Publishers Press/Publishers Printing Company Label-Pak Division has been awarded the Flexographic Technical Association’s (FTA) Gold Award, as well as the Best of Show Award for Narrow Web Publication for Alpinist Magazine’s “Autumn 2010” cover.The Label-Pak Division was awarded one of six Best of Show Awards given annually. Over 900 submissions were entered for consideration.Other Narrow Web Gold Publication winners include MPI Label Systems’ Bushdoctor Microbe Brew (label) and Labeltronix’s NLN Chocolate Banana Lean Pro Matrix and Eagle Castle 2009 Chardonnay (both labels).The 145-year-old Publishers Press/Publishers Printing Company is based in Louisville, Kentucky. The company currently offers magazine, catalogue and full service commercial printing, distribution, email marketing and label printing.
2019 Porsche Macan S: As lovely as ever 24 Photos 78 Photos Land Rover Range Rover VelarStarting at $49,950, the 2019 Velar is clearly not cheap. But this compact SUV feels far more expensive than it really is. Its minimalist design inside and out makes it one of the most aesthetically pleasing vehicles on the road.The Velar delivers a comfortable, quiet ride with reasonable athleticism and more off-road ability than any owner will ever ask of it. More From Roadshow SUVs Crossovers 62 Photos 2018 Nissan Kicks: Appreciating the unconventional Subaru CrosstrekThe 2019 Subaru Crosstrek starts at $21,895, offers standard all-wheel drive, and hey — you can even get it with a six-speed manual transmission. Equipped with the continuously variable transmission, the Crosstrek is pretty darn efficient, returning 27 miles per gallon city and 33 mpg highway.The 2019 Crosstrek now comes with automatic emergency braking on all models, as well as adaptive cruise control, lane-departure warning and lane-keeping assist. Apple CarPlay and Android Auto are standard across all trims. 2019 Honda CR-V is one of our favorite compact crossovers 12 Photos 24 Photos Subaru ForesterThe Forester still has enough of its tall-wagon DNA to make it enjoyable to drive, yet it offers all the practicality of an SUV, starting at $24,295.Subaru’s EyeSight driver-assistance package is among the best available in this category, and while the rest of the Forester’s in-cabin tech is a little tame, support for both Android Auto and Apple CarPlay mostly mitigates that.A number of 2019 model-year upgrades make the Forester an even better offering in the compact SUV space.Originally published March 21. 2018 Subaru Crosstrek: Just as good as before, only better The 2019 Subaru Crosstrek is rugged, practical and affordable 50 Photos Enlarge Image Honda Boy howdy, do American consumers love their crossovers and SUVs. And because many are looking to utility vehicles in place of compact sedans and hatchbacks, we’ve seen a big rise in the number of pint-size SUVs on offer.To that end, we’ve put together a list of some of our favorite small-ish crossovers and SUVs. From cheap-and-cheerful compacts to luxury and performance offerings, these are the best small utility vehicles on sale in America today.. And these days, compact offerings seem to be all the rage. To that end, we’ve gathered up some of our favorite small utility vehicles that hit all the sweet spots, from utility, efficiency, luxury and yes, even sportiness. Subaru Meet Roadshow’s long-term 2019 Volvo XC40 Mazda CX-5Starting at under $25,000, the Mazda CX-5 offers tons of style, inside and out.A 2.5-liter four-cylinder engine offers 187 horsepower and 186 pound-feet of torque. Whether in all-wheel or front-wheel-drive guise, the Mazda serves up genuine driving fun and excitement behind the wheel.The post-refresh model is now considerably quieter and better riding than earlier models, with blind-spot monitoring and rear cross-traffic alert standard across all trim lines. Volvo XC40A brand-new Volvo for $32,000? Sign us up.The XC40 doesn’t scrimp on kit, even though it’s the cheapest Volvo, coming standard with the Sensus Connect infotainment system and the always-great “Thor’s Hammer” LED headlights.Not only does it look good, the XC40 drives really well, too. Fuel economy isn’t the best, but life is full of tradeoffs. Mazda CX-3The smaller Mazda CX-3 can be had starting at $20,390. While it’s been updated for 2019, Mazda didn’t mess with the good stuff. The same 2.0-liter engine is under the hood as last year, though power and torque have increased just slightly.The rear glass is thicker, which should result in a quieter cabin. The LED taillights out back get a new design as well. Blind-spot monitoring and rear cross-traffic alert are standard on all trims. The base Sport model can be had with an i-ActivSense Package, including such active driving aids as full-range adaptive cruise control, lane departure warning and rain-sensing wipers. Volkswagen Tiguan is bigger all around, but still compact 2019 Mazda CX-5 Diesel review: Was it really worth the wait? Volkswagen TiguanThe Volkswagen Tiguan is softer than it used to be, but it’s also much more spacious, comfortable and quiet than the previous generation. The new look is bold, but also very classic VW.VW’s Car-Net and Digital Cockpit are basically scaled-down versions of the amazing Audi tech we’ve been raving about for years.A rare thing among compact SUVs, the Tiggy can actually be optioned with a third row of seats. 