One Shot TM TOP10 chemically competent E. As a negative control, the same reaction was performed without insert addition. The digested products were purified with Gel/PCR DNA Fragments Extraction kit, dephosphorylated with rAPid Alkaline Phosphatase kit (Roche, Basel, Switzerland) through incubation at 37 ☌ for 10 min followed by 2 min at 75 ☌ and then ligated using Rapid DNA Ligation kit (Roche, Basel, Switzerland), with samples incubated for 5 min at 20 ☌. Plasmid vector and insert were first digested with NheI and XbaI restriction enzymes (New England Biolabs, Ipswich, MA, USA) through incubation for 1h at 37 ☌. Primer sequences are reported in Table 1. EFEMP1 3′UTR sequence to be cloned was amplified by PCR using ThermoScientific Phusion Hot Start High-Fidelity DNA polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). However, since only EFEMP1 downmodulation was validated in public datasets comparing tumor vs normal stroma of breast cancer patients, we decided to focus our efforts on studying EFEMP1 contribution to the observed phenotype.ĮFEMP1 3′UTR was cloned into pmirGLO vector plasmid (Promega, Medison, WI, USA), designed to perform luciferase reporter assay and carrying β-lactamase coding region (Ampicillin resistance). These molecules were selected for further analyses since they are known to be involved in the crucial pathways of ECM synthesis and remodelling. Gene expression profile of miR-9 overexpressing NFs revealed EFEMP1, collagen type1 alpha1 (COL1A1) and matrix metalloproteinase-1 (MMP1), as the most significantly modulated genes, being the first two transcripts predicted miR-9 targets. We also showed that these CAF-like cells can increase, in turn, tumor cell aggressiveness. Accordingly, in our previous work by Baroni S et al., we demonstrated that TNBC cells are able to induce the acquisition of CAF-like properties in NFs by releasing the known breast metastamiR miR-9, packaged into exosomes. Indeed miRNAs, small non-coding RNAs involved in post-transcriptional gene regulation, have been proven to act as “messages” to induce the acquisition of malignant traits in stromal cells. Overall, our results demonstrate that miR-9/ EFEMP1 axis is crucial for the conversion of NFs to CAF-like cells under TNBC signaling.Īn increasing body of evidence supports the involvement of microRNAs (miRNAs) in the interaction between tumor and stroma. Moreover, TNBC cells conditioned with the supernatant of NFs transfected with miR-9 or si- EFEMP1 became more resistant to cisplatin. In particular, EFEMP1 siRNA-transfected (si- EFEMP1) fibroblasts have an increased ability to migrate and invade. The luciferase reporter assay showed that miR-9 directly targets EFEMP1 and its silencing recapitulates miR-9-induced pro-tumoral phenotype in fibroblasts. Indeed, upon miR-9 overexpression in NFs, EFEMP1 resulted downmodulated, both at RNA and protein levels. Here we explored the impact of EFEMP1 downmodulation on fibroblast’s acquisition of CAF-like features, and how this phenotype influences neoplastic cells to gain chemoresistance. EGF containing fibulin extracellular matrix protein 1 ( EFEMP1), which encodes for the ECM glycoprotein fibulin-3, emerged as a miR-9 putative target upon miRNA’s exogenous upmodulation in NFs. We previously demonstrated that miR-9, a known metastamiR, is released by triple negative breast cancer (TNBC) cells to enhance the transition of normal fibroblasts (NFs) into cancer-associated fibroblast (CAF)-like cells. MicroRNAs are thought to act as tumor messengers to “corrupt” stromal cells. Tumor growth and invasion occurs through a dynamic interaction between cancer and stromal cells, which support an aggressive niche.
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To avoid spoilers, I’ll hold off on saying anything more than that. You’ll quickly discover that something is wrong…surprise, surprise. In this case, you are Lauren, a woman who is visiting her friend Kate. These six stages do, however, tie into one larger story. While it’s refreshing to see something other than pages to collect, it’s still a variation on the same theme. You might have to turn on a set number of generators, for example, before progressing to the next stage. New to the series is the idea that the other stages won’t necessarily include pages to collect. Those who are able to speed run through “The Eight Pages” will be scrambling to find that next page before the mystery man himself catches up with them. The first mission after the prologue, for example, will task you with collecting eight pages in a forest-like setting…sound familiar? Not to worry, the layout of the level is completely different, as is the structures and objects you’ll see. The game is broken up into six different levels, each one tasking you with collecting or interacting with a set number of objects. This menu also provides the time it took for you to complete a particular level, allowing you to go back and attempt to complete them faster. The extras menu is place where you can view your scrapbook, which consists of various notes that you’ll be picking up along the way. The graphics settings include your common toggles like screen resolution, vsync, fullscreen, texture quality, bloom, anti-aliasing, and more. The options menu is fairly meaty, allowing you to adjust the difficulty, mouse sensitivity, gamma, key bindings, audio sliders, and various graphics settings. The main menu allows the user to start a new game, select a stage, view extras, and adjust game options. If you're looking for an unblocked version of Flappy Bird, there are plenty available online. 