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 Law enforcement removed three students out of a Louisiana State University Faculty Senate meeting on Tuesday after the students began shouting demands for the school to divest from corporations linked to Israel.  All three of the students were reportedly members of the leftist Students for a Democratic Society (SDS).  the president of Students for a Democratic Society at LSU told The Reveille that he wasn’t surprised by the activists’ removal; the chapter put in a request for comment at the Faculty Senate meeting but it was denied since it apparently did not correspond with the meeting’s agenda.  [RELATED: UMich student government criticized by pro-Israel org for withholding student funds in divestment demans, calls it a ‘gross abuse of power’]] Spalt also said his organization understood why its request was denied SDS members reportedly rushed to the front of the room The group was also said to have been protesting Resolution 24-01 titled “Supporting Academic Freedom and Inquiry through Institutional Neutrality on Social and Political Issues.”  It’s Spalt’s belief that by approving this neutrality resolution LSU is taking Israel’s side in the Hamas-Israel war.  [RELATED: New school administrators override student senate decision to halt funding to student organizations in latest anti-Israel action]] “LSU is having its pockets lined by people from Israeli companies who are participating or upholding genocide,” Spalt told The Reveille “Our goal is ultimately to negotiate with faculty for divestment.”  Faculty Senate President and political science professor Daniel Tirone insisted that the resolution did not indicate that LSU was supporting Israel in the conflict.  “It’s meant to protect the freedom of the faculty and the students to engage in scholarship and protect their academic freedom,” he told the outlet Tirone also told The Reveille that the Faculty Senate was not the appropriate place for students to stage a demonstration.  Senate Vice President and physics professor Parampreet Singh wanted to be clear that the students weren’t denied comment because of their political stance.  “If the other group would’ve come they would have also been denied public comment,” said Singh “We have to be fair to everyone and we have to follow the law.”  Campus Reform has contacted Louisiana State University and LSU’s chapter of Students for a Democratic Society for comment Mary Stevens is a student at Michigan Technological University in Houghton She is also the media director for her school's chapter of Turning Point USA Mary has years of experience in journalism and writing Metrics details Butterfly eyespot colour patterns are a key example of how a novel trait can appear in association with the co-option of developmental patterning genes co-opted genes function in eyespot development Here we use CRISPR/Cas9 genome editing to determine the roles of two co-opted transcription factors that are expressed during early eyespot determination are sufficient to reduce or completely delete eyespot colour patterns thus demonstrating a positive regulatory role for this gene in eyespot determination deletions in Distal-less (Dll) result in an increase in the size and number of eyespots illustrating a repressive role for this gene in eyespot development Altogether our results show that the presence absence and shape of butterfly eyespots can be controlled by the activity of two co-opted transcription factors A major challenge in eyespot development work Although candidate gene expression patterns are clearly associated with eyespots both developmentally and evolutionarily these genes function in colour pattern formation Because of this it has not been possible to assess the morphological or adaptive significance of eyespot-associated gene co-option Here we use CRISPR/Cas9 genome editing to assess the developmental functions of spalt and Dll in eyespot colour pattern development to show that the co-option of these transcription factors has played a role in the evolution of butterfly wing patterns (a) Location of sgRNAs relative to the V Sequences of selected deletion alleles from eggs and the butterflies shown (M2 (b) Ddc deletions result in depigmentation of larvae (c) An example of mosaic melanin-specific depigmentation in adult wings Sequences of spalt alleles from the animals shown confirm lesions in target regions Green: novel sequences not observed in wild-type alleles Somatic deletions in spalt cause complete loss of forewing eyespots in J coenia: (b) ventral forewing; (c) dorsal forewing; (d) dorsal hindwing cardui ventral hindwings (top) and forewings (bottom) Ectopic wing veins (dashed lines) resulting from spalt deletion subdivide eyespot patterns in (f) J coenia (also showing a missing posterior eyespot) All comparisons shown are left–right asymmetrical phenotypes from individual injected butterflies ‘aberrant’ phenotypes are non-wild type and vary asymmetrically from the normal patterns on the same animal we tentatively speculate that these colour pattern defects are due primarily to vein-related pattern disruption since spalt expression has not been observed in discal spot patterns Altogether our results show that spalt is a positive regulator of eyespots and that it is required for eyespot determination in J Sequences of Dll alleles from the animals shown confirm lesions in target regions (b) Dll deletion causes multiple effects on distal colour patterns of the V including an increase in size and number of eyespots Both wings shown are from the same individual (Vc M15) where the ‘normal’ wing shows a wild-type phenotype while the ‘aberrant’ wing shows several abnormalities as annotated to the right coenia Dll mutant (Jc M3) shows distal elongation of a hindwing eyespot The white dotted circle shows the size of the wild-type eyespot on the opposing wing of the same individual coenia Dll deletion result in patches of dark pigmentation on both the forewing (left) and hindwing (right) The individual on the left also shows disruption of margin patterns and anterior eyespot size asymmetry our strong Dll deletion phenotypes in both species clearly show that Dll plays a variety of roles in colour pattern development organizing parafocal and margin colour patterns In this study we used CRISPR/Cas9 somatic mosaics to show that the transcription factor genes spalt and Dll play key roles in butterfly wing pattern development that the co-option of these genes has played a role in colour pattern evolution the loss and/or reduction of eyespots—clearly shows that this gene plays a role in promoting eyespots spalt is one of the earliest known factors to presage eyespots in imaginal disc development; therefore it is reasonable to hypothesize that this gene is a positive upstream regulator of eyespot determination as is consistent with earlier models based on expression associations (a) Dll expression in line patterns in a mid-last instar V The red box highlights the area of the wing shown in subsequent panels (b) Eyespot markers Notch and spalt are expressed in foci at a time point when Dll is only expressed in a line extending from the eyespot focus to the wing margin (c) A time series of Dll expression shows the progression of line Future models of wing pattern development must consider the complex role of Dll as both a promoter and repressor of multiple colour pattern traits our results show that the early stages of butterfly eyespot determination involve the activity of both promoting and repressing regulators shape and colour of eyespots can be modulated by the activity of two co-opted genes Our surprising finding that Dll is a repressor of eyespot determination requires a significant revision of current models of eyespot development and provides a cautionary tale about interpreting correlations between traits and candidate gene expression levels In addition to providing the first unambiguous functional validation of the role of specific genes in butterfly wing pattern determination our study also demonstrates the power and potential of CRISPR/Cas9 somatic mosaics for work in new model systems cardui caterpillars were obtained from Carolina Biological Supply (Burlington coenia eggs were obtained from Fred Nijhout (Duke University All butterflies were reared on artificial diet at 16/8-h light/darkness at 28 °C and 70% humidity sgRNA template was generated by PCR using Phusion polymerase (New England Biolabs In vitro transcription was conducted using Megascript T7 Kit (Ambion USA) and purified by phenol–chloroform extraction and isopropanol precipitation Butterfly eggs were collected from host plant leaves within an hour of being laid and arranged on double-sided adhesive tape on a microscope slide coenia eggs were dipped in 5% benzalkonium chloride (Sigma-Aldrich and then washed in water for 2 min before mounting on microscope slide Eggs were then dried for 15 min in a desiccant chamber USA) and 250 ng of each sgRNA were mixed in a 2.5 μl volume and injected into eggs using a 0.5-mm borosilicate needle (Sutter Instruments To rule out non-CRISPR variation when scoring phenotypes we only called an individual as having an aberrant deletion phenotype if a trait showed a strong left–right asymmetry Although there is a small degree of individual wing pattern variation in species we surveyed mosaic variation only in individuals injected with sgRNAs and Cas9 Images of all wings from butterflies showing presumptive deletion phenotypes. along with a reference spreadsheet, are available on Dryad: http://dx.doi.org/10.5061/dryad.tj45p Genome editing in butterflies reveals that spalt promotes and Distal-less represses eyespot colour patterns The Development and Evolution of Butterfly Wing Patterns Smithsonian Institution Press (1991) The origin and evolution of animal appendages Pattern formation and eyespot determination in butterfly wings Gene co-option in physiological and morphological evolution A single origin for nymphalid butterfly eyespots followed by widespread loss of associated gene expression Evolutionary history of the recruitment of conserved developmental genes in association to the formation and diversification of a novel trait Cryptic variation in butterfly eyespot development: the importance of sample size in gene expression studies Butterfly wing pattern evolution is associated with changes in a Notch/Distal-less temporal pattern formation process Distal-less regulates eyespot patterns and melanization in Bicyclus butterflies Contribution of Distal-less to quantitative variation in butterfly eyespots Distal-less induces elemental color patterns in Junonia butterfly wings Highly efficient Targeted mutagenesis of Drosophila with the CRISPR/Cas9 system Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies The genetics of dopa decarboxylase in Drosophila melanogaster Isolation and characterization of dopa-decarboxylase-deficient mutants and their relationship to the a-methyl-dopa-hypersensitive mutants The Spalt transcription factors regulate cell proliferation survival and epithelial integrity downstream of the Decapentaplegic signalling pathway The homeobox gene Distal-less induces ventral appendage development in Drosophila The evolution-development interface and advances with the eyespot patterns of Bicyclus butterflies Power tools for gene expression and clonal analysis in Drosophila The developmental physiology of color patterns in Lepidoptera The generation and diversification of butterfly eyespot color patterns Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease Identification of regulatory regions driving the expression of the Drosophila spalt complex at different developmental stages The development of crustacean limbs and the evolution of arthropods Molecular interactions between the protein products of the neurogenic loci Notch and Delta Download references This work was supported by National Science Foundation grant DEB-1354318 to R.D.R the University of Iowa Developmental Studies Hybridoma Bank for Notch antibody and F Wolfner for discussions and comments on the manuscript Department of Ecology and Evolutionary Biology and L.Z.: designed the study and wrote the paper The authors declare no competing financial interests Supplementary Figures 1-2 and Supplementary Table 1 (PDF 2724 kb) Download citation Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Sign up for the Nature Briefing newsletter — what matters in science Read today's Portuguese stories delivered to your email a gifted engineer artisan and talented artist Michael and Tania Spalt have recently made the move to the Algarve with their family where they deem it the “ideal mini version of California” noting that what attracted them to the Algarve is its positive cosmopolitan nature and its sense of community they shared with Central Magazine that they have adapted well with Michael and Tania turning their home into a guitar workshop haven which houses a treasure trove of trinkets “It was always my dream to have a place where I have the space for the workshop at home.” Michael’s guitars are beautifully crafted and captivating where you might think that they are simply true works of art but Michael affirmed that the guitars do in fact play and sound wonderful It was interesting to hear all about Michael’s career and where his love for guitar-making stemmed from “I was heavily into film and music and then I went to school in San Francisco It was just about film and I attended the San Francisco Art Institute so I did a lot of painting and photography I went back to Austria to do compulsory military service I also continued to study art at the Academy for Applied Arts in Vienna “A friend of mine from my San Francisco days said he was making a movie and I should come and help him so I went back to LA and I ended up staying there for more than 25 years doing all sorts of things including special effects work and that is how I got into working with a lot of different materials such as metal I was working in screenwriting where you are mostly working by yourself and I found that I needed to do something with my hands as a balance Michael’s background in the arts field led him to develop the ‘resinTop’ line of guitars called Totemguitars which are particularly striking as they perfectly balance functionality with aesthetics Reliability and playability are fairly objective criteria repairing and modifying numerous instruments has allowed me to achieve a high standard in this regard.