Stack Media relied on a regional freelance network to reach a few hundred thousand viewers per month when it started its integrated video campaigns in 2008.Within two years, a Stack Studios crew had been assembled and was traveling the country filming for sponsors like Gatorade and Nike. They’re now getting upwards of 15 million views per video and generating 60 percent of the company’s revenue.Stack Magazine, the flagship of the larger media group, was established in 2005 as a training and fitness title centered on professional athletes. Video came in 2008 with the launch of the company’s digital network—a “natural extension” of the content they were already producing, according to Nick Palazzo, the company’s co-founder and CEO.Is It For Your Audience?The demographics worked for Stack. The majority of its audience is comprised of 16 to 24-year-old males—among the most active online video viewers, according to several recent reports. Overall, 18 to 24-year-old men and women with regular access to the Internet spend close to 11 hours watching web videos per month, or nearly one-third of their total time online, says Nielsen’s Cross Platform report from the second quarter of this year. A Pew Research survey from this fall indicates that the group is highly engaged, as well, with nearly a quarter of all adult American males sharing videos.All told, comScore has tallied more than 100 million Americans watching video online each day, up 43 percent from 2010.How Do You Make It?Pricing for the content is highly variable, Palazzo says, with the number of shoots being the key determinant. A single session—requiring travel, equipment, venue, lighting, etc. (along with the personnel to manage it all)—can run up to $10,000. On the low end, a shoot can be arranged for around $1,500. It’s up to the advertiser.The same goes for concept. The project is usually handed off to Stack right from the start, but sponsors can play a larger role.“Sometimes they can be very involved in storyboarding, bringing the concept to life and providing assets,” Palazzo says. In Stack’s case, those “assets” are usually famous athletes, but they can be products, venues, signage, memorabilia, logos, uniforms or anything else they want included.The bigger the asset, the bigger the response. Timing plays a role, but the golden-rule for luring readers to the newsstand applies to attracting viewers online: recognition.“What really drives traffic and engagement is getting the biggest star possible,” Palazzo says.How Do You Sell It?Palazzo estimates that as much as 85 percent of Stack’s content is now video-based, so naturally it’s their lead story to tell potential advertisers. From there it’s a matter of selling the complete distribution and media package.“It’s hard to sell just straight video. You have to package it with other media assets that you already have—that’s really the most important thing,” he says. “In a lot of what we do, you’ll get the video content, the distribution across our platforms, and media—banner and print—all together in one package. It’s much more efficient to buy everything rather than just piecing it out. We make it an offer they can’t refuse.”Integrated Sales Still Difficult To Do With AgenciesIn a divided print and digital world, Stack can run into difficulties trying to manage a deal across the multiple ad agencies running those respective arms. Whenever possible, Palazzo will try to deal directly with the company.“It’s sometimes hard to do integrated ad sales if they have separate agencies for digital and print,” Palazzo says. “One of the most important things to focus on is working directly with the client. They’re going to be the ones that have the ability to do multiplatform deals, as well as the foresight and knowledge base to help make a video program successful.”How Do You Measure It?Stack’s integrated video campaigns don’t feature any product links or buy options—they’re straight branding enterprises—so measuring success can be a challenge. In the absence of conversions, Palazzo relies on engagement metrics like video plays and average time spent with the video to let him know how they performed.“If you get someone to watch a minute or minute-and-a-half of video, that’s a pretty valuable impression,” he says. “If you can get them to click-through, then great, that’s something you can quantify. But that’s not the key metric that we focus on.”
A special exclusive ticket presale for Citi card holders will run from Tuesday, April 10, at noon local time through Thursday, April 12 at 10 p.m. local time. Tickets for the general public will go on sale on April 13 at 10 a.m. local time via LiveNation.Catching Up On Music News Powered By The Recording Academy Just Got Easier. Have A Google Home Device? “Talk To GRAMMYs”Read more The Offspring, 311 Plot 2018 Arena Tour News Email Facebook Twitter The legends of ’90s alternative radio will be joined by special guests Gym Class HeroesBrian HaackGRAMMYs Apr 9, 2018 – 4:00 pm Summer 2018 is shaping up to be a bit of a victory lap for ’90s alternative, hard rock and skate punk bands and their fans. Already announced are the anticipated return of System Of A Down, a three-day all-punk campout festival hosted by NoFX’s Fat Mike, and a unique “tri-headlining” tour featuring Stone Temple Pilots, Bush and Cult making for an increasingly packed summer festival and tour schedule. Now added to mix is a brand new co-headlining tour featuring proto-pop punk hitmakers 311 and the Offspring, joined by special guests Gym Class Heroes.The “Never-Ending Summer Tour” will hopscotch through 29 cities across North America for a series of area shows and festival appearances kicking off on July 25 at Mountain View, Calif., Shoreline Amphitheater and run through Sept. 9, where the tour will close out in Wichita, Kan. The Offspring And 311 Plot 2018 Arena Tour offspring-311-plot-2018-arena-tour
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.
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.