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Mayor, however, is careful to point out that not all stories of dragons and giants are inspired by fossils and notes that Scandinavia has many stories of dragons and sea monsters, but has long "been considered barren of large fossils." In one of her later books, she states that, "Many dragon images around the world were based on folk knowledge or exaggerations of living reptiles, such as Komodo dragons, Gila monsters, iguanas, alligators, or, in California, alligator lizards, though this still fails to account for the Scandinavian legends, as no such animals (historical or otherwise) have ever been found in this region." In China, a region where fossils of large prehistoric animals are common, these remains are frequently identified as "dragon bones" and are commonly used in traditional Chinese medicine. She argues that the dragon lore of northern India may have been inspired by "observations of oversized, extraordinary bones in the fossilbeds of the Siwalik Hills below the Himalayas" and that ancient Greek artistic depictions of the Monster of Troy may have been influenced by fossils of Samotherium, an extinct species of giraffe whose fossils are common in the Mediterranean region. In her book The First Fossil Hunters: Dinosaurs, Mammoths, and Myth in Greek and Roman Times (2000), Adrienne Mayor argues that some stories of dragons may have been inspired by ancient discoveries of fossils belonging to dinosaurs and other prehistoric animals. Dragons are usually said to reside in "dank caves, deep pools, wild mountain reaches, sea bottoms, haunted forests", all places which would have been fraught with danger for early human ancestors. Jones therefore concludes that dragons appear in nearly all cultures because humans have an innate fear of snakes and other animals that were major predators of humans' primate ancestors. The earliest attested dragons all resemble snakes or have snakelike attributes. He cites a study which found that approximately 39 people in a hundred are afraid of snakes and notes that fear of snakes is especially prominent in children, even in areas where snakes are rare. Jones (anthropologist) suggests a hypothesis that humans, like monkeys, have inherited instinctive reactions to snakes, large cats, and birds of prey. In his book An Instinct for Dragons (2000), David E. Nonetheless, scholars dispute where the idea of a dragon originates from and a wide variety of hypotheses have been proposed. Famous prototypical draconic creatures include the mušḫuššu of ancient Mesopotamia Apep in Egyptian mythology Vṛtra in the Rigveda the Leviathan in the Hebrew Bible Grand'Goule in the Poitou region in France Python, Ladon, Wyvern, and Kulshedra in Albanian Mythology and the Lernaean Hydra in Greek mythology Jörmungandr, Níðhöggr, and Fafnir in Norse mythology and the dragon from Beowulf. Stories about storm-gods slaying giant serpents occur throughout nearly all Indo-European and Near Eastern mythologies. Draconic creatures are first described in the mythologies of the ancient Near East and appear in ancient Mesopotamian art and literature. Several bones purported to belong to the Wawel Dragon hang outside of Wawel Cathedral, but actually belong to a Pleistocene mammal.ĭraconic creatures appear in virtually all cultures around the globe and the earliest attested reports of draconic creatures resemble giant snakes. The Greek word probably derives from an Indo-European base * derḱ- meaning "to see" the Sanskrit root दृश् ( dr̥ś-) also means "to see". This is thought to have referred to something with a "deadly glance," or unusually bright or "sharp" eyes, or because a snake's eyes appear to be always open each eye actually sees through a big transparent scale in its eyelids, which are permanently shut. The Greek word δράκων is most likely derived from the Greek verb δέρκομαι ( dérkomai) meaning "I see", the aorist form of which is ἔδρακον ( édrakon). The Greek and Latin term referred to any great serpent, not necessarily mythological. The word dragon entered the English language in the early 13th century from Old French dragon, which, in turn, comes from the Latin: draconem (nominative draco) meaning "huge serpent, dragon", from Ancient Greek δράκων, drákōn (genitive δράκοντος, drákontos) "serpent, giant seafish". An early appearance of the Old English word dracan in Beowulf 5/1/2023 0 Comments Garageband for mac osArtists from Radiohead to Kendrick Lamar have used the app to demo, produce and sometimes even finalize master recordings. Musicians’ applause for