“ The guitar certainly has iconic qualities which go well beyond its role as a simple musical instrument More than any other instrument it has a distinctly visual impact it contributes to the image of the musician and his persona in a way that goes far beyond the purely tonal aspects of the music The guitar has become a cultural signpost and the sheer variety of its myriad embodiments is a testament to the central role it plays in our imagination Tania went on to explain how they co-founded an association in Germany for small guitar companies and out of this came a guitar show in Berlin which showcased 135 exhibitors from over 30 countries who we became friends with and he invited us to participate in The Guitar Barrel Project in Lisbon in 2018 Becoming part of that project ultimately led Michael to say to Tania For more information or to request a custom build, please visit https://www.spaltinstruments.com/ and https://www.michaelspalt.com The project was a collaboration of six European luthiers who have each built a guitar using wood originally belonging to the Marquês de Pombal in the form of wine barrels. The creation of the collection of six guitars has been documented in a feature-length documentary, titled The Guitar Barrel Project, which can be watched here: https://www.theguitarbarrelproject.com/the-documentary/ Adriano and German luthier Ulrich Teuffel purchased the wood together comprising some of the best and most celebrated guitar makers in Europe Adriano (Ergon Guitars) and Ulrich (Teuffel Guitars) were joined by Michael and Tania Spalt (Spalt Instruments) Claudio and Claudia Pagelli (Pagelli Guitars) Andy Manson (Andy Manson Custom Guitars) and Nik Huber (Nik Huber Guitars) have created a very special Carcavelos Wine utilising two barrels fully reconstructed from the same wood The collection was showcased to the public at Villa Oeiras / Palácio do Marquês de Pombal in Lisbon in May 2023 and subsequently during a special exhibition at Loulé Criativo It is currently on display at The Quake Museum in Lisbon http://www.theguitarbarrelproject.com Following undertaking her university degree in English with American Literature in the UK Cristina da Costa Brookes moved back to Portugal to pursue a career in Journalism where she has worked at The Portugal News for 3 years Cristina’s passion lies with Arts & Culture as well as sharing all important community-related news We appreciate that not everyone can afford to pay for our services but if you are able to we ask you to support The Portugal News by making a contribution – no matter how small You can change how much you give or cancel your contributions at any time Send us your comments or opinion on this article Reaching over 400,000 people a week with news about Portugal The cohort of Austrian noblemen and one priest had gathered to discuss an idea radical in 1819: a modest bank so craftsmen and small entrepreneurs of their ilk could put money aside and withdraw when necessary they created the Erste Oesterreichische Spar-Casse Over 200 years since that historic event, Erste Group has far outgrown the vision of its founders The multibillion-dollar bank services clients across seven countries in Central and Eastern Europe While it has retained a strong focus on retail banking it has also branched out into corporate banking infrastructure development financing and capital markets But CEO Bernd Spalt is quick to point out Erste Group hasn’t outgrown its purpose “Everybody wants to have a purpose these days We have one which is very old and still very relevant.” Erste Group’s founding document states that the bank offers access to greater prosperity to all “That was a radically modern approach at that time and it’s even more modern today,” Bernd says “It’s not so important that we’re 200 years old Erste Group has endured its fair share of upheaval: two world wars we’ve been able to ensure that our customers’ savings are safe.” the bank has enjoyed the kind of prosperity it seeks to provide to its clients “What we are is essentially a bank that wants to create prosperity in the Eastern part of the European Union,” Bernd explains “We have a balance sheet north of €300 billion we employ nearly 45,000 people and we have around 16 million customers very good mix of digital competence and physical proximity.” It’s a challenge to stay appealing to the next generation This clarity of vision and strength of personality has helped it to weather the storms Erste Group was able to act as a partner easily able to bridge temporary problem situations for clients Although Erste Group is not active in Ukraine the bank seeks to help both refugees from the country and its clients who have been impacted by second-round effects Bernd says it’s not possible to do anything less: “This is who we are.” The Group’s strong identity also inspired Bernd to pursue a career in a field he never originally considered “I was a student of law and economics and had zero dreams of becoming a banker,” he says “But I wanted to be financially independent so when a colleague asked if I wanted a part-time job at a bank Starting on the bottom rung wasn’t Bernd’s dream job but it did give him a deep understanding of the ins and outs of the financial industry small and medium-size enterprises and large corporate customers We’re exposed to political risk and economic cycles; we see ups and downs and we move along with that very colourful and you never stop learning I’m devoted to Erste Group and this profession.” That means Bernd and his team are constantly keeping on top of industry developments to make sure Erste Group remains at the top of its game “It’s always important to stay alert and understand how and why you’ll be relevant tomorrow,” he says a large part of that has been digital competence.” provides a standardised money management experience for eight million customers in six countries a way to create a platform experience,” Bernd says “It was a very important step in our development.” That development has been a lengthy process and one shared by many within the industry ‘What kind of role can a traditional bank like us play?’” I want Erste bankers recognisable no matter where they are not by their clothes or any corporate design That’s something customers will and do appreciate The conclusion the Erste Group’s upper echelons came to was that the bank had lasted as long as it had because it had a clear purpose and it could make a difference in terms of behaviour anybody can sell a loan or take deposits,” he says “But where we can make a very meaningful difference is through behaviour that’s different from others – transparency understandable language free from small print.” closely aligned to the original purpose intended by Erste Group’s founders “I want Erste bankers recognisable no matter where they are not by their clothes or any corporate design That’s something customers will and do appreciate.” Bernd is determined that Erste Group remains attractive to potential employees “It’s a challenge to stay appealing to the next generation One area of enduring appeal is Erste Group’s societal relevance “We’re a large business partner for the NGOs and we work in deep cooperation with them,” he says “People see that this commitment to playing a societal role changes things and that this matters This sense of responsibility again harkens back to the intent of Erste Group’s forefathers and is something that Bernd says provides much comfort in an unstable world when fragile supply chains are stressed and when political risks are rising large players like ourselves have a responsibility to society when it comes to the economy,” he says “And here in particular is where we want to be visible in how we run our shop.” A year before the upcoming renewal of his contract as CEO of Erste Group Bernd Spalt has informed the nomination committee of the banking group’s supervisory board that he has decided not to renew his contract The CEO Magazine is more than a business title; it’s a source of information inspiration and motivation for the world’s most successful leaders Learn all about The CEO Magazine at TheCEO.com ' + scriptOptions._localizedStrings.webview_notification_text + ' " + scriptOptions._localizedStrings.redirect_overlay_title + " " + scriptOptions._localizedStrings.redirect_overlay_text + " Emergency personnel rushed to the scene of a partial barn collapse with fire Saturday morning off Country Club Lane at North Rogers Street near the Waterloo Country Club initial on-scene investigation shows that at 9:35 a.m. a 911 call was placed by a worker on scene stating that the dilapidated barn they were working to tear down at 166 Country Club Lane had collapsed on two of his co-workers Firefighters were eventually able to reach both victims with one man transported by ambulance to Mercy Hospital South in St Louis County for treatment of injuries not considered life-threatening Monroe County Deputy Coroner Theresa Hitzemann pronounced the other trapped individual Assisting the Waterloo and Columbia fire departments and coroner’s office on scene were Monroe County EMS the Waterloo Police Department and Monroe County Sheriff’s Department The fire was reported to be under control within a short amount of time “They had a fire burning in the back of the structure that was not a factor in the collapse,” Waterloo Fire Chief Mike Lloyd told the Republic-Times on Monday part of the structure fell onto the fire.” Hill said an autopsy was conducted Sunday morning to assist in the investigation the Monroe County Sheriff’s Department said a deputy arrived within two minutes of the 911 call to confirm the subjects were trapped and smoke and flames were showing from the collapsed barn Monroe County EMS personnel and an assisting Waterloo police officer located a 42-year-old man from Kirkwood “The first responders were able to lift and break part of the roof to free the male and pulled him to safety,” MCSD Sgt Deputies then located the second trapped worker “Deputies and other first responders attempted to free the victim the weight of the roof that had collapsed on (Spalt) was too much,” Biggs said The Waterloo Fire Department was able to free Spalt “Our investigation revealed three workers were hired to take the barn apart and save the barn wood so the wood could be sold at a later time,” Biggs said “The workers were on the property for approximately two hours before the barn collapsed Two workers were inside the barn during the collapse The third was outside and was one of the individuals who called 911.” Biggs said the workers had cut and were taking down support beams when the collapse occurred.Biggs added that first responders put their own lives at risk in efforts to save the trapped men trying to break some of the roof to get to those trapped,” Biggs said the part of the roof that had collapsed on Mr Spalt was too much weight for the first responders to move by hand.” The MCSD asks the public to remain off the property as the structure is still unsecured and pending investigation by the Occupational Safety and Health Administration due to it being a work-related incident Cobden’s Logan Spalt is The Southern Illinoisan’s baseball player of the year Cobden pitcher Logan Spalt winds up for a pitch in the first inning against Fulton during the Class 1A state baseball semifinal There probably wasn’t a whole lot more Logan Spalt could have done to have a better year on the baseball field than he did this past season The Cobden junior finished 14-1 on the mound with a miniscule 0.49 ERA He also hit a robust .456 with two homers and 42 RBIs His 14 pitching wins are tied for second in the nation Javascript is required for you to be able to read premium content scott.mees@thesouthern.com PEORIA -- Before Cobden's bats won the IHSA Class 1A state championship on Saturday PEORIA -- Logan Spalt shook off a bases-loaded balk to lead Cobden's baseball team to the Cl… Logan Spalt has transformed himself into a complete baseball player for Cobden this season Each year The Southern's sports staff selects players of the year in various prep sports Here are all of 2015's athletes of the year Email notifications are only sent once a day Your browser is out of date and potentially vulnerable to security risks.We recommend switching to one of the following browsers: Account processing issue - the email address may already exist Invalid password or account does not exist Submitting this form below will send a message to your email with a link to change your password An email message containing instructions on how to reset your password has been sent to the email address listed on your account HYANNIS - Ice hockey started in the Spalt family with James the first Spalt to play at Barnstable High School she’s a three-year varsity ice hockey and field hockey player and her ice hockey team hits the Valley Sports ice in Haverhill on Tuesday to face Masconomet in what could be her final high school game but I guess that comes along with it,” Caroline said has been playing ice hockey since she was 7 taking part in the Learn to Skate program at the old Kennedy Rink in Hyannis the younger ones quickly took up the sport “The hockey community here is just so good A big family meant Caroline spent a lot of time playing against boys something she kept doing once she joined competitive teams “When my dad dropped me off for hockey when I was 8 I had to figure out how to tie my own skates because he didn't stay,” she said “It was almost a way for me to advocate for myself and figure out how to do things It was kind of a rule for us: If you want to play hockey carry your own bag and tie your own skates.’” Caroline left the coed level once she reached Pee Wee but going against boys larger than her as a kid has paid off at the high school level She’s only 5-foot-2 and is often one of the smallest players on the ice but she goes after bigger players without a moment’s hesitation while playing left or right wing Ice hockey has been part of Spalt’s life for a decade Field hockey came much more recently when Caroline tried out as a freshman and liked it enough to stick with it She still thinks of herself first and foremost as an ice hockey player But it’s impossible to ignore her impact as a center-midfielder in field hockey where she was a two-time Old Colony League All-Star She helped Barnstable win the OCL in 2013 and make the playoffs all three years she was on the varsity “She is one of the most natural field hockey players I’ve ever coached,” Barnstable coach Ashley Bishop said “When she graduates it will be kind of the end of the Spalt legacy here at BHS.” The Red Raiders (10-7-3) also made the ice hockey playoffs all three of Caroline’s years and won their conference this season She said she works out almost every day during the offseason usually in Hyannis at Fitness 500 with trainer Jeff Handler Handler also worked with older sister Carly Spalt and Caroline’s parents and Caroline started training with Handler in the seventh grade She was disappointed after failing to make varsity as a freshman in either of her two sports and Handler increased her training regimen to help make sure it never happened again “Great athletes are made both physically and mentally,” Handler said and I think the maturity level has increased over the last two seasons.” Spalt also works at Clam Shack and Seaside Saloon in Hyannis in the spring and summer working her way up from busser as an eighth-grader to hostess and hopefully waitress this year She also spends time at home with her mother as it’s just the pair of them now in a house that once was home to nine people Kristen and James Spalt are still together but James moved to Louisiana five years ago to start Cape Coastal Marine an offshore oil transportation company that sons Andrew and James are also involved with Caroline said she sees her father about once a month and travels to Louisiana to visit for big holidays “It kind of makes me grow up a little faster (develop) my independence,” Caroline said.”Being alone Familiarity with Louisiana is one of the biggest reasons Caroline wants to go to school in the South preferably at Louisiana State University but potentially at the University of Mississippi or Texas Christian University Caroline doesn’t yet know what she wants to study but her favorite subjects skew more toward history and she said she’s hoping to join intramural or club teams so she can stay active (none of those three schools offer varsity ice hockey or field hockey) Caroline also wants to