This is the best such database I have found. Windows 10 Enterprise 2016 LTSB N (圆4) - DVD (English-United Kingdom) ![]() title).įor posterity, here's a screenshot of what the interface looks like.Īnd here you will find the metadata for the file you used in your example, along with a short description of what it is. At the time of this writing, the metadata for 67253 files is browseable and searchable (by SHA1 or SHA256 hash values or by text, i.e. ![]() It is much better than the yooneed.one site you linked to in that it not only provides you with hash values for a handful of Windows 10 images, but the hash values for the entire MSDN catalog. However, if you are willing to trust a third party provider, the same information is readily available at. This way you don't have to worry if Microsoft or any other party you download your files from kills the service that provided this bit of inforamtion, or if the company itself goes out of business.Īs it has been noted, Microsoft has decided to remove this information from public view when they revised their MSDN program. That way you can always remind yourself what the correct value is, then compute the corresponding hash value (MD5, SHA1 or SHA256) of the file anew and compare it to the original hash value that you have on record. Ideally, you would make a note of the hash value that you store alongside the ISO file from where you downloaded it (presumably Microsoft). Get-FileHash Win10_2004_English_圆4.iso | Format-List Navigate to the folder with the iso image.Right-click Start and select Windows PowerShell.Select Verify Your Download to view and copy the hash for your selected iso.Select the version and language of your choice.After changing your user agent, navigate to the Microsoft ISO download site.Restart Firefox by closing all windows for the changes to take effect.Instructions for other browsers can be found at HowToGeek. ![]() Use the user agent of a non-Windows browser such as WebKit/Blink as the setting value.Select String and save with the + button.Enter into the search preference name box.Type about:config in the address bar and press Enter.Without this step, Microsoft will force you to download the Media Creation Tool.Ĭhange User Agent Using Mozilla Firefox Browser: To download Windows 10 ISOs using Windows 10, you will need to change your browser's user agent. Download ISOs and Hashes directly from Microsoft
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![]() ° T-Mobile iPhone 15 Special Deal: Monthly price reflects net monthly payment, after application of T-Mobile trade-in credit applied over 24 months with purchase of an iPhone 15 Pro, iPhone 15 Pro Max, iPhone 15, or iPhone 15 Plus and trade-in of eligible smartphone. Purchase, financing, other limits, and restrictions apply. May not be combinable with other offers, discounts, or credits. To get all credits, device must remain on agreement for entire term and you must keep eligible service on device for entire installment term. Installment agreement starts when device is shipped. Wireless line must be on an installment agreement, active, and in good standing for 30 days to qualify. ![]() Will receive catch‑up credits once credits start. Bill credits are applied as a monthly credit over the 36‑month installment plan. If you cancel eligible wireless service, credits will stop and you will owe the remaining device balance. $0 down for well‑qualified customers only, or down payment may be required and depends on a variety of factors. Requires upgrade of an existing line (or activation of a new line) and purchase on qualifying 36‑month 0% APR installment plan, subject to carrier credit qualification. Buy an iPhone SE 256 GB and get $184.36 in bill credits applied over 36 months. Buy an iPhone SE 128 GB and get $264.36 in bill credits applied over 36 months. Activation required.ĪT&T iPhone SE 3 Special Deal: Buy an iPhone SE 64 GB and get $358.36 in bill credits applied over 36 months. Will receive catch-up credits once credits start. Bill credits are applied as a monthly credit over the 36-month installment plan. $0 down for well-qualified customers only, or down payment may be required and depends on a variety of factors. Requires upgrade of an existing line (or activation of a new line) and purchase on qualifying 36-month 0% APR installment plan, subject to carrier credit qualification. Buy an iPhone 14 512 GB and get $274.68 in bill credits applied over 36 months. Buy an iPhone 14 256 GB and get $254.52 in bill credits applied over 36 months. Activation required.