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One of the most in-demand services that you can offer to fit the active-income bill online, is software development. Software development, or coding as it's generally referred to nowadays, is in high demand and will continue to be so for the foreseeable future as the reach of technology continues to unravel its tentacles further and further into our lives.
You know those top-down cooking or craft videos you just can't seem to get away from these days? There are people out there making a living from them. 78% of B2C companies depend on user-generated content, like those videos, for their marketing campaigns. You can sign up as a creator on a site like Darby Smart and potentially work with brands like Nordstrom, Mattel, and BarkBox. Or, use them to build your YouTube following and monetize through ads and views.
One of the easiest ways to make money online is to register for paid survey websites where you can earn money for taking surveys. You can also search online for "website testing jobs" and get paid to record yourself testing websites. If you want something that pays better, become an online tutor or start an online freelance writing business. For more tips from our reviewer, like what kind of items you can sell online to make money, keep reading!
Similar to e-commerce websites such as Amazon and OLX, a peer-to-peer (P2P) platform is a marketplace for money lending activities. You can lend money to others in a more organised and structured manner using the P2P lending platform. The P2P platforms have the recovery process in place and one should understand it before using the services of the platform. Since this is an unsecured loan where there is no face-to-face interaction, a P2P lender needs to be aware of the risks involved. Currently, the interest rate earned can be anywhere from 13 percent to 30 percent.
Take advantage of marketing tools whenever possible. Many affiliate programs offer useful resources to their affiliates aimed at helping affiliates make more sales. These range from monthly newsletters highlighting upcoming sales, tips or affiliate case studies, Facebook Groups just for affiliates, private webinars explaining marketing strategies in depth and more. Ultimate Bundles does this exceptionally well.
Clearly, there's a lot of demand on Amazon, and if any product is going to sell, it's going to sell well on Amazon. But the goal here is to source the right products that will easily sell at the world's largest online retailer. Generally, products between $10 and $50 sell very well here. Just be sure to do the right market research before jumping on this bandwagon.
There's an excellent chance that you have one or several musical instruments sitting in storage that haven't been used in years. Perhaps it's a leftover from when you were in school, or even from your days playing in a band. Whatever the reason that you have it, it's probably worth money if you can sell it. Brand-new instruments are ridiculously expensive, so people often look to buy used equipment instead, particularly if they are novices.
By quite a large margin Amazon has the largest affiliate marketing program out there, with products from more than 1.5 million sellers. Amazon has the most easy-to-use technology of all the affiliate programs I will be reviewing today. Beginners to affiliate marketing with even the most limited technical expertise will have no problems in getting up and running with the Amazon associates program, while more experienced marketers can create custom tools and websites with the APIs and advanced implementations available to them. The great thing about Amazon is that anything from kids toys to laptops can generate sales if they are purchased through any Amazon affiliate link.
MemberPress is a feature-rich plugin that will enable you to turn your WordPress blog into a fully functional membership site. Depending on how you want to run your membership site, you could have a mixture of free and premium membership plans. Premium plans could include no adverts, free downloads, access to extra content, membership forums, and much more. However, it is important to remember that if people are going to pay to view your content then it must be high quality. Otherwise your members won't renew their subscriptions when the time comes.
Individual sellers and companies offering products or services have to deal with their consumers and ensure they are satisfied with what they have purchased. Thanks to the affiliate marketing structure, you'll never have to be concerned with customer support or customer satisfaction. The entire job of the affiliate marketer is to link the seller with the consumer. The seller deals with any consumer complaints after you receive your commission from the sale.
Is an affiliate program right for your business? Only you can decide. Break out the ol' pro/con list. One of the easiest ways for you to determine whether or not it's a good idea is by checking to see if any of your competitors have one and seeing what commission they're offering. Simply look for a link in their website's footer that says something like "Affiliates" or "Become an Affiliate."
Once you have decided what type of product you are going to sell, you need to decide where to sell them. Selling merchandise on Amazon or eBay aren't your only options. Creating your own eCommerce store is another way to promote your products and generate sales. Once you have decided what you are going to sell, whether it is white labeled products, your own designs, or other people's merchandise, you can set up an eCommerce website to display these products.
Active income requires your time and you can only produce as much money as your time allows. For example, a doctor or a lawyer who charge high per-hour or per-consultation fees and make exorbitant amounts of cash are stifled by the amount of time they have in a day. We all have the same 24 hours. Not one person on this earth gets more time than the next. It's the greatest equalizer in life. When the time you work directly correlates to your income, it's called active income.
Sponsored posts work much in the same way as paid guest posts, but they are posted by big businesses instead of individual bloggers. Therefore, the scope for fees is much higher, as businesses have larger marketing budgets than humble bloggers. Having sponsored posts by large companies will also help promote your site as reputable and as a leader in its field.
Build up a following on your Instagram account and you could quickly be making extra money online. Major brands, gear companies, and even startups are willing to shell out $500-$5,000+ per post to get in front of your audience. While it's getting harder and harder to build a massive Instagram audience, if you already have a solid niche and are posting quality content regularly, with a few small tweaks you can make yourself an influencer. Check out this awesome article from Shopify on how to build and grow your Instagram following to get started.
As you can see the two types of programs are different. Even so, there are some similarities. The biggest one, as you can guess, is that incentives drive people to promote. Whether it be promoting a link in a piece of content, like in affiliate marketing. Or sharing a referral link with a friend, as with referral marketing. The incentive isn't the only thing on the line, however…
Creating and selling courses on your blog can be a successful way to make money online. The course itself should be relevant to your blog, and based on a topic that will benefit your audience. Although it may take time to create the course in the first instance, once completed, you can then repeatedly make money from the course each time you sell it.
One thing that I look for in an affiliate program is a good transparent analytics system. I want to be able to see the clicks and transactions, so that I can track my stats. Otherwise, I get a little nervous about the legitimacy of the program. Other things to look for is to see how transparent they are when answering questions and how the responsiveness of the affiliate manager.
Now, making money online should seem like a pleasurable activity. Why wouldn't we do just about anything to see things through, since it would be a major source of pleasure, right? Wrong. In the beginning, like anything else, we might get really excited about it. We might also set our hopes very high. But that all comes crashing down when we begin to fail.
So many people dream of writing a book, but never go through with it. Yet no matter what, I'm sure you have experience and value you could give through writing a book. By packaging your skills and knowledge into a downloadable eBook that helps people learn a skill, advance their careers, or start a businesses, you can change someone's life and even make good money online.
Normally you'll be asked to test a few websites by visiting them and to document and record your reactions and thoughts as you go through it. It's really easy to get set up making extra money online by testing websites. All you need to do is sign up to the following services: UserTesting.com, Userlytics, TryMyUI, Userfeel, TestingTime (UK only), or Side Income Jobs.
I have 3 VAs. I couldn't function without them. It doesn't matter how big or how small a business is, they all need help running their day-to-day administrative tasks. Virtual assistants perform a variety of tasks that a traditional assistant or secretary would normally do, including making travel arrangements, paying bills, or managing expense reimbursements. I use them to upload content, optimise images, brief writers etc.