2019 Mazda CX-3 is brimming with personality 33 Photos Nissan KicksStarting at just under $18,000, the Nissan Kicks is a lot of car for the money, and can be had with a truly outstanding Bose Personal Plus audio system.The Kicks is a great, honest little crossover — and it’s not too shabby to drive, either.Aside from moderate noise over bumps, this budget crossover is a lot quieter than many of its competitors. 42 Photos Honda CR-VThe latest-generation Honda CR-V features a lot more style, has a roomier backseat, boats class-leading cargo space, and starts at a reasonable $25,000.A strong engine lineup includes a 2.4-liter naturally aspirated base engine with 180 horsepower and 180 pound-feet of torque, or a punchy 1.5-liter turbo with 190 horses and 179 pound-feet. The good thing about the turbocharged engine is that peak torque is available from just 2,000 rpm.The CR-V provides a well-damped ride quality, while also being competent through corners. 2019 Subaru Ascent review 24 Photos Porsche MacanThe Porsche Macan does the small-sporty-SUV thing better than pretty much any of its rivals. It’s handsome, spacious and super fun to drive. Plus, that Porsche badge ain’t for nothin’ — the Macan will outdrive any other sporty crossover on the road today.For 2019, the Macan S got a brand-new turbocharged V6 engine, and some slightly refreshed style. It’s an SUV we’d love to drive every day — and on some of our favorite roads, too. 2019 Mazda CX-5 adds premium style with new Signature trim Tags Share your voice 7 Comments 22 Photos Range Rover Velar SVAutobiography packs 550 hp, high-end trimmings 2019 Subaru Forester: Welcome improvements for an already solid SUV Land Rover Honda Mazda Nissan Porsche Subaru Volkswagen Volvo
Enlarge ImageShots of cars in sand dunes never get old. Land Rover We’re slowly approaching the point where Land Rover will finally pull back the veil on its hotly anticipated 2020 Defender SUV. However, today isn’t that day, which means we’ll have to live with more cool pictures of a camouflaged Defender for now. Shucks.Land Rover on Friday released a new smattering of pictures showing the 2020 Defender tackling the sand dunes in Dubai. Part of the Defender’s testing program ahead of its official unveiling, Land Rover teamed up with the International Federation of Red Cross and Red Crescent Societies to give this little ute the ol’ what-for in an area where its buyers are likely to travel.Out in Dubai, the Defender didn’t just whip around the dunes. It also tackled hairpin curves on the Jebel Jais highway. Temperatures shot past the triple-digit mark, which makes it a great place to suss out the Defender’s high-temp behavior, as well. It also tackled altitudes well above the one-mile mark, ensuring its engine doesn’t choke itself out when the air gets thin. Share your voice Land Rover Tags 2018 Range Rover Velar: Effortless SUV elegance on- and off-road Land Rover Post a comment 33 Photos SUVs Luxury cars Future Cars 2020 Land Rover Range Rover Evoque review: Style, now with more substance Land Rover first announced the return of the Defender in late 2018, and it revealed that the model would finally return to the US, as well, following a 23-year absence. Its boxy good looks have remained hidden by camouflage, but Land Rover hasn’t been shy about throwing out a whole lot of teasers ahead of its anticipated debut, which could come as early as the Frankfurt Motor Show in September. Even rumors have been few and far between. The most notable one we’ve heard about, though, is that the Defender will be available in three sizes — 90, 110 and 130, with the smallest being a three-door model. We’ve also seen what appears to be the Defender’s interior thanks to some clever spy shots. If you can’t afford one, don’t worry, because Lego has a Technic Defender coming out, which should be significantly easier on the wallet. 2020 Land Rover Defender hits the dunes of Dubai More From Roadshow 0 2019 Land Rover Range Rover P400e review: A hard hybrid to recommend
Vidyadhar Joshi gets eliminated from Bigg Boss Marathi 2?Colors MarathiBigg Boss Marathi season 2 has once again reached its elimination phase where among 5 of the nominated contestants – Surekha Punekar, Parag Kanhere, Vidyadhar Joshi, Shiv Thakre and Veena Jagtap – will have to leave the house in tonight’s Weekend Cha Daav episode.On Saturday, Mahesh Manjrekar announced that among the nominated contestants, Veena has been saved from this week’s elimination while Parag has landed in the danger zone.And though Manjrekar had decided to reveal the name of the Bigg Boss Marathi 2 contestant who got the least votes from the viewers on tonight’s episode, according to The Khabri, Vidyadhar Joshi aka Bappa has been eliminated from the show. Bigg Boss Marathi 2 contestantsColors Marathi#EXCLUSIVE #BBMarrathiFirst only on THE KHABRIVIDYADHAR has been ELIMINATED— The Khabri (@TheKhbri) June 22, 2019Meanwhile, Abhijit Bichukale has been sent to judicial custody at Central Jail in Satara for his involvement in 2012 extortion case after he was granted bail in another cheque bouncing case that has been going on in the court since 2015. It now remains to be seen if Abhijit would be replaced with a new contestant and it will be interesting to know who will take his place in the Bigg Boss Marathi 2 house.