ĪT&T iPhone 14 Special Deal: Buy an iPhone 14 128 GB and get $334.44 in bill credits applied over 36 months. Price for iPhone 15 and iPhone 15 Plus includes $30 AT&T connectivity discount. Limits: one trade-in per qualifying purchase and one credit per line. ![]() Will receive catchup credits once credits start. Bill credits are applied as a monthly credit over the 36 month installment plan. Trade in device may not be on existing installment plan. ![]() Requires activation on eligible unlimited plan. $0 down for well qualified customers only, or down payment may be required and depends on a variety of factors. AT&T Installment Plan with Next Up is not eligible for this promotion. Requires upgrade of an existing line or activation of a new line and purchase of a new iPhone 15 Pro, iPhone 15 Pro Max, iPhone 15, or iPhone 15 Plus on qualifying 36 month 0% APR installment plan, subject to carrier credit qualification. Max bill credits will not exceed the cost of the device. Receive credit with purchase of an iPhone 15 or iPhone 15 Plus of either $800 or $350 (based upon the model and condition of your trade-in smartphone). Receive credit with purchase of an iPhone 15 Pro or iPhone 15 Pro Max of either $1000, $830, or $350 (based upon the model and condition of your trade-in smartphone). Δ AT&T iPhone 15 Special Deal: Monthly price reflects net monthly payment, after application of AT&T trade-in credit applied over 36 months with purchase of an iPhone 15 Pro, iPhone 15 Pro Max, iPhone 15, or iPhone 15 Plus and trade-in of eligible smartphone. Each whiff carries hints of berries and grapes up front, followed by notes of soil and wood.įlavor: Although full of grassy flavors and heavy floral notes if not given the proper time to grow and cure, Purple Urkle can taste like a skunky glass of Welch’s grape juice when done correctly. Smell: At this point in breeding, plenty of strains take on the syrupy grape smell that made Purple Urkle so popular, but there’s something to be said for trying one of the originals. The strain’s buds are largely round or cone-shaped, while their structure is very dense. ![]() Looks: A true purple heavyweight, with dark, velvety calyxes and spots of forest green contrasting against rust-orange pistils and speckles of trichomes. I’ve seen it at Alternative Medicine on Capitol Hill, Altitude the Dispensary, Denver Kush Club, Doc’s Apothecary, Euflora, Green Tree Medicinals, Kind Meds, Lyon’s Finest, Medicine Man, Native Roots, New Amsterdam Organics, Oasis Cannabis Superstore, Silver Stem Fine Cannabis and Urban Dispensary. Purple Urkle has been a fixture of the Denver dispensary scene since the early medical days, and - part of the black market long before that. Those predictable relaxing effects make the strain a great candidate for a litany of medical issues, though, and a reliable way to help you fall asleep when closing your eyes just isn’t enough. The body’s unraveling isn’t far behind, either, as you’re pulled into a pit of munchies and lethargy before ultimately going off to dreamland.Īnyone smoking Purple Urkle before the sun goes down should expect a nap in the near future, or a very early bedtime, at the least. One session with Purple Urkle will quickly showcase the strain’s powerful sedating qualities, instantly cloaking the brain in a lazy fog. The only argument against that theory is that anyone who’s high on Purple Urkle is far from nerdy, because being nerdy requires a modicum of thought. The prevailing theory is that Purple Urkle was named for the strain’s potent high, which often leads to bumping into walls, irritating behavior and falling and not being able to get up - all hallmarks of everyone’s favorite nerdy annoyance in the ’90s, Steve Urkel. The history behind the fruity, tranquilizing indica’s name is cloudy. In fact, I’m pretty sure I’m in the majority. ![]() I can’t be the only person who instantly thinks of Family Matters the minute Purple Urkle makes an appearance on a dispensary shelf. If you’re not sure what’s causing your Battery Tender to flash yellow, check the troubleshooting section of the manual. The temperature may be too cold for the charger to operate properly. There may be something wrong with the battery or charger, such as a loose connection. The battery may be nearly fully charged and the charger is reducing its output to avoid overcharging the battery. This could be due to a number of factors, including: If your Battery Tender is flashing yellow, it means that the charger is outputting a lower voltage than normal. With this knowledge in hand, you should be able to get your battery tender to stop flashing yellow and green. Possible explanations for the flashing include a malfunctioning circuit board, low battery voltage, or a shorted output wire.įortunately, you can fix this issue by checking the voltage of your battery, replacing the circuit board or output wire if necessary, and ensuring that everything is properly connected. In conclusion, if you have a battery tender that is flashing yellow and green, it could be due to several causes. If the output voltage is below 13V, then the charger needs to be replaced as it cannot charge properly anymore. In order to do this, connect a multimeter between the terminals of the charger and check the voltage output. If this does not work, then it could be that the Battery Tender needs to be tested to see if it is faulty or not. Once this is done, the light should turn off and begin charging again. To fix this issue, it is necessary to disconnect the charger from the battery for a few minutes, then reconnect it and press the “reset” button. But still, the battery is not working then it could be caused by a faulty battery Tender. ![]() If the Battery Tender is flashing yellow and green, this indicates that the battery has been fully charged to its maximum capacity. To fix this issue, you will need to replace your old batteries with new ones. This happens when there is an internal fault in the cells of your lead acid or AGM type batteries, which prevents