It was uploaded to the official YouTube Channel on 4 January 2018. TV 3.2M views 1 year ago F.R.I.E.N.D. Top 5: Breakdowns is the fifth video in the series. Friends Funniest Moments Friends Warner Bros. Top 5 Scary Moments Thomas & Friends Top 5: Breakdowns Marion gets frightened by the dinosaur models ( Marion and the Dinosaurs).Henry gets repainted with luminous paint ( Henry in the Dark).The dynamite catches fire ( Sodor's Legend of the Lost Treasure).Percy's spooky mail run ( Tale of the Brave).Henry meets the "abominable snowman" ( The Beast of Sodor).Only child asks her friends what its like to grow up with siblings. We're counting down Thomas & Friends Top 5 most frightening moments of all time! viral video funny videos funny parenting videos parenting motherhood. It was uploaded to the official YouTube Channel on 28 December 2017. Top 5: Scary Moments is the fourth video in the series. Top 5 Re-Builds Thomas & Friends Top 5: Scary Moments The Railway Series book Thomas the Tank Engine is acknowledged, and the Rev.Glynn's restoration ( The Christmas Coffeepot).James' repaint and repairs ( The Adventure Begins).Stephen's restoration ( King of the Railway).Gordon gets streamlined ( The Great Race) The One With Ross Tan (season 10, episode 3) Ross decides to get a spray tan but, thanks to some mishaps in the booth, ends up with very tanned front and a very pale back.Thomas is painted blue ( The Adventure Begins).We are counting down the top 5 engine re-builds! Can you remember these moments? It was uploaded to the official YouTube Channel on 21 December 2017 and is included as a Bonus Feature on the Danger at the Docks DVD. Top 5: Re-builds is the third video in the series. Top 5 Derailments Thomas & Friends Top 5: Re-builds Hiro crashes into the pipes ( Helping Hiro).
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When a business is Agile means that it can adapt to changes swiftly and move forward the projects with the right priorities. 1995), and its role may be to release these complexes from endosomes as well as to facilitate fusion of the outer cell membrane with liposome/nucleic acid complexes. DOPE is considered a “fusogenic” lipid (Farhood et al. It is unclear how the nucleic acids are released from the endosomes and lysosomes and traverse the nuclear membrane. Following cellular internalization, the complexes appear in the endosomes and later in the nucleus. Entry of the liposome complex into the cell may occur by endocytosis or fusion with the plasma membrane via the lipid moieties of the liposome (Gao and Huang, 1995). For cultured cells, the overall net positive charge of the liposome/nucleic acid complex generally results in higher transfer efficiencies, presumably because it allows closer association of the complex with the negatively charged cell membrane. The cationic portion of the lipid molecule associates with negatively charged nucleic acids, resulting in compaction of the nucleic acid in a liposome/nucleic acid complex (Kabanov and Kabanov, 1995 Labat-Moleur et al. Often, the cationic lipid is mixed with a neutral lipid such as L‑dioleoyl phosphatidylethanolamine (DOPE Figure 3), which can enhance the gene transfer ability. 1995).Ī lipid with overall net positive charge at physiological pH is the most common component of liposomes developed for gene delivery (Figure 2). Unlike DEAE-dextran or calcium phosphate chemical methods, liposome-mediated nucleic acid delivery can be used for in vivo transfer of DNA and RNA into animals and humans (Felgner et al. Cells transfected by liposome techniques can be used for transient expression studies and long-term experiments that rely on integration of DNA into the chromosome or episomal maintenance. Liposome-mediated delivery offers advantages such as relatively high efficiency of gene transfer, the ability to transfect certain cell types that are resistant to calcium phosphate or DEAE-dextran, in vitro and in vivo applications, successful delivery of DNA of all sizes from oligonucleotides to yeast artificial chromosomes, delivery of RNA, and delivery of protein (reviewed in Kim and Eberwine, 2010 Stewart et al. The cationic head group of the lipid compound associates with negatively charged phosphates on the nucleic acid. The next advance in liposomal vehicles was the development of synthetic cationic lipids (Felgner et al. Artificial liposomes were first used to deliver DNA into cells in 1980 (Fraley et al. The term “liposome” refers to lipid bilayers that form colloidal particles in an aqueous medium (Sessa and Weissmann, 1968). In addition, small pH changes (± 0.1) can compromise transfection efficiency (Felgner, 1990).ĭue to their shortcomings, chemical reagents have been largely superseded by synthetic lipid-based reagents. However, calcium phosphate co-precipitation is prone to variability and is not suited for in vivo gene transfer into whole animals. Calcium phosphate transfection is routinely used for both transient and stable transfection of a variety of cell types. The precipitate is taken up by cells via endocytosis or phagocytosis. The controlled addition generates a precipitate that is dispersed onto the cultured cells. The protocol involves mixing DNA with calcium chloride, adding this mixture in a controlled manner to a buffered saline/phosphate solution and incubating the mixture at room temperature. The method is still popular because the components are easily available and inexpensive, the protocol is easy to use, and it is effective with many different types of cultured cells. 