get involved with Right to Play International a charity that tries to use sports to positively impact the lives of children in underdeveloped parts of the world It wouldn’t be her first foray into community service - she’s also helped the Raiders organize Skate Nights and veterans outreach events and she’s also involved with the Our Lady of Victory Christmas Fair “She doesn’t sit back and let things happen to her … I am totally going to miss having a Spalt to work with.” She will takes the ice Tuesday for what could be her final high school game Caroline Spalt has already written a wonderful tale both as part of a family legacy and as an individual Please select what you would like included for printing: Copy the text below and then paste that into your favorite email application This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply Service map data © OpenStreetMap contributors History: We end the Hidden Park series with a park that was first dedicated in 1884 making it among the oldest parks in the city It is located across the street from what was once Jefferson Elementary School and is now St using the same basic design as those used at Riverside and Centennial parks a couple of years earlier The cost was estimated at about $26,000 at the time it was renamed to honor the sister city arrangement St A sign was placed there during a visit by more than 70 Spalt residents Spalt Mayor Udo Weingart was pleased by the gesture which make the park a shady oasis surrounded on all four sides by residential streets a climber and a swing set with four swings (two for smaller children) on a bed of sand Sidewalks run around all four of the park's sides What it means: Quite a bit to those who live in the northeast St "I wouldn't like living here if we didn't have a park like this right across the street," said Gene Bottomley who has lived across from the park for over 40 years Follow Frank Rajkowski on Twitter at @rajko1973 or like him on Facebook at www.facebook.com/sctimesfrank Here are links to earlier stories in the Hidden Parks series: Overview story Goldthorpe Park BBC Park Barden Park Colonial Gardens Centennial Park Central Park Crestview Park Southwood Heights Park Carlin Park Beck (nee Spalt) 81 of Oak Crest Retirement Community; beloved wife of Jerome R Beck; devoted mother of Theresa Wydra and her husband Phil Barbara Trentler and her husband Matt and the late Deborah Ann Beck; cherished grandmother of Thomas Wydra and his wife Kristin Daniel Wydra and his girlfriend Amy Johnston Dominic Pedri and his fiancé Courtney Pons Nate Trentler and Nick Trentler; great grandmother of Aubrey Austin and Aria Wydra; also survived by many family and friends Family and friends will honor and celebrate Marie’s life at Peaceful Alternatives Funeral and Cremation Center 2325 York Road (Timonium) on Friday December 17 from 3-5 pm & 7-9 pm with a funeral mass at St contributions may be sent to the American Heart Association or American Diabetes Association in her memory Online condolences may be sent to the family at www.evansfuneralchapel.com Family requests if you are not fully vaccinated please use a face mask to attend viewing and funeral American Heart Association7272 Greenville Ave., Dallas TX 75231Tel: 1-800-242-8721Web: https://www.heart.org/?s_src=22U5W1AEMG&s_subsrc=evg_sem&gclid=CjwKCAiA0KmPBhBqEiwAJqKK4-SbV_dNdXtkD American Diabetes Association2002 Clipper Park Street Ste. 110, Baltimore MD 21211Tel: 1-410-265-0075Web: http://www.diabetes.org ST. CLOUD (WJON News) -- A school on the east side of St Cloud is adopting the city park in its neighborhood Cloud Christian School has announced it is joining the effort to breathe some new life into the East Side by forming a partnership with the city Cloud Christian School is currently running a capital campaign to raise the necessary funds for a brand new playground that also includes the removal of the old inactive and inoperable wading pool Spalt Park is located directly across the street from the school and will be used as the school's playground during school hours and available for use by the community year-round A new multi-use grass recreation area will take the place of the pool The school says utilizing the park for student recreation will free up the space behind the school to add classrooms  The school is asking the community to help support the effort through donations of any size You might remember, that it was just last month the St. Cloud Christian School announced a 10-Day Holiday Lights Festival that will be held from December 1st through the 10th in that same park Cloud Christian School has been in its current building since 2003.  The K-12 school enrolls over 265 students from 16 communities and more than 35 churches St. Cloud School Adopting Neighborhood Park\nRead More ST. CLOUD (WJON News) -- A school on the east side of St You might remember, that it was just last month the St. Cloud Christian School announced a 10-Day Holiday Lights Festival that will be held from December 1st through the 10th in that same park Haunted Rooms America considers it one of the six most haunted hotels in New York City, while the New York Post called it \"the city's spookiest destination for Halloween\" based on legends of the ghosts that roam its halls Guests can still book an overnight stay in one of the hotel's ship-like \"cabin\" rooms and look for ghostly figures loitering the long hallways or in one of the communal bathrooms Accessibility | Terms of Use | Privacy Policy | Modern Slavery Statement chief executive Bernd Spalt’s exit over disagreements with his board comes as a shock to the banking community Russia’s invasion of Ukraine has only served to emphasize how well Erste is doing versus its regional peers Staff at the Austrian lender are unused to frequent changes at the top While Spalt was previously deputy CEO and spent his entire career at Erste he has only held the chief executive job since early 2020 One insider at Erste says Spalt is unlikely to serve out his contract Spalt is not the only European bank chief executive to announce an exit this May the long-standing chief executive of Societe Generale announced he would not renew his contract next year Erste and SocGen have big businesses in central and eastern Europe but Erste stands out among the leading regional players because it is the only one without a large Russian business You have reached the limit for gifting for this month Metrics details The Drosophila wing imaginal disc is a sac-like structure that is composed of two opposing cell layers: peripodial epithelium (PE also known as squamous epithelia) and disc proper (DP also known as pseudostratified columnar epithelia) The molecular mechanism of cell morphogenesis has been well studied in the DP but not in the PE Although proper Dpp signalling activity is required for proper PE formation the detailed regulation mechanism is poorly understood we found that the Dpp target gene sal is only expressed in DP cells Increasing Dpp signalling activity cannot activate Sal in PE cells The absence of Sal in the PE is essential for PE formation The ectopic expression of sal in PE cells is sufficient to increase the PE cell height Down-regulation of sal in the DP reduced DP cell height We further demonstrated that the known PE cell height regulator Lines is mediated by sal mis-activation in PE because sal-RNAi and lines co-expression largely restores PE cell morphology we demonstrated that Lines- and Sal-heightened PE cells are morphologically similar to the intermediate cell with cuboidal morphology the downstream mediators of Dpp signalling remain unknown The absence of sal but not omb insures correct PE morphogenesis The ectopic expression of sal in the PE is sufficient to elongate PE Lin requires sal function to elongate the PE because suppressing sal can rescue the phenotype induced by lin ectopic expression When the microtubule distribution is visualized Sal-elongated PE cells are morphologically similar to cubically intermediate cells that normally link the PE and DP layers sal is not induced by Dpp signalling in the PE otherwise developmental stage is indicated x-y views are oriented with dorsal up and anterior left Cryosections (x-z views) along the dorsal-ventral boundary are oriented with apical up and anterior left In x-y views of (B–G) panels are focused on the PE plane (A) Schematic drawing of wing disc structure in x-z view The wing disc is a sac-like structure composed of two opposing cell layers with apical sides facing an internal lumen Note that pMad is detected in the PE (arrowheads) and the DP Boxed region in B′ is presented at a higher magnification in B″ (C–C″) brk expression pattern in the wing disc Note that brk is transcribed in both the PE (arrowheads) and the DP as reveal by the brk-lacZ reporter Boxed region in C′ is presented at a higher magnification in C″ (D–D″) omb expression pattern in the wing disc Note that omb is only transcribed in the DP Boxed region in D′ is presented at a higher magnification in D″ (E-E″) sal expression pattern in the wing disc Note that sal is only transcribed in the DP Boxed region in E′ is presented at a higher magnification in E″ (F–F″) Expression of brk-RNAi does not induce Sal in the PE (arrowheads) Boxed region in F′ is presented at a higher magnification in F″ (G) Expression of tkvQD does not induce Sal in the PE (arrowheads) Boxed region in G′ is presented at a higher magnification in G″ (H-I″) Expression of tkvQD in clones induced Sal in the DP (arrow) but not in the PE (arrowhead) Boxed regions in I are presented at higher magnifications in I′ (a DP clone (A,C,E,F,I) are x-y views and the remaining images are x-z views (A) Ectopic expression of sal in the dpp-Gal4 domain induces elongation of lateral PE height (A′ (C) Ubiquitous expression of sal elongates PE height (C′ Note that the density of DAPI-stained cell nuclei in the PE is considerably increased compared with control We compared central (flat) cells in C765-Gal4 experiments (C) and lateral (less flat) cells in dpp-Gal4 experiments (A) (E) Expressing omb in the dpp-Gal4 domain does not elongate lateral PE height (E′ (F) Ubiquitous expression of omb does not elongate PE height (F′ (G) The statistics of Image-J program calculated ratio between PE and DP cell heights within one wing disc (H) The statistical diagram of normalized PE height ratio between gene manipulations and corresponding control In both (G,H) means ± SEM indicated by *** are significantly different (pairwise comparison of t-tests (I) Clones co-overexpressing sal and CD8-GFP cell membrane marker Clones in the PE exhibit overgrowth (arrowheads) and elongation of PE height (I′ (K) Expression of lin in the PE leads to the activation of sal transcription in the PE (arrowhead) as revealed by the sal-lacZ reporter (L) Co-expression of salm-RNAi with lin largely rescues PE height (M) Suppression of omb expression in the posterior DP has a non-autonomous effect on PE height (N) Expression of dad in DP induces non-autonomous PE elongation (arrowhead) (O) Expression of brk in DP induces non-autonomous PE elongation (arrowhead) we conclude that sal is sufficient to elongate PE height potential non-autonomous effects between DP and PE can not be ruled out (B) Expression of UAS-tkvDNreduces DP height (C–C″) Expression of salm-RNAi (nub>salm-RNAi) reduces Sal levels in the DP (C and C′) and slightly reduces DP height (C″) (D) Expression of salr-RNAi (nub>salr-RNAi) reduced Sal levels in the DP (D and D′) and slightly reduces DP height (D″) The changes in PE height are not apparent but are statistically significant based on the PE/DP height ratio (E,F) arrowheads) in the medial DP undergo apical retraction (G′ (H) sal-overexpressing clones (absence of GFP arrowheads) in the medial DP undergo apical retraction (H′ Microtubule dynamics during cell height change Red arrowheads indicate the apical side of DP cells Blue arrows indicate the intermediate cells Green bars in (D–G) indicate the cell height of PE (A–C) Time course of microtubule dynamics in wild type control wing discs PE height starts to flatten at the early L3 stage (B Microtubules are polarized to the apical side of the DP (C,D red arrowheads) compared with the even distribution in PE (C,D white arrowheads) and intermediate cells (C (E) Microtubules are enhanced in the PE (white arrow) when sal is expressed (F) lin expression induces similar microtubule changes upon sal expression Note that the cell height and microtubule levels of the PE (E,F white arrows) are similar to that of intermediate cells (C (G) Co-expression of sal-RNAi and lin rescues PE height and microtubule levels similar to that noted in wild type For the efficient expression of RNAi and UAS transgenes driven by the weak C765-Gal4 y w hs-flp; ubi-GFP FRT40/Df(2 L)32FP-5 FRT40 y w hs-flp; act5c>CD2>GAL4/UAS-GFP; UAS-tkvQD/+ y w hs-flp; act5c>CD2>GAL4/UAS-CD8-GFP; UAS-salm/+ y w hs-flp; act5c>CD2>GAL4/UAS-CD8-GFP; UAS-dad/+ Dissected wing imaginal discs were fixed and stained with antibodies according to standard procedures The following primary antibodies were used: rabbit anti-pMad 1:200 (Cell Signalling); mouse anti-α-Tubulin 1:2000 (Sigma); rabbit anti-β-galactosidase 1:500 (a gift from Coralia Pérez Fernández); mouse anti-EGFR 1:200 (abcam); and mouse anti-Wg 1:200 (DSHB) Secondary antibodies (diluted 1:200) included goat anti-mouse DyLight 488 and goat anti-mouse DyLight 549 (Agrisera) and goat anti-rabbit DyLight Cy5 (Jackson ImmunoResearch) F-actin was visualized with Rhodamine-phalloidin Images were collected using a Leica TCS SP2 AOBS confocal microscope Cell height was calculated based on high resolution confocal images using the Image-J program discs were re-fixed for 30 minutes in 4% paraformaldehyde washed and stored in 30% sucrose solution at 4 °C overnight Discs were oriented in Tissue-Tek (Sakura Finetek) frozen and cut into 20-μm sections on a cryostat (YD-1900 Asymmetric distribution of Spalt in Drosophila wing squamous and columnar epithelia ensures correct cell morphogenesis Drosophila gastrulation: from pattern formation to morphogenesis Apical constriction: a cell shape change that can drive morphogenesis Human neural tube defects: developmental biology Strategies of vertebrate neurulation and a re-evaluation of teleost neural tube formation Shaping the actin cytoskeleton using microtubule tips Two distinct mechanisms for long-range patterning by Decapentaplegic in the Drosophila wing Different mechanisms initiate and maintain wingless expression in the Drosophila wing hinge Developmental analysis and squamous morphogenesis of the peripodial epithelium in Drosophila imaginal discs Dpp signalling promotes the cuboidal-to-columnar shape transition of Drosophila wing disc epithelia by regulating Rho1 In The development of Drosophila melanogaster Vol Novel signalling from the peripodial membrane is essential for eye disc patterning in Drosophila Extrusion of cells with inappropriate Dpp signalling from Drosophila wing disc epithelia Extrusion and death of DPP/BMP-compromised epithelial cells in the developing Drosophila wing Spatial discontinuity of optomotor-blind expression in the Drosophila wing imaginal disc disrupts epithelial architecture and promotes cell sorting Fat and Wingless signalling oppositely regulate epithelial cell-cell adhesion and distal wing development in Drosophila Wingless signalling and the control of cell shape in Drosophila wing imaginal discs Lumenal transmission of decapentaplegic in