them from charging properly. The last cause for a flashing yellow and green LED on a battery tender is due to a faulty battery. This should resolve any issues with Battery Tender flashing yellow and green. In addition, if there are any loose connections or corroded cables on either end of the charger or battery posts then these need to be cleaned and re-tightened or replaced altogether as necessary. To fix this issue, be sure to check that all connections are correct before connecting them to each other. ![]() This occurs when a reverse polarity connection is made between the positive terminal of the charger and the negative terminal of the battery. Incorrect connection is another common cause of the flashing yellow and green LED on a battery tender. To fix this issue, you will need to recharge the battery using an external charger or an alternator from any other battery charger before reconnecting it to the battery tender. This usually happens when the battery has been discharged to a level below what the battery tender can charge. Low voltage is the most frequent cause of battery tender flashing yellow and green. The most common causes are low voltage, incorrect connection, and faulty battery. Problem 1: Low Voltage, Incorrect Connection, And Faulty Battery The battery tender flashing yellow and green is an indication of a few problems with the battery tender. Battery Tender / USB Charger AdaptorĬauses & Fixes Of The Battery Tender To Flash Yellow And Green ![]() In this article, we will discuss what causes the battery tender to flash yellow and green and how to fix it. However, if both the green and yellow lights are on, this may indicate an underlying problem that needs to be addressed before racing can resume. ![]() Patients who are clearly overusing multiple drugs for acute or symptomatic treatment of headache but cannot give an adequate account of their names and/or quantities are coded 8.2.7 Medication-overuse headache attributed to unspecified or unverified overuse of multiple drug classes until better information is available.Patients who use multiple drugs for acute or symptomatic treatment of headache may do so in a manner that constitutes overuse even though no individual drug or class of drug is overused such patients should be coded 8.2.6 Medication-overuse headache attributed to multiple drug classes not individually overused.The exception occurs when patients overuse combination-analgesic medications, who are coded 8.2.5 Combination-analgesic-overuse headache and not according to each constituent of the combination-analgesic medication. For example, a patient who fulfils the criteria for 8.2.2 Triptan-overuse headache and the criteria for one of the subforms of 8.2.3 Non-opioid analgesic-overuse headache should receive both these codes. Patients should be coded for one or more subtypes of 8.2 Medication-overuse headache according to the specific medication(s) overused and the criteria for each below.Not better accounted for by another ICHD-3 diagnosis.Regular overuse for >3 months of one or more drugs that can be taken for acute and/or symptomatic treatment of headache 1 2 3.Headache occurring on ≥15 days/month in a patient with a pre-existing headache disorder.It usually, but not invariably, resolves after the overuse is stopped. Headache occurring on 15 or more days/month in a patient with a pre-existing primary headache and developing as a consequence of regular overuse of acute or symptomatic headache medication (on 10 or more or 15 or more days/month, depending on the medication) for more than 3 months. Patients who meet criteria for both 1.3 Chronic migraine and 8.2 Medication-overuse headache should be given both diagnoses. Patients with a pre-existing primary headache who, in association with medication overuse, develop a new type of headache or a significant worsening of their pre-existing headache that, in either case, meets the criteria for 8.2 Medication-overuse headache (or one of its subtypes) should be given both this diagnosis and the diagnosis of the pre-existing headache. 8.3.4 Headache attributed to withdrawal from chronic use of other substanceĨ.2 Medication-overuse headache (MOH) Previously used terms:ĭrug-induced headache medication-misuse headache rebound headache.8.3 Headache attributed to substance withdrawal. ![]() 8.2.8 Medication-overuse headache attributed to other medication.8.2.7 Medication-overuse headache attributed to unspecified or unverified overuse of multiple drug classes.8.2.6 Medication-overuse headache attributed to multiple drug classes not individually overused.8.2.5 Combination-analgesic-overuse 1 headache.8.2.3.3 Other non-opioid analgesic-overuse headache.8.2.3.2.1 Acetylsalicylic acid-overuse headache.8.2.3.2 Non-steroidal anti-inflammatory drug (NSAID)-overuse headache.8.2.3.1 Paracetamol (acetaminophen)-overuse headache.8.2.3 Non-opioid analgesic-overuse headache.8.1.11 Headache attributed to use of or exposure to other substance.8.1.10 Headache attributed to long-term use of non-headache medication.8.1.9 Headache attributed to occasional use of non-headache medication.8.1.8 Headache attributed to exogenous acute pressor agent.8.1.7.1 Immediate CGRP-induced headache.8.1.7 Calcitonin gene-related peptide (CGRP)-induced headache.8.1.6.2 Delayed histamine-induced