Mirza Fakhrul Islam Alamgir. File PhotoBNP secretary general Mirza Fakhrul Islam Alamgir on Tuesday termed ‘baseless’ different media reports on their chairperson Khaleda Zia’s release on parole saying that there has been no decision yet in this regard.”Several newspapers have been publishing news with different headlines on the issue (of parole) for a few days. I would like to clearly say Khaleda Zia didn’t give any decision to go (abroad) for treatment on parole,” he said.The BNP leader came up with the remarks while speaking at the annual general meeting of a faction of Dhaka Union of Journalists (DUJ) at the National Press Club.About their meeting with Khaleda at Bangabandhu Sheikh Mujib Medical University (BSMMU) on Sunday, he said as prisoners are allowed to meet their relatives and friends on the special occasions like New Year, they took the chance and met their chairperson. “But there’s nothing to invent from such a meeting.”Reacting to a media report on Khaleda’s release on parole for going to the UK, Fakhrul said an English newspaper ran news with specific date and day of their chairperson’s release on parole. “It’s very unfair.””It’s also regrettable that they (the English daily) contacted me for comment, and I said it’s completely baseless and untrue. Even after that, they published it prominently. We urge all to refrain from exercising such yellow journalism and not to confuse people,” he said.The BNP secretary general also called upon journalists to talk to responsible BNP leaders before running any news on important issues relating to their party.He also observed that social media platforms are now being used for the character assassination of people, including the politicians. “It’s a social crisis, and the media people should think about it.”The BNP leader alleged that a ruling party-backed quarter is controlling the media in a planned way and working to build society as per the government’s desire.He said the media is being controlled in such a way by creating an appalling situation that many popular intellectuals now cannot join television talk shows. “Now, it has become difficult to provide authentic information.”Fakhrul said many journalists who tried to present accurate information lost their jobs after the national election. “A section of journalists is now in a good position, but not majority ones while many are jobless.”He called upon all to mobilise public support to change the current situation and protect press freedom and people’s rights.The BNP leader alleged that the law-and-order situation has badly deteriorated in the country as the current government has no accountability to people. “The law enforcers have taken a position against people.”Fakhrul also bemoaned that politics has now got polluted so badly that it is difficult for good people to survive in politics.