The phrase, "Affiliates are an extended sales force for your business", which is often used to explain affiliate marketing, is not completely accurate. The primary difference between the two is that affiliate marketers provide little if any influence on a possible prospect in the conversion process once that prospect is directed to the advertiser's website. The sales team of the advertiser, however, does have the control and influence up to the point where the prospect either a) signs the contract, or b) completes the purchase.
While this won't get you rich by any measure, you can sell products online using sites like Craigslist, Alibaba, eBay and even the Amazon Marketplace. In fact, these are some of the top online destinations where you'll be able to easily reach the masses. Instead of building your own ecommerce shop, selling products on these marketplaces, can be an instant boon for you.
Now most affiliate programs have strict terms and conditions on how the lead is to be generated. There are also certain methods that are outright banned, such as installing adware or spyware that redirect all search queries for a product to an affiliate's page. Some affiliate marketing programs go as far as to lay out how a product or service is to be discussed in the content before an affiliate link can be validated.
If you are more confident in your skills, you can also market directly to websites and blogs. You can contact the sites by email to market your services. That will also enable you to select the specific types of sites that you are more comfortable working with. Since there are literally thousands of websites and blogs on the web, the potential market is limitless.
Hi Jamie, awesome content that is very helpful esp with the resources, links and the rich discussions. Want to start e-commerce and blog for money…selling others products, want to go full on with this, tired of the daily routine crunch working for others. I live in a developing country (PNG) that has high internet costs (work still in progress with getting rates down…) so will see how I go with your posts. Any advise? Don't have a website yet, have to build one I guess….
This is something like Experts Exchange, except that it's more general in nature. As a professional in just about any field, you can offer your expertise on JustAnswer. They are looking for professionals in various fields who will provide answers to reader's questions. You can make money online by answering these questions and pointing people in the right direction to get the solutions that they need.
The easiest thing to do is to sign up for an affiliate marketing network like Commission Junction. They provide a marketplace where your affiliate program will be advertised to other affiliate marketers. They also provide the tracking software for your affiliates so you don't have to build your own tracking system. In some ways this is better because it takes care of the trust issues. Affiliates are always suspicious of whether or not they're getting credited for the sales they generate. By having an intermediary take care of transaction tracking and payments, the fear of being cheated is alleviated.
Signing up with Google AdSense will enable you to make money from advertising on your blog. Google AdSense will place relevant ads around your site, and if your site visitors click on these ads, you will earn a fee. Although each click only amounts to small change, if your blog has high levels of traffic, this will quickly amount to a reasonable sum.
Not only will this multiply the money you're bringing in in a serious way, but it protects you against any sudden changes in the market or in your business. Remember that old saying about putting all your eggs in one basket? A few hours a week committed to just one or two of the following opportunities will put you in a much stronger position to be financially safe and independent.
Jump up ^ Shashank SHEKHAR (2009-06-29). "Online Marketing System: Affiliate marketing". Feed Money.com. Archived from the original on 2011-05-15. Retrieved 2011-04-20. During November 1994, CDNOW released its BuyWeb program. With this program CDNOW was the first non-adult website to launch the concept of an affiliate or associate program with its idea of click-through purchasing.
If you want to sell products online, you may can it by creating your own website. As there's lot of competition and several existing websites in place already catering to this market, an attempt to create a niche in terms of products may be considered. Or, you can use the platform such as Amazon, Flipkart to sell. One can increase reach and visibility through affiliate marketing.
"The Rakuten Marketing team developed and executed an online marketing performance strategy that far exceeded our target goals for brand awareness, new customer acquisition and ROAS in our first-ever partnership with Disney. The campaign was such a success, we're aiming to replicate the results with the launch of our Incredibles Collection launch this year. We look forward to continued, long-term program success with Rakuten Marketing's expertise and proven technologies."
As someone who's been immersed in a number of online industries for quite some time, I know a thing or two about what it takes to succeed in this arena. However, just like you, I started at ground zero with little knowledge, but a great deal of passion. What I learned along the way were some invaluable lessons from failure that hurt at the time, but helped immensely in the grand scheme of things.
There are some key differences between affiliate marketing and the new world of so-called "influencer marketing" with "brand ambassadors." Influencer marketing, born with the rise of social media, refers to finding and reaching out to individuals you deem to be "influencers" in your space — bloggers, vloggers, journalists, experts, etc. — who have a strong online presence in your target market and who fit your brand well. By partnering with influencers, brands hope to increase exposure and subsequent sales via the influencers' followings.
Infographics are currently an extremely popular type of media. An infographic showing information or data in an interesting and fun way can help engage an audience and generate numerous likes and shares. As infographics take more time to create than just simply uploading an image, busy website owners are prepared to pay for custom infographics. And don't worry, an infographic can be created using simple design software such a Canva. So extensive graphic design skills are not necessary, just a creative eye.
Posting ads on Craigslist is technically easy, but people often have fears about posting their personal information on the site. I communicate mostly through email when doing business on Craigslist, and I've never run into any issues. I've never been ripped off, nor have I been murdered or raped for using the site. It takes common sense, so use your best judgment, but don't assume someone is a thief just because of their preferred communication method. For an extra bonus, google "funny Craigslist ads" to see some delightful examples of guerilla and grassroots marketing.
If you are building a site that has the potential for information that will never age and remain useful for your audience, you have the opportunity to create what is known as evergreen content. It's important to carry out extensive keyword research before planning any evergreen content for a site like this, as your site could hugely benefit from the proper usage of keywords within such content.
You can earn above average rates of return because peer-to-peer lending eliminates the bank function. That means that you participate in nearly the entire interest rate being paid by the borrower, rather than the less than 1% that you will typically earn on certificates of deposit. And you can reduce your risk by investing in slices of hundreds of different loans.
Some good tips. I fully agree that it's all about trust and it is always good to recommend an affiliate product that you have actually used. Why risk your reputation on a product you don't know if it's any good or not? If people buy one dodgy product from your site, they will probably never make that mistake again. That it why my blog strongly advocates ethical affiliate marketing. It's the best long term stragey for affiliate marketing success.
There are many ways to get people onto your list. Lead magnets are one such resource. For example, you can build ebooks, checklists and cheat sheets. But you can also do content upgrades, such as PDF versions of an article with added resources in them, four-part video training series, and more. Think about your audience and what you can offer them to better serve them, then treat them with some respect and you'll eventually reap the rewards.
To create your own blog, you don't even need a web domain. You can start a completely free blog on either WordPress or Blogger. Each of these sites has its ups and downs, but you can't beat the price. You generally want to keep blog posts between 150-500 words until you have a few dozen under your belt. Links between your blog posts encourage people to stay on your blog once they find it, increasing traffic. Once your blog is up, promote it on social media for the greatest effect.
A site called User Testing will actually pay you a fee to evaluate websites. It typically pays you $10 for each video that you review – which typically takes about 20 minutes. If the work is there, and you are particularly good at it, you could earn up to $30 per hour. That's a pretty solid pay rate for a work-at-home job. You wouldn't have to work a whole lot of hours to generate a decent part-time monthly income.
Many affiliate programs will often run promotions with good discounts or giveaways that might be attractive to your audience. For example, if you're an Amazon Associate and the site have a big Holiday Sale, it would be the perfect opportunity for you to promote discounts to your website visitors. This is a great way to promote your offers while also providing good value to your audience.
