handicap ramp Three people were taken to a local hospital with minor injuries according to Martin County Fire Rescue spokesperson Cory Pippin The woman passed a sobriety test performed after the police were called and she told officials she did not know how to drive the golf cart and that it was not hers "According to the lady that was driving it she in fact did not know how to drive a golf cart," Romero said and her statements are that when she turned it on it immediately just started going forward." Gianna Montesano is TCPalm’s trending reporter. You can contact her at gianna.montesano@tcpalm.com, 772-409-1429, or follow her on X @gonthescene Please select what you would like included for printing: Copy the text below and then paste that into your favorite email application Enter your phone number above to have directions sent via text This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply Service map data © OpenStreetMap contributors Tako CA-based AI search engine for visualizing and sharing the world’s knowledge The round was led by Kevin Hartz at A* and Ramtin Naimi at Abstract Ventures with participation from Stanley Druckenmiller and Kevin Warsh The company intends to use the funds to expand its partnerships hire talent and expand its platform’s capabilities Co-founded by Alex Rosenberg and Bobby Abbott Tako is an AI search engine that delivers interactive up-to-date and shareable knowledge cards built exclusively from the world’s most authoritative data sources visualize and share factual information to make informed decisions based on data available Tako has already partnered with Perplexity its search and visualization tools were integrated into Perplexity’s answer engine the company expanded its capabilities further with a Polymarket tie-in allowing for visual display of dynamic data like election trends Try a different filter or a new search keyword Get all 19 MUJU RECORDS releases available on Bandcamp and save 20% Streaming and Download help Redeem code Etienne de la Sayette tribute to Korean pop songs from the 60ies MUJU RECORDS Great groovy/crazy stuff made in Paris - France MUJU RECORDS supported by 10 fans who also own “HIRSUTES FARFELUS - LP” supported by 9 fans who also own “HIRSUTES FARFELUS - LP” Los Pirañas = Siempre triunfando <3 Guachímxn Afro-jazz ensemble Jarabi Band explore the stories of contemporary African and Australian musical culture Bandcamp Daily  your guide to the world of Bandcamp The Stories Behind Big Crown Records’ Soulful Singles Composer Meara O’Reilly Brings Hockets Into the Future The Merch Table: Samana Give Listeners a Map to Locate an Obelisk Hidden in Wales Backxwash joins the show to discuss her recent release El Sueño’s sushi is so popular that the owners decided to convert the restaurant’s second story into a brand-new sushi and crudo-focused concept called Tako Vibrant Sushi It’s slated to soft open at 2836 Juan Street on Tuesday Tako will offer Mexican-Japanese fusion and rotate based on what fish and other seafood are in season and Trattoria Don Pietro with his father Sal and partner Gustavo Rios says guests should expect cocktails with an “over the top” presentation He says the food will be just as eye-popping “The crudo and sushi at Tako is fresh, playful and bursting with flavor,” he promises. “As with our other restaurants, we’ll be focusing on quality, service and the full experience.” Head chef Eric Steadman (Hane, Barbusa) is behind Tako’s menu development “He also developed El Sueño’s popular inaugural crudo menu we decided to expand and let him do his thing,” says Busalacchi The 1,100-square-foot space, designed by Busalacchi, features floral and jungle-themed décor, with a DJ booth, a 12-seat bar, and what he says is a small and intimate atmosphere that feels vibrant and fun. “[Tako] feels more like a speakeasy or intimate supper club,” he explains Tako’s hours will run Tuesdays, Thursdays, and Sundays from 6 p.m. to 11 p.m.; Fridays and Saturdays from 6 p.m. – 11:30 p.m.; and closed Mondays and Wednesdays. Reservations aren’t open yet, but locals should watch before the tourists catch wind. (No word on whether any of the Whaley ghosts are sushi fans.) College Area’s food selections are firing up. Now, the best of the best will showcase their stuff at the inaugural Taste of College Area on Sunday, September 29. From Ultreya Coffee to Corbin’s Q, taste over 25 local vendors from 11 a.m. to 3 p.m. Buy your ticket ahead of time and save $5. View this post on Instagram A post shared by 𝐌𝐈𝐆𝐑𝐀𝐍𝐓 𝐒𝐎𝐂𝐈𝐄𝐓𝐘 (@migrant.society) Have breaking news, exciting scoops, or great stories about new San Diego restaurants or the city’s food scene? Send your pitches to [email protected] Beth Demmon is an award-winning writer and podcaster whose work regularly appears in national outlets and San Diego Magazine By clicking subscribe you're confirming that you agree with our Terms and Conditions Email: [email protected] By clicking Subscribe you’re confirming that you agree with our Terms and Conditions Email: [email protected] © Copyright 2023 San Diego Magazine 1230 Columbia Street Movies Here's what you need to know about the Israeli stars of the new film about the mother of Jesus The movie “Mary,” directed by D.J. Caruso (“Disturbia”), about Jesus’ mother is now out on Netflix and its titular character is played by an Israeli actress while Anthony Hopkins plays the antagonistic King Herrod there has been a backlash to the casting of Israeli Jewish actors in these roles from activists who claim that Mary and Jesus were Palestinians Mary was a Jewish woman who lived in what was at her time known as Judea and Caruso told Entertainment Weekly that “it was important to us that Mary be selected from Israel to ensure authenticity.” Mary’s parents are also played by Israeli actors The movie’s Hebrew-speaking cast members attended its Netflix premiere in Los Angeles wearing yellow ribbons in a show of solidarity for the plight of the Israeli hostages Here’s everything you may want to know about the Israeli cast of Netflix’s “Mary.” has had an incredibly prolific career in Israel She’s been a mainstay on Israeli TV for over a decade and professional model since she was 7 having booked campaigns with Smashbox and Essence she starred in her first TV show — the Israeli Children’s Channel reality competition “The Boys and the Girls.” She became a TV host at the Children’s Channel and then in the Israeli late-night show “HaTzinor,” when she became the show’s youngest host Her first acting project was a guest role in the 2017 kids’ sitcom “My Sister Skipped a Grade.” Since then she’s starred in teen comedies and dramas like “My Nephew Benz” and “Infinity,” and most recently had a role in the army thriller “8200” (titled “North Wind” in Hebrew) she starred in the spy thriller “Silent Game” as Shiryn a young Jewish woman in Iran who speaks sign language The movie shot in Azerbaijan and Cohen and the multinational cast Cohen has also worked as a Hebrew voice actress on movies like “Fireheart.” A post shared by Noa Cohen (@noacohen_official) Cohen has spoken vocally about being socially ostracized as a kid and teen and became a source on social media for young people who experienced the same She’s participated in a campaign for at-risk youth and for animal rescues and the Israeli actress seemed to have a blast shooting the film in Morocco “This project has been an unforgettable journey and I’m beyond excited to share it with the world,” she wrote on her Instagram Ido Tako is one of the busiest young actors in Israel and has been acting in TV and movies since 2020 He was inspired to start acting by a trip to LA as a teen Tako lost his mother to breast cancer when he was 6 years old His first role was in the 2020 TV show “Kibbutznikim,” about young teens who live in a kibbutz He starred in the Israeli short “By His Will,” playing a religious gay teen struggling with his identity which was later featured in “Boys on Film 23: Dangerous to Know.” You can also see him in this 2021 Dennis Lloyd music video for “The Way”: He has starred in quite a few shows for teens to the supernatural TV drama “Sky.” He played Raffa in the drama “Black Space,” which is currently streaming on Netflix and had a role in the latest season of the very popular Israeli comedy “Sisters.” His feature debut was in the 2023 “The Vanishing Soldier,” where he played Shlomi a soldier who escapes to his girlfriend’s home in Tel Aviv only to discover that he is believed to be missing He plays Nati in the film “Come Closer,” Israel’s contender for the Oscar this year inspired by the life story of its director the daughter of iconic Israeli director Avi Nesher who lost her brother in a hit-and-run accident — Tako’s character is based on that brother A post shared by Ido Tako (@idotako) is one of the most successful and prolific Israeli actors who many have never heard of — even though he’s been in movies with some of Hollywood’s biggest stars including “World War Z,” “Munich,” “Don’t Mess With the Zohan” and “Hackshaw Ridge.” The actor’s career has been split between Israel and the U.S since he came to America after serving in the IDF to study acting He had small guest roles in “NYPD Blue,” the “CSI” franchise and “Alias.” He was in the Israeli movie “Princess,” directed by “Tattooist of Auschwitz” director Tali Shalom Ezer where he acted opposite Shira Haas in her first movie role He’s been in TV shows like the limited series “The Spy” opposite Sasha Baron Cohen He played Alon in Netflix’s “Messiah” and had a major role as Detective Golan Cohen in the Gideon Raff show “Dig” opposite Jason Isaacs and the late Anne Heche The Harley Davidson-riding Jerusalem-born actor is married to Israeli model Goldman came with him to the “Mary” premiere A post shared by oripfeffer (@oripfeffer) Lior Zaltzman is the deputy managing editor of Kveller By submitting I agree to the privacy policy the mill would shut down its