– / 5Please Sir, I want some more.Even for people unfamiliar with the story of Oliver Twist, many still know that famous line.Now, imagine someone saying it in sign language.Recently, Theatre Under the Stars gave a performance of the musical Oliver!. But on this night at the Hobby Center, there aren’t just actors onstage. On the floor to the right, in the dimly-lit performance hall, there’s another spotlight on two people dressed in black and gray, acting out the scenes in American Sign Language, or ASL.A musical for people who can’t hear? It may seem contradictory at first.“Deaf people get music,” says the Hobby Center’s Audience Services Manager Judi Stallings. “Just because they can’t hear everything, it doesn’t mean they don’t understand it. They get the rhythm, the flow, the fluidity of the music.”The Hobby Center is partnering with TUTS and the University of Houston’s American Sign Language Interpreting program to make the arts more accessible to Houston’s deaf community. It’s the first time they’re trying something like this.Brittany Best is majoring in American Sign Language Interpreting at UH and is one of the seven seniors responsible for translating the lines of the 58 characters in Oliver!. It’s pretty demanding.“I’ve got to know the actors’ lines; I’ve got to know my interpretation of those lines; and I’ve got to know what they’re doing onstage, because I’m not looking at them,” Best says. “So I’ve got to know all three and keep in line with the music (and) the tempo of the song.”The preparation began weeks in advance.“They engaged in complete script analysis and, along with me, worked through the script line, by line, by line,” says Sharon Hill, Program Coordinator and faculty member for UH’s ASLI program. She’s been instrumental in getting the project started.ASL is its own language and doesn’t have the same sentence structure as English, which means that translating word for word doesn’t always work. That’s also why subtitles aren’t usually ideal when trying to translate dialogue and music.“As Long as He Needs Me is really a challenging song,” says UH senior and ASL interpreter Barae Frizzel. “I almost spent a day just trying to study that piece. (There are) no signs that you can match to it. It’s more of a feeling and emotion that you have to connect with your audience.”When the lights come up at the end of the show, some people up front are waving their hands in the air. (Think jazz hands, but with more excitement.) That’s the deaf version of applause.Kristina Rodriguez, another student who was signing in the performance, has a unique experience with sign language. Both her parents are deaf and she grew up speaking ASL. This was her mom’s first time to a TUTS show.When she was asked what it was like to see her daughter in a live performance.Kristina translated the question and answer: “I’ve never seen my daughter act that way and have so much expression on her face. It was really exciting to watch. I’m just really happy to see her here.”It was also the first time for Robyn Brittan, another member of the deaf community.Sharon Hill translated her comment: “I see, as a deaf person, that I have access in my language. I miss not one piece of the show.”Stallings says this is a test-run to see if it’s something the Hobby Center can begin to offer more often as part of their accessibility initiative they began this year.“People that I know in the deaf community have assured me, ‘Don’t leave me out just because you think I can’t hear the music,’” she says. “There’s still something there for them.” 00:00 /04:01 Listen X To embed this piece of audio in your site, please use this code: Share
Nathalie Dieterle/for NPRThe Resilience Builder Program teaches young students techniques for handling tough emotions, like visualizing a remote control for thoughts so they can switch from negative to happier feelings.Sometimes 11-year-old B. comes home from school in tears. Maybe she was taunted about her weight that day, called “ugly.” Or her so-called friends blocked her on their phones. Some nights she is too anxious to sleep alone and climbs into her mother’s bed. It’s just the two of them at home, ever since her father was deported back to West Africa when she was a toddler.B.’s mood has improved lately, though, thanks to a new set of skills she is learning at school. (We’re using only first initials to protect students’ privacy.) Cresthaven Elementary School in Silver Spring, Md., is one of growing number of schools offering kids training in how to manage emotions, handle stress and improve interpersonal relationships.At Cresthaven, some fifth-graders like B. get an intensive 12 weeks of such training, a course called the Resilience Builder Program. Created by psychologist Mary Alvord, it’s a form of group therapy designed to help students who are struggling with trauma or cognitive disorders — or everyday anxiety caused by things like bullying or moving schools..