Apple’s Garageband - which celebrates its 15th birthday this year, humbly, still living in the media shadow of many of the tech giant’s more glittering products - is similar across genres and skill levels. “I just started recording, without having to learn a new program, which was always one of the scariest things about music.” While other programs he’d tried in the past were sophisticated enough, including the one on the tour bus that day, Stump says, they were glitch-prone and impossible to use without frustration. “But I opened it that first time and never looked back,” says Stump, who talks about the software with a particular fondness, as if remembering his meeting with an old friend. While he’d heard of Garageband, a piece of free software shipped with all Mac computers, he’d thought it was more toy than tool - and no one else was giving it much attention then, in the early 2000s. Madly clicking around on his laptop in search of a new route, Stump happened to open one of its pre-loaded programs. I thought: I’m not going to be able to do this.” When you’re composing, time is everything, because you’re thinking the second guitar has to do this and the background vocals are going to do this and you just want to get it all out as quickly as possible. “When you’re being creative, you just want to get your idea out. “I just lost it, screaming in the back of a bus,” Stump tells Rolling Stone, a decade and a half later. Stump can still precisely recall the panic in the moment he finally finished the rough sketch of a song only to see the whole apparatus glitch and crash on his computer. On a lurching tour bus rigged with a wobbly Jenga tower of recording equipment, the singer and Fall Out Boy frontman had been trying to lay down demos for the band’s second album - it’d been hours, fiddling with rubber cords and finicky software - and nothing was working well together. 5/1/2023 0 Comments Filehippo ccleanerOther settings let you specify browser cookies that you always want to keep, add custom folders for CCleaner to analyze, or exclude other folders. The free version of CCleaner doesn't bug you about paying for the premium version, nor does its interface try to trick you into looking at a sales pitch. If you believe this comment is offensive or violates the CNET's Site Terms of Useyou can report it below this will not automatically remove the comment. They're not supposed to get anywhere near this large, and old logs just tend to sit unused and take up space, but we'd prefer to keep at least the most recent one by default, in case something goes haywire later. If you're more security minded and want to fully erase unwanted files, CCleaner's Analyze writes over them with random ones and zeroes, up to 35 times. Sort Date Most helpful Positive rating Negative rating. Provided excellent service como limpar seu pc com ccleaner MP3 free Como fazer a limpeza do seu PC com o faça o download do CCleaner e instale-o no seu computador com Qual o melhor programa para limpar o seu PC Kit. Muitos usuários já ouviram falar ou possuem instalados em seu computador o programa Ccleaner mas não sabem para que servem exatamente ou como utilizar. para limpar todo o seu A Restauração do sistema do Windows permite que você "volte no tempo" com o seu PC para. The limpid como limpar seu pc com ccleaner launchedĬomo por exemplo "Backup do Ccleaner". 5/1/2023 0 Comments Morgan wallen the way i talk(Among the eight songs that do not, is Wallen’s reverent take on “Cover Me Up,” Jason Isbell’s signature meditation on fresh, frightful sobriety. He sings the word “whiskey” 21 times throughout the course of the record, just one less than the number of its 30 songs (22) that reference drinking. Wallen also understands that he sounds best when reveling in whiskey worship, to the point where he risks overdoing it. Wallen falls into that trap as often as he impressively avoids it, and there are entire stretches of this record that are as unconvincing (“Outlaw”) as they are dull (“Whatcha Think of Country Now”). The flaws of Dangerous, apart from being 17 songs too long, is that Wallen does not always seem up to the heavy task of pumping fresh life into well-worn topics. The singer’s Tennessee twang is supple and dynamic enough to make the words Bud Light sound poignant on “This Bar,” and to earn sympathy during his heartbroken “sunburnt Silverado” joyrides after getting ditched on album opener “Sand in My Boots.” Twenty-six songs later, after an hour-plus of drunk dials and heartbroken pleas, it’s strangely moving to discover that the most emotional phone call Wallen makes throughout this feature-film length album is to the local newspaper to place a backpage ad for that same beloved truck on “Silverado for Sale.” In his teens, the family relocated south to Knoxville, where he graduated from Gibbs High School. 