operations later in 2025 Finnish packaging company Metsä Board has revealed plans to shut down its Tako mill in Tampere and enhance the efficiency of its Kyro mill in Kyröskoski has initiated change negotiations at both mills and associated operations The talks may result in up to 215 job cuts and are expected to last at least six weeks the Tako mill’s operations will cease in 2025 In October 2024, the company revealed its plans for change negotiations on potential temporary layoffs at its Finnish mills impacting 1,100 employees due to low projected demand in 2025 The Tako and Kyro mills have struggled with underutilisation and temporary layoffs due to market conditions Both mills have been loss-making through 2023 and 2024 with the Tako mill’s cost competitiveness further hampered by high energy costs and the limited scale of its production lines Its central location in Tampere also restricts development opportunities Metsä Board assures that customer deliveries will remain unaffected Don’t let policy changes catch you off guard Stay proactive with real-time data and expert analysis The Tako mill houses two folding boxboard machines with a combined annual capacity of 210,000 tonnes which is approximately 13% of Metsä Board’s total annual folding boxboard capacity of 1.6 million tonnes Metsä Board anticipates that the proposed measures could enhance the company’s comparable earnings before interest and amortisation by around €30m ($30.8m) annually The measures would also necessitate asset write-downs of roughly €30m The company will detail the one-off costs related to potential redundancies and restructuring at a later stage These expenses are expected to be reflected in the first quarter of 2025 operating result as an item affecting comparability In November 2024, the company announced an investment of €60m to upgrade its Simpele Give your business an edge with our leading industry insights View all newsletters from across the GlobalData Media network This website is using a security service to protect itself from online attacks The action you just performed triggered the security solution There are several actions that could trigger this block including submitting a certain word or phrase You can email the site owner to let them know you were blocked Please include what you were doing when this page came up and the Cloudflare Ray ID found at the bottom of this page Sign In Subscribe Now Incumbent Conservative MP Tako Van Popta has won Langley Township-Fraser Heights With 191 of 193 polls reporting Monday night preliminary results show Van Popta had 32,025 votes or 51.5 per cent with Liberal John Aldag second with 26,292 or 42.3 per cent NDP candidate Holly Isaac was third with 2,474 or 4.0 per cent Van Popta told cheering supporters at his campaign office that the win was a "huge relief."  "Every day I prayed that the volunteers would have success," Van Popta told supporters. "Indeed but primarily for all the volunteers who have worked for me." Tako Van Popta has won re-election. pic.twitter.com/92uCfHL14I Much of the reconfigured riding was "new to us," Van Popta remarked "It was very sad with the redistribution of the riding to lose Brookswood and Aldergrove, but I'm very happy to have kept Fraser Heights," Van Popta said.  and national results came in at the Van Popta campaign headquarters through the evening there were groans at Liberal gains and cheers at Conservative wins Van Popta said the party was affected by "a lot of events completely outside of our control Certainly outside of our control here locally like what's happening in the United States Or Trudeau resigning and Carney coming in and "adopting a number of conservative policies," Van Popta added "A lot of Canadians are giving Carney credit for adopting conservative policies that we were going to be running." the result of a "collapse" in NDP support.  which recorded one of the largest advance turnouts in the province during early voting Three other candidates won one per cent or less of the vote each: Green Party's Debora Soutar People's Party of Canada candidate Sepehr Haghighat In the last election in 2021 in what was then the Langley-Aldergrove riding the final count gave Van Popta 28,643 votes out of 62,635 ballots cast or 45.7 per cent of the vote while second-place Liberal Kim Richter had 16,565 votes or 26.4 per cent and third-place NDP candidate Micahel Chang had 12,288 votes or 19.6 per cent.  who served as Cloverdale-Langley City MP from 2015 – 2019 and 2021 – 2024 took over as the Liberal candidate in Langley Township-Fraser Heights just weeks after he went through major heart surgery a first vice president of the Canadian Union of Postal Workers Sign up for a free account today and start receiving our exclusive newsletters Dialogue and debate are integral to a free society and we welcome and encourage you to share your views on the issues of the day. We ask that you be respectful of others and their points of view, refrain from personal attacks and stay on topic. To learn about our commenting policies and how our community-based moderation works, please read our Community Guidelines Metsä Board has concluded the change negotiations that began in January to improve the company's profitability and cost competitiveness Metsä Board will permanently close Tako board mill and enhance the operational efficiency of Kyro board mill.  The measures will lead to a total reduction of 208 jobs CEO Mika Joukio: "The change negotiations with the personnel were conducted in a constructive spirit Finding new jobs for the people being laid off is our primary goal and we also aim to help with employment by offering reemployment training and support." Production at the Tako board mill is planned to end by the end of the second quarter of 2025 at the latest The closure of the mill will not affect customer deliveries as Metsä Board's total annual folding boxboard production capacity is sufficient to meet current demand Metsä Board's annual folding boxboard capacity will be approximately 1.4 million tonnes Metsä Board estimates that the measures now decided will improve the annual comparable EBITDA by approximately EUR 30 million A one-off profit impact of approximately EUR -28 million will be recorded in connection with the closure of the Tako mill mainly related to the impairment of assets It will be recorded in Metsä Board's operating result for the first quarter of 2025 as an item affecting comparability The one-off cash impact is approximately EUR -1 million Kimberly-Clark Corporation recently announced plans to invest over $2 billion over the next 5 years.. International Paper announces the groundbreaking of its new state-of-the-art sustainable a leading full-service packaging solutions supplier for the world’s biggest beauty SPM International announces Bill Partipilo as the new CEO of SPM Instrument boxboard production in Q1 2025 fell 1% compared to Q1 2024 Smurfit Westrock plc announced it will permanently close its coated recycled board (CRB) mill in.. Valmet will deliver a disc filter refurbishment and reinstallation to Holmen Paper’s Braviken Paper Mill.. By Belinda Kelly According to new research Nordic Paper has today decided to approve environmental investments of up to SEK 450 million.. Together with its Turkish partner company Labtek the German testing equipment manufacturer emtec Electronic will.. Mondi has successfully started up its Duino mill This milestone further strengthens Mondi’s position as.. Gulf Paper Manufacturing and Toscotec started up PM2 after a forming section upgrade at their.. Södra delivered a stable result for the first quarter of 2025 VPK Group announces the acquisition of Open Imballaggi a well-established sheet plant specializing in corrugated.. a leading full-line supplier of technologies and services for the paper industry Amcor and Berry Global Group recently announced the European Commission (EC) has granted unconditional approval.. Stora Enso plans to implement a new organisation with seven P&L responsible business areas reflecting.. Georgia-Pacific’s Savannah River mill in Rincon now accepts polyethylene (PE)-coated paper cups in its.. a global leader in sustainable chemical solutions for water-intensive industries announces that it has invested in Kinki Danboru a privately-owned business located in Zellwood John Hardwick Twitter Facebook JENSEN BEACH, Fla. — A woman was hospitalized after she was hit by a golf cart that ended up in the middle of a busy Jensen Beach bar Saturday night. Tako Tiki owner Troy Pulley was out of state when his manager called. He said she sounded panicked. "A golf cart just drove through the front door and hit a table," Pulley recalled. One woman had to be taken to the hospital with a serious pelvic injury. “Great lady, wonderful customer… a local," said Pulley. Captain Ruben Romero with the Martin County Sheriff's Office says it looks like a case where a person didn’t know how to operate the vehicle correctly. Romero added the golf cart the woman was operating actually belonged to the Ocean Breeze Resort, about half a mile away from the bar. “The driver was probably one of the employees and the female that attempted to drive the vehicle when the accident happened may have not been a person who was supposed to be operating it," said Romero. WPTV's Jon Shainman went to the resort and asked if anyone knew what had happened. No one could answer my questions and I never heard back from a manager. Investigators are thankful this wasn’t as bad as it could have been. The sheriff’s office is still investigating this case and say it’s a good bet the driver of that golf cart will face some traffic citations. Astro Bot Tentacle System Boss 100% Walkthrough This will unlock a new stage: Bot of War No part of this website or its content may be reproduced without the copyright owner's permission Rock Paper Shotgun is a registered trademark of Gamer Network Limited Lucie County Watchdog