“I think it’s so critical that kids know they have the power to make changes. While we can’t control everything about our lives, we can control many facets,” Alvord says.If students can learn this kind of resilience, the ability to adapt to emotional challenges, she says, “I think the whole world gets better.”The idea of teaching social and emotional skills in school is more than 20 years old. Research has shown this kind of intervention is effective and has a lasting impact. One analysis published last year in the journal Child Development reviewed dozens of programs with similar approaches. Participants were 11 percent more likely to graduate from college and less likely to have mental health problems or be arrested than were students who never went through these programs.In Australia, Canada and the U.K., social and emotional learning in schools is already being implemented on a large scale. Here in the U.S. it has spread, but not as quickly as some would hope. With all the mandates that schools have to keep up with, social emotional learning gets moved to the back burner.But at Cresthaven, the school counselor, Marina Sklias, and the school principal were hungry for it.Sklias says getting help for students dealing with trauma and emotional problems has been tough. In a high-poverty school such as Cresthaven, with a lot of immigrant families, she says there is only so much she can do.“Oftentimes I refer students for counseling and parents request counseling, but due to financial situations or transportation issues, parents can’t always follow through,” she says. At school, she is already stretched thin meeting with students or giving classroom presentations.When Alvord offered to bring the Resilience Builder Program to Cresthaven pro bono as part of a research project, Sklias selected a group of students she thought could benefit from it. It has been used especially with students dealing with attention deficit hyperactivity disorder or anxiety or trauma — officially, students with “social competence deficits.” She met with parents, and many agreed to sign up their kids.The program focuses on fifth-graders, Alvord explains, to prepare them for middle school, where pressures like dealing with sex or substance use really kick in. It’s a time that can be especially hard for kids already struggling with social and emotional issues.“It’s a big transition, big change,” Alvord says.For 12 weeks, small groups of Cresthaven students learned about topics like leadership skills, stress management, problem-solving, and empathy. The emotional-problem solving techniques they learned were based in cognitive behavioral therapy — adapted for kids.They also drew on the whiteboard together, did role-playing and yoga, had snack time, and played lots of charades.“I was thinking we’ll just sit and just do work,” B. says. “Instead we would play games and do things that were fun and do things that you usually don’t do in regular class.”Some of the time they worked on simple social skills, like making eye contact during conversation, greeting people in the morning, respecting personal space.The kids also learned handy techniques for working with negative thoughts, like visualizing a special kind of remote control. Alvord shows a drawing with buttons that say things like “happy place” and “things thankful for.”“You can switch channels in your head,” she explains. “Instead of ‘that math test was really hard,’ if we think, ‘I got through it and I’m proud myself.’ “Alvord developed the Resilience Builder Program in her private practice decades ago, and now she is working with researchers including Dr. Brendan Rich at Catholic University to measure how well the program works in schools with underserved students. Early pilot studies on the program show it is effective.For kids with ADHD, parents and teachers reported that after the program, students were more social, were able to handle their emotions, and weren’t as hyperactive as before. Parents also reported that kids with anxiety were able to manage their emotions and seemed less depressed.The new research compares students who have done the program with those who haven’t. Researchers have collected data from 119 kids at four schools in the Washington, D.C., area, including those at Cresthaven. They just had a paper accepted by the International Journal of Group Psychotherapy, which found that students who went through the program reported better emotional control than students who hadn’t. They plan to publish more in the coming months, and Alvord is excited by some early results that show the program is helping kids academically.“It all goes together,” she says. “If you’re not struggling with relationships or teasing and bullying, you have more head space to give to study, and you’re also just more positive.”After each Resilience Builder Program wraps up, Alvord organizes a showcase for the students and their parents. The kids get to share what they’ve learned and get certificates of achievement, and their parents cheer them on.On a warm spring evening, the handful of Cresthaven fifth-graders who had just finished the program gathered in the school library for their showcase.Most of the families are immigrants from all over the globe, East Africa, Latin America. Alvord is first generation herself and grew up speaking Russian and Armenian. She tells these families that they already know about how important and hard it can be to adapt.“You have had to make many changes and learn languages and customs — that’s resilience,” she says. There are nods of agreement in the room.The students take turns coming up to the front to get their certificates and share their favorite takeaways.When it’s her turn, B. — the 11-year-old who was being bullied — says that she learned how to solve “friendship problems.”“It helped me not get as mad at my friends as I used to,” she says.B.’s mother is thrilled with the changes she has seen in her daughter. Her eyes well up as she talks about how proud she is. B. seems less nervous; she doesn’t come home in tears as often.B. is pleased, too. She especially likes the relaxation skills they learned — things like breathing in and then slowly out again, and clenching fists and then letting go. She can take her new set of problem-solving tools with her to middle school next year and beyond.Your Turn: Share Your Parenting StoryParents make mistakes. It comes with the job. What do you wish you had known about raising kids before becoming a parent?Read this post for inspiration, then share your story on Instagram or Twitter using the hashtag #HowToRaiseAHuman. We are collecting stories until June 30. We may feature your post on NPR.Copyright 2018 NPR. To see more, visit http://www.npr.org/. Share