In the past, many affiliate marketers focused on a catch-all approach, offering traffic up to hundreds or thousands of sites, even if they had little authority or traffic to give. But in 2018, advertisers will laser their focus into smaller groups of highly credible, targeted, and popular influencers. To map your strategy, focus on a few key influencers in your industry. Start with them, then nail your niche before expanding outward.
If it's not clear yet, information and expertise are two of the most consistent and lucrative ways to make extra money online. If you don't like talking on the phone, you can sign up to get paid to answer professional questions on JustAnswer. There are thousands of questions being asked every month from people looking for help from lawyers, doctors, mechanics, vets, and more. To apply, you'll need to supply your professional verification, resume, and a form of identification.
Drop shipping is another great 'hands-off' way to sell products. Firstly you will need to find businesses that sell products in your niche that offer a drop shipping service. Then you will need to create a website promoting and selling the products. When you make a sale, you take the payment on your site and then the manufacturer ships the goods to the buyer. The profit comes from charging a higher rate than the manufacturer, and if you are selling a high number of products this can quickly add up to a healthy revenue.
If you don't mind doing other people's chores, then TaskRabbit is a great option for making money online. Earn extra income by walking your neighbor's dog or mowing Mr. Smith's lawn. It might seem like not the most lucrative option, but the top taskers reportedly earn as much as $7000 a month, making this a full-time way to make money online for some.
If you have a knack for organization, you can make money online as a virtual assistant helping people to keep their days in order. A virtual assistant will do everyinthing from bookkeeping to research, database entry, booking travel, and managing email. It can also be an awesome way to rub shoulders with some very important people, build up your professional network, and of course grow another stream of income. You can find great gigs on UpWork, Fiverr, Indeed, and Remote.co.
If you're a musician, writer, artist, tech nerd, pundit, or can produce any type of audio, video, or text worth consuming, you may be able to sell some stuff in Apple's flagship iTunes store. By selling your work here, you're able to stand next to the marketing clout of big business. You can make a healthy living off the iTunes store, and there's no better time than now. Learn more about iTunes with this lifehack.
Most marketers track and analyze a variety of metrics when it comes to their digital marketing efforts, especially for e-commerce. Some of the metrics are largely useless, others are very interrelated, and maybe only a few are the critical KPIs. Here, we'll examine the most important metric of them all when assessing your e-commerce website. Specific to digital marketing and e-commerce, I'll argue that the most important (and often most misunderstood) metric is Revenue Per Visitor, or RPV.
No. Before you get too excited, you must understand one very important thing. Affiliate marketing for beginners can be hard. It is, after all, a business. It is not a get rich quick scheme, and no honest affiliate manager will ever tell you it is. It requires patience, perseverance and heaps of learning. Sometimes it can take years to achieve the results you really want to see, although with the help of our affiliate management team, we hope you would see them much sooner than that!
Ever since the idea of online auctions came into existence, the online selling market has been on the rise. Many are interested, but don't know how to get started. There are still all kinds of ways to make money by selling online, whether you're selling what you already have or buying and selling like a store. Before we get started, here are a few general tips when selling anything online:
Still, much of the world is engaged in active-income work. They work so that they can earn. When they don't work, they don't earn. Yet, there's almost zero chance that you'll get rich or make any significant amount of money when you're solely reliant on active income. Now, don't get me wrong. It's possible. People do it by slowly saving and investing over the course of 30 or 40 years, but we're not talking about that here.
Take advantage of marketing tools whenever possible. Many affiliate programs offer useful resources to their affiliates aimed at helping affiliates make more sales. These range from monthly newsletters highlighting upcoming sales, tips or affiliate case studies, Facebook Groups just for affiliates, private webinars explaining marketing strategies in depth and more. Ultimate Bundles does this exceptionally well.
Do you see all of those articles, tutorials and guides all over the Internet? Somebody wrote every one of them! If you have decent writing ability (no, you don't need a journalism degree!), and knowledge in a few specific topic areas, you can be one of those writers. It's an opportunity to make money online and without ever leaving your home. It's also the kind of venture that can start out as a small side business, but grow into a full-time career.
If you have a passion project, Kickstarter may be exactly what you need to get it off the ground. Celebs like Zach Braff and Melissa Joan Hart have used this site in attempts to fund their movie projects. Some crack dealers in Canada inspired a "crackstarter" campaign from Gawker for pictures of the Toronto mayor smoking crack with them. All you need is a dream, a goal, a budget, and a great pitch to convince people to come to the site and donate to your Kickstarter campaign. Panhandling isn't dead – it's gone digital…
If you've got experience and expertise in a specific area of business, there are hundreds if not thousands of business owners who would love to jump on a phone call and pick your brain. This can be an easy way to make extra money online and to hone your skills to use in your consulting or freelance business. Sites like Clarity.fm let you create a profile where entrepreneurs and business owners can pre-pay to book a phone call with you for a set amount of time. Whether you're an expert in raising money for startups, building apps, or running a restaurant, you can make extra money selling a few minutes of your time to people in need.
My 10-year-old son brought home a book from our park's free library box. It was a biology textbook – teachers edition. He said it looked interesting and hey, it was free (having no idea you could sell it). I scanned it in my Amazon seller app and realized it was worth around $150. He was so excited. We listed it for sale for $130 and it sold! Going to tell him, he just made $130!
Said to be the largest affiliate marketing network in North America (though it operates globally) and claiming the number-two spot in the 2012 Blue Book of Top 20 Affiliate Networks for being "the best at balancing the relationship between the merchants, the network, and the affiliates," California-based Commission Junction, owned by ValueClick, Inc., offers affiliate, media, and tracking services and provides either a self-management or company-managed option for your affiliate relationships.
If you seriously want to make money online, you need passive income. You need your income to come automatically so that you can use the precious little time you have to produce more streams of income that will pay you on autopilot. That's the holy grail of income-producing activities. Now, it's not easy to do, so I'm not going to sit here and try to insult your intellgience. It takes an enormous amount of effort and very little financial gains at the outset.
Find a profitable niche. Starting with your interests, write down as many niche ideas as you can. Think about topics people might search online. Ideas include passions (like surfing or body building), fears (like spiders or speaking in front of crowds) and problems (like getting out of debt). Do keyword research to see it others are interested in the topic. Find out if a domain name is available that matches the keyword 100 percent. [7]
The initial setup and implementation of a successful, fully developed affiliate program takes some time. Similarly, huge results from affiliate marketing do not happen overnight; it is a long-term investment. Thinking it's a one-time, set-and-forget strategy is the wrong approach. Affiliate marketing can provide massive results, but only if you're willing to put in the time.
Re-selling web hosting will enable you to make money hosting your own client's websites. Large hosting companies like HostGator allow you to white label their hosting services. You could start your own hosting business or, if you are a web designer, include hosting into your web design packages. And the best of it is the hosting companies take care of the hosting for you, so all you need to worry about is selling.