Reporter Wicker Perlis recommends Tako Tiki in Jensen Beach It opened in 2019 at 3340 NE Pineapple Ave A: Tako Tiki features a rotating live music schedule It also is within walking distance of my house so it is nice to be able to walk there when it is not too hot out I sometimes even can hear the music in the distance from my backyard The food and drinks are creative and consistently well-executed It is also a great place to start a night out before venturing a couple of blocks away to some of the bars on Jensen Beach Boulevard You can also go on a Thursday before exploring Jamin' Jensen A: The carne asada fries are a great appetizer that are also more than filling enough to make them your full meal They are French fries topped with Monterey cheese there are a number of good draft beer options but beyond that I would recommend the watermelon jalapeno margarita The strawberry basil margarita is also great Q: Is there something unique on the menu you recommend trying A: The octopus taco (or "tako," as they style it) is unique and delicious lime and sea salt is grilled and topped with pickled onions A: Tako Tiki is an outdoor restaurant with a few different dining areas I have enjoyed sitting in the area closest to the train tracks when my focus is on conversation as the music is still enjoyable but much quieter I have really enjoyed being up close and taking it all in There is also a dining area between the stage and the further-away area expect to see people dancing throughout the restaurant Wicker Perlis is TCPalm's Watchdog Reporter for St You can reach him at wicker.perlis@tcpalm.com plans to close its Tako mill as well as to improve the efficiency of its Kyro mill and launches change negotiations at both mills and related operations The negotiations will involve a total of approximately 360 employees and may lead to a reduction of up to 215 employees The negotiations are expected to last at least six weeks The aim of the change negotiations at the Kyro mill is to improve the profitability of the mill and safeguard its operational conditions Any decisions on personnel reductions and mill closure will only be taken after the change negotiations have been concluded the production of folding boxboard at Tako and Kyro mills has remained well below capacity due to the market situation and the mills have had to implement several temporary layoffs Due to weak market situation and rapidly rising costs both mills have been loss-making in 2023 and 2024 The cost competitiveness of Tako board mill is particularly weakened by its high energy costs and the small size of its production lines The development of the mill is limited by its exceptional location in the centre of the city of Tampere The possible closure of Tako board mill would have no impact on customer deliveries as Metsä Board’s overall capacity for folding boxboard production is sufficient to meet current demand Metsä Board estimates that the planned measures would improve the company's comparable EBITDA by approximately EUR 30 million per year the measures would lead to a write-down of assets of approximately EUR 30 million The one-off costs related to possible redundancies and restructuring would be specified later Items related to write-down and other costs would be recognised in the first quarter 2025 operating result as an item affecting comparability Tako board mill has two folding boxboard machines with annual production capacities of 70,000 and 140,000 tonnes This represents approximately 13% of the company's current annual production capacity of approximately 1.6 million tonnes of folding boxboard CEO Mika Joukio: "The planned measures published today Our employees at our Pirkanmaa mills are highly skilled and have shown a lot of flexibility in the difficult market situation of recent years we need to consider alternatives to improve our competitiveness and profitability We currently have a number of ongoing and planned investments to improve the competitiveness of our mills and products which will require capital and a strong financial position in the coming years." Focus Check podcast ep62: Next-Gen Efficiency: Sony’s a7S III FW 4.0 & the Rise of Smarter Gear - WATCH or LISTEN now Focus Check podcast ep62: Sony's a7S III FW 4.0 | Insta360 X5 & much more it can also be equipped with a fluid video head and a 1/4″ mounting thread with locating pins it will come with either a ball head or a video head The carbon fiber tripod should be available by the end of 2024 for around $300 The Chinese company YC Onion does not have a very wide product portfolio, but I feel like by focusing on very few products, they usually put a lot of thought into each design. My colleague Nino recently reviewed the YC Onion Pineta Peak tripod for filmmakers At this year’s IBC we talked to Sandy from YC Onion about yet another tripod but this time a more lightweight version for photographers Let’s take a look at the new YC Onion Tako tripod the new Tako is primarily focused on photographers who are looking for a lightweight tripod that is very easy and quick to set up The tripod features the company’s new one-way single lock mechanism which is in a semi-locked state by default (when locked) so you do not have to unlock each leg in order to extend them for further adjustments (and for retracting the legs back) you can unlock each leg by twisting the single lock lever This design should enable an even faster deployment of the tripod in the field There will be two versions of the Tako tripod – one for photographers and one for filmmakers that will feature a video head Sandy from YC Onion further told us that the tripod is weather-sealed preventing dust from entering each section and reducing the need for maintenance the tripod can be lowered to approximately 17cm/6.7in above the ground The good thing is that the tripod features an integrated leveling bowl This is especially handy in combination with the video head YC Onion Tako has a 1/4″ thread mount with locating pins at the top for mounting accessories The YC Onion Tako tripod is yet to be released The company informed us that it is expected to launch at the end of 2024 (around November or December) with a price of approximately $300 We will inform you once the YC Onion Tako becomes available What do you think about the YC Onion Tako tripod Do you use any YC Onion gear for your productions Let us know your thoughts in the comments section underneath the article Δdocument.getElementById( "ak_js_1" ).setAttribute( "value" Δdocument.getElementById( "ak_js_2" ).setAttribute( "value" Stay current with regular CineD updates about news You can unsubscribe at any time via an unsubscribe link included in every newsletter. 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For further details, see our Privacy Policy Jakub Han is a freelance filmmaker based in Vienna He is interested in new tech and trends in filmmaking and passionate about action sports and short documentaries Jakub has over 10 years of experience with camera work and post-production Metsä Board has finalized change negotiations initiated in January to enhance profitability and cost efficiency the company will permanently close the Tako board mill and improve the operational efficiency of the Kyro board mill Production at the Tako board mill is scheduled to cease by the end of the second quarter of 2025 customer deliveries will remain unaffected with Metsä Board's annual folding boxboard production capacity at approximately 1.4 million tonnes Metsä Board projects the measures will improve its annual comparable EBITDA by around Euro 30 million predominantly starting from the second half of 2025 The company will record a one-off profit impact of approximately Euro -28 million related to asset impairment in its first-quarter 2025 operating result The one-off cash impact is estimated at Euro -1 million Republication of Lesprom Network content is prohibited without the prior written consent of Lesprom Network General Terms and Conditions and Privacy policy It has recently been revealed that two Israeli actors were cast as leads in Netflix’s biblical coming-of-age epic “Mary,” which also stars Hollywood legend Sir Anthony Hopkins the film tells the story of “Mary’s courageous journey to birth baby Jesus,” and their escape “from the relentless pursuit of King Herod,” played by Hopkins “to save Jesus’ life at all costs.” It is set to be released on the streaming platform on December 6 The film is directed by D.J. Caruso, director of the 2007 hit thriller “Disturbia.” Caruso told Entertainment Weekly the decision to cast Israeli actors stemmed from the desire to “to ensure authenticity.” and somewhere in the middle of that process From the moment we witnessed her first cold audition “Noa has a unique blend of grace and presence that is striking able to access the deep emotional core of Mary’s character Her ability to convey both vulnerability and strength was exactly what we needed for this role.” Cohen previously starred in “Unit 8200,” a TV series about the elite Israeli Intelligence Corps unit The cast of “Mary” also features Israeli actors Ori Pfeffer Keren Tzur and Hilla Vidor in episodic roles Tako no Himitsu has already surpassed its initial Kickstarter funding goal Tako no Himitsu (literally "Secret of the Octopus") has particularly gorgeous art as well as soundtrack contributions from GBA and SNES vets Motoi Sakuraba (Golden Sun Dark Souls) and Masanori Hikichi (Terranigma And what's here is really interesting mechanically: Tako no Himitsu feels a lot like Secret of Mana with real time combat and light puzzle solving mixed up with a tactical menu where you select special moves and spells tied to magical octopi you collect throughout the story The little buddies can be mixed and matched with different party members to unlock synergies and combos but in the demo portion I played I was limited to a projectile attack and enemy scan The octopus system's