Re-engineering the cell membrane for improved biofunction is an emerging, powerful tool in cell biology to develop next-generation cell therapies. The process can allow users to supplement cells with added therapeutic functionalities. Additional functionalities can include cell homing, surface adhesion or resistance to hypoxia for enhanced cellular capabilities. However, the number of such examples on re-engineered plasma-membranes to activate membrane-bound enzymes that promote the assembly of extracellular matrix (ECM) proteins to promote cell functionalities are limited. 3D projection of fibrin gel containing fibrinogen stained with Alexa 594 (red) fibrinogen and hMSCs incorporating sc_thrombin [ox890] stained with Hoeschst 33342, imaged after 60 min of cell associated fibrin formation in cell culture. Credit: Nature Communications, doi: 10.1038/s41467-019-09763-0. They synthesized the artificial membrane binding thrombin complex using a two-step process to generate an active supercationic thrombin construct (sc_thrombin). Deller et al. also generated a polymer surfactant corona or green halo using electrostatic coupling of glycolic acid ethoxylate 4-nonylphenyl ether (ox890) to sc_thrombin to engineer a third variant sc_thrombin [ox890]. The scientists controlled the reaction conditions (pH, temperature and chemical composition) carefully and monitored the reaction progress using zeta potentiometry across a period of two hours. They observed the activity of thrombin by monitoring/characterizing the increasing turbidity of the fibrinogen solution. They then obtained MALDI-TOF spectra (matrix-assisted laser desorption ionization time of flight mass spectroscopy) of the native and modified thrombin to show full cationization of the construct. When Deller et al. conducted compression testing of the resultant self-supporting structures, the Young’s moduli were similar to soft fibrin hydrogels, indicating consistency. To investigate thrombin adhesion to cell membranes, the scientists chose a monolayer of bone marrow derived hMSCs (with well-characterized adipogenic, chondrogenic and osteogenic pathways). First, they incubated a monolayer of hMSCs with fluorescently-labelled analogues of thrombin; thereafter, they labeled the hMSCs with a plasma membrane-specific dye and imaged immediately to confirm the thrombin-plasma membrane binding. Using time-lapse confocal fluorescence microscopy, they showed the nucleation and fibrin growth from the cells thereafter. Welding with stem cells for next-generation surgical glues Advanced cell therapies are currently approaching clinical translation in response to an increasing demand for newly modified, cell-specific matrices (scaffolds) for biocompatible therapeutic performance. However, the rational design of matrices is extremely challenging since the cell phenotype and cell fate can be intertwined to a wide-range of scaffold-dependent factors; including cell adhesion, surrounding chemical composition, cell receptor stimulation, surface micro-/nano-morphology and mechanical stiffness. These factors immediately impact cells during in vitro tissue engineering, typically when cells are seeded to adhere on biocompatible and biodegradable scaffolds in the lab, where the scaffolds act as a surrogate extracellular matrix (ECM). Eventually, when the cells grow and differentiate, they can produce natural ECM of their own, to gradually replace the biomimetic scaffold material and form a structurally self-supporting biological entity. Deller et al. also completed cell growth assays to determine relative metabolic activities of labelled hMSCs-thrombin to show the modified cells were without observable cytotoxicity in varying concentrations of thrombin (1 µm to 25 mM). Using confocal microscopy again, the scientists showed the arrangement of fibrin structures emanating from the plasma membrane of the hMSC monolayer, in contrast to hMSCs without thrombin. The work protocol thereby generated a 3-D fibrin hydrogel construct with dense cellular aggregates surrounded by a 3-D fibrin matrix. The scientists also investigated the ability of the fibrin hydrogel system to sustain 3-D cultures for long-term growth; a requisite for tissue engineering, to show hMSC differentiation via adipogenic and osteogenic pathways. To verify the results, the scientists conducted extensive biomechanical tests on the cell types and tested for the upregulation of specific genes SOX9 and RUNX2 involved in chondrogenesis and osteogenesis respectively, using RT-PCR (reverse transcription polymerase chain reaction), to substantiate the fibrin hydrogel system sustained long-term hMSC proliferation in vitro.After confirming the membrane re-engineering approach for in lab tissue engineering applications, Deller et al. investigated the ability to produce thrombin coated cells in lab for their injection at a site of injury to initiate a healing response for tissue-engineering applications in vivo. For this, the scientists used an in vivo Zebrafish model system to perform preliminary cell transplant studies. Zebrafish is a model organism, established for fluorescently labelled live cell