11 Tommy served for a time as a local church pastor, 12 while Lesli worked as a teacher. The album’s most moving moment may be the chorus of “865,” which consists almost entirely of Wallen rattling off a Knoxville phone number he cares about enough to have memorized. Morgan Wallen was born to Tommy and Lesli Wallen on May 13, 1993, in Sneedville, Tennessee. When Wallen sings, “for some folks a back road gets old, but for me it just can’t,” it feels like he’s singing about Nashville songwriting as much as two-lane dirt roads.ĭangerous is most affecting when Wallen’s husky, emotive voice does the heavy lifting. Morgan Wallen still wants to talk about the way Morgan Wallen talks, and he spends the radio-friendly backwoods party ode to anyone who identifies “more podunk than pop” defensively advocating for the same dated party-country formula he is working within. Halfway through Dangerous: The Double Album, Morgan Wallen’s 30-song collection - part album, part playlist, part content dump - Wallen begins “Still Goin’ Down,” the LP’s most compelling moment, by reciting the title of that same inaugural 2016 single. On 2018’s mega-hit “Whiskey Glasses,” he made it seem like he was the first country singer to discover Jack Daniel’s. Wallen has a penchant for making classic Nashville themes his own, adding dashes of winking self-awareness, aggrieved insecurity, and flirty playfulness to otherwise worn-out formulas. Like countless country singers before him, Wallen employed his small-town drawl, which feels so resonantly country that it can seem like the product of a Music Row laboratory, as the focal point of his prideful rural identity. Morgan Wallen’s debut single, 2016’s “The Way I Talk” was a fitting introduction to the former Voice contestant from East Tennessee. 5/1/2023 0 Comments Accupix mybud 3d developThe controls are also kind of difficult to operate while graphics are off-putting. The models don't interact with each other and exploring the categories gets repetitive after a while. I'd imagine if you were selling something like a coffee maker, this feature would come in handy for customers to see if the appliance matched their kitchen.Īugment app gives you directions on how view the model.īut if you're just a regular person or a kid, this app is, frankly, going to seem boring. I couldn't find any real objects, only Google Image results that led me to weird YouTube clips. You can also scan in objects to try them with the Augment QR code. And then you can place the virtual item anywhere in your space. Other options let you bookmark models, create static backgrounds and create your own models with an account. You can turn off the AR with a button and the app will redirect you to a 3D view of the model in your screen, which you can rotate fully to see the product or design. The graphics looked a bit blocky and sometimes glitchy. I then tried placing the cat from "Alice in Wonderland" and that worked better, if not kind of creepy. I tried moving a store kiosk around my backyard and it just did not work well. The finger motions were a bit rough at first as the object in my screen did not move easily. Some shoes are named simply "TF 17" while a rabbit model had a name that spelled out an entire phrase.Īugust AR app lets you flip through categories to see what you can create. These categories could obviously be organized better-my theory is that anybody could create a 3D model and push it out to the public. The arts and entertainment option gave me model sorceresses as well as shoes, while other options showed me anything from Walmart's display shelves to the cat from "Alice in Wonderland." Public models come in all different categories and themes. To resize, you simply pinch the model like you would with a regular photo. You move and pan the model with one finger while rotating it requires two fingers. When you first open the app, there's a short tutorial on how to work with the models. The app lets customers try out products and other objects before they purchase them while also letting distributors make models for their customers. Updated OctoAs much for distributors as it is for consumers, Augment 3D is a free augmented reality (AR) app, that's free for iOS and Android. What do you make of the movie’s being retooled for children, while still carrying so much horror, and so many mildly risque gags for adults? (Okay, go ahead and get me started: That dance is hilarious.) And while I hate Disney’s habit of putting characters through fake deaths and instant revivals, I still have to admit that the lovingly painted image of Pinocchio’s “corpse” floating face-down is pretty unsettling. And don’t get me started on Stromboli’s Dance Of The Disturbingly Well-Defined Buttocks. It’s a little off-color, too: Jiminy Cricket is quite the little skirt-chaser, gawping at salacious can-can puppets, manhandling the ass of a big-bustled ceramic doll, and flirting with clockwork women. The whole business with the donkeys on Pleasure Island is unnerving and nightmarish. Monstro the whale is animated as a terrifying force of nature. That said, Pinocchio has some surprisingly dark material. The company’s first animated feature, Snow White, stuck close to the source material (apart from streamlining out the wicked witch’s earlier, redundant attempts to kill Snow White), but Pinocchio was its second feature, and