finishing touch is a home base fountain where your little guys hang out and you can take care of the critters to build up your bond with them⁠—it reminds me a lot of the Chao Garden from Sonic Adventure 2 My favorite thing here is definitely the aesthetic Tako no Hitsume really nails the soft lines and vibrant colors of GBA pixel art and the sound effects also have a distinctly GBA je ne sais quoi that I dig the sort of thing to really evoke a long car ride sat in the back seat playing Golden Sun Keep up to date with the most important stories and the best deals, as picked by the PC Gamer team. Associate EditorTed has been thinking about PC games and bothering anyone who would listen with his thoughts on them ever since he booted up his sister's copy of Neverwinter Nights on the family computer He is obsessed with all things CRPG and CRPG-adjacent When he's not playing or writing about games you can find Ted lifting weights on his back porch showing you where to find all the collectibles throughout this Astro Bot level Login | Sign Up Push Square Guest Login or Sign Up Looking for a Wako Tako collectibles walkthrough in Astro Bot? This page will show you where to find all the collectibles in Wako Tako, a level found in Tentacle System. As part of our Astro Bot guide our Wako Tako collectibles walkthrough will show you where to find all Bots in the stage Did you find this Wako Tako walkthrough in Astro Bot helpful? Let us know in the comments section below, and check out our Astro Bot guide for lots more Stephen has been part of the Push Square team for over six years bringing boundless enthusiasm and a deep knowledge of video games to his role as Assistant Editor Having grown up playing every PlayStation console to date He's also our go-to guy for Sonic-related matters Hold on there, you need to login to post a comment.. Clair Obscur: Expedition 33: The Continent Walkthrough - Lost Gestrals, Music Records, Outfits, Journals All collectibles locations in The Continent Clair Obscur: Expedition 33 Trophy Guide: All Trophies and How to Get the Platinum How to unlock all Trophies in Expedition 33 Clair Obscur: Expedition 33 Guide: A 100% Collectibles Walkthrough Clair Obscur: Expedition 33: All Journals Locations Where to find every Journal in Expedition 33 Clair Obscur: Expedition 33: All Outfits and How to Get Them Where to find every Outfit for all characters Show More © 2025 Hookshot Media, partner of IGN Entertainment | Hosted by 44 Bytes | AdChoices | Do Not Sell My Personal Information has announced plans to close its Tako board mill and enhance operational efficiency at the Kyro board mill as part of a strategy to address ongoing financial difficulties and improve competitiveness Metsä operates two board machines with a total capacity of 210,000 tpy of folding boxboard (FBB) The board machine at the Kyro mill can produce up to 190,000 tpy of FBB The company informed that it initiated change negotiations at both facilities Up to 215 jobs may be cut as a result of the announced measures The change negotiations are expected to span at least six weeks with final decisions on closures and layoffs made afterward Production at the Tako mill is slated to cease in 2025 if the plan proceeds Metsä Board assures customers that their FBB production capacity would remain sufficient to meet market demand Economic and Operational Challenges The Tako and Kyro mills have been operating under capacity due to weak market conditions and rising costs Metsä already implemented several temporary layoffs at the sites Both facilities reported losses in 2023 and 2024 including high energy costs and limited scalability due to its location and small production lines Metsä Board estimates these measures could bolster its annual Ebitda by approximately €30m the company anticipates a €30 million asset write-down with further one-off restructuring costs to be detailed later Service Customer Service+49 7224 9397-701servicenoSpam@GO-AWAYeuwid.de Editorial Team+49 7224 9397-0papernoSpam@GO-AWAYeuwid.com Get the latest news about developments and trends in the industry sent to you once a week free of charge by newsletter Sign up for our newsletter We use cookies and external services on our website others enhance your user experience or help us improve this website You can change your privacy settings any time by clicking privacy policy Necessary cookies are required for the correct functioning of the website Content from video platforms and map services is blocked by default. 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Or sign-in if you have an account The Conservative party retained its hold on Langley on Monday night winning both Langley Township-Fraser Heights and Cloverdale-Langley City Subscribe now to read the latest news in your city and across Canada Create an account or sign in to continue with your reading experience We apologize, but this video has failed to load.Try refreshing your browser, ortap here to see other videos from our team.Play VideoArticle contentLangley Township-Fraser Heights incumbent Tako Van Popta secured victory early in the night Van Popta had 32,025 votes to Aldag’s 26,292 votes who in 2024 abandoned his federal Liberal seat of Cloverdale-Langley City to run as a B.C where he was defeated by the provincial Conservative candidate Start your day with a roundup of B.C.-focused news and opinion By signing up you consent to receive the above newsletter from Postmedia Network Inc The next issue of Sunrise will soon be in your inbox Interested in more newsletters? Browse here. Van Popta said he was proud to be heading back to Ottawa He said his riding boundary had changed and that he had lost voters in South Surrey who had traditionally supported him “We are looking at a Liberal minority government but with not the same numbers of support from the NDP as before It will be a weak minority government,” he said adding the minority government will likely not last longer than two years The race was tighter in Cloverdale-Langley City where Conservative incumbent Tamara Jansen led from the start of counting Jansen was ahead with 22,942 votes (48 per cent) to Liberal Kyle Latchford’s 22,019 (46.1 per cent) dcarrigg@postmedia.com transmission or republication strictly prohibited This website uses cookies to personalize your content (including ads), and allows us to analyze our traffic. Read more about cookies here. By continuing to use our site, you agree to our Terms of Use and Privacy Policy You can manage saved articles in your account KRQE NEWS 13 - Breaking News, Albuquerque News, New Mexico News, Weather, and Videos NEW MEXICO (KRQE) – Check out community events around New Mexico from May 2 to 8 Albuquerque New Mexico Albuquerque Notices 🔶 Celebrate Mother’s Day at the ABQ BioPark Zoo – The ABQ BioPark Zoo is celebrating all moms during its Mother’s Day at the Zoo on May 11 2024 at 5:22 pm CDTExpandJuan Martín Muñoz and Frederick Abarca two of the owners from Tako Sushi Food Truck are shown standing in front of their food truck Aug DeKALB – A new food truck dubbed Tako Sushi promises its own take on the traditional Japanese-style rolls with a Mexican twist Tuesday marked the food truck’s first day in business at its home in the Lowe’s parking lot, 2050 Sycamore Road, DeKalb Owner Juan Martin Munoz said he’s excited to get his food out there but everything is going good so far,” Munoz said El Senor Roll was prepared by those at Tako Sushi Food Truck and is shown Aug The food truck’s menu features familiar favorites such as a California roll and shrimp tempura roll The menu also features Mexican-style highlights Munoz said the concept behind Tako Sushi is simple “[I’ve] always liked Japanese food,” Munoz said Munoz runs and operates the food truck with Jacqueline Abarca and Frederick Abarca A plate of sushi bomb was prepared by those at Tako Sushi Food Truck and is shown Aug This item is a season dish available to patrons Munoz said he thinks patrons are happy to see a new addition to the business community “Everyone’s saying it’s exciting to have something new,” Munoz said “They’ve never heard of Japanese and Mexican food combined in one The food truck intends to set up shop in the parking lot outside Lowes from 10 a.m Tako Sushi Food Truck also is open to covering events on Saturdays and Sundays The Daily Chronicle is committed to keeping readers up to date with business happenings in the area. Much of our reporting relies on what we see and hear, but we’re also reaching out to readers for tips on business items. If you have a tip to share for Eyes on Enterprise, email news@daily-chronicle.com Megann HorsteadMegann Horstead writes about DeKalb news, events and happenings for the Daily Chronicle - Shaw Local News Network. Support my work with likes, clicks and subscriptions. Metsä Board has finalised change negotiations for the Tako and Kyro mills initiated in January 2025 resulting in the permanent closure of its Tako board mill in Finland the operational efficiency of the Kyro board mill will be improved These measures will lead to a total reduction of 208 jobs Production at Tako board mill is set to cease by the end of the second quarter of 2025 at the latest Metsä Board assures that customer deliveries will not be impacted as the company’s total annual folding boxboard (FBB) production capacity remains sufficient to meet existing demand Metsä operates two board machines with a total capacity of 210,000 tpy FBB Metsä Board’s annual FBB capacity will stand at approximately 1.4 million t The company expects the restructuring to enhance its annual comparable Ebitda by around €39m a one-off negative profit impact of approximately €-28m will be recorded in the first quarter of 2025 as an item affecting comparability a one-off cash impact of approximately €-1m is anticipated Genome Engineering and Neurologic Disorders Volume 2 - 2020 | https://doi.org/10.3389/fgeed.2020.598522 This article is part of the