imaging and thrombolytic and hemostatic processes; suited for the present work. The scientists isolated, labelled and delivered fluorescently labelled primary Zebrafish fibroblasts, labelled with sc_thrombin[ox890] conjugate via microinjection to show cell survival after 3 days at a site of incisional injury. The synthesis and characterisation of the supercationic thrombin-polymer surfactant conjugate. a Schematic showing the electrostatic surface potential of native and supercationic thrombin (sc_thrombin) (PDB; 1UVS) at pH 7, highlighting the anionic (blue) and cationic (red) charged regions. Generation of the polymer surfactant corona (green halo) via electrostatic coupling of glycolic acid ethoxylate 4-nonylphenyl ether (ox890) to sc_thrombin gives [sc_thrombin][ox890]. b Zeta potential (ca. pH7; n = 3) of thrombin as a function of cationization times (0–120 min). Data reported as means ± standard deviation (s.d.). c Rate of fibrinogen solution (3.125 mg mL−1) gelation as measured by changes in turbidity (600 nm) catalysed by sc_thrombin subjected to various cationization times (0–120 min). Data shown as one-phase association curves of raw data. d MALDI-TOF MS spectra (m/z = 3) of native and sc_thrombin (60 min). Credit: Nature Communications, doi: 10.1038/s41467-019-09763-0 Schematic diagram showing in situ fibrin hydrogel formation from the membranes of bone-marrow derived human mesenchymal stem cells (hMSCs). Artificial membrane binding thrombin constructs comprising supercationic thrombin molecules (white) surrounded by a polymer surfactant corona (yellow) that associates with surface exposed cationic (red) residues spontaneously insert into bilayer regions of hMSC plasma membranes. In the presence of fibrinogen, the membrane-immobilised thrombin catalyses fibrin formation (blue fibres) within the interstitial spaces between the cells giving rise to a self-supporting hydrogel monolith. Credit: Nature Communications, doi: 10.1038/s41467-019-09763-0 More information: Robert C. Deller et al. Artificial cell membrane binding thrombin constructs drive in situ fibrin hydrogel formation, Nature Communications (2019). DOI: 10.1038/s41467-019-09763-0 D. E. Discher. Tissue Cells Feel and Respond to the Stiffness of Their Substrate, Science (2005). DOI: 10.1126/science.1116995 Tamer A.E. Ahmed et al. Fibrin: A Versatile Scaffold for Tissue Engineering Applications, Tissue Engineering Part B: Reviews (2008). DOI: 10.1089/ten.teb.2007.0435 © 2019 Science X Network In a recent study, Robert C. Deller and co-workers at the interdisciplinary departments of Cellular and Molecular Medicine, Engineering, Functional Nanomaterials and Pharmacology, and Neuroscience in the UK, bioengineered a self-contained cell matrix-forming system. In the experiments, they modified the plasma membrane of human mesenchymal stem cells (hMSCs) to integrate a new thrombin construct, which gave rise to spontaneous fibrin hydrogel nucleation and growth when supplemented with human plasma concentration levels of fibrinogen in cell culture media. The scientists bioengineered the cell membrane by synthesizing a membrane-binding supercationic thrombin-polymer surfactant complex. Thereafter, they observed cell differentiation in the resulting robust, stem cell-containing fibrin hydrogel constructs to form osteogenic and adipogenic cell lineages. The differentiated cells could eventually secrete fibrin to form self-supported bioengineered cellular monoliths that exhibited Young’s moduli as expected of the native extracellular matrix. The results are now published in Nature Communications. A range of natural biocompatible polymers have produced such transient hydrogel scaffolds for tissue engineering; including chitosan, gelatin and fibrin. Fibrin hydrogels are the most popular among them, since they can be produced readily at room temperature using proteolytic cleavage. Biological fibrin formation occurs in response to injury, culminating from a biochemical cascade of proteolytic cleavage, which converts prothrombin to thrombin and forms a fibrin-hydrogel clot. Fibrin hydrogels can therefore mediate cellular biomolecular functions and regulate the osteogenic and chondrogenic differentiation of human stem cells such as hMSCs. They can also be conveniently delivered using syringes, albeit with complications related to reduced cell viability.In the present work, Deller and co-workers first described a method to synthesize supercationic thrombin-polymer surfactant complexes that spontaneously bound to the plasma membrane of hMSCs to drive in situ fibrin hydrogel nucleation and growth. The resulting self-supporting hydrogel construct allowed high levels of metabolic activity as an artificial matrix for effective differentiation of stem cells to form adipogenic or osteogenic cell lineages. The scientists then showed the feasibility of the method of cell functionalization by injecting thrombin-labelled GFP-expressing fibroblasts (GFP: Green fluorescence protein) into a zebrafish (Danio rerio) skin wound model to demonstrate their in vivo biocompatibility for hemostatic applications. Explore further Journal information: Nature Communications In vivo zebrafish injury and [sc_thrombin][ox890] labelled GFP + fibroblast addition. Schematic representation of the in vivo adult zebrafish injury model. a Wildtype (non-transgenic) recipient zebrafish were anaesthetized and a 4 mm incisional injury made on the ventral upper thorax. A lateral view is shown. b Unlabelled or [sc_thrombin][ox890] labelled, FACS sorted GFP+ fibroblasts were injected at six sites around the edge of the incisional