Affiliate marketing programs typically work by having the merchant handle all the logistics involved in selling products or services, processing customer orders and payments, and shipping merchandise—all while the affiliate sits back and collects a commission for each agreed-upon action completed by the visitors the affiliate sends to the merchant's website via an affiliate link. As long as the affiliate has done her homework and chosen a trustworthy affiliate program, she needn't worry about non-payment.
You could also opt to use existing websites for making money. These include both active income and passive income methods. For example, you could sell some used items or invest in creating some digital designs that then can be sold on merchandise. Again, devote a sizable portion of your time to passive income so that you can slowly build up earnings that will arrive on autopilot without any extra added effort.
Despite its older origins, email lists are still a viable source of affiliate marketing income. Some affiliates have email lists they can use to promote the seller's products. Others may leverage email newsletters that include hyperlinks to products, earning a commission after the consumer purchases the product. Another method is for the affiliate to cultivate email lists over time. They use their various campaigns to collect emails en masse, then send out emails regarding the products they are promoting.
Creating and selling courses on your blog can be a successful way to make money online. The course itself should be relevant to your blog, and based on a topic that will benefit your audience. Although it may take time to create the course in the first instance, once completed, you can then repeatedly make money from the course each time you sell it.
Create a writer website or blog. A website not only demonstrates your technical ability, but it also creates an online hub that allows clients to connect to you. Keep the design of your website clean and uncluttered. Include examples of your work that demonstrate the kind of writing you do. Make the samples easy to find and to read, and make it easy for visitors to figure out how to contact you.[23] . A blog highlights your technical ability and showcases your ability to write blog posts. Your blog can be about different topics than those you write about for your clients. In fact, it should be on a topic that interests you. Visitors will see that you can not only write, but you can also build an online community. A good blog has the potential to earn you many referrals for more clients.[24]
Video is growing like crazy. And more and more people are looking for professional help cutting their raw footage into viral-worthy content. If you have the right software and a bit of skill, you can easily make money online as a video editor. Check out these article of Fstoppers on how to become an online video editor and then look for relevant jobs on Mandy.com, Creative Cow Job Search, or ProductionHub.
If writing books interests you, an option you have is to self-publish eBooks and paperbacks with Kindle Direct Publishing, and reach millions of readers on Amazon. Publishing takes less than 5 minutes and your book appears on Kindle stores worldwide within 24-48 hours. One can earn up to 70 percent royalty on sales to customers in the US, Canada, UK, Germany, India, France, Italy, Spain, Japan, Brazil, Mexico, Australia, and so on. One can keep control of one's rights and set their own list prices and also make changes to one's books at any time. BooksFundr and Pblishing.com are two other places to get your book published and earn money.
Although it differs from spyware, adware often uses the same methods and technologies. Merchants initially were uninformed about adware, what impact it had, and how it could damage their brands. Affiliate marketers became aware of the issue much more quickly, especially because they noticed that adware often overwrites tracking cookies, thus resulting in a decline of commissions. Affiliates not employing adware felt that it was stealing commission from them. Adware often has no valuable purpose and rarely provides any useful content to the user, who is typically unaware that such software is installed on his/her computer.
There's an excellent chance that you have one or several musical instruments sitting in storage that haven't been used in years. Perhaps it's a leftover from when you were in school, or even from your days playing in a band. Whatever the reason that you have it, it's probably worth money if you can sell it. Brand-new instruments are ridiculously expensive, so people often look to buy used equipment instead, particularly if they are novices.
When creating a membership site, always offer different plans and pricing tiers to appeal to your different categories of audience. Access to specific types of content is then dependent on the plan selected by a member. Membership fees can be a one-off payment or a repeat subscription. And you can even sell products on your membership site to boost your income.
Lauren McManus is a well-known Pinterest expert from Create and Go as well as other successful websites and courses. I asked her to share her best Pinterest tips as a bonus for my course because I am a huge believer in Pinterest and how it can help you to improve your affiliate income. This strategy has allowed her to receive hundreds of thousands of visitors from Pinterest, significantly improving her income.
Find your niche partners, collaborators, and champions: As you're creating your course, look for notable people who are also creating content in the space. Look at how their businesses operate and incorporate that into your own plan. You can also reach out to any influencers and make them affiliates for your own course. This way, they'll be incentivized to share your content with their own audiences (which can be a major way to generate your first sales and start building your own community!)


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Back To The Bible
A Pardon Not Accepted
John Collins
Though it is a good, touching story, the "Pardon not received" story told so many times is not historically accurate.
You remember the pardon that the man was shot, that I sometimes tell, during the civil war when he was a good man? He was innocent, and they found him guilty, although he was guilty in a way, that he run away in time of battle. And they found him guilty and was going to shoot him. And a man went to President Lincoln and said, "Mr. Lincoln, this is a Christian man. He was scared, the boy. I know his people. He was just afraid. He didn't mean no harm. He run away." Said, "Mr. Lincoln, it's in your hands. You're the only one can pardon him." Mr. Lincoln picked up a piece of paper and his pen and signed, "Pardon this So-and-so. Abraham Lincoln."
He ran back to the jail, and he said, "Here it is. I got your pardon."
And the man said, "I refuse to look at it. It would have a big seal on it. It would be everything. You're only trying to make me a laughing stock. It is not Abraham Lincoln. Anybody could sign his name, but it would have to be documented by his seal and so forth if it comes from him." And the man persuaded him, though the man in the prison thought he was kidding, and just walked away. The next morning he was shot. And then after he was shot, then there was a Federal Court trial, because Abraham Lincoln, twenty-four hours before the man was shot, signed his name that this man was pardoned. And then the government shot him anyhow. Then what? Then the Federal Court of the United States said-come to this decision of the Federal Court, said, "A pardon is not a pardon unless it be received as a pardon."
61-0730M Gabriels Instructions To Daniel
The first time this issue came to the Supreme Court was in 1833 during Andrew Jackson's presidency. Some interesting things to note:
*The defendant was not innocent. He was caught stealing and reading US postal mail, and endangering the postal worker.
*There is no record that the president knew his family, or that he even was a Christian
*The pardon was not hand-written on a hand-written note, it was an official document with president's seal.
*In 1927, the Supreme Court ruled that a pardon did not have to be accepted by the recipient, it was the decision of the courts.
Supreme Court Ruling:
United States v. Wilson, 32 U.S. 7 Pet. 150 150 (1833)
The defendant was indicted for robbing the mail of the United States, and putting the life of the driver in jeopardy, and the conviction and judgment pronounced upon it extended to both offenses. After this judgment no prosecution could be maintained for the same offense, or for any part of it, provided the former conviction was pleaded.
The power of pardon in criminal cases had been exercised from time immemorial by the executive of that nation whose language is our language, and to whose judicial institutions ours bear a close resemblance. We adopt their principles respecting the operation and effect of a pardon, and look into their books for the rules prescribing the manner in which it is to be used by the person who would avail himself of it. A pardon is an act of grace, proceeding from the power entrusted with the execution of the laws, which exempts the individual on whom it is bestowed from the punishment the law inflicts for a crime he has committed. It is the private though official act of the executive magistrate, delivered to the individual for whose benefit it is intended and not communicated officially to the court.