Drosophila imaginal discs Reciprocal roles for bowl and lines in specifying the peripodial epithelium and the disc proper of the Drosophila wing primordium revealing a differential requirement for dpp signalling in initiation and propagation of morphogenesis in the Drosophila eye Injury-induced BMP signalling negatively regulates Drosophila midgut homeostasis Hedgehog creates a gradient of DPP activity in Drosophila wing imaginal discs A gene complex acting downstream of dpp in Drosophila wing morphogenesis Control of the gene optomotor-blind in Drosophila wing development by decapentaplegic and wingless brinker is a target of Dpp in Drosophila that negatively regulates Dpp-dependent genes Conversion of an extracellular Dpp/BMP morphogen gradient into an inverse transcriptional gradient Transducing the Dpp morphogen gradient in the wing of Drosophila: regulation of Dpp targets by brinker The Drosophila gene brinker reveals a novel mechanism of Dpp target gene regulation The orthologous Tbx transcription factors Omb and TBX2 induce epithelial cell migration and extrusion in vivo without involvement of matrix metalloproteinases function and potential origin of the Drosophila gene spalt adjacent which encodes a secreted protein expressed in the early embryo Visualization of gene expression in living adult Drosophila Short-range cell interactions and cell survival in the Drosophila wing Control of growth and positional information by the graded vestigial expression pattern in the wing of Drosophilamelanogaster Roles for Rac1 and Cdc42 in planar polarization and hair outgrowth in the wing of Drosophila A developmentally regulated two-step process generates a noncentrosomal microtubule network in Drosophila tracheal cells Transiently reorganized microtubules are essential for zippering during dorsal closure in Drosophila melanogaster Control of cell flattening and junctional remodeling during squamous epithelial morphogenesis in Drosophila A dynamic microtubule cytoskeleton directs medial actomyosin function during tube formation The Dorsocross T-box transcription factors promote tissue morphogenesis in the Drosophila wing imaginal disc Regulation of the spalt/spalt-related gene complex and its function during sensory organ development in the Drosophila thorax Ectopic eye development in Drosophila induced by directed dachshund expression Direct and long-range action of a DPP morphogen gradient Egfr/Ras pathway mediates interactions between peripodial and disc proper cells in Drosophila wing discs Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis The Drumstick/Lines/Bowl regulatory pathway links antagonistic Hedgehog and Wingless signalling inputs to epidermal cell differentiation Synergistic signalling by two BMP ligands through the SAX and TKV receptors controls wing growth and patterning in Drosophila Daughters against dpp modulates dpp organizing activity in Drosophila wing development dorsalizes both the eggshell and the embryo Induction of Drosophila eye development by decapentaplegic spalt encodes an evolutionarily conserved zinc finger protein of novel structure which provides homeotic gene function in the headand tail region of the Drosophila embryo Analysis of genetic mosaics in developing and adult Drosophila tissues Targeted gene expression as a means of altering cell fates and generating dominant phenotypes Download references the Bloomington Stock Center and VDRC Stock Center for fly stocks and Adi Salzberg and Coralia Pérez Fernández for the Sal antibody This research was supported by the National Natural Science Foundation of China (NSFC31372255) the 973 program (2013CB127603) and the Chinese Universities Scientific Fund (2016QC079) developed the concept and designed the experiments analysed the data and wrote the manuscript Download citation Metrics details Locusta has strong fly wings to ensure its long distance migration but the molecular mechanism that regulates the Locusta wing development is poorly understood To address the developmental mechanism of the Locusta flying wing we cloned the Dpp target gene spalt (sal) and analyzed its function in wing growth in the Locusta The Locusta wing size is apparently reduced with vein defects when sal is interfered by injection of dsRNA indicating that sal is required for locust wing growth and vein formation This function is conserved during the Drosophila wing development To better understand sal’s function in wing growth we then used Drosophila wing disc as a model for further study We found that sal promotes cell proliferation in the whole wing disc via positive regulation of a microRNA bantam Our results firstly unravel sal’s function in the Locusta wing growth and confirm a highly conserved function of sal in Locusta and Drosophila little is known about the molecular mechanism of how the Locusta wing develops into such delicate structure sal may play a conserved role in insect wing development to generate the fast oscillating wings and whether sal functions through ban remains to be illustrated To better understand the developmental mechanism of Locusta flying wings we cloned Locusta sal cDNAs and investigated the role of sal during wing blade and vein formation we found that sal is required for wing growth and vein formation which is conserved between the Locusta and Drosophila using the Drosophila wing disc model we dissected sal function in wing growth and found that sal promotes cell proliferation rate during larval wing development mircoRNA bantam mediates the role of sal in cell proliferation regulation Locusta migratoria was raised at 28 °C with 60% relative humidity migratoria sal genes were confirmed by searching the BLASTX algorithm against the non-redundant NCBI nucleotide database using a cut-off E-value of 10−5 Based on the Drosophila Spalt protein sequences, the deduced Locusta Spalt protein domains were determined by using DNAMAN software (Lynnon Biosoft). Other insect Spalt and Spalt-like proteins are obtained from NCBI (https://www.ncbi.nlm.nih.gov/protein) Global protein alignment and neighbor-joining phylogenic tree were performed by Geneious R9 The primers to make the double strand RNA probes were as follows: Lmsal411-F 5′-ATGTTGCAGCGGCGTGCACAAGAGG-3′ Lmsal411-R 5′-TTGTCCCAGTTGTGCCAGCAGTGGA-3′ Lmsal468-F 5′-GCCATAGACCCTGCTAAGGACCCAG-3′ Lmsal468-R 5′-GTCCTTCACCTCTGCAGCTTGTATC-3′ and GFP-dsRNA were injected into the 4th instar nymphs using 20 μg dsRNAs per insect Nymphs were raised at 28 °C for 10 days until they grew up to the 5th instar Imaging the whole body and wing discs using SONY DSC-HX1camera before the wing discs were lysed Total RNAs were extracted with Trizol reagent from the 5th nymph wing discs and reverse transcribed using the FastQuant cDNA Synthesis kit (Tiangen Biotech) then quantitative real-time RT-PCR was performed using Go Taq qPCR Master Mix (Promega) The primers for quantitative RT-PCR were as follows: Relative transcript levels were assessed using the Comparative CT method Dissected third instar wing discs were co-cultured with BrdU (1:100) in Schneider’s medium for 40–50 min at 25 °C samples were fixed in 4% formaldehyde and washed in PBT before immunostaining The primary antibodies used were mouse anti-BrdU 1:100 (MBL) and mouse anti-β-galactosidase1:2000 (Promega Z3783) and the secondary antibody was anti-mouse DyLight 549 (1:200 Images were obtained using an Olympus FV10-ASW laser scanning confocal microscope and processed with Adobe Photoshop 8.0 Image-J program was used to measure the wing areas and vein distances High resolution images were opened in Image-J program a straight line was drawn at the wanted sites between L2 and L4 veins to calculate the distance; and a wanted region as outlined along the L2 and L4 veins (for Drosophila wing) or the wing margin (for Locusta wing) to calculate the areas Two unigenes named Lmsal411 and Lmsal468 are found in locusts. Protein sequence alignment of Locusta and Drosophila Sal shows highly conserved sequences in the ZnF-C2H2 domains. Stars indicate the identical amino acids. Underlines show the putative Drosophila Salm ZnF-C2H2 domains. and potential Sal from other insects are used to build the neighbor-joining tree the regional growth effects of Sal on wing growth and cell proliferation rate have not yet intensively investigated (A and B) Relative expression of the two genes in the wing discs of 5th instar nymphs after Lmsal411 dsRNAi (A) or Lmsal468 dsRNAi (B) was injected into 4th instar nymphs for 10 days (C) The body and wing size are normal after GFP-dsRNA control injection (D) The wing is smaller in Lmsal411-dsRNA injected locusts (E) The wing is much smaller in Lmsal468-dsRNA injected locusts (C’,D’and E’) show the higher magnification in (C,D and E) five nymphs were injected which give similar results as shown ** and *** represent significant difference with the GFP-dsRNA control (pairwise comparison of t-tests (A) Adult wing of ci-Gal4 control shows the normal wing L1‒L5 indicates the five longitudinal veins The dashed line from the distal end of L2 crossing L3 and extending to L4 indicates the distance between L2 and L4 (B) The wing is larger in salm overexpressed flies (C and D) Wings are smaller when salm or salr is knocked-down and the L2 and L3 veins are fused (E) Adult wing of sal-Gal4 control shows the normal wing The dashed line circles the whole area between L2 and L4 (F) The anterior wing is larger in salm overexpressed flies (G and H) Wings are smaller when salm or salr is knocked-down and the L2 and L3 veins are fused (I) Relative distance between L2 and L4 represented by dashed line in (A) (J) Relative areas circled by L2 and L4 represented by dashed line in (E) The number in (I) and (J) shows the quantification number ***Represents significant difference (pairwise comparison of t-tests (A) The cell proliferation rate is increased when salm is overexpressed in ci-Gal4 region (B) The cell proliferation rate is increased when salm is overexpressed in ap-Gal4 region (C and D) The cell proliferation rate is repressed when salr is knocked-down (E) The cell proliferation rate is repressed when salm is knocked-down White boxes define the area of fluorescence quantification of BrdU staining (A) The expression pattern of br-C12-lacZ reporter in the wild-type wing disc (B) ban is elevated in salm overexpression cells (GFP positive) in both pouch and notum regions (C) salm overexpression clones show upregulated ban level in the whole wing disc (C”) is the magnification of the arrow-indicated clone in (C) spalt is functionally conserved in Locusta and Drosophila to promote wing growth Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Ancient trans-Atlantic flight explains locust biogeography: molecular phylogenetics of Schistocerca The locust genome provides insight into swarm formation and long-distance flight Approaches to the structural modelling of insect wings De novo transcriptome analysis of wing development-related signaling pathways in Locusta migratoria manilensis and Ostrinia furnacalis (Guenee) Geographic variation in wing size and shape of the grasshopper Trilophidia annulata (Orthoptera: Oedipodidae): morphological trait variations follow an ecogeographical rule MicroRNAs of the mesothorax in Qinlingacris elaeodes an alpine grasshopper showing a wing polymorphism with unilateral wing form Veins improve fracture toughness of insect wings Numerical investigation of insect wing fracture behaviour doi: 10.1016/j.jbiomech.2014.10.037 (2015) Morphogen gradients: new insights from DPP A Hedgehog activity gradient contributes to AP axial patterning of the Drosophila wing The bantam gene regulates Drosophila growth The brinker gradient controls wing growth in Drosophila bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila Dpp signaling inhibits proliferation in the Drosophila wing by Omb-dependent regional control of bantam Regulation of spalt expression in the Drosophila wing blade in response to the Decapentaplegic signaling pathway Spalt mediates an evolutionarily conserved switch to fibrillar muscle fate in insects Function of the spalt/spalt-related gene complex in positioning the veins in the Drosophila wing The Spalt Transcription Factors Generate the Transcriptional Landscape of the Drosophila melanogaster Wing Pouch Central Region Spalt major controls the development of the notum and of wing hinge primordia of the Drosophila melanogaster wing imaginal disc Comprehensive microarray-based analysis for stage-specific larval camouflage pattern-associated genes in the swallowtail butterfly Regulation of the Drosophila transcription factor A gain-of-function screen identifying genes required for growth and pattern formation of the Drosophila melanogaster wing Cooperative regulation of growth by Yorkie and Mad through bantam The Decapentaplegic morphogen gradient: from pattern formation to growth regulation Extrusion of cells with inappropriate Dpp signaling from Drosophila wing disc epithelia Comment on “Dynamics of dpp signaling and proliferation control” The Drumstick/Lines/Bowl regulatory pathway links antagonistic Hedgehog and Wingless signaling inputs to epidermal cell differentiation Ultrabithorax regulates genes at several levels of the wing-patterning hierarchy to shape the development of the Drosophila haltere Ultrabithorax is required for membranous wing identity in the beetle Tribolium castaneum Download references the Bloomington Stock Center and Vienna Drosophila RNAi Center for fly stocks This research was supported by the National Natural Science Foundation of China [NSFC31372255] and the 973 Program [2013CB127603] analyzed the data and wrote the manuscript Download citation Metrics details Here we identify the transcription factor Spalt major (Salm) as a master regulator of fibrillar flight muscle fate in Drosophila salm is necessary and sufficient to induce fibrillar muscle fate salm switches the entire transcriptional program from tubular to fibrillar fate by regulating the expression and splicing of key sarcomeric components specific to each muscle type Spalt function is conserved in insects evolutionarily separated by 280 million years We propose that Spalt proteins switch myofibres from tubular to fibrillar fate during development a function potentially conserved in the vertebrate heart—a stretch-activated muscle sharing features with insect flight muscle Prices may be subject to local taxes which are calculated during checkout The changes in power requirements and muscle efficiency during elevated force production in the fruit fly Drosophila melanogaster Molecular genetic analysis of muscle development Systematic genetic analysis of muscle morphogenesis and function in Drosophila Regulation and function of Spalt proteins during animal development Founder myoblasts and fibre number during adult myogenesis in Drosophila Twist and Notch negatively regulate adult muscle differentiation in Drosophila spalt-induced specification of distinct dorsal and ventral domains is required for Drosophila tracheal patterning Two-step process for photoreceptor formation in Drosophila Flightin is essential for thick filament assembly and sarcomere stability in Drosophila flight muscles a protein in the thick filaments of insect muscle Two muscle-specific LIM proteins in Drosophila Control of apterous by vestigial drives indirect flight muscle development in Drosophila The Vestigial and Scalloped proteins act together to directly regulate wing-specific gene expression in Drosophila Shaping leg muscles in Drosophila: role of ladybird a conserved regulator of appendicular myogenesis A troponin switch that regulates muscle contraction by stretch instead of calcium Control of Drosophila paramyosin/miniparamyosin gene expression Differential regulatory mechanisms for muscle-specific transcription a novel Drosophila indirect flight muscle protein has both myosin dependent and independent isoforms Characterization of the gene for mp20: a Drosophila muscle protein that is not found in asynchronous oscillatory flight muscle Expression and function of the Drosophila ACT88F actin isoform is not restricted to the indirect flight muscles Phylogenomic analysis reveals bees and wasps (Hymenoptera) at the base of the radiation of holometabolous insects Larval RNAi in Tribolium (Coleoptera) for analyzing adult development Loss of the Sall3 gene leads to palate deficiency Murine homolog of SALL1 is essential for ureteric bud invasion in kidney development Myofibril assembly visualized by imaging N-RAP The Frank–Starling mechanism in vertebrate cardiac myocytes Wingless signaling induces nautilus expression in the ventral mesoderm of the Drosophila embryo SCORE: a computational approach to the identification of cis-regulatory modules and target genes in whole-genome sequence data is required for myoblast fusion in Drosophila Analysis of Drosophila photoreceptor axon guidance in eye-specific mosaics spalt encodes an evolutionarily conserved zinc finger protein of novel structure which provides homeotic gene function in the head and tail region of the Drosophila embryo Development of the indirect flight muscle attachment sites in Drosophila: role of the PS integrins and the stripe gene Download references the Bloomington and the VDRC stock centres for fly stocks Vogt for critical comments on the manuscript This work was supported by the Max-Planck-Society a Career Development Award by the Human Frontier Science Programme to F.S. a Doc-fForte predoctoral fellowship from the Austrian Academy of Sciences to C.S. Friedrich-Alexander-University Erlangen-Nuremberg analysed the data and created most of the figures acquired the time-lapse movies and performed western blots and M.F conducted the Tribolium RNAi experiments were involved in the initial characterisation of the salm mutant phenotype conceived and supervised the project and wrote the manuscript with input from C.S The file contains Supplementary Figures 1-8 with legends legends for Supplementary Table 2 and Supplementary Movies 1-3 and Supplementary Data for the Tc’sal fragment for RNAi injections 3320bp The table shows top 500 salm targets and IFM specific genes of microarray analysis (see Supplementary Information for full legend) The movie shows early IFM development in a wild-type pupa (see Supplementary Information for full legend) The movie shows early IFM development in a UAS-salm-IR pupa (see Supplementary Information for full legend) The movie shows salm expression in developing IFMs (see Supplementary Information for full legend) Download citation The indirect flight muscles of flying insects have evolved to power insects' wings through as many as 1,000 oscillations per second and to produce extreme mechanical forces These exquisitely specialized muscles contain fibrillar stretch-activated myofibres that are very different from the tubular muscles found elsewhere in the insect's body A genome-wide RNA interference screen for muscle morphogenesis in Drosophila has identified the transcription factor Spalt major (Salm) as a master regulator of fibrillar flight-muscle fate Salm switches the structure of muscles from tubular to fibrillar during development by regulating gene transcription and splicing The spalt gene is conserved in insects that are separated by 280 million years of evolution and the fact that mutations in the human spalt-like gene SALL1 cause heart abnormalities in Townes–Brocks syndrome suggests that spalt function might also determine fibrillar stretch-activated muscle in vertebrates Metrics details MicroRNAs are a class of small non-coding RNAs which act as oncogenes or tumor suppressors through targeting specific mRNAs Colorectal cancer (CRC) is one of the most common malignancies worldwide MiR-3622a-3p is found to be decreased in colorectal cancer (CRC) by analyzing data from TCGA database and there are few reports about the role of miR-3622a-3p in cancers Our research aimed to explore the effects of miR-3622a-3p on CRC MiR-3622a-3p was found to be down-regulated in CRC tissues and cells by qRT-PCR The effect of miR-3622a-3p on proliferation migration and invasion of CRC cells were investigated by a serious of biological function assays and the results revealed that miR-3622a-3p could inhibit the malignant biological properties of CRC RNA immunoprecipitation (RIP) assay and pull-down assay to confirm the interaction between miR-3622a-3p and spalt-like transcription factor 4 (SALL4) Western blot was carried out to determine the effects of miR-3622a-3p and SALL4 on stemness features and EMT We found that miR-3622a-3p suppressed stemness features and EMT of CRC cells by SALL4 mRNA degradation MiR-3622a-3p could inhibit CRC cell proliferation and metastasis in vivo with tumor xenograft model and in vivo metastasis model The CRC organoid model was constructed with fresh CRC tissues and the growth of organoids was suppressed by miR-3622a-3p the results of our study indicate miR-3622a-3p exerts antioncogenic role in CRC by downregulation of SALL4 The research on miR-3622a-3p might provide a new insight into treatment of CRC One of the reasons for CRC patients’ poor prognosis is lack of early diagnosis biomarkers and effective treatment targets It is urgently required to uncover the molecular mechanisms underlying CRC the effect of miR-3622a-3p on CRC has not been elucidated so far we studied the biological functions of miR-3622a-3p and the underlying molecular mechanism The results obtained from our study support the hypothesis that miR-3622a-3p suppresses progression and metastasis of CRC by SALL4 mRNA degradation and inactivation of Wnt/beta-catenin signaling pathway a The volcano plot of the differentially expressed miRNAs in CRC from TCGA database b The cluster heat map of the top 20 up-regulated and down-regulated miRNAs in CRC from TCGA database c The top 10 up-regulated and down-regulated miRNAs in CRC were listed d MiR-3622a-3p was the most down-regulated miRNA in CRC based on the analysis of TCGA miRNAs-seq data e MiR-3622a-3p was confirmed to be down-regulated in CRC by qRT-PCR performed on 80 pairs of CRC tissues and adjacent normal tissues f Overall survival (OS) analysis revealed that high miR-3622a-3p expression was an advantage for CRC patients’ prognosis All data are from three independent experiments and are presented as the means ± SD (*p < 0.05 a Expression level of miR-3622a-3p was detected in CRC cell lines and NCM460 cell line by qRT-PCR b Expression of miR-3622a-3p was increased in DLD-1 by miR-3622a-3p mimics transfection c MiR-3622a-3p expression level was reduced in HCT116 by miR-3622a-3p inhibitor transfection e The effect of miR-3622a-3p on proliferation of CRC cells was evaluated by CCK-8 cell proliferation assay g Colony forming ability of CRC cells was negatively corelated with miR-3622a-3p expression level i The results of EDU assay suggested overexpression of miR-3622a-3p suppressed proliferation of CRC cells while knockdown of miR-3622a-3p promoted CRC cell proliferation b The effect of miR-3622a-3p on cell apoptosis was examined by flow cytometric analysis d The effect of miR-3622a-3p on cell cycle was detected by flow cytometric analysis f MiR-3622a-3p inhibited migration ability of CRC cells h Transwell invasion assay was performed to assess the influence of miR-3622a-3p on CRC cell invasion ability a The predicted binding of miR-3622a-3p at 3′UTR of SALL4 by online databases b The luciferase reporter assay was performed to validate the interaction between miR-3622a-3p and SALL4 c SALL4 protein expression of stable transfected CRC cells was determined by western blot d SALL4 was negatively regulated by miR-3622a-3p by qRT-PCR e SALL4 was up-regulated in CRC cell lines compared with normal colon epithelial cell line by qRT-PCR f SALL4 protein expression was detected by western blot in 6 pairs of CRC tissues and adjacent normal tissues g qRT-PCR was performed on 80 pairs of CRC tissues and adjacent normal tissues for SALL4 detection h SALL4 expression level was negatively corelated with miR-3622a-3p expression level in CRC specimens i Western blot was used to verify protein expression of SALL4 in stable transfected CRC cells j qRT-PCR was performed to determine the expression level of SALL4 in stable transfected CRC cells a–d The effect of SALL4 on CRC cell proliferation was evaluated by colony formation assay The rescue experiment for miR-3622a-3p overexpression was performed by ectopic expression of SALL4 in DLD-1 The rescue experiment for miR-3622a-3p silencing was performed by inhibition of SALL4 in HCT116 e–h The promoting role of SALL4 in CRC cell proliferation was further validated by the EDU assay and the rescue experiments were performed i–l Flow cytometric analysis was employed to explore the impact of SALL4 on apoptosis of CRC cells and the rescue experiments were performed a–d Flow cytometric analysis was used to examine the percentage of CD133(+) CRC cells e–h Sphere formation assay was performed to investigate the stem-cell like properties of stable transfected CRC cells and CD133 were determined by qRT-PCR in stable transfected CRC cells l Protein levels of CSC-related biomarkers and pluripotency-related genes were detected by western blot in stable transfected CRC cells b Protein levels of EMT markers were determined by western blot in stable transfected CRC cells d The effects of miR-3622a-3p and SALL4 on Wnt/beta-catenin signaling pathway were explored by TOPflash/FOPflash luciferase reporter assay f Expression levels of Wnt/beta-catenin signaling pathway related proteins were detected by western blot MiR-3622a-3p was demonstrated to suppress CRC in vitro in the above experiments and we were interested in whether miR-3622a-3p could exert similar influences in vivo a Representative xenograft tumors derived from nude mice of different groups b The growth curves of the xenograft tumors were drawn according to tumor volumes which were measured every 4 days c The weight of the harvested tumors was recorded on day 24 d The results of ki67 staining and TUNEL assay revealed that overexpression of miR-3622a-3p inhibited proliferation and promoted apoptosis of DLD-1 xenograft tumors SALL4 protein expression level was confirmed by IHC and western blot to be down-regulated by miR-3622a-3p overexpression e Knockdown of miR-3622a-3p promoted proliferation and inhibited apoptosis of HCT116 xenograft tumors SALL4 protein expression level was elevated by miR-3622a-3p knockdown g Overexpression of miR-3622a-3p suppressed lung metastasis of CRC cells in vivo while silencing of miR-3622a-3p exerted the opposite influence i Hematoxylin and eosin staining were performed on the harvested lungs from nude mice and the number of metastatic foci was counted k High miR-3622a-3p expression level was a favorable factor for OS of nude mice while low miR-3622a-3p expression level was a risk factor l The effects of miR-3622a-3p overexpression and knockdown on growth of CRC organoids m MiR-3622a-3p expression in CRC organoids was detected by qRT-PCR following transfection with miR-3622a-3p mimics or inhibitor lentivirus n The expression level of SALL4 in CRC organoids was determined by qRT-PCR Then miR-3622a-3p expression was confirmed to decrease in CRC by performing qRT-PCR on 80 pairs of CRC tissues and adjacent normal tissues We also analyzed the overall survival of the CRC patients recruited to the study and found miR-3622a-3p was a positive factor for diagnosis of the patients According to the clinicopathological features of the CRC patients miR-3622a-3p was negatively corelated with CRC tumor size and lymph node metastasis we hypothesized that miR-3622a-3p might act as a tumor suppressor in CRC and further research needed to be done RIP assay and pull-down assay were adopted to confirm the interaction between miR-3622a-3p and SALL4 Expression level of SALL4 was determined by qRT-PCR and western blot to be negatively corelated with miR-3622a-3p expression level The results of qRT-PCR and western blot also revealed SALL4 was up-regulated in CRC cells and tissues The expression level of SALL4 was observed to decrease with increasing of miR-3622a-3p in CRC tissues by linear correlation analysis Rescue experiments were conducted to find out whether functions of miR-3622a-3p could be mediated by regulation of SALL4 overexpression of SALL4 could reverse the effect of miR-3622a-3p on CRC cell proliferation The influence of miR-3622a-3p knockdown could also be reversed by silencing of SALL4 SALL4 is not only essential for pluripotency and self-renewal of ESCs, but also important for regulation of CSCs maintenance46 hence we studied the effects of miR-3622a-3p and SALL4 on stemness features of CRC MiR-3622a-3p decreased the percentage of CD133(+) CRC cells and reduced the sphere forming ability of CRC cells the effects of miR-3622a-3p on EMT and Wnt/beta-catenin signaling pathway were explored miR-3622a-3p resulted in inhibition of EMT and inactivation of Wnt/beta-catenin signaling pathway The suppressive effects of miR-3622a-3p on stemness features and Wnt/beta-catenin signaling pathway might account for inhibition of organoid growth after being transfected with miR-3622a-3p mimics The application of the organoid model in our study was only limited to laboratory research We will try to construct more human CRC organoids and develop individual treatment plans for CRC patients based on the responses of organoids to chemotherapeutic drugs Our data revealed that miR-3622a-3p was aberrantly down-regulated in CRC and inhibited progression and metastasis of CRC cells in vitro and in vivo High expression level of miR-3622a-3p was an advantage for prognosis of CRC patients MiR-3622a-3p exerted anticancer influences by decreasing SALL4-mediated stemness features and inactivating EMT and Wnt/beta-catenin signaling pathway in CRC Our research on miR-3622a-3p might help understanding the molecular mechanisms underlying CRC and provide new insights into therapeutic strategies for CRC The 80 pairs of CRC tissue samples and adjacent normal tissue samples were obtained from Department of General Surgery the First Affiliated Hospital of Nanjing Medical University The patients recruited to our study did not receive preoperative chemotherapy and radiotherapy All the patients signed written informed consents before the study The research was approved by theInstitutionalEthical Board of the First Affiliated Hospital of Nanjing Medical University The clinical stages of the CRC patients were determined according to the International Union Against Cancer (UICC) on Tumor-Node-Metastasis (TNM) staging system (7th edition) The collected tissues samples were stored in liquid nitrogen before use LoVo and DLD-1 were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai Human normal colon epithelial cell line NCM460 was obtained from American Type Culture Collection (ATCC Cell lines were authenticated using Short Tandem Repeat (STR) analysis and tested for mycoplasma contamination All of the cell lines were cultured in DMEM medium (Wisent 100 U/ml penicillin