injury. At the desired time-point, fish were sacrificed and the tissue surrounding the incision was fixed, imaged and embedded for sectioning. A ventral view is shown. Ventral view of the area of tissue surrounding the incision at 3 dpi following transfer of c unlabelled or d [sc_thrombin][ox890] labelled GFP+ fibroblasts. Similar numbers of cells were retained at all wounds. The red line depicts the approximate position of the incisional injury which is fully re-epithelialized at this stage. Sections through the injury region at 3 dpi following transfer of e unlabelled or f [sc_thrombin][ox890] labelled GFP+ fibroblasts. No obvious differences were observed between wounds containing labelled or unlabelled cells. Arrows indicate the position of the incision. Credit: Nature Communications, doi: 10.1038/s41467-019-09763-0. Citation: Engineering artificial cell membranes to drive in situ fibrin hydrogel formation (2019, May 6) retrieved 18 August 2019 from https://phys.org/news/2019-05-artificial-cell-membranes-situ-fibrin.html Using macroscopic observations and histological assays, the scientists further revealed that there were no adverse effects between fish injected with engineered or native fibroblasts. However, Deller et al. expect to complete further investigations to understand the precise effects on the specific wound healing response using bioengineered cells in the future.In this way, Deller et al. synthesized and characterized a new membrane active thrombin construct and demonstrated its application to drive in situ fibrin formation in the plasma membranes of stem cells. The scientists showed that thrombin-based fibrin hydrogel constructs prepared using the new protocol supported high levels of cell growth and viability to eventually produce a self-supporting tissue engineered construct. The stem cells were also able to differentiate along the well-defined adipogenic and osteogenic pathways while demonstrating Young’s moduli similar to the native cells to indicate high levels of integration of the modifications. Deller et al. propose to optimize the protocols for further experiments in vitro prior to in vivo translation, to gain further insight to the enzymatic activity of cell membrane bound bioengineered proteins to develop biocompatible, hemostatic products. Evaluating rhodamine (rh) labelled rh_thrombin, rh_sc_thrombin and [rh_sc_thrombin][ox890] on hMSC plasma membrane affinity. Cells labelled with CellMask (green) and rhodamine labelled thrombin (magenta) visualized with confocal microscopy. Video shows the rh_sc_thrombin [ox890] labelled hMSCs supplemented with fibrin gel conjugating with fibrinogen (green) to highlight fibrin formation emanating from the bioengineered cell surface. Credit: Nature Communications, doi: 10.1038/s41467-019-09763-0 , Science This document is subject to copyright. Apart from any fair dealing for the purpose of private study or research, no part may be reproduced without the written permission. The content is provided for information purposes only.
Hear from Polar Explorers, ultra marathoners, authors, artists and a range of other unique personalities to better understand the traits that make excellence possible. How Success Happens 3 min read My son has just started middle school, and along with becoming oriented to an unfamiliar school environment, he now has “homework like never before,” and it is not just the amount of homework.His teachers no longer send home printed homework sheets. Students are expected to visit the teachers’ individual websites to download assignments, study guides, and watch lectures. And while going online is not normally a problem for technophile middle schoolers, it becomes a problem when the websites are poorly designed.Visitors come to a website to satisfy goals, to perform tasks, and to get answers to questions. If users cannot find what they’re looking for on a website, they will go elsewhere. But my son can’t go elsewhere. He has no choice but to muddle through the unclear navigation and bad design to find what he needs. It is maddening.Still, you can learn a lot from being forced to use a poorly designed website, including ways to improve your own or a client’s.1. Do everything you can to reduce the number of clicks.From the “Sixth grade homework” page, where he was told all assignments would be linked, he has to click through three pages to reach the math assignments. Those math assignments should be directly linked from the main page. When navigating, site users don’t want to stop and read along the way. They want to keep moving until they find the right link.2. Make the links meaningful.Links should be descriptive. Don’t tell readers to “Read more,” “click here,” or “more.” Tell them what they will read if they click. “Math homework October 6-10” is an example of a descriptive link.3. Make copy easy to scan with subheads and bullets.Lists make information easy to grab and help people skim through the information. If possible, keep lists short. But if your list must be long, use white space to break it up.4. Heading content should be concise and descriptive.It should stand out from the rest of the text. Well-written headings facilitate scanning so users can find exactly what they need. They can also make the information less dense and more readable, allowing users to get a quick overview of the page.5. Keep in mind that less is often more on the Web.Eliminate distracting site features such as flash animation or scrolling text. Use animation where it helps, not just for show. Listen Now October 17, 2014 This story originally appeared on PR Daily