Pardon from Andrew Jackson:
"Andrew Jackson, President of the United States, to all who shall see these presents, greeting:"
"Whereas a certain George Wilson has been convicted before the Circuit Court of the United States for the Eastern District of Pennsylvania of the crime of robbing the mail of the United States, and has been sentenced by the said court to suffer the penalty of death on 2 July next, and whereas the said George Wilson has been recommended as a fit subject for the exercise of executive clemency by a numerous and respectable body of petitioners, praying for him a remission of the sentence of death, inasmuch as, in such a case, sentence of imprisonment for twenty years may yet be pronounced against him on the indictments to which he has pleaded guilty in the circuit court of the United States for the said district, and a still more severe imprisonment may be awarded him for the same acts in the criminal courts of Pennsylvania, now therefore, I, Andrew Jackson, President of the United States of America, in consideration of the premises, divers other good and sufficient reasons me thereunto moving, have pardoned and do hereby pardon the said George Wilson the crime for which he has been sentenced to suffer death, remitting the penalty aforesaid, with this express stipulation, that this pardon shall not extend to any judgment which may be had or obtained against him in any other case or cases now pending before said court for other offenses wherewith he may stand charged."
"In testimony whereof I have hereunto set my hand and caused the seal of the United States to be affixed to these presents. Given at the City of Washington this 14 June, A.D. 1830, and of the independence of the United States the fifty-fourth."
Commentary on Biddle v. Petrovich (1927):
There is, however, an apparent limitation on the Burdick rule as applied in the law of commutations. In Biddle v. Petrovich (1927), the recipient of a Presidential commutation of sentence from death to life imprisonment claimed it was invalid because he had not consented to the commutation. The Supreme Court ruled that his consent was not required. Mr. Justice Holmes wrote: "A pardon in our days is not a private act of grace from an individual happening to possess power. It is a part of the Constitutional scheme. When granted it is the determination of the ultimate authority that the public welfare will be better served by inflicting less than what the judgment fixed...Just as the original punishment would be imposed without regard to the prisoner's consent and in the teeth of his will, whether he liked it or not, the public welfare, not his consent determines what shall be done." This logic could theoretically be applied to pardons as a whole, but the Supreme Court has not yet overruled Burdick.
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Matthew Faulkner
East Palestine High Class of 2007
Personal Info
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Real Estate in Live Oak/Pleasure Point Santa Cruz
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Most people call this beatiful creation Bylochki but I call them Piroshky, we always have in our family.
We Slavic people love our piroshky with poppy seed, the more poppy seed the better. We grew some in our garden in Ukraine, mom would use it in very small potions for piroshky as it was treated like a delicacy. Out of all piroshky she made one or two rolls with poppy seed and we would eat it on Sunday. We are lucky, here in USA poppy seed are cheap, available in most stores and often come already premade with sugar for filling.
My sister calls them Lazy piroshky. She would always bring her piroshky loaded with poppy seed and ohh man… they are good! I made them recently and they are easy to make. The most surprising part to me was the fact that you don't need to wait for dough to rise. So next time I have unexpected guest, I am making this.
Other Piroshki Recipes you Must Try:
- 2 1/2 cups
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- 3/4 cups
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- Flour
- 1
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- 16 ounces
- Poppy Seed Filling; I purchase my from local Russian store
Note: Almost seems like step is skipped, they do not have to rise even after you will roll them all out. Piroshky do double in size while they bake so don't worry if they look tiny when you pot them inside the oven.
Tip: You can do twice the portion and freeze them for busy day. To warm them up you can use oven or microwave. If you plan to use oven, you should under cook them for about 5 minutes.
Frozen piroshky were really helpful once for me when I needed to host unplanned lunch.


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The holiday season is a time that's ripe with traditions, and for nearly three decades, the Eagle community has ushered in December with its Christmas on Broadway celebration.
The 29th annual event is planned for Saturday, Dec. 1, in downtown Eagle.
A nighttime lighted parade is the centerpiece of the celebration and participants will march down Broadway at 5:30 p.m. The parade starts at Second Street and concludes at Town Park, where kids can visit Santa's workshop set up at the Fifth Street ECO bus shelter – sponsored by Alpine Bank.
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Following the parade, a bonfire, live music and other entertainment is planned at Eagle Town Park, downtown businesses staying open for special shopping hours.


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Public Private Partnerships
The Billion Dollar Question…to P3 or Not to P3
Royal Columbian Hospital is undergoing 3-phase $1.35 billion redevelopment, the largest government funded health care capital project in the province's history. Phase 1 was approved and procured as a Design Build and is now in construction. Phases 2 and 3 were approved as a DBFM-with-private-sector hospital-wide-maintenance. While the project was preparing for Phase 2/3 procurement the shift focused away from P3.
Our team undertook an extensive analysis of enhanced DB, enhanced DBF, and DBFM-with-public-sector-hospital-wide-maintenance while still keeping the procurement process and timelines on track. In this session, we share our lessons learned and the findings of an extensive procurement options analysis undertaken by Partnerships BC, Ernst and Young, and Fraser Health, as we answered our billion dollar question, to P3 or not to P3.


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Opened 11 years ago
Last modified 4 years ago
#2417 reopened enhancement
New Trac does not allow to cc by username
|Reported by:
|Owned by:
|Blocked By:
|Has a Patch:
The old Trac system allowed to edit the Cc: field of a ticket to add one's username.
The new Trac only allows to edit the full Cc: field for a new ticket and to add one's email address to the Cc field of an existing ticket via checkbox.
The username is advantageous in not exposing the email address on the ticket's Web page and for allowing to change one's email address in the future.
Attachments (1)
Change History (18)
comment:1 Changed 11 years ago by
|nielx added
I think you lost me. What exactly is happening right now and how would you like it to be?
Changed 11 years ago by
|add_cc.png added
image of new "add cc" functionality
comment:2 Changed 11 years ago by
Mine shows it at the bottom now as a checkbox (see attached image).
comment:3 Changed 11 years ago by
|umccullough@… added
Oh, i didn't read the whole description - the email is not shown - just like in the old version :)
comment:4 follow-up: 9 Changed 11 years ago by
I get it.
Don't worry, your username is not exposed. You can only see your own email and not someone else's. Only administrators can see emails. That's the way it is currently set up.
You can check it out by adding yourself to the CC field and then click 'Log out' at the top of the page. That is what it looks like to innocent bystanders.
If you are satisfied with the situation I'd suggest we close this ticket.
comment:5 Changed 11 years ago by
I'm not the originator of the ticket, I was just commenting on what I see here for his sake :D
comment:6 Changed 11 years ago by
Ah, but let me point out - it *does* show the full email address in the notification that is emailed...
Niels, is there any way you can fix that to anonymize in a similar way?
comment:7 Changed 11 years ago by
Okay, it may not be visible to the outside but I'd still prefer the checkbox to add/remove my username rather than my email address.
Niels, what I did previously was I added in the Cc field ", andreasf"; now, via the checkbox, ", andreas.faerber (at) ..." is added. I would prefer if instead the username were used, to have the Trac preferences as the central place to administer my email address. (I have no idea if this is easy to change or not.)
comment:8 Changed 11 years ago by
comment:9 Changed 11 years ago by
Replying to nielx:
Don't worry, your username is not exposed. You can only see your own email and not someone else's. Only administrators can see emails. That's the way it is currently set up.