Research TopicMethods, Applications and Developments of Genome Editing ToolsView all 5 articles The development of CRISPR/Cas9 technology has vastly sped up the process of mammalian genome editing by introducing a bacterial system that can be exploited for reverse genetics-based research generating homozygous functional knockout (KO) animals using traditional CRISPR/Cas9-mediated techniques requires three generations of animals A founder animal with a desired mutation is crossed to produce heterozygous F1 offspring which are subsequently interbred to generate homozygous F2 KO animals This study describes an adaptation of the CRISPR/Cas9-mediated method to develop a cohort of homozygous gene-targeted KO animals in one generation A well-characterized ethanol-responsive gene was chosen as a candidate gene for generation of KO mice as proof-of-concept Previous studies have reported changes in ethanol-related behavioral outcomes in MyD88 KO mice One-cell mouse embryos were simultaneously electroporated with four gRNAs targeting a critical Exon of MyD88 along with Cas9 protein DNA and RNA analysis of founder mice revealed a complex mix of genetic alterations all of which were predicted to ablate MyD88 gene function Behavioral testing confirmed the hypothesis that successful one-generation KO of MyD88 would reproduce the decreased ethanol-induced sedative/hypnotic effects and increased ethanol consumption in males that were observed in previous studies This study additionally compared responses of Mock treatment control mice generated through electroporation to controls purchased from a vendor No substantial behavioral changes were noted between control cohorts which we call CRISPR Turbo Accelerated KnockOut (CRISPy TAKO) will be useful for reverse genetic in vivo screens of gene function in whole animals Clustered regulatory interspaced short palindromic repeats (CRISPR) paired with CRISPR associated protein 9 (Cas9) is currently the dominant and preferred gene editing tool in scientific research. CRISPR-based screens of gene function in vitro have been tremendously useful for identifying genes involved in tumor suppression (Michels et al., 2020), mitochondrial function (Khan et al., 2020), and dendritic development (Muir et al., 2020) High throughput CRISPR loss-of-function reverse genetic screens allow for the rapid identification of genes involved in phenotypes of interest in vitro screens are limited by phenotypes that can be readily assayed in cell culture the acquisition of transcriptome data has greatly outpaced our capacity to functionally study genes of interest particularly those that pertain to dysfunction of the central nervous system where behavioral abnormalities are the primary phenotype of interest in vivo tests of behavior must be employed Because behavior is the phenotype of interest we sought to develop a method with moderately high throughput that could be used in vivo to screen genes for effects on behavior CRISPR reagents are introduced to one-cell embryos that develop into founder (F0) animals that are screened for the desired mutation F0 animals are typically an eclectic mix of wild-type (WT) and mutant animals and most mutant alleles differ in the individual mutations they harbor in the target gene of interest A founder animal that harbors a desirable mutation (typically a frameshift or a large deletion) is then mated to WT mice to produce heterozygous F1 offspring heterozygotes are interbred to produce homozygous F2 mutant KO offspring These F2 mutant animals have both alleles of the gene of interest inactivated they all harbor the same mutation in the gene of interest and they can be compared to WT littermate controls for relevant phenotypic changes Although this CRISPR approach for creating gene KO animals represents a dramatic savings in time and expense compared to traditional embryonic stem cell-based gene targeting approaches the CRISPR approach still requires considerable time and expense because three generations of animal production is time consuming and results in substantial animal care and housing expenses This process also requires a considerable amount of personnel time for colony maintenance and genotyping Comparison of the timeline required to produce knockout (KO) mice for behavioral testing using the traditional and CRISPy TAKO approaches (A) Traditional CRISPR/Cas9-mediated method to create a stable KO line Founder (F0) animals are an eclectic mix of wild-type (WT) A founder with an inactivating mutation is selected for breeding to establish a KO line of mice First generation offspring (F1) are heterozygous and must be interbred to produce an F2 generation A subset (~25%) of the F2 generation are homozygous KO mice and can be compared for behavioral phenotypes with WT littermates (B) CRISPy TAKO method for creating functional KO mice By using multiple gRNAs that target a small but functionally critical part of the gene of interest most F0 mice harbor biallelic mutations that functionally inactivate the gene of interest and are suitable for behavioral phenotyping We endeavored to establish a one-generation CRISPR KO approach in which F0 animals could be directly used to test for the behavioral consequences of gene inactivation. We reasoned that a very high efficiency CRISPR mutagenesis approach could be used to efficiently create F0 animals in which both alleles of the gene of interest are mutated and are functionally inactivated (i.e., gene KOs) (Figure 1B) Although each F0 animal may have different mutations they would all be functionally and phenotypically equivalent if a critical part of the gene were sufficiently mutated Single generation F0 MyD88 KO animals were hypothesized to exhibit decreased ethanol-induced sedative/hypnotic effects decreased sensitivity to ethanol-induced motor ataxia and a male-specific increase in ethanol consumption relative to controls To further streamline this accelerated KO mouse protocol we reasoned that for first pass screening of genes for behavioral phenotypes isogenic animals purchased directly from a vendor could be used as a control group for comparison to KOs one concern is that the CRISPR and/or electroporation procedures could hypothetically exert uncharacterized deleterious effects we also created in-house Mock treatment controls that were produced under an identical protocol to the KOs except that the Mock-treated animals were created with procedures that lacked custom CRISPR RNA (crRNA) This Mock-treated control group was directly compared to isogenic C57BL/6J WT mice (Jax controls) purchased from the Jackson Laboratory (JAX) We hypothesized that these two control groups would not differ on behavioral endpoints of interest we describe implementation and validation of a novel technique for the accelerated production of CRISPR KO mice in one generation Animals produced via this protocol are herein affectionately referred to as CRISPR Turbo Accelerated KnockOuts (i.e. We report that our CRISPR protocol can reliably produce a large number of F0 KO animals and that the ethanol phenotype of MyD88 CRISPy TAKOs largely recapitulates results previously reported for traditional MyD88 global KOs vendor purchased mice and Mock treatment controls did not differ substantially these results establish the CRISPy TAKO method for screening gene function in vivo This method has moderately high throughput and will be especially useful for phenotypes All experiments were approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh and conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals C57BL/6J male and female mice used to generate embryos for electroporation and the purchased control group were procured from The Jackson Laboratory (Bar Harbor CD-1 recipient females and vasectomized males were procured from Charles River Laboratories Mice were housed under 12-h light/dark cycles and had ad libitum access to food (irradiated 5P76 ProLab IsoProRMH3000; LabDiet Guide RNAs (gRNAs) were generated using a commercially available two-piece system termed ALT-R™ CRISPR/Cas9 Genome Editing System (IDT DNA, Coralville, IA). This system combines a custom CRISPR RNA (crRNA) for genomic specificity with an invariant trans-activating crRNA (tracrRNA) to produce gRNAs (Homanics, 2019). crRNAs were designed using the computational program CCTop/CRISPRator (Stemmer et al., 2015; Labuhn et al., 2018) which gauges candidate gRNAs for efficiency and specificity the four crRNAs were annealed separately with tracrRNA in a 1:2 molar ratio then combined into a single solution Embryo CRISPy TAKO genotypes for 4930425L21Rik and Gm41261 (A) Sequence results for the major product(s) of TAKO embryos targeting gene 4930425L21Rik electroporated with 100 and 200 ng/μL Cas9 protein Individual embryo numbers are presented on the left The gRNAs and PCR primers used are shown as blue and yellow arrows (B) Agarose gel electrophoresis of PCR amplicons for 4930425L21Rik in embryos Samples 1.1 through 1.8 were electroporated with 100 ng/μL Cas9 protein while samples 3.1 through 4.2 were electroporated with 200 ng/μL Cas9 protein (C) Sequence results for the major product of TAKO embryos targeting gene Gm41261 with 100 and 200 ng/μL Cas9 protein (D) Agarose gel electrophoresis of PCR amplicons for Gm41261 in embryos Samples 5.1 through 5.6 were embryos electroporated with 100 ng/μL Cas9 protein while samples 7.3 through 8.1 were electroporated with 200 ng/μL Cas9 protein (A) Sequencing results from mice selected for behavioral experimentation CRISPR/Cas9 mutagenesis of MyD88 Exon 3 was performed and animals used for experimentation were sequenced to confirm successful deletion Individual animal tag numbers are presented on the left (B) PCR results from DNA of MyD88 CRISPy TAKO mice used for experimentation (C) Random sample subset RT-PCR results from cerebellar brain tissue showing abnormal MyD88 RNA transcripts in TAKOs RT-PCR using Beta-Actin was used as an internal