1996).Ĭalcium phosphate co-precipitation as a transfection method was introduced in the early 1970s (Graham and van der Eb, 1973). 1995) and dendrimers (Haensler and Szoka, 1993 Kukowska-Latallo et al. Other synthetic cationic polymers have been used to transfer DNA into cells, including polybrene (Kawai and Nishizawa, 1984), polyethylenimine (Boussif et al. However, DEAE-dextran is not generally useful for stable or long-term transfection studies that rely upon integration of transferred DNA into the chromosome. This method successfully delivers nucleic acids into cells for transient expression that is, for short-term expression studies of a few days. Uptake of the complex is presumably by endocytosis. An excess of positive charge, contributed by the polymer in the DNA:polymer complex, allows the complex to come into closer association with the negatively charged cell membrane. DEAE-dextran, a cationic polymer, was one of the first chemical reagents used to transfect cultured mammalian cells (Vaheri and Pagano, 1965 McCutchan and Pagano, 1968). Visitors can make use of the park's five multi-use fields, soccer fields, sand volleyball pits, basketball courts, two picturesque ponds, and a small playground with an enjoyable climbing rock. The park's enchanting features include a delightful butterfly garden, inviting play areas, serene fishing lakes, and versatile multi-use fields. Freestone Park stands as a truly amazing destination, catering to the diverse needs and interests of individuals and families alike.ĭiscovery Park is a stunning public park spanning an impressive 48 acres. 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Freestone Park is a remarkable community park that spans over 88 acres of pristine parkland. However, they’re not all for fine noodles. This 10-blade spiralizer comes with more blades than the others in our selection, meaning you can make everything from veggie angel-hair noodles to pappardelle. Brieftons 10-Blade Spiralizer: The best spiralizer you can buy Key specs – Type: Manual Number of blades: 3ģ. All in all, though, this is a fantastic spiralizer for the price and perfect for your first foray into the world of veggie spaghetti, chips and more. The only niggle we have with it is that it’s not as easy to use with smaller veggies, so if you find yourself buying these a lot, you may want to consider a smaller handheld version instead. Better still, you’ll be adding tonnes more fibre and vitamins into your diet.Īs well as interchangeable blades, this manual spiralizer also has non-slip feet and an easy-turn handle to make shredding veggies a breeze. With three blades for fine noodles, thick noodles and tight, curly ribbons, you can recreate some of your favourite carb-heavy dishes, like pasta or curly fries, without having to worry about the extra calories or fat. It is a tad big, but don’t let that put you off. If you want something sturdier than a handheld spiralizer but don’t want to splash out on anything too complex before you’re certain you’ll use it, this cheap and cheerful option from Salter is ideal. Salter BW04294 Healthy 3-Blade Spiralizer: Best simple spiralizer READ NEXT: Check out our guide to the best blenders you can buy The best spiralizers to buy 1. Some of the cheaper models may struggle with harder veg, such as sweet potatoes and carrots, so if that’s what you plan to prepare, invest in a model that’s built to last. The general robustness is, of course, something to consider if you want a spiralizer that can handle the firmest of vegetables and slice them neatly for years to come. Whatever you’re going to spiralize, choosing a machine with good suction pads on the bottom will help keep it in place while you work – and avoid any nasty accidents. That said, many spiralizers are now dishwasher safe, so if you own a dishwasher it’s worth keeping an eye out for one with parts that you can throw in with the rest of the dishes. Most spiralizer blades can be rinsed under the tap, but sometimes they will require a bit more graft, especially with softer vegetables like courgettes and cucumbers. While most models come with removable blades that won’t add much time to your washing up routine, some are quicker and easier to clean than others. If you want to experiment with different vegetables and shapes or make salads with intricate garnishes, those extra blades will come in handy. Most spiralizers come with changeable blades for everything from thin curls to chunky ribbons, so you can create more than just spaghetti-like vegetables. However much you decide to spend on your spiralizer, these are the key features you’ll want to look for. The exception to this is handheld spiralizers, which cost a little less – and have fewer features as a result – but we’ll come back to that later. You can pick up a basic spiralizer for around £10-15, although by doubling your budget you can bag some extra features and a more robust machine, which