I doubt that I am an admin, and today I received a notification from Trac that showed somebody else's email as being added to the CC list. This was for ticket:
The email is obscured in Trac, but the email notification is sending out the full email address as is, and that is IMO undesired.
comment:10 Changed 11 years ago by
Yes, it is. Due to illness I did not feel like browsing through the code this week. I will do this, but it will have to wait for a bit. Sorry.
comment:11 Changed 10 years ago by
|umccullough@… removed
comment:12 Changed 10 years ago by
Test whether email addresses are still visible on the mailing list.
comment:13 Changed 10 years ago by
|niels.reedijk@… added; nielx removed
comment:14 Changed 10 years ago by
|→ fixed
|new → closed
Seems to be fixed now.
(Note that people with 'higher' Trac privileges are able to see the email addresses, but that is intended).
comment:15 Changed 9 years ago by
|closed → reopened
Actually the issue I opened this ticket for is not fixed:
In the "Change Properties" section below, as a regular user, I still see "Add to Cc: myemail@… [ ]", and it does add that specific email address to the CC list.
I was asking for "Add to Cc: andreasf [ ]" or similar, so that I can have my username added to the CC list in place of an email address, whether displayed partially or fully. Originally, I was able to manually type into the list of CCs, so that I could make this choice myself.
comment:16 Changed 9 years ago by
Note to self:
The email is retrieved from trac/ticket/web_ui.py:
In theory by changing this line to retrieve the username instead of the email associated with the session, it might work again.
comment:17 Changed 4 years ago by


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Welcome to Sapporo International
SAPPORO International is a well-known and unique educational consultancy and language school in Kathmandu, registered under the Act of Nepal Government. It is a leading institution which provides Japanese language trainings, consultancy service for students willing to go to Japan for their destination of higher education. Moreover, we are committed to provide quality education to all the learners Read More
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Patricia Matz
Fort Benton High Class of 1959
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Yesterday I posted my lonely dinner menu. I wanted to rep my veggie friends, so I decided to post another dinner for one, this time vegetarian, that I ate earlier in the week.
Just like when I serve salad for dinner, The Hubs doesn't love when I make these stuffed peppers for dinner either. He never feels like he gets enough to eat so I always try to reserve these for when he's out of town. I love them, and they are a great portion size for me!
I have made these with many different types of beans; from black beans, refried beans, pinto beans, you name it. I have also used a variety of onion/veggie/cheese combos. The ones shown have black beans, red onion, red pepper and sharp cheddar. Just make do with whatever you have in your fridge or pantry!
First, get yourself a poblano pepper or two
Halve them, seed them, then char them on a grill for a few minutes
Make your filling mix and put the peppers in a shallow baking dish
Stuff the peppers and top them with cheese. Broil them in the oven for a few minutes until the cheese melts!
If you have any leftover the next day, chop them up and make a mexican scramble!
As yummy as these were, I'm happy to not be eating dinner for one any more this week! The conference is over, my friend Krista arrived from Chicago last night, and my other friend Katie is flying in today! Looking forward to an AWESOME weekend catching up with old friends and enjoying this GLORIOUS, warm, Southern California winter weather! Happy Friday to all!


text:
Host factors render cells susceptible to viral infection. One family of susceptibility factors, the tetraspanin proteins, facilitate enveloped virus entry by promoting virus-cell membrane fusion. They also facilitate viral egress from infected cells. In this Gem, we discuss recent insights into how tetraspanins assemble viral entry and exit platforms on cell membranes, and we speculate that tetraspanins contribute to nonviral membrane fusions by similar mechanisms.
For enveloped viruses, host factors determining susceptibility to infection include several "activators" which operate at particular subcellular sites to stimulate virus-cell membrane fusion. Depending on the infecting virus, activators may be cellular receptors, cations, or proteases which engage viral fusion proteins and trigger their refolding into forms that catalyze membrane coalescence. For many viruses, multiple activators are utilized in rapid succession. In these cases, activators must be in close proximity to each other, and to the incoming viral fusion proteins, to allow for rapid and efficient viral entry. Recent research identified the role of cellular proteins, including tetraspanins, in coalescing these activators. In this Gem, we discuss how tetraspanins facilitate efficient viral infection by organizing activators on host cell membranes.
The tetraspanins are membrane proteins; they have four (tetra) transmembrane spans linked extracellularly by one large and one small extracellular loop (termed LEL and SEL, respectively) (Fig. 1, inset). They are ubiquitously present in eukaryotes (1). Their conserved structures and interacting partners suggest that they perform central roles in controlling membrane architecture. For example, they induce positive (outward) membrane curvatures (2), potentially through their cone-shaped transmembrane domains (3). Tetraspanins further influence membrane architecture by interacting more directly with lipids and with other integral membrane proteins. The tetraspanin CD81 binds cholesterol, which incorporates into a pocket between the tetraspanin transmembrane domains and alters LEL conformations (3). Several other tetraspanins associate with palmitic acid, which covalently links to cytoplasmic domains (4, 5). In conjunction with cholesterol and palmitate, tetraspanins interact with several so-called "partner" proteins. Distinct LEL- and cytoplasmic tail-dependent interactions with integrins (6, 7), adhesion molecules (8), and other tetraspanins and their associated partners (9) give rise to web-like tetraspanin-enriched microdomains (TEMs) on cellular membranes (10, 11). It has been proposed that these TEMs might be platforms for virus entry (12, 13), particularly for viruses that directly use tetraspanins as receptors (14). Yet there have been several reports that both enveloped and nonenveloped viruses preferentially enter cells through TEMs, presumably without directly binding a tetraspanin molecule (15–18). These intriguing reports stimulated our interest in the mechanisms by which viruses use TEMs for cell entry.
Our inroads into mechanisms by which tetraspanins facilitate virus entry came from investigations of coronaviruses (CoVs). These are enveloped viruses that cause respiratory and enteric infections in humans and animals (19); life-threatening severe acute respiratory syndrome CoV (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV) are notable members. CoV infections are driven by spike (S) fusion glycoproteins. Extending from virions, these S proteins bind target cell receptors and then encounter cellular activators. The activators are proteases which cleave S proteins in ways that liberate domains catalyzing virus-cell fusion (20). Frequently, and in MERS-CoV infections, this proteolytic activation process takes place in stages. Furin and related proprotein convertases cleave S proteins during virus egress from producing cells (a "priming" step) (21) and then, after secreted viruses attach to target cells, serine or cysteine proteases cleave S proteins again (a "triggering" step) (22–25). Here, at the second triggering step, is where the tetraspanins come into play (Fig. 1, left panel).
TETRASPANINS PROMOTE CoV ENTRY BY LINKING CoV RECEPTORS AND PROTEASES
Since receptor-bound CoV S proteins are susceptible to triggering cleavages (26, 27), we surmised that receptors and S-cleaving proteases must be in close proximity to trigger membrane fusion. Tetraspanins appeared to be reasonable candidates for bringing receptors and proteases together. Indeed, there were good suggestions that the CoV receptors and proteases are coalesced within TEMs (28–31). Therefore, we hypothesized that tetraspanins condense CoV entry factors into localized positions whereby effective spatiotemporal activation of viral fusion takes place.