control Female C57BL/6J mice were superovulated with 0.1 mL of CARD HyperOva (CosmoBio followed by 100 IU of human chorionic gonadotropin (Sigma Donor females were caged overnight with C57BL/6J males starting 4–6 h post-gonadotropin injection and allowed to mate Embryos were harvested from oviducts between 9 and 10 a.m cumulus cells were removed using hyaluronidase and embryos were cultured under 5% CO2 in KSOM medium (Cytospring Embryos were electroporated in 5 μL total volume of Opti-MEM medium (ThermoFisher #31985088) containing 100 ng/μL of each sgRNA and 100 or 200 ng/μL Alt-R® S.p #1081060) with a Bio-Rad Gene-Pulser Xcell in a 1 mm-gap slide electrode (Protech International #501P1-10) using square-wave pulses (five repeats of 3 msec 25V pulses with 100 msec interpulse intervals) Two different concentrations of Cas9 protein were used to assess which produced greater mutagenesis in embryos targeting genes 4930425L21Rik and Gm41261; only the 200 ng/μL concentration was used for MyD88 embryos Electroporated embryos were placed back into culture under 5% CO2 in KSOM embryos were cultured for three days until the morulea/blastocyst stage and subsequently analyzed for mutations one- or two-cell embryos were implanted into the oviducts of plug-positive CD-1 recipients (20–40 embryos per recipient) that had been mated to a vasectomized male the previous night Mock-treated controls were manipulated in parallel as described above except that the electroporation mix lacked the MyD88-specific crRNAs (i.e. For 4930425L21Rik and Gm41261, DNA was amplified from individual embryos using a Qiagen Repli-G kit (Qiagen, #150025) and subject to PCR genotyping under the following settings: 95°C for 5 min (1×); 95°C for 30 s, 60°C for 30 s, 72°C for 1 min (40×); 72°C for 10 min (1×). Primers for PCR amplification of 4930425L21Rik and Gm41261 are listed in Supplementary Table 1 respectively) were analyzed by agarose gel electrophoresis and Sanger sequencing For MyD88, DNA was isolated from tail snips of MyD88 CRISPy TAKO and Mock-treated control offspring using Quick Extract (Lucigen, #QE09050). Primers for MyD88 genotyping are listed in Supplementary Table 1 PCR amplicons (WT = 494 bp) were analyzed by agarose gel electrophoresis and Sanger sequencing Samples that did not produce clear chromatograms were subcloned to identify allelic variants The TOPO™ TA cloning kit (ThermoFisher Scientific #K457501) was used according to manufacturers instructions sample PCR product was incubated at room temperature for 15 min with TOPO reagents then the TOPO vector mixture was incubated with chemically competent DH5α (ThermoFisher Scientific #18265017) cells on ice for 30 min Cells were then heat-shocked for 45 s in a 42°C water bath then immediately placed back on ice for 2 min #15544034) was added and cells were incubated in a bacterial shaker at 37°C for 90 min at 225 rpm Cells were plated on kanamycin-resistent LB plates and incubated at 37°C for 16–18 h Single colonies (n = 10 per sample) were collected and their DNA was used for PCR Colonies that produced a single PCR band were then Sanger sequenced A reaction that lacked reverse transcriptase was used as a negative control for each sample tested All mice were moved into a reverse light-cycle housing/testing room (lights off at 10 a.m.) at 5 weeks of age and allowed to acclimate for 2–3 weeks before the start of experiments Experiments were performed in the housing room (ethanol drinking) or an adjoining room (loss of the righting response (LORR) Mice were group-housed 4 to 5 per cage based on genotype and sex The same mice were sequentially tested on the rotarod Injectable ethanol solutions were prepared fresh daily in 0.9% saline (20% Inc.) was injected intraperitoneally (i.p.) at 0.02 mL/g of body weight In order to assess ethanol-induced motor ataxia mice were trained on a fixed speed rotarod (Ugo Basile Training was considered complete when mice were able to remain on the rotarod for 60 s i.p.) and every 15 min mice were placed back on the rotarod and latency to fall was measured until mice were able to remain on the rotarod for 60 s Sensitivity to the sedative/hypnotic effects of ethanol was determined using the LORR assay they were placed in the supine position in V-shaped plastic troughs until they were able to right themselves three times within 30 s LORR was defined as the time from being placed in the supine position until they regained their righting reflex Body temperatures were maintained using a heat lamp throughout the assay v/v) and water for 24 h sessions every other day for 12 days starting at 12 p.m Purchased drinking bottles were 15 mL with 3.5-inch sipper tubes (Amuza The side placement of the ethanol bottles was switched with each drinking session to avoid side preference Ethanol solutions were prepared fresh daily Bottles were weighed before placement and after removal from the experimental cages Empty cages with sipper bottles were used to control for leakage and leakage amount was subtracted from amount consumed by the mice was calculated as g/kg body weight per 24 h Preference was calculated as amount ethanol consumed divided by total fluid consumed per 24 h Statistical analysis was performed with GraphPad Prism (GraphPad Software, Inc., La Jolla, CA) for two-tailed Mann–Whitney test and two-way ANOVA (with mixed-effects analysis (i.e., when technical failures were present), multiple comparisons, and repeated measures when appropriate). Significant main effects were subsequently analyzed with Benjamini, Krieger, and Yekutieli two-stage linear step up procedure post-hoc analysis (Benjamini et al., 2006) Technical failures were approprietely removed from analysis The two control groups were first compared to one another; if no difference was found between control groups these groups were pooled and tested against the MyD88 KO group Graphs show control groups plotted separately even though they were analyzed together Because of well-known sex differences on the behaviors of interest and because male and female mice were tested on separate days Statistical significance was defined as p < 0.05 and q < 0.05 All data are presented as mean ± S.E.M Preliminary testing of the CRISPy TAKO method occurred in vitro using embryos electroporated at the one-cell stage, cultured until the blastocyst stage, then genotyped (Figure 2) each gene was targeted simultaneously with four gRNAs that were tiled across a small section of the gene we tested two concentrations of Cas9 protein (100 and 200 ng/μL) that were higher than the minimum amount we typically use (i.e. none of the embryos electroporated with 200 ng/μL Cas9 harbored WT amplicons that were visible on the agarose gel or detectable by amplicon bulk sequencing five of six embryos electroporated at 200 ng/μL Cas9 protein did not harbor detectable WT amplicons by agarose gel or amplicon bulk sequencing Because the higher 200 ng/μL Cas9 concentration showed greater mutagenic activity in both 4930425L21Rik and Gm41261 this concentration was utilized in targeting MyD88 the mice used for phenotyping presented variable deletions mainly ranging from 200 to 300 bp therefore all mice were expected to manifest the same behavioral phenotypes It is important to note that 100% of all crRNA-electroporated pups contained a MyD88 indel a subset of MyD88 CRISPy TAKOs (n = 15/sex) containing MyD88 inactivating mutations were selected for ethanol-related behavioral interrogation Cerebellar tissue from a random subset of MyD88 CRISPy TAKOs and controls were used for RT-PCR analysis using PCR primers that bind to Exons 2 and 4 to examine MyD88 mRNA. This analysis revealed the expected 280 bp fragment in both WT control samples (Figure 3C) none of the six MyD88 CRISPy TAKO mice examined produced a fragment of this size Five of the six samples produced a predominant band of ~99 bp as would be expected for MyD88 mRNA that lacked Exon 3 One sample produced a major band of ~210 bp and may represent a splicing defect MyD88 CRISPy TAKO mice are likely to be functional KOs Duration of LORR induced by ethanol (3.5 g/kg i.p.) injection in male (A) and female (B) MyD88 KOs Mock controls and Jax controls did not differ and therefore were pooled and compared to same sex MyD88 KOs (data are plotted separately for completeness) KO males had reduced duration of LORR compared to the combined control group (**p < 0.01) no difference was observed between MyD88 KOs and the combined control group Data were analyzed using a two-tailed Mann–Whitney test Post-hoc comparisons revealed that male Mock control mice recovered more quickly than Jax controls at the 15 min timpoint (q = 0.0042) Recovery from ethanol-induced motor incoordination in MyD88 KO and control mice (A) A significant time x genotype interaction was observed for male Mock controls and Jax controls Jax controls recovered more slowly compared to Mock controls at the 15 min timepoint (##q < 0.01) (B) Female Mock controls and Jax controls did not differ and were combined for comparison to MyD88 KOs (data are plotted separately for completeness) Post-hoc analysis did not reveal any significant differences between groups at any time points Data represent time in seconds on the rotarod after injection of ethanol (2 g/kg Data were analyzed by repeated-measures two-way ANOVA with multiple comparisons followed by Benjamini Post-hoc analysis did not reveal any significant differences (C,F) Total fluid intake (g/kg/24 h) in control (Jax C57BL/6J Data were analyzed by repeated-measures two-way ANOVA mixed-effects analysis with multiple comparisons followed by Benjamini and Yekutieli post-hoc tests (*q < 0.05 and ***q < 0.001 between MyD88 KO and combined controls ethanol intake and preference did not differ between female MyD88 KOs and the combined control group The current study reports on a CRISPR/Cas9-mediated mutagenesis protocol that is suitable for rapid screening for the phenotypic effects of