and streptomycin (15140148 USA) in a moist incubator with 5% CO2 at 37 °C the lentiviral vector containing SALL4 DNA sequencing (LV-SALL4) and the lentiviral vector containing SALL4 shRNA sequence (sh-SALL4) were purchased from GenePharma (Shanghai Aldrich) to screen the transfected CRC cells to establish stable transfected cell lines according to the manufacturer’s protocol Total RNAs were extracted with Trizol Reagent (15596018 Japan) and New Poly(A) Tailing Kit (ThermoFisher China) were used for mRNA and miRNA reverse transcription respectively into cDNA followingthemanufacturer’s instructions We carried out qRT-PCR with a 7500 Realtime PCR System (Applied Biosystems USA) and SYBR Green Master Mix (4913914001 Relative expression of miR-3622a-3p was normalized to snRNA U6 Beta-actin was taken as internal control for pri/pre-miR-3622a-3p The 2−ΔΔCT analysis method was used to calculate the relative expression of miR-3622a-3p and SALL4 The primers for qRT-PCR were listed below: has-miR-3622a-3p forward 5′-AACGCTTCACGAATTTGCGT-3′; pri-miR-3622a-3p forward 5′-TCGTGAGCTGCTTGATGACTGAT-3′; pri-miR-3622a-3p reverse 5′-AGGAAGCCCAGGAAACCCTTTG-3′; pre-miR-3622a-3p forward 5′-ACCTGACCTCCCATGCCT-3′; pre-miR-3622a-3p reverse 5′-TATGCTTGTTCTCGTCTCTGTGTC-3′; SALL4 forward 5′-GAGCGGACTCACACTGGAGA-3′; beta-actin forward 5′-GCATCGTCACCAACTGGGAC-3′; beta-actin reverse 5′-CTGGGGCTGCTGTTTATTATTCTG-3′; CD133 reverse LoVo cell lines and modified with bisulfite The CpG islands were amplified by PCR and the products were separated by agarose gel electrophoresis and cloned into the pUC18 T-vector (Sango DNA sequencing on 10 clones was performed after bacterial amplification of the cloned PCR fragments To assess proliferation of stable transfected cells 2 × 103 cells were seeded into each well of a 96-well plate The cells in each well were incubated with 10 μl CCK8 reagent for 2 h at 37 °C Absorbance at 450 nm was measured by a microplate reader at the same time point for 5 days DNA synthesis of stable transfected CRC cells was measured with an EDU assay kit (C00052 The cells were seeded at a density of 2 × 104 cells per well into a 24-well plate and cultured in DMEM containing 10% FBS for 24 h After incubation with 50 uM EDU reagent at 37 °C for 2 h the cells were fixed and permeabilized with 4% formaldehyde and 0.5% TritonX-100 respectively at room temperature (RT) Then we added 1 × Apollo R reaction cocktail (400 μl) to each well 400 μl Hoechest33342 was added to stain the nuclei of stable transfected cells Red and blue signals were observed and taken by a Nikon microscope (Nikon The stable transfected cells were seeded into a six-well plate (500 cells/well) and cultured in DMEM (10% FBS) for 15 days the colonies were stained with crystal violet (Beyotime We washed the colonies with phosphate buffered solution (PBS) for three times and counted the number of colonies The stable transfected cells were digested with trypsin and then collected After being washed with PBS twice and fixed with 75% ethyl alcohol incubated with RNase and stained with a Cell Cycle Staining Kit (CCS012 The cell-cycle was analyzed with a FACScan flow cytometer (BD To evaluate apoptosis of stable transfected cells the cells were collected and stained with a PE Annexin V Apoptosis Detection Kit I (559763 The ratio of apoptotic cells was determined by a FACScan flow cytometer (BD The collected stable transfected cells were incubated with PE-conjugated CD133 antibody (372804 Then the cells were placed on ice for 10 min and washed with precooled PBS before flow cytometric analysis for CD133(+) cells detection To assess the migration ability of stable transfected cells 2 × 104 stable transfected cells were seeded into the upper chamber and cultured in 200 ul DMEM without FBS 500 μl DMEM medium containing 10% FBS which acted as chemoattractant was added to the lower chamber Part of the cell migrated to the underside of the membrane The cells which did not penetrate the membrane were removed and the cells on the lower surface of the membrane were fixed and stained with 75% alcohol and crystal violet respectively The stained cells were counted with a microscope USA) was coated on the upper side of the membrane before cell plating The remaining steps were similar to those in migration assay The 3’-UTR sequences of SALL4 containing wild-type (WT) or mutated (MUT) miR-3622a-3p binding site were designed and synthesized by GeneScript (Nanjing The sequences were cloned into a a pGL-3 luciferase reporter vector (Promega DLD-1 was co-transfected with miR-3622a-3p mimics or miR-NC and pGL3-WT-SALL4 or pGL3-MUT-SALL4 HCT116 was co-transfected with miR-3622a-3p inhibitor or miR-NC and pGL3-WT-SALL4 or pGL3-MUT-SALL4 The luciferase activity was determined with a Dual Luciferase Reporter Assay System (Promega The ratio of firefly luciferase to renilla luciferase was defined as the relative luciferase activity RNA immunoprecipitation (RIP) assay was carried out with a Magna RNA immunoprecipitation kit (Millipore Stable transfected CRC cells were lysed with RIP buffer The cell lysis was incubated with magnetic beads conjugated with anti-Ago2 antibody (03–110 the immunoprecipitated RNA was extracted and followed by qRT-PCR Biotinylated-miR-3622a-3p (Bio-miR-3622a-3p) and biotinylated-miR-NC (Bio-NC) were purchased from GenePharma (Shanghai DLD-1 and HCT116 were transfected with the constructs The cell lysis was incubated with streptavidin-coated magnetic beads (Invitrogen the biotin-coupled RNA complex was pulled down and followed by qRT-PCR China) was used to extract total proteins from paired CRC tumors and adjacent normal tissues and CRC cells The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore After being blocked in 5% non-fat milk at RT for 2 h the membranes were incubated with primary antibodies at 4 °C all through the night The membranes were washed with TBST buffer for three times and incubated with secondary antibodies at RT for 2 h the next day we washed the membranes for three times using TBST buffer and the proteins on the membranes were visualized by an enhanced chemiluminescence (ECL) detection system (Millipore The primary antibodies included: anti-SALL4 (ab57577) anti-beta-catenin (ab32572) and anti-Cyclin D1 (ab16663) from Abcam (Cambridge anti-TCF1 (#2203) and anti-c-myc (#5605) from Cell Signaling Technology (Boston The secondary antibodies used in our study including anti-rabbit IgG-HRP (ab6721) and anti-mouse IgG-HPR (ab6789) antibodies Stable transfected cells were seeded into a six-well ultra‐low attachment surface plate (Corning The cells were cultured in serum-free DMEM/F12 (Gibco The culture medium was supplemented with B27 (Invitrogen The spheres were observed and photographed 10 days later The Topflash/FOPflash reporter plasmids were obtained from Upstate Biotechnology (NY The cells were transfected with the plasmids using Lipofectamine 3000 (L3000015 The luciferase activity was detected by a Dual Luciferase Reporter Assay System (Promega The results were shown as normalized TOPFlash/FOPFlash values The BALB/c nude mice (aged 5 weeks) used in our study to build tumor xenograft model and tumor metastasis model were purchased from Animal Center of Nanjing Medical University (NJMU) The animal experiments were approved by NJMU Animal Ethics Committee 1 × 106 CRC cells stably transfected with miR-3622a-3p mimics or inhibitor were injected into the flanks of nude mice (6 mice/group) The volume of the tumors was measured every 4 days with a vernier caliper following calculation formula: volume = (width2 × length)/2 1 × 106 stable transfected CRC cells suspended in 100 ul PBS were injected into lateral tail veins of nude mice the distant metastases of 8 nude mice in each group were visualized with an IVIS Imaging system (Caliper life Sciences The remaining of the nude mice were kept to analyze the effect of miR-3622a-3p on survival of nude mice with 12 weeks as cutoff All animal experiments were performed in accord with the National Institutes of Health Guide for the Care and Use of Laboratory Animals Standard of blinding and randomization was complied with in this study The subcutaneous tumors of nude mice were obtained and fixed in 4% formaldehyde Then they were embedded in paraffin and cut into 4 μm thick sections The sections were incubated with primary antibodies such as anti-ki67 (ab156956) and anti-SALL4 (ab57577) overnight at 4 °C After being washed with PBS for three times the sections were incubated with HRP-polymer-conjugated secondary antibody at RT for 1 h We used 3,3′-Diaminobenzidine (DAB) solution to stain the sections for 3 min and hematoxylin to counterstain nuclei The percentage of positive cells was determined based on three random fields of the sections We used a TUNEL apoptosis detection kit (C1091 China) for this assay according to manufacturer’s protocol The subcutaneous tumor sections were rehydrated in the ethanol and fixed in 4% formaldehyde The sections were then incubated with proteinase K at RT for 20 min and 3% hydrogen peroxide was used to inactivate endogenous peroxidases Working solution and chromogenic agent were prepared following manufacturer’s instructions Hematoxylin was used to stain nuclei of the cells The percentage of apoptotic cells in randomly selected fields was determined with a microscope (Nikon The lungs of the nude mice were fixed in 4% formaldehyde and embedded in paraffin The sections prepared from paraffin mass was incubated with hematoxylin for 3 min and washed with deionized water Then we used eosin Y solution to dye the sections and 95% alcohol followed by absolute ethanol to dehydrate the specimens xylene was adopted for alcohol extraction and neutral balsam was used to seal the sections The CRC organoid model was constructed based on the protocols published previously39 The organoids were transfected with miR-3622a-3p mimics or inhibitor lentivirus the diameters of the organoids were measured Then the organoids were collected from Matrigel for miR-3622a-3p and SALL4 detection with qRT-PCR All the statistical analyses adopted in the study were performed with Statistical Product and Service Solutions (SPSS) 20.0 software The data was shown as mean ± standard deviation (SD) All the experiments were carried out at least three times Linear correlation analysis was performed to analyze the correlation between miR-3622a-3p expression and SALL4 expression χ2 test was used to determine the relationship between miR-3622a-3p expression level and the CRC patients’ clinicopathological features The Caplan-Meier method was adopted in survival analysis Two-tailed Student’s t-test and one-way analysis of variance (ANOVA) were performed to analyze the data obtained from experiments P < 0.05 (*) and P < 0.01 (**) were considered statistically significant Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries Colorectal cancer in 2017: practice-changing updates in the adjuvant and metastatic setting Control of translation and mRNA degradation by miRNAs and siRNAs miR-3622a promotes proliferation and invasion of bladder cancer cells by downregulating LASS2 A novel microRNA regulator of prostate cancer epithelial-mesenchymal transition Sall4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1 Sall4 interacts with Nanog and co-occupies Nanog genomic sites in embryonic stem cells Genome-wide analysis reveals Sall4 to be a major regulator of pluripotency in murine-embryonic stem cells is constitutively expressed in human acute myeloid leukemia (AML) and induces AML in transgenic mice SALL4 is a key regulator of survival and apoptosis in human leukemic cells The transcription factor SALL4 regulates stemness of EpCAM-positive hepatocellular carcinoma SALL4 as an epithelial-mesenchymal transition and drug resistance inducer through the regulation of c-Myc in endometrial cancer SALL4 is essential for cancer cell proliferation and is overexpressed at early clinical stages in breast cancer a novel marker for human gastric carcinogenesis and metastasis Inhibition of SALL4 suppresses carcinogenesis of colorectal cancer via regulating Gli1 expression SALL4 as a new biomarker for early colorectal cancers Cancer stem cells: current status and evolving complexities STRAP promotes stemness of human colorectal cancer via epigenetic regulation of the NOTCH pathway Kinesin family member 15 promotes cancer stem cell phenotype and malignancy via reactive oxygen species imbalance in hepatocellular carcinoma EMT and inflammation: inseparable actors of cancer progression Wnt signalling and its impact on development and cancer WNT signalling pathways as therapeutic targets in cancer Signaling mechanisms of the epithelial-mesenchymal transition MiR-92a promotes stem cell-like properties by activating Wnt/beta-catenin signaling in colorectal cancer Downregulation of microRNA-362-3p and microRNA-329 promotes tumor progression in human breast cancer A feedback loop between miR-30a/c-5p and DNMT1 mediates cisplatin resistance in ovarian cancer cells Downregulation of miR-130b~301b cluster is mediated by aberrant promoter methylation and impairs cellular senescence in prostate cancer SALL4 - KHDRBS3 network enhances stemness by modulating CD44 splicing in basal-like breast cancer Upregulation of SALL4 by EGFR activation regulates the stemness of CD44-positive lung cancer MicroRNA-137 reduces stemness features of pancreatic cancer cells by targeting KLF12 Inhibition of SALL4 reduces tumorigenicity involving epithelial-mesenchymal transition via Wnt/beta-catenin pathway in esophageal squamous cell carcinoma Knockdown of Sall4 inhibits intrahepatic cholangiocarcinoma cell migration and invasion in ICC-9810 cells Oligomeric proanthocyanidins (OPCs) target cancer stem-like cells and suppress tumor organoid formation in colorectal cancer Organoid models of human and mouse ductal pancreatic cancer Global burden of colorectal cancer: emerging trends Therapeutic inhibition of miR-4260 suppresses colorectal cancer via targeting MCC and SMAD4 MicroRNA-494 promotes cancer progression and targets adenomatous polyposis coli in colorectal cancer miRNA-99b-5p suppresses liver metastasis of colorectal cancer by down-regulating mTOR Spalt-like transcription factor 4 as a potential diagnostic and prognostic marker of colorectal cancer SALL4: an emerging cancer biomarker and target Organoid culture of human cancer stem cells Modeling development and disease with organoids Plasma MicroRNA signature validation for early detection of colorectal cancer Hedgehog signals mediate anti-cancer drug resistance in three-dimensional primary colorectal cancer organoid culture Download references The work was financially supported by the Key Project of Administration of Traditional Chinese Medicine of Jiangsu Province (JD2019SZXZD01); the National Nature Science Foundation of China (81904203) These authors contributed equally: Shuchen Chang The Affiliated Hospital of Nanjing University of Chinese Medicine The First Affiliated Hospital of Nanjing Medical University The authors declare that they have no conflict of interest Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41419-020-02789-z Article GENDER R25 will provide funding for courses and curricula to improve gender-focused education and equity in pharmacy A new program from the Office of Research on Women’s Health (ORWH) in the National Institutes of Health (NIH) is aiming to improve sex- and gender-specific training in science and other health care professions by supporting curricula and courses that