We first explored this hypothesis by using biochemical approaches. We isolated TEMs from cells containing CoV receptors and activating proteases and determined whether they localized to TEMs. Indeed, two CoV-activating proteases and the receptors of MERS-, SARS-, 229E-, and murine hepatitis virus (MHV)-CoVs were all found in TEMs (32). TEMs, while containing only ∼20% of all plasma membrane proteins, contained 50% to 90% of the CoV receptors and proteases, indicating that CoV activators are targeted to TEMs. Notably, isolated TEMs activated CoV S proteins in vitro, allowing these viruses to enter cells in a manner independent of proteases on target cells. TEM activating potential also correlated with protease abundance (32). These findings indicated that TEMs act as platforms for fusion-activating CoV S protein proteolysis.
We next used genetic manipulation to consider whether individual tetraspanins are required for fusion activation and virus entry. Given the specificity of tetraspanin-partner protein interactions (33, 34), it was likely that individual tetraspanins are responsible for ferrying particular CoV receptors into TEMs. In support of this hypothesis, the MERS-CoV receptor dipeptidyl peptidase 4 (DPP4) and the 229E-CoV receptor APN localized to TEMs only in the presence of a particular tetraspanin, CD9 (35). Furthermore, cells lacking CD9 were resistant to MERS and 229E pseudovirus entry, making it clear that these viruses needed CD9, and probably TEM-localized receptors, for their entry. In contrast, deletion of a related CD81 tetraspanin did not confer resistance to virus entry. Other CoVs, however, apparently use distinct tetraspanins, because omission of CD9 did not relocalize SARS-CoV and MHV receptors, nor did it affect susceptibility to SARS and MHV pseudoviruses. With these findings, tetraspanins were further elucidated as partners for ferrying of specific CoV receptors into TEMs.
The idea that tetraspanins bring CoV receptors to TEM-associated proteases was further supported by the discovery that a well-known CoV-activating protease, type II transmembrane protease serine subtype 2 (TMPRSS2), was adjacent to the MERS-CoV receptor DPP4 as measured by proximity ligation assays (35). Yet in cells lacking CD9, this colocalization was not observed, and even though CD9 knockout cells retained cell surface DPP4 and TMPRSS2, they were resistant to virus entry. Only when excess TMPRSS2 was supplied through transfection was virus susceptibility restored. Therefore, these findings demonstrated that CD9 was required to coalesce DPP4 and TMPRSS2, with this juxtaposition of entry factors being necessary for robust MERS-CoV membrane fusion and infection (Fig. 1, left panel). It is conceivable that all CoVs utilize closely juxtaposed entry factors. This may explain, in part, why most known CoV receptors are transmembrane peptidases, even though their peptidase activities are not used to activate CoV fusion proteins (36, 37). The subcellular localization of transmembrane peptidases appears to be tightly controlled (38), and the CoV receptors we surveyed all localized to TEMs (32). It is possible that CoVs have selected peptidases so that virus entry is localized to protease-rich TEMs.
TETRASPANINS DICTATE CoV ENTRY ROUTES
The proteases that cleave CoV S proteins and activate virus cell entry are present at several subcellular locations. On CoV-susceptible cells, there may be serine-class TMPRs at or near plasma membranes (39, 40), and there may also be cysteine-class cathepsin proteases within endosomes (22, 39). Therefore, depending on the abundance of these proteases, a CoV can be activated for entry at different locations along the endocytic pathway, either early (TMPR mediated) or late (cathepsin mediated) after virus endocytosis (Fig. 1, right panel). Notably, in the presence of CD9, we found that MERS pseudovirus entry was early and was suppressed by serine protease inhibitors, while in the absence of CD9, entry was late and was unaffected by serine protease inhibitors but rather was blocked by cathepsin inhibitors and endosome-neutralizing agents (35). These findings fit well with the identification of CD9 as an agent of DPP4 and TMPR colocalization: by linking the receptors and TMPR proteases at or near the plasma membrane, the CD9 tetraspanin facilitated early CoV entry into target cells. Note that the early entry route is far more likely to result in productive infection, as demonstrated by Shirato and colleagues in separate studies of HCoV-229E, -OC43, and -HKU1 (41, 42). Late entry, at least for some CoVs, appears to be a lower-efficiency, last-chance infection route before virus destruction in lysosomes.
TETRASPANINS CONTRIBUTE TO CoV INFECTION AND PATHOGENESIS IN VIVO
The in vitro finding that CD9 facilitated the highly infectious early entry route suggested that CD9 and other tetraspanins might also be in vivo CoV susceptibility factors. We expected that this would be the case, as strong evidence from protease inhibitor studies indicated that early-acting TMPRs might be central for respiratory CoV infections (43). Indeed, we did find key roles for CD9 and TMPRSS2 in MERS-CoV infections. Using adenoviral vectors expressing short hairpin RNAs (shRNAs), we transiently depleted either CD9 or TMPRSS2 in mouse lungs and then determined their susceptibility to MERS-CoV infection. Depleting CD9 and TMPRSS2 reduced MERS-CoV lung titers by ∼90% and ∼80%, respectively (35). Clearly, the vast majority of MERS-CoVs enter lung epithelial cells through the early entry route, and late entry pathways cannot compensate for early entry blockade. Given that humans express 33 tetraspanins and 19 serine TMPRs, it is remarkable that MERS-CoV is so highly dependent on a single tetraspanin and a single protease to infect the lung. Further investigation is needed to determine whether other CoVs utilize particular tetraspanins and proteases to enter cells in vivo.
The importance of early entry for MERS-CoV pathogenesis was further highlighted by the mutations that occur upon adaptation of this virus to the mouse lung environment. MERS-CoVs can infect mice when the human DPP4 (hDPP4) receptor is present, but these infections usually do not cause overt respiratory disease. Serial passage of MERS-CoVs in hDPP4 mouse lungs, however, generated mouse-adapted (MA) MERS-CoVs that caused severe lung disease characteristic of acute respiratory distress in humans (44). Notably, MA viruses accumulated S protein mutations that facilitated rapid, CD9-dependent early entry (35). In contrast, the avirulent cell culture-adapted (CCA) MERS-CoVs took 3 times longer to enter cells, and their infection was not supported by CD9. It is noteworthy that Vero cells, in which CCA MERS-CoVs were adapted, have low TMPRSS2 expression levels (39), as do several other cell lines routinely used to investigate CoV entry mechanisms in vitro (27, 39). TMPR+ cell culture models may better reflect the in vivo environment of CoV entry, since CoVs utilize TMPR-mediated "early" entry routes in the lung (35, 43).