decades of sweetening up children’s stories started here, with a puppet protagonist who’s naïve and easily led rather than malicious, and who behaves like a cheerful young child rather than an amoral id-monster. Walt Disney Animation has a longtime habit of basing animated movies on classic fairy tales and children’s novels, but redrafting them considerably to make them more whimsical, innocent, and sentimental. Most strikingly, he straight-up murders the Jiminy Cricket equivalent who tries to give him good advice, and is subsequently haunted by its ghost. He befriends Lampwick and sees him become a donkey, as in the film-but he also re-encounters donkey Lampwick later, just in time to watch him die of overwork. Later, she and the fox end up blind, lame, and starving, and Pinocchio-in spite of his own constant moral failings and repentances-laughs at them and tells them it serves them right. (Like a movie cop, killed by the villain one week from retirement.) Pinocchio bites off one of the cat’s paws, when she attacks him while dressed as a bandit, and she repays him by hanging him, leading to a morbid sequence where he’s menaced by black rabbit undertakers. He deliberately betrays his father and the good fairy over and over, including at one point when he’s behaved himself for a year and is literally one day away from becoming a real boy. Pinocchio is a selfish little beast, aggressive and violent and constantly defying authority. It originated as a newspaper serial, so it’s made up of short episodes, almost all of which involve death, mutilation, or some grotesque betrayal. But Collodi’s version is so much darker and more disturbing. Many of the basics we see in the movie are there-Pinocchio is carved from wood by an old man who wishes for a son, he comes to life, he runs away at the behest of a scheming fox and cat, he ends up in a puppet show, he goes to a place where boys enjoy themselves until they turn into donkeys, he rescues his creator from the belly of a giant fish, and he becomes a real boy. Tasha: So Nathan, have you read Carlo Collodi’s 1883 novel The Adventures Of Pinocchio, the basis for Disney’s animated film? It’s an incredibly disturbing book. TWO DISSOLVE WRITERS KEEP THE PINOCCHIO CONVERSATION GOING… 5/1/2023 0 Comments Arduino camera sketchThis article will show how to use this camera in two different Arduino projects: the first creates a timing system to take pictures on specific intervals, storing them on the SD-card, while the second is an automatic surveillance system activated only if the special PIR sensor (Passive Infrared Radar) detects a warm object moving in the camera view field. In practice, this second application is a time-lapse video surveillance device that records what happens in a room, activated by the detection of moving people or vehicles. Of course, the PIR detection area must match the same camera angle so that the captured images effectively show the triggering “foe”. The basic concept of these cameras is that you can send all the setup commands, take screenshots and viewing images through appropriate bytes sequences via their serial port. It is quite easy to understand that using a composite or USB interface camera requires a considerable effort, on both hardware and software sides. If you decide to bet on a serial interface camera, everything gets easier, because all the prototyping boards (Arduino in particular) and all microcontrollers (Microchip, Atmel etc.) have at least one serial port as a standard feature. Recently, miniaturized cameras with serial communication interface have been launched on the market. Compared to traditional cameras, they offer easier integration with Arduino or other microcontroller-based boards. Also check out woojay's instructable, he has good wiring diagrams.Let’s equip Arduino with a serial-interface JPEG Camera with IR and try two applications: the first one saves shots (on a SD-card) at regular intervals while the second uses a PIR sensor to detect intrusions and photograph what happens. Find the calibration program and wiring diagram attached. The the value you get out of you chip must be edited in to your specific program for the gimbal.įor the mpu6050 to work you also need I2Cdev and the MPU6050 Libraries installed.įor this sample to work, the I2Cdev and the MPU6050 libraries need to be installed. Everyone of these sensor are unique in there own way, therefor you need to run the calibration program stand alone on your MPU6050 first to read its characteristics. The only thing to be clear about here is the calibration of the MPU6050. So i went ahead and built his system with his code and it worked without any issues. This really helped a lot since I am not a programmer in any ways. It was a prototype of the system made by woojay. But there was one instructable that made it all happen. And i couldn't find much info, so I assumed that this wouldn't even work since the internet wasn't full of info. So i searched a lot around the web to find anyone else making a gimbal with micro servos. |
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