address the intentional integration of sex and gender considerations into NIH research as breakthroughs in women’s health research continue to occur at every stage of the biomedical research continuum the paradigm shift from inclusion of women to the intentional integration of sex- and gender-related differences and information requires the development of new skills and innovative methodologies an ORWH social and behavioral scientist administrator explained further in an interview with Pharmacy Times that ORWH welcomes applications from pharmacy professionals as pharmacists are critical members of patient health care teams with direct impact on managing adverse effects (AEs) and polypharmacy—all of which can impact women more than men due to the limited nature of data on these subjects in relation to sex and gender-related differences sex and gender remain underexplored in curricula within biomedical educational programs “As each health profession has its own discipline-specific language and accreditation standards pharmacy professionals are ideally positioned to develop curricula to fill the gap within their field; we are hopeful that pharmacy professionals will consider submitting applications for the GENDER R25.” With the understanding of the clear impacts sex and gender have on health using clear terminology for sex and gender is essential to facilitate a more accurate study of human health and the delivery of health care by professionals in the field The classification of sex—which is a multidimensional construct based on clusters of anatomical and physiological traits—can include external genitalia is often misunderstood as being an extension of or synonym for sex; instead gender is a social and cultural variable that includes a variety of domains “Understanding the influence of gender on health...can facilitate the delivery of equitable care by all biomedical professionals,” Barr said “Pharmacy professionals could all benefit from an enriched understanding of the domains of sex and gender [in their practices].” By understanding the differences in terminology and the impact of both sex and gender on patient health pharmacy professionals can better provide more effective individualized health care professionals themselves can impact patient health care outcomes based on the results of 1-on-1 communication with patients regarding the patients experience of symptoms gender can impact patient health on a relational and structural level as well gender may have an impact on whether or not a patient may be experiencing intimate partner violence in their home setting while structural impacts of gender can include varying levels of access to health care based on geographic location for certain female-specific conditions Federal law and NIH policy require clinical research funded by the NIH to include women this does not only include enrolling women in studies but must also require considering research design Studies must be designed to detect the influence of sex on efficacy and safety If women are included but sex-specific results are not collected and reported women and health care professionals lack crucial knowledge that can influence patient health sex differences can also have impacts on treatment outcomes such as through varying levels of absorption women typically have different amounts of adipose tissue and may have different kidney and liver functions compared with men all of which can influence dosages and AEs women are often underrepresented in clinical trials leading to results that may not be disaggregated by sex with potential drug distribution differences remaining unidentified Gender can also impact whether and how patients report AEs as well as how those effects are perceived by health care professionals Women have frequently reported that they experienced the dismissal of their reported symptoms in health care settings with this experience confirmed in research published in professional journals This dismissal of symptom experience can result in delayed dose adjustments and even discontinuation of effective therapies Sex and gender issues also remain underexplored in biomedical education programs Due to their work with multiple specialists and health experts on patient care teams pharmacists are perfectly positioned to have a critical role in impacting discussions of sex and gender differences and the use of appropriate terminology for patients by filling in gaps during communication with these professionals GENDER R25 can help support pharmacists in these efforts by improving their education on these issues in pharmacy programs which can also be utilized across other disciplines For pharmacists interested in learning more the technical assistance webinar on Wednesday September 21 will provide potential applicants a chance to learn about the scope and requirements for the GENDER R25 program and members from the NIH Center for Scientific Review will also be participating in the discussion to provide further details about the program and answer applicants’ questions For those interested in applying for funding non-AIDS-related applications are due by 5:00 pm EDT on October 27 while HIV/AIDS-related applications are due January 7 “Discipline-specific training on sex and gender such as what will be supported through the GENDER R25 provides pharmacy professionals with an opportunity to advance pharmacy education and practice—and therefore improve human health,” Barr said “The benefits of improved inclusion in clinical studies and enhanced integration of sex and gender in all aspects of pharmacy study and care are impressive and attainable.” Additionally, ORWH will be offering other resources and programs for career development and interprofessional education Developed in partnership with the FDA Office of Women’s Health the course explores sex- and gender-related differences in key disease areas The 6 modules included provide biomedical researchers and students in health care professions with knowledge they can use when designing and conducting research and/or interpreting evidence for clinical practice Developed with funding support from the National Institute of General Medical Sciences the course helps investigators understand and apply the NIH Policy on Sex as a Biological Variable (SABV) in research design The 6-module course is designed for the biomedical research community to account for and appropriately teach the SABV policy A self-paced introductory training aimed at researchers the downloadable slide deck and accompanying facilitator’s guide can be used by individuals or teams to initiate a dialogue about how—and why—it may be beneficial to incorporate a sex-and-gender lens into research and clinical care GPRC5D-Targeted CAR T-Cell Therapy Shows Promise After BCMA Treatment Failure Insights from a Pharmacy Times Peer Exchange: ADC Drug Design and HER2 Ultra-Low Breast Cancer Indiciations Transformative Pharmacy Fellowships Can Shape Future Leaders The RIVAWAR Trial and ACS Guidelines for LV Thrombus Emerging Efforts to Expand Access to Psilocybin Therapy: A Critical Opportunity for Underserved Communities Facing Mental Health and Addiction Crises Expert: Understanding OTC Allergy Medications and Symptom Management 609-716-7777 HYANNIS — An 18-year-old Barnstable man pleaded innocent at his court arraignment this morning in connection with a homemade bomb that exploded in a Shaw's supermarket parking lot last night Andrew Spalt was arraigned in Barnstable District Court on a charge of possession of explosives He will continue to be held on $2,500 bail Spalt turned himself in to police at the scene of the explosion where police and fire officials had taped off a large section of the parking lot around the second bomb a large soda bottle filled with a mixture of common household cleaning products Two other companions were interviewed by police but were not charged with a big flash and a loud bang that could be heard inside the Shaw's and neighboring stores The blast was witnessed by an employee at the Shaw's supermarket who also identified the suspect's vehicle and license plate number The witness also provided a description of Spalt Barnstable police called in state police explosives experts to destroy the unexploded crude homemade bomb in the parking lot off Route 132 an unidentified 18-year-old man and at least one companion purchased the bomb-making materials at the supermarket early in the evening using instructions they had obtained from the Web site YouTube Store security cameras show the suspects purchasing the ingredients common household products that they later mixed in two large soda bottles Kenney said acid in the bottles could have severely hurt someone walking nearby The materials also produce a noxious gas when they explode Both rigged bottles were left in a section of the parking lot but close to busy entrance roads to that section of the mall although the bottle had expanded and distorted from gases produced by the chemical reaction inside Technicians from the State Police Hazardous Devices Unit in Stow fired a projectile into the soda bottle that vented it Mcguire said Spalt was cooperative at the scene and was arrested by Officer Nicole Provitola Doug Fraser can be reached at dfraser@capecodonline.com Please enable JS and disable any ad blocker Logging out of EU Login will log you out of any other services that use your EU Login account Use the CORDIS log out button to remain logged in on other services This is a machine translation provided by the European Commission’s eTranslation service to help you understand this page. Please read the conditions of use This website may no longer be available or may not contain the original content Please confirm that you want to visit {{ url }} Scientists in Austria and Germany have discovered a genetic switch that regulates the formation of flight muscles in flies creatures with very small wings in relation to their bodies suggests that spalt proteins switch myofibres fr.. Permalink: https://cordis.europa.eu/article/id/34128-scientists-discover-what-makes-flies-fly Your booklet {{ title }} generated on {{ timestamp }} is available for download The file will remain available for {{ hours }} hours £3750 worth of beautifully conceived and crafted guitar Europe is teeming with great guitar makers and if you haven’t come across Michael Spalt then you really should and with a diverse background in art and film Spalt has been making guitars professionally for some 15 years He creates some pretty ‘out there’ sculptural designs from wood and metal but all are fantastically crafted with an ever-present artistic eye guiding the lines This Custom 624 is one of Spalt’s more conventional designs his take on the Les Paul – albeit with a flat front and Fender-like scale length The top here is zebrawood and gives a striking appearance that’s contrasted by the sand-cast aluminium scratchplate though – not least due to its chambered back It’s such a beautifully conceived and crafted piece – and certainly recalls the work of Tony Zemaitis – that we’re really not sure whether it should be in an art display or strapped on and plugged in If you choose the latter route you’ll be rewarded with a sonorous sustaining single-cut voice played clean and throaty girth with the wick turned up But there’s a semi-like lively snarl in there no doubt helped by the wenge fingerboard and that exotic wood top too; it’s a work of art that sounds the part Available in the UK from The North American Guitar: 0207 835 5597 www.spaltinstruments.com Switching is conventional with a three-way toggle plus pull/push coil-splits for each pickup A Bigsby B-5 not only provides its classic shimmer and tonality but fits the aesthetic perfectly bone nut and ABR tune-o-matic-style bridge intonation and tuning stability are both excellent although the scale length is Fender-like at 25.5 inches Old stock Honduran mahogany is used for both the back and neck wood "I switched to a Stratocaster - I got a really good deal in the middle of the night for $120 - and then in 1972 Lowell George showed me his MXR compressor pedal”: Bonnie Raitt on how she developed her famous slide guitar style “The melodies work so well in this CGDGAD tuning I’ve absolutely fallen in love with this arrangement”: Magic Mike Dawes is at it again – he’s turned a Sleep Token track into a work of solo acoustic guitar genius It was unbelievable”: The making of Robert Palmer's Addicted To Love A beautiful guitar that will only get better with age Microscopic attention to detail; fascinating use of timbers; great playability and vibrato system Some might prefer raunchier tones from this 'rock' style guitar Patrick James Eggle 96 Spalt Drop TopPatrick James Eggle 96 Spalt Drop TopPatrick James Eggle is one of the UK's most highly respected luthiers having turned out superlative quality guitars for three decades Although more recently associated with extravagant flat-tops "I just haven't built too many over the last few years But I think we have a lot of good things to offer based on past experience and also the way we operate - as a small team using a blend of clever technology hands-on skill and hopefully some inspired ideas." Our instrument today certainly looks inspired: a striking bolt-on 96 Spalt Drop Top ('Spalt' referring to spalted maple 'Drop Top' to the way the maple elegantly drapes across the forearm contour and '96' to Georgia's east-west state highway) The instrument features familiar and exotic woods Bog oak is used for the 96's neck; it's an ancient and extremely dense material that's literally dug out of the peat The 96's back is the more traditional flamed maple and swamp ash but timber that's been affected this way is highly sought after in woodworking circles and looks fabulous as the 96's body facing sonically and structurally," affirms Patrick "I want the bodies to be light and resonant The neck has the lightest of sealing coats and is a limed but matt acrylic that reveals the swamp ash's grain beautifully so the internal surfaces have been painstakingly finished - a mini work of art in itself It features Marc Ransley's Mojo pickups - a humbucker at the bridge and a P-90 in neck position "I first used Marc's pickups about two years ago," explains Pat "I needed some custom wound pickups for one of my own guitars and he absolutely nailed it The P-90 and humbucker are balanced so they work perfectly together - you've got to be careful because P-90s can be a bit overpowering in the neck position." It's just as comfy in the palm and there's no need to rethink playing style even though this is a Fender-ish 648mm (25.5-inch) scale The 96's curved body heel aids upper-fret access and its four-bolt neck plate is neatly inlaid into the ash for a tidy The modified Gotoh vibrato is slick in operation but the tonal balance between the flutey neck P-90 and darker humbucker at the bridge is just right on each instrument full of woody goodness and equally at home on clean rhythm as on crunchy riffs or searing solos Coil splits on the bridge pickups add even more to the sonic stew by way of spanky clean leads and funky rhythms or arpeggios 'openness' and 'articulation' describe this guitar well but not just on clean tones - pile on the overdrive and that clarity remains along with great string-to-string separation We can't get over the sublime attention to detail on display here This instrument owes a nod to tried-and-tested formulae but Patrick's eye and his team's ability as woodworkers mean that the 96 stands as unique and rather wonderful instrument "My radical bent gets me into trouble sometimes Guitarists love what is classic and familiar I like to balance classic old-school with something fresh because if we don't bring something new to the table it can become a little dull." There's nothing dull about this gorgeous guitar It offers an interesting take on the familiar but brims with Eggle's understanding of what's 'right' in a guitar build - even when things like timber choices hardware and electrics are not your usual meat and two veg "This is a guitar that proves affordability can still bring looks that stun": Gretsch G5230T Electromatic Sparkle Jet FT review here comes the pitch and you can either knock it out or you can strike’”: Dylan's live sound engineer Jason Frankhouser on mixing front-of-house for The Never Ending Tour “There are very few artists whose best work was the very last thing they ever did in their lives”: Ronnie James Dio is the exception that proves the rule WATERTOWN – The Barnstable girls hockey team prides itself on its physical style of play doesn’t come without its fair share of penalties but it is something Peter Nugnes and his coaching staff are willing to accommodate The Red Raiders took it to another level Tuesday in the Div Barnstable was called for 11 penalties in their showdown with the Colonials and the Red Raiders’ season came to a close with a 3-1 loss in Watertown we haven’t a big crowd like this all year,” Nugnes said We just got out of our game and we don’t usually act like that.” It marks the third consecutive season the Red Raiders (16-2-4) ended their year against the Colonials A-B needed overtime to dispatch Barnstable “We have a monkey on our back with them,” Nunges said A-B coach] has our number and I don’t know what to do about it.