Accumulating evidence indicates that tetraspanins promote the entry of multiple viruses, including influenza A virus (IAV) (15, 32), human cytomegalovirus (HCMV) (45), human papillomavirus (HPV) (17, 18), hepatitis C virus (HCV) (14, 46, 47), Lujo virus (48), and several alphaviruses (49, 50). In some of these cases, the tetraspanins may be proviral because they coalesce the multiple host factors required to catalyze membrane fusion and infection. For example, IAV hemagglutinin (HA) proteins require proteolytic cleavage to undergo fusion-catalyzing structural transitions, and these proteolytic cleavages occur in TEMs (32). Depleting the tetraspanin CD81 from target cells restricts ∼90% of IAV entry (15), suggesting that this tetraspanin may aid in localizing IAV-activating proteases with sialate receptors. Tetraspanins can also condense virus-associated membrane proteins into complexes prior to endocytosis. The tetraspanin CD151 provides an example of this, as it drives HPV endocytosis by coalescing receptor-bound viruses and integrin coreceptors targeted for endocytosis. HCV and the tetraspanin CD81 set another illustration. CD81 acts as the primary HCV receptor, binding directly to the HCV E2 glycoprotein (14), yet CD81 also coalesces the HCV coreceptors claudin-1, syndecan-1, and scavenger receptor B1 into a complex required for subsequent HCV endocytosis (46, 47, 51, 52). We speculate that many viruses have evolved to utilize entry factors that reside in close juxtaposition and that tetraspanins act to aggregate these entry factors into TEMs.
Cells have evolved to target tetraspanins to restrict viral infection. Among the antiviral type I interferon-stimulated gene products are the interferon-induced transmembrane proteins (IFITMs), which are small alpha-helical proteins composed of ∼130 residues that inhibit several virus entry processes at the level of virus-cell membrane fusion (53, 54). Intriguingly, IFITMs intercalate into TEMs, disrupting tetraspanin interactions with their partner proteins (55). One can speculate that as these intruding IFITMs accumulate during innate immune responses, they might separate or disorganize virus entry factors on target cell membranes. Notably, it is known that IFITM1 alters HCV coreceptor interactions, in correlation with restricted HCV-cell membrane fusion (56). It is possible that TEM disruption is a common mechanism by which the IFITMs restrict viral entry and infection.
In addition to promoting virus entry into target cells, tetraspanins also facilitate viral assembly and egress (Fig. 2). Many enveloped viruses, including IAV (15), human immunodeficiency virus (HIV) (16, 57), and herpes simplex virus 1 (HSV-1) (58), bud out of TEMs. Viruses may utilize these TEMs to facilitate vesicle budding and release. One possible mechanism by which this takes place is through tetraspanin-mediated membrane remodeling. The four tetraspanin transmembrane domains form two pairs of antiparallel helices, which adopt a cone-shaped structure opening toward the outer leaflet of the lipid bilayer (3). This architecture may facilitate membrane curvatures that may promote enveloped virus vesicle formation and membrane fission (2). Investigations into IAV egress support this speculative role for tetraspanins in virus budding. IAV HA localizes to TEMs containing CD81, and this tetraspanin is required for successful virion release (15). In the absence of CD81, virions bud from the plasma membrane but do not undergo fission to be released from the cell (15). They instead become elongated, with no fission at a membrane neck. These findings suggest that tetraspanins facilitate the membrane curvatures required for membrane fission and virus release.
Another way in which tetraspanins facilitate viral egress is by aggregating viral structural proteins. For example, the HSV-1 capsid protein VP26 interacts with the tetraspanin TSPAN7, leading to its accumulation on the nuclear membrane (58). This interaction is required for viral budding and egress from the nucleus. In the absence of TSPAN7 or upon mutation of VP26, HSV-1 capsids are sequestered in the nucleus, unable to exit (58). In addition, filamentous influenza virus strains accumulate CD81 at the ends of budding virions (15), along with the viral M2 protein, which mediates scission of IAV membranes from host cells (59). It is conceivable that CD81 facilitates M2 accumulation at the site of virus-cell membrane scission (15). These investigations highlight the ability of tetraspanins to facilitate multiple stages of the viral life cycle.
During viral biogenesis, tetraspanins incorporate into nascent virions (15, 57, 60, 61), suggesting that they may function in the context of the extracellular virion, perhaps at the virion cell entry stage. Intriguingly, virus-incorporated tetraspanins suppress rather than promote membrane fusion and virus entry (62, 63). In the context of HIV, overexpressing the tetraspanins CD9, CD63, and CD81 in virus producer cells inhibits HIV envelope-mediated membrane fusion (62–65), while depleting tetraspanins by use of small interfering RNAs (siRNAs) promotes membrane fusion (63, 65). We speculate that the high abundance of tetraspanins in HIV membranes may lead to these inhibitory effects. HIV Gag actively recruits tetraspanins to viral assembly sites, leading to the aggregation of large TEMs in HIV membranes (16). Notably, overexpressed tetraspanins inhibit HIV membrane fusion only in the presence of Gag (63). It is possible that Gag concentrates tetraspanins and cholesterols to rigidify HIV membranes, rendering them incompatible with membrane merger. Antibody-mediated tetraspanin cross-linking aggregates TEMs and suppresses CoV entry (32), supporting this notion. Further investigation is needed to determine whether the relative abundance of tetraspanins may dictate their pro- or antifusion activities.
Tetraspanins and assembled TEMs contribute to a wide variety of biological processes, including those involved in cell metabolism, signal transduction, and intercellular adhesion. One of their additional critical roles is in controlling nonviral cellular membrane fusions. Cell-cell fusions generate many biologically important structures, including fertilized zygotes (66), skeletal muscles (67), placentae (68), and multinucleated macrophages (69). Notably, tetraspanins are known regulators of many cell-cell fusion reactions (70–73). Their role is most striking during fertilization, which absolutely requires the tetraspanin CD9 (70). CD9 knockout mice are infertile, and eggs lacking CD9 are unable to fuse with sperm (70). CD9-laden extracellular vesicles, however, can compensate for loss of CD9 by bridging sperm and egg cells and subsequently fusing them together, in a process known as "fusion from without" (71). CD9 overexpression stimulates monocyte cell-cell fusions (72), which lead to the formation of multinucleated osteoclasts that resorb bone. Similarly, antibodies targeting CD9 suppress the fusion of myoblasts into the large syncytia required to form skeletal muscle (73). While the mechanisms behind tetraspanin-facilitated cell-cell fusions are not fully elucidated, we speculate that tetraspanins generally regulate membrane fusion events through their ability to bend membranes and cluster the factors required for membrane fusion catalysis. Surprisingly, many eukaryotic cell-cell fusion catalysts remain unidentified. Proteomic analysis of TEMs from fusogenic cells, along with molecular genetic approaches, may identify these elusive endogenous cellular fusogens.
- Received 21 November 2018.
- Accepted 11 December 2018.
- Accepted manuscript posted online 19 December 2018.
- Copyright © 2019 American Society for Microbiology.


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When running a business, you need to be wary and make certain every small interest of business is blanketed. When you manufacture a product or float a design or think of a suggestion for the betterment with the you are promoting, it is very important for you to copyright it. Without a correct copyright in the situation, any person can make use of recommendations or replicate your designs. It will power the gains away also as in addition, take the exclusivity out of your group. So that you could avoid this kind of factor from taking place, make certain you get yourself a copyright registration accomplished. Several advantages of copyright registration, read on to grasp extra.
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How Does a Copyright Help You may?
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