could potentially save significant time and effort down the line. To learn more about how and for what purposes Amazon uses personal information (such as Amazon Store order history), please visit our Privacy Notice.How to choose the best spiralizer for you You can change your choices at any time by visiting Cookie Preferences, as described in the Cookie Notice. Click ‘Customise Cookies’ to decline these cookies, make more detailed choices, or learn more. Third parties use cookies for their purposes of displaying and measuring personalised ads, generating audience insights, and developing and improving products. This includes using first- and third-party cookies, which store or access standard device information such as a unique identifier. If you agree, we’ll also use cookies to complement your shopping experience across the Amazon stores as described in our Cookie Notice. We also use these cookies to understand how customers use our services (for example, by measuring site visits) so we can make improvements. We use cookies and similar tools that are necessary to enable you to make purchases, to enhance your shopping experiences and to provide our services, as detailed in our Cookie Notice. However, some people believe that there are issues with the phrase, "hope you're doing well." For instance, many people argue that the phrase is so overused that it has become insincere and even "spammy." Others, such as Elisa Doucette in her article in The Startup, mention that the phrase can be presumptive and even pressure the recipient to pretend everything is going well even if it isn't. Of course, it’s acceptable to begin a message with "hope you are doing well." It's friendly and succinct, and it's more professional than "Happy Tuesday!" or "TGIF!" Should You Say 'Hope You Are Doing Well'? Tip: If your email or text recipient recently lost a loved one, our post-loss checklist can help them sort through the complicated tasks and challenges they might be facing. So how do you check in with the people you know without using the standard phrase, “hope you’re doing well," particularly in emails and text messages? Here are a few ideas you can use in any situation. It’s important to ask about each other’s physical and mental well-being. In today’s world, it’s more important than ever before that you check in with your work and personal contacts and wish them good health and happiness. More Tips for Saying 'Hope You're Doing Well'.How to Reply to 'I Hope You're Doing Well'.How to Say 'I Hope You're Doing Well and Staying Safe'. How to Say 'I Hope You’re Doing Well' After a Loss or Tragedy.How to Say 'I Hope You're Doing Well' to a Friend or Family Member.How to Say 'I Hope You're Doing Well' in a Work Email.Should You Say 'Hope You Are Doing Well'?.Saying that you hope someone's doing well is a meaningful and important sentiment, so finding alternative ways to convey that message in emails and texts is a great idea. You likely use this phrase often in your emails and texts, and you might feel like it's time to mix things up and try a different message. "Hope you're doing well" is a time-honored phrase that's useful when checking in on loved ones, colleagues, and friends. As an Amazon Associate, we earn from qualifying purchases. We also may earn commission from purchases made through affiliate links. We follow a strict editorial process to provide you with the best content possible. views import mycoolapp create application def createapp (debugTrue): app Flask (name, instancerelativeconfigTrue) app.debug debug. You need to use currentapp: import sqlite3 from flask import Flask, g, currentapp from. any help where i can find the latest deployment packages. Your mycoolapp is an instance of Blueprint which doesnt have a config attribute. db files is not updated wiht latest rows only the initial files during deployment was present. i am downloading this and when verify by unzipping it the. import sqlite3 from flask import g DATABASE /path/to/database. i am downloading the folder from azure-advanced tools->go->Site. Lets see an example of SQLite3 with flask. db file should have all recent rows updated using the hosted api link and thought of using it again for redeployment so that the continuity is maintained and i have to repeat this same methodology in client implementation too for trouble shooting and database correction. pls helpĮxpected solution: the downloaded. Thought of using the downloaded db for trouble shooting but it got only the initial rows. db file but it contains only the initial rows which i upload during the initial deployment. Now i downloaded the wwwroot from app editor from azure page assuming all the recent rows inserted via hosted api can be seen in the download file ( wwwroot folder).Using hosted API I performed many crud operations and verified it to via postman all rows are updated. it has got all rows from the local which is fine. I deployed the project to azure using vs code. all crud operations works fine in local env. db file along wiht app.py for it to access it. I am using azure webservices for hosting my python flask app (os- linux), used SQLite DB which will do all crud operations from code. |
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