gene KO in vivo Traditional CRISPR mouse KO procedures require three generations of animal breeding and genotyping with the CRISPy TAKO protocol described here first generation gene-targeted F0 mice can be rapidly produced and screened for phenotypic effects Although individual F0 mice harbor a variety of mutant alleles for the gene of interest careful project design ensures that each F0 animal is functionally equivalent F0 animals can be directly screened for phenotypes of interest the gene is rapidly eliminated from further consideration a F0 animal can be bred as in traditional approaches to establish a true breeding line that can first be tested to confirm the phenotype This will ensure rigor and reproducibility in the experimental pipeline the line can be maintained long-term and more detailed rigorous mechanistic studies can be conducted The CRISPy TAKO approach can save valuable time and minimize animal numbers and financial resources CRISPy TAKO and C-CRISPR differ in a few subtle we used electroporation whereas C-CRISPR used microinjection of individual embryos large numbers of embryos can be transfected with minimal time and effort and electroporation does not require expensive embryo microinjection equipment and embryo micromanipulation skills we used high fidelity Cas9 protein whereas C-CRISPR used Cas9 mRNA we also tested and validated the use of animals procured directly from a vendor as a control group for comparison to F0 KO animals in phenotypic screens These subtle differences provide the CRISPy TAKO approach with significant savings of time and financial resources We conclude that mice purchased directly from a vendor can be used as a control group for screening CRISPy TAKO mice for behavioral alterations provided the controls are the same genetic background as those animals that served as embryo donors Using a single vendor-derived control group will substantially increase throughput and reduce expenses Comparison of MyD88 cohort results with previous findings The approach described here should be very useful for a first pass screening method to identify genes with a large effect on a phenotype of interest The usefulness of this approach for detecting subtle genotypic differences requires further evaluation we propose using the CRISPy TAKO approach for rapidly screening large numbers of genes in vivo to identify those that have large effects on a phenotype of interest an individual animal that harbors a confirmed KO allele should be mated to establish a true breeding mutant KO line A true breeding line will be useful for future studies to (a) confirm the phenotype of interest (b) to test for and rule out the potential impact of off-target mutations (c) to enable the rigorous testing of control and KO littermates derived from heterozygous matings and (d) to provide an unlimited source of uniform animals for further in-depth analyses and long-term line maintenance We conclude that the CRISPy TAKO method can be used for efficient in vivo screens to identify genes that impact whole animal responses when ablated This method avoids the extensive animal breeding and resources required with traditional CRISPR animal KO approaches This method should find widespread use in studies where moderate to large numbers of genes must be rapidly screened for effects that cannot be interrogated in vitro The raw data supporting the conclusions of this article will be made available by the authors The animal study was reviewed and approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh Project conception and gRNA design devised by GH SP and AS managed the behavioral experimentation together All authors contributed to writing and editing of the manuscript This work was supported by the National Institutes of Health Grants No The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest The authors would like to acknowledge Carolyn Ferguson for expert technical support, Tanya Kenkre, Ph.D for statistical consultation, and members of the INIA-Neuroimmune consortium for helpful discussions and constant encouragement. This manuscript has been released as a pre-print at bioRxivs (Plasil et al., 2020) The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fgeed.2020.598522/full#supplementary-material Adaptive linear step-up false discovery rate controlling procedures CrossRef Full Text | Google Scholar Sedative and motor incoordination effects of ethanol in mice lacking CD14 Highly efficient mouse genome editing by CRISPR ribonucleoprotein electroporation of zygotes Genetics of mouse behavior: interactions with laboratory environment Genome-wide off-target analysis in CRISPR-Cas9 modified mice and their offspring Electroporation of Cas9 protein/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse Gene-edited CRISPy critters for alcohol research PubMed Abstract | CrossRef Full Text | Google Scholar Toll-like receptor-mediated dendritic cell-dependent and-independent stimulation of innate and adaptive immunity CrossRef Full Text | Google Scholar No unexpected CRISPR-Cas9 off-target activity revealed by trio sequencing of gene-edited mice Mitochondrial carrier homolog 2 (MTCH2) is necessary for AML survival Refined sgRNA efficacy prediction improves large-and small-scale CRISPR–Cas9 applications Pooled in vitro and in vivo CRISPR-Cas9 screening identifies tumor suppressors in human colon organoids Efficient mouse genome engineering by CRISPR-EZ technology Bi-allelic loss-of-function variants in NUP188 cause a recognizable syndrome characterized by neurologic CRISPR turbo accelorated knock out (CRISPy TAKO) for rapid in vivo screening of gene function CrossRef Full Text | Google Scholar flexible and reliable CRISPR/Cas9 target prediction tool Stability of inbred mouse strain differences in behavior and brain size between laboratories and across decades Delivery of Cas9 protein into mouse zygotes through a series of electroporation dramatically increases the efficiency of model creation Gene editing in mouse zygotes using the CRISPR/Cas9 system Mutation frequency is not increased in CRISPR–Cas9-edited mice Inhibiting the TLR4-MyD88 signalling cascade by genetic or pharmacological strategies reduces acute alcohol-induced sedation and motor impairment in mice One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs Seth A and Homanics GE (2020) CRISPR Turbo Accelerated KnockOut (CRISPy TAKO) for Rapid in vivo Screening of Gene Function Received: 24 August 2020; Accepted: 05 October 2020; Published: 21 October 2020 Copyright © 2020 Plasil, Seth and Homanics. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Gregg E. Homanics, aG9tYW5pY3NnZUB1cG1jLmVkdQ== Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher. 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish. 2024Astro Bot - Wako Tako Boss Walkthrough (All Collectibles)Here's the Astro Bot walkthrough with all collectibles for Wako Tako in World 2 (Tentacle System) Already a Honolulu Star-Advertiser subscriber? Log in now to continue reading Here’s where you can enjoy this delicious starter: Don Don Donki Kapolei (500 Kamokila Blvd.) opened Feb 15 and is jam-packed with everything you could ever need customers can also find an assortment of sandwiches The latter includes options like unagi ($13.99) Visit donquijotehawaii.com/store/don-don-donki-kapolei 100) is known for serving Japanese fare in a casual atmosphere skewers and bento boxes are all popular options The latter includes a hoso-maki sashimi box ($35) a nigiri hand roll box ($42) and a deluxe nigiri box ($70) crispy rice with spicy tuna ($14 for three pieces) and takoyaki (contact for price) Enjoy authentic Japanese cuisine at Tanto Gyoza & Ramen Izakaya (1035 University Ave including the Tanto tonkotsu black ($17.75) Tanto tonkotsu red ($18.75) and Tanto shoyu ramen ($16.75) The latter is the biz’s original ramen and boasts aged shoyu The biz also carries the gamut of small plates including its popular gyoza ($8 for eight pieces) one-bite tonkatsu ($9.75) and takoyaki ($9.75) Its takoyaki comes with six pieces of grilled octopus topped with bonito flakes Privacy Policy | About Our Ads won the Tako Division at the 36th annual Larry Libres Fishing & Diving Tournament Sato is the oldest tako champion in the 36-year history of the event He was the fifth oldest of the 201 divers who dove in the north shore waters for the event A total of 90 octopuses were brought to the scale by 56 divers a Pukalani resident who started diving 20 years ago brought his winning octopus to the scale at 9:40 a.m. The top 10 tako placers receive awards and prizes This was on my bucket list,” Sato said “I always used to pick up a pretty big tako but then I would always be out of the running So this time when I found this one I was really surprised Sato described how he found and bagged his 5 pound past the lifeguard station close to the stone house I see this little old hole which was kind of big I looked inside and I could see he was home [It] kind of clung to the side of the rocks I grabbed it [and] had a hard time putting it into the bag.” Twenty-four minutes before Sato came to the scale Darrell Naeole was the first diver to weigh his octopus “I was on the Hoʻokipa side of the lifeguard stand I look in a hole… all I see is this rock [It] looked kind of disturbed but maybe it wasn’t It was camouflaged so I figure I [would] just tap it and sure enough that was the four pound kept his string of competing in every Libres contest alive The 79-year-old was able to bring two takos to the scale caught the longest oama in the 119-person 13 and over oama division with the help of her father,topped the 65 keiki in the 12 and under division The free event was sponsored by the Maui Boardsailing Association Dumplings are making their way to the restaurant spot adjacent to The Mill & Mine just north of the Old City The former Tako Taco location on Depot Avenue will soon serve Asian-style dumplings in a new restaurant called