“ Acton-Boxboro advances to the semifinals against Arlington Catholic or Duxbury The story of the game nearly was the Red Raiders penalty kill unit which killed off the first nine power plays of the game including two separate five-on-three opportunities With the game tied at one in the second period Caroline Spalt was called for a major boarding penalty As Spalt was locked into the box for four minutes the Colonials struck twice in a 53-second span to jump out to a 3-1 lead University of Michigan commit Megan Barrett and Leah Cardarelli each scored who was ejected from the game for accumulating seven penalty minutes after two minors earlier in the game Spalt was moved to the Barnstable’s top line and had been a big reason behind its recent offensive success Nugnes was forced to re-arrange his lines on the fly “It hurt big time losing Spalt and she’s a big part of our penalty kill “It’s one of those games you wish you could do over again leading point-scorer Callie Rogorzenski and Lindsey Phelan were moved to the top line alongside Danielle Derosier but the new lines failed to close the gap in the third as the Colonials held on “It’s very disheartening and the girls have worked so hard all year long,” Nugnes said “They felt real good about this game and we know Acton-Boxboro They don’t run a lot of players and they ran nine skaters tonight so we thought we would run them down.” Senior captain Morgan Richard scored the lone Red Raider goal at the midway point of the game blasting a puck from the point through traffic to tie the game at one freshman goalie Olivia Sollows kept her team in the game with 12 first period saves and an additional six in the second before the Colonials scored on their final two shots of the period The Red Raiders will once again be a threat next season However those seniors were a staple of this Barnstable team Kylie Hallam and Nicole Derosier all move on after the season Bernhard Spalt is to join the Board of the Commercial Bank of Romania (BCR) as Chief Risk Officer starting 1 January 2017 who tool on an executive position with a company that is not part of Erste Group “We would like to thank Jon Locke for his dedication and hard work with BCR specifically for the general recalibration of the risk management We wish him all the best and are confident that we will continue towards the sustainability of the BCR business,” said Sergiu Manea a manager of caliber and a recognized leader in matters relating to risk management within the Erste Group His appointment with the BCR shows strong commitment to Romania and to the BCR “Bernhard knows well Romania and the BCR, therefore we are certain that his leadership in risk management will contribute significantly to the future development of the BCR,” Manfred Wimmer, President of the BCR Supervisory Board, said. Bernhard Spalt joined Erste Group in 1991 and held various management positions in the field of operational and strategic risk management From 1999 to 2002 he was responsible for the management of non-performing loans in the Czech Republic he set up the Directorate of Strategic Risk Management and between 2006 and January 2012 was Chief Risk Officer of Erste Group From 2012 to 2015 he was responsible for risk management as a Board member in Hungary before joining the Slovak subsidiary We use cookies for keeping our website reliable and secure providing social media features and to analyse how our website is used Sam Jones selects her favourite stories in this weekly newsletter Central and eastern Europe economies will outpace their wealthier western counterparts in the post-pandemic recovery according to the head of Austria’s largest bank who believes they will benefit from strong economic fundamentals and better government decision-making “The economy has rebounded very strongly all over our region,” says Bernd Spalt one of central and eastern Europe’s biggest lenders “We are back in a situation where we have almost full employment Insolvencies are at record lows and credit demand is rising It is a picture he does not see elsewhere in Europe: “Everyone has built their own little ringfence around their country.” Central and eastern Europe economies are very likely to do better partly because of extensive government support for sectors hit hard by Covid 80 per cent of the sector’s revenues were subsidised by the government We are back in a situation where we have almost full employment full order books for the corporates and consumption has returned The region’s underlying economic skew towards industry and manufacturing which have largely continued during the pandemic will also give it an advantage over more developed western economies which are dominated by service-oriented businesses “The desire of people and corporates to get back to a normal state of affairs here is huge,” Spalt explains Less stringent lockdowns also mean economic life in the region has not been as painfully affected as in the west Erste has now revised its earlier 2021 gross domestic product growth forecast of 4.5 per cent up to 7 per cent Speaking from Erste’s headquarters in Vienna between the city’s main train station and the 18th century Belvedere Palace — famed for its collection of Klimt paintings — Spalt says the bank is forecasting significant revenue growth as it is known — a hub of sinuous low-rise glass office blocks intersected by a public piazza — may finally be in keeping with the bank’s potential The project for the headquarters was conceived in 2007 when the breakneck expansion of Austrian banks into eastern Europe suggested a glittering future for Vienna as an eastern European financial counterpart to Frankfurt or London It soon became a symbol of hubris. The financial crisis hit Erste hard. In 2008, the Austrian government took a €2.7bn stake in the bank to stabilise its balance sheet. Erste continued to rack up huge losses in the years that followed as the aftershocks of the crisis rippled through the eurozone Sinuous: the flashiness of Erste’s headquarters may finally be in line with its prospects © Christian WindThe 53-year-old Spalt — a life-long Erste Group employee and lawyer by training — took over as leader of the company in January last year who had held the post for more than two decades As the pandemic took hold within months of his appointment Spalt began to provision for “massive” losses and to shore up Erste’s balance sheet But it became clear relatively early on that liquidity would not dry up and that central and eastern Europe was positioned to weather the crisis economically Erste began an aggressive expansion in eastern Europe The bank — including its subsidiaries — is the largest lender in several key economies in the region as lenders and markets fearful of eastern European exposure withdrew liquidity In its first-half results reported at the end of July revenues at the bank were up 9.2 per cent year-on-year to €3.8bn The bank’s shares were almost 40 per cent higher at the end of August than at the start of the year “We went into this crisis from a position of strength,” Spalt says “This wasn’t a financial crisis which turned into an economic crisis [like 2008] but a health crisis that turned into an economic crisis banks were identified not as a part of the problem but the solution.”  My fear still is that we are going into the second half of this year much too naive The rate of vaccine hesitancy in eastern Europe concerns him “Maybe it's a little early to declare victory,” he says “Everyone says there will be no more further lockdowns but my fear still is that we are going into the second half of this year much too naive and missing out on what we need to prepare for.”  rising inflation is also a worry — specifically that it will not be acted on fast enough by the European Central Bank Central and eastern Europe is already seeing rapidly rising costs “If you have inflation but not interest rate hikes [has] access to experts and [economic] knowhow but I think we need to have more attention to inflation and more understanding of how fundamental drivers of it will change over time,” he says “I am more sceptical [about writing off inflation risks]  the temporary nature of inflation now might quickly turn into something more permanent.”  The economics – and the enjoyment – of overhauling his 21-year-old Mako appealed to this boat owner more than buying new Though new-boat prices have softened during the recession some boaters still aren’t comfortable laying down big money for a new vessel in a struggling economy Rather than spend $100,000 on a new center console in the 25- to 28-foot range for fishing and day cruising Spalt spent about half that to overhaul his 1989 Mako 261 named Dauntless which replaced a pair of 200-hp Yamaha 2-strokes accounted for nearly half of the renovation cost “Ask yourself what you get for 55 grand new today and you don’t get a 26-footer with a 350-hp 4-stroke The addition of a custom Armstrong engine bracket to the Mako freed up space in the cockpit and created new storage areas The bracket became part of the hull’s running surface mimicking its shape along the centerline and stretching the overall length from 26 to 29 feet “It’s twice the ride that we used to get out of the boat,” says Spalt 6-inch beam and a 23-degree transom deadrise “I was in probably 3- to 4-foot seas and going about 25 and I wasn’t getting a pound – there was no pound In addition to adding the bracket-mounted engine the project included installing a hardtop and replacing the fuel tank used to maintain a fleet of boats in a commercial fishing business Spalt operated in the 1980s and ’90s “There were no discounts involved because he is my brother-in-law,” says Spalt “The 2-strokes were probably not maintained well by the previous owner,” says Spalt and then when ethanol was introduced into the gasoline With the 4-stroke it’s more of a turnkey situation I can go down to the boat with my family and not worry about it starting and not running right.” which includes electronic throttle and shift and power-assist steering consumes much less fuel than the twin 2-strokes I would be burning 25 or 26 gallons per hour at around 30 knots,” he says “That’s tremendous fuel economy,” says Spalt “It has exceeded what we thought it would do And I don’t have to buy 2-stroke oil.” Top speed exceeds 46 mph The Mako topped out at 44 mph with the 2-strokes a V8 with a displacement of 325 cubic inches But the twin 200-hp Yamaha 2-strokes weighed a combined 870 pounds and the new power weighs roughly 70 pounds less Though the weight of the 4-stroke wasn’t a problem I thought that it would float much lower in the water,” says Gallagher because of the positive buoyancy of the bracket the boat ended up floating about an inch higher.” The aluminum bracket is hollow Gallagher hired a local fiberglass shop to replace Dauntless’ wood-cored transom with epoxy-coated marine plywood and reglass the structure he took care of the most difficult and involved part of the project himself: replacing the aluminum fuel tank installed on centerline under the cockpit sole because it was bonded in place with adhesive foam “I tried different methods of removal,” says Gallagher “In the end I had to just get in there with my Sawzall and cut it out chunk by chunk.” we realized the existing hull was a better hull than any new hull its size on the market,” says Spalt That’s when they decided to keep the Mako and “do a complete facelift starting in August 2009 and finishing in late December Spalt sealed the deal with a new twin-axle trailer for his Mako which Gallagher hauled to Florida for the winter Gallagher ran the boat through its shakedown logging 20 hours around the Keys and other areas Returning to Cudjoe Key after fishing near the Dry Tortugas Gallagher and two friends ran into 4-foot seas “It was just like a Nantucket Sound chop,” says Gallagher “The wind was running against the Gulf Stream It was comfortable for the people on board.” Spalt traveled to Florida to drive his “new” boat “The ride that we get with the bracket is phenomenal,” he says and Spalt is itching to get back to his boat “It’s in Jupiter [Fla.] just waiting for me,” says Spalt who enjoys boating on Nantucket Sound with his wife We take it out around Monomoy and Nantucket Sound and fish the rips.” With its freshly painted Corinthian blue hull and prominent outboard Dauntless should get plenty of second looks this summer “I think it’s a good-looking boat,” says Spalt This article originally appeared in the May 2010 issue Many products featured on this site were editorially chosen Soundings may receive financial compensation for products purchased through this site Copyright © 2025 Firecrown Reproduction in whole or in part without permission is prohibited One of the Dragon Quest X Online weapons designed by Bleach manga artist Tite Kubo will return to the game. The one-handed sword Spalt Edge is one of the three collaboration weapons designed by Kubo released in 2015 for DQX Online, and it will make a reappearance soon. [Thanks, ryokutya2089!] The weapons were originally available through download codes included in the August 2015 issue of the V Jump magazine as well as the DQX Online third anniversary fan book The other two Spalt weapons are the two/handed Spalt Blade and the Spalt Claws knuckle weapon The Spalt weapons were originally made available as part of several collaboration events in 2015 The Spalt Blade appeared in Dragon Quest X Online on July 2015 and the Spalt Claws on October of the same year Bleach author Tite Kubo also collaborated by drawing an illustration for the Dragon Quest X Online third anniversary fan book depicting three members of the Ogre species wielding the Spalt weapons