Otsu Otsu is an abbreviation for "Otsukaresama," a Japanese greeting that translates to either "thank you for your hard work" or "you did a great job." It's a nod that ties directly into a service mindset at the restaurant "We want guests to come in and we want to treat them," partner and chef Jesse Newmister said "We want to create an area where people can reward them themselves and help people reward themselves." be open in downtown Knoxville?The opening date is somewhat up in the air but Newmister said the goal is mid- to late-September after a handful of soft openings The restaurant will be start out serving dinner only Wednesday through Saturday as well as curated Japanese-style cocktails and small plates Newmister is still working out the details but said there will be five or six culturally traditional core dumplings and a couple from Thailand that even foodies might not know will have Japanese-style charcoal grills for skewers Vegetarian and vegan dishes also are in the works connection to The Mill & Mine music venueNewmister and his wife Stolfi also serves as general manager of The Mill & Mine and helped oversee Tako Taco prior to closing the couple knew that they would be the ones to open the next restaurant instead of leasing out the space Tako Taco already had a well-trained staff Stolfi played a large role in developing the aesthetic and handling finances while Newmister will focus on food preparation The inside of the venue and the outside patio are undergoing a renovation is planning to close its Tako board mill while improving the efficiency of its Kyro board mill and has launched change negotiations at both mills and related operations which the company said are a result of prolonged weak profitability will involve xapproximately 360 employees and may lead to a reduction of up to 215 employees the production of folding boxboard at the Tako and Kyro mills has remained well below capacity due to the market situation Due to the weak market situation and rapidly rising costs Metsä Board said both mills have been loss-making in 2023 and 2024 while the development of the mill is limited by its location in the centre of the city of Tampere Metsä Board has estimated that the planned measures would improve the company's comparable EBITDA by approximately €30m (£25m) per year the measures would lead to a write-down of assets of approximately €30m The Tako mill has two folding boxboard machines with annual production capacities of 70,000 and 140,000 tonnes This represents approximately 13% of the company’s current annual production capacity of approximately 1.6 million tonnes of folding boxboard “Our employees at our Pirkanmaa [Tampere Region] mills are highly skilled and have shown a lot of flexibility in the difficult market situation of recent years which will require capital and a strong financial position in the coming years.” and the company has around 2,300 employees whose parent company Metsäliitto Cooperative is owned by more than 90,000 Finnish forest owners The sales of the whole Metsä Group in 2023 were €6.1bn Printweek welcomes informed debate, but please read our House Rules before posting Printweek is the premier website for the print industry You can reach her at igarfinkel@thejc.com  Imogen Garfinkel is a reporter at the Jewish Chronicle  Imogen Garfinkel 2 min readA Netflix trailer has sparked outrage and boycott calls following the announcement that an Israeli Jew will be playing Jesus’ mother Mary in an upcoming biblical epic will star 21-year-old Noa Cohenin the starring role alongside 22-year-old Israeli actor Ido Tako as Joseph The release of the trailer on social media on Tuesday sparked a storm among users on X/Twitter who immediately criticised the film for its casting of Cohen as the Virgin Mary who some claim should be depicted by an actress of Palestinian descent 🧵 1) Israeli actress Noa Cohen is starring in the new film “Mary” The dumb and racist Anti-Israel crowd are losing their minds because a Jewish woman is playing… a Jewish woman. Here are some examples of their racism. Some hilariously dumb takes. Some quite frightening. pic.twitter.com/6PSVhQt4m2 The cast contains many other Israeli actors and features British film and theatre veteran Anthony Hopkins as King Herrod but needless to say avoid this like the plague,” tweeted one account Half the cast is Israeli, including Noa Cohen who plays Mary. Looks like shit anyway, but needless to say avoid this like the plague https://t.co/bRVNm72Ek6 with one account tweeting: “This is a joke and a slap in the face to all Christians How dare you let that disgusting jew play Mary.” This is a joke and a slap in the face to all Christians How dare you let that disgusting jew play Mary Another account described the casting of Cohen in the titular role as blasphemous “There is something deeply blasphemous about casting an Israeli to play the role of Mary while Israel is carrying out a genocide against Palestinians killing some of the oldest Christian populations in the area and destroying their heritage sites.” a different social media user took issue with the film because the titular roles would be played by “white Europeans” “The actors in the show are both Ashkenazi Israeli Jews The closest people to Mary’s race are Palestinians There is something deeply blasphemous about casting an Israeli to play the role of Mary, the mother of Jesus, while Israel is carrying out a genocide against Palestinians, killing some of the oldest Christian populations in the area and destroying their heritage sites. https://t.co/hnifzq1re7 The actors in the show are both Ashkenazi Israeli Jews, white Europeans. The closest people to Mary’s race are Palestinians. Middle easterners always get whitewashed, blackwashed or indianwashed. Also this movie is disrespectful anyway. https://t.co/0cYy7FgRqy pic.twitter.com/mcM2NHDfxC Another claimed that “Jesus is a Palestinian man described his film to Entertainment Weekly as a “coming of age” story following a “smart strong-willed young woman facing monumental challenges: overcoming social stigma evading a jealous king and bearing the weight of a world-changing destiny” Explaining his decision to cast Israeli actors in the lead roles be selected from Israel to ensure authenticity” Cohen has previously starred in many Israeli films and TV shows including the “Silent Game” (2022) and “8200” (2024) putting her in front of international audiences for the first time Netflix Film Jesus Posted in: Games, Indie Games, Video Games | Tagged: , , Check out the latest free demo for Tako no Himitsu: Ocean of Secrets on Steam right now Indie game developer Christophe Galati and publisher Deneos have released an updated demo for their latest game, Tako no Himitsu: Ocean of Secrets The game has been running a crowdfunding campaign and recently revealed it scored 220% of its funding so it's basically a matter of time before we get a release date the devs gave players an updated free demo on Steam showing off the Game Boy Advance-inspired gameplay Tako no Himitsu: Ocean of Secrets is an indie Action RPG made as a tribute to the Game Boy Advance era of gaming play in a world where Octopuses were erased from history Following centuries of peace after the last war between Octopuses and Humans the world must now face the secrets of its past tensions between countries are brewing following the rise of the mysterious Temple of the Order and shadow voids are appearing across the lands won the Tako Division at the 36th annual Larry Libres Fishing & Diving Tournament Sato showed the young divers how it’s done as he was the oldest tako champion in the 36-year history of the event He was the fifth oldest of the 201 divers who dove in the north shore waters Ninety octopus were brought to the scale by 56 divers topped the 65 keiki in the 12 and under division brought his winning octopus to the scale at 9:40 am an hour and 40 minutes after the 8 am shotgun start The top ten tako placers receive awards and prizes “Twice I was just outside the top ten,” Sato said “I always used to pick up a pretty big tako Sato described how he found and bagged his five-pound had hard time putting it into the bag.” “I was on the Hookpa side of the lifeguard stand Look kind of disturbed but maybe it wasn’t It was camouflaged so I figure I just tap it and sure enough that was the four-pound one ounce The 79-year-old former champion was able to poke two tako which he brought to the scale Copyright © 2025 Maui News Publishing Company LTD | https://www.mauinews.com | 100 Mahalani Street Egli Tako will return to "Big Brother VIP Albania" The news was announced by the production itself through a post on the official Instagram of "BBVA" What will happen?" is written above the video showing Egli staying at home A post shared by Big Brother Alb VIP Edition (@bigbrotheralb_vip) It is not yet clear whether Egli will be reintroduced as a resident or will simply be interviewed by the show's moderator we recall that Egli made a reaction the day before regarding her exit from "BBVA" and the motives that forced her to make this decision She also emphasized that she does not want to have a meeting with him Created by Egli Tako has left the "Big Brother VIP Albania 4" competition The news was announced to the residents through a black envelope explaining that "Big Brother" made this decision due to her health condition Big Brother has made the decision that the contestant will immediately discontinue the competition for health reasons the residents wished Egli all the best in the future A post shared by Big Brother Alb VIP Edition (@bigbrotheralb_vip) due to situations created at home and mostly due to arguments with Gjesti which has not gone unnoticed by viewers of this format She has also often spoken about her 'battle' with anxiety while in conversations with other residents she has stated that she has taken medications to calm her anxiety even during her stay at "BBVA"