headache.8.1.6.1 Immediate histamine-induced headache.8.1.4.2 Delayed alcohol-induced headache.8.1.4.1 Immediate alcohol-induced headache.8.1.3 Carbon monoxide (CO)-induced headache.8.1.2 Phosphodiesterase (PDE) inhibitor-induced headache.8.1.1.2 Delayed NO donor-induced headache.8.1.1.1 Immediate NO donor-induced headache.8.1.1 Nitric oxide (NO) donor-induced headache.8.1 Headache attributed to use of or exposure to a substance.Headache attributed to a substance or its withdrawal ![]() Unfortunately, archiving a backup doesn't make it impervious to time. That way, you can restore your iPhone to the way it was at any time. In the future, whenever your iPhone that's running iOS 14.5 backs up, Finder or iTunes will save it as an independent file, leaving your archive backup untouched. Instructions: How to Create an Archived Backup of Your Current iOS Version.Archiving prevents your computer from overwriting the backup in the future, which just adds a little security to your valuable data. However, we suggest backing up and archiving it, which requires you to go through either Finder if running macOS Catalina and later or iTunes for older systems. But that's a lot of waiting.īefore installing the beta profile for iOS 14, you can back up your iPhone with iCloud. You can delete the beta profile from your iPhone, restart the device, then wait until Apple releases the next stable version of iOS 14.x. You don't have to revert to iOS 14.4 to escape the beta. Even dependable iCloud won't be your friend here since a backup there will only work with the iOS 14.5 or iOS 14.5 beta. If you jump right into iOS 14.5 and something goes wrong, downgrading back to iOS 14.4 (while it's still being signed) will result in you losing all of your data if you don't have a proper backup. There's nothing more exciting as an iPhone user than beta testing cool new features before anyone else, but you can't rush into things. These instructions assume you're updating an iPhone to iOS 14.5 beta, but consider each step the same if updating an iPod touch to iOS 14.5 beta or an iPad to iPadOS 14.5 beta as well. For context, here's the full list of iPhones that supports iOS 14 and all future iOS 14.x versions: ![]() While iOS 13 and iPadOS 13 ditched a few devices, namely the iPhone 5 S, 6, 6 Plus, iPad Air, and mini 3, iOS 14 and iPadOS 14 support all of the same devices that iOS 13 does. Install Dev Beta Restore Image on Mojave.Install Dev Beta Restore Image on Catalina & Later.Install Dev Beta Profile for OTA Updates. ![]()
![]() There's also a feature to hear notifications and Caller ID announcements as well as enable/disable auto pause. The app also lets you adjust in-call volume, including how loud your voice will sound. It's a nice feature, but we were disappointed to learn we couldn't disable the feature when using the earbuds in Active mode. In case you'll be using the buds outdoors, there's the HearThrough feature for ambient noise. It's nice, but we wish Jabra had added more info, like heart rate and calories burned. The buds have an integrated accelerometer, which turns them into a reliable fitness tracker that monitors steps per minute and the time elapsed. Jabra didn't skimp on the features in the Elite Active. You can choose between volume, quick ambient sound or voice assistant. Lastly, the app gives you the ability to configure what launches during a long press. You can also toggle how much outside noise can get in via Ambient Sound. Offering five presets (Bass boost, Soft, Dynamic, Clear and Treble boost), the Galaxy Buds' equalizer can be tuned to your specific listening tastes. High- and mid-levels are still pleasant, creating a profile which is warm and immersive.You access even more functions in the Galaxy Wearable app, including a Find My Earbuds feature and an equalizer. As sports in-ears they're clearly not balanced for audiophiles, but we like that the Elite's bass doesn't muddy the rest of the frequencies. There's lots of the low-end, which will definitely please those who want some bang to their beats. Like the first version, there's a generous amount of bass in the Elite Sports sound profile. Four external mics for noise cancelling/Heathrough.Jabra also comfortably beats the Bragi Dash Pro in this department by more than double. While the first-generation Elite Sport wasn't terrible in terms of longevity, the newer model is an improvement. In our testing we listened to an hour of music in one sitting, then placed them back in the case, and did this five times in addition to a 40 minute session while running and listening to music, and still the case was able top the earphones' battery back up to 100 per cent. The long and short of it is that the users should comfortably be able to make these last a full work-week's worth of commutes. You can track running, running on treadmill, cycling, walking, hiking, spinning, skating and walking on a treadmill, as well as a selection of circuit/weight-training programmes. When it comes to training and activity tracking, the Jabra app has a selection of all the basics. There's also the Cooper test to see how far you can run in 12 minutes, and the Orthostatic Heart Rate test to see whether or not you're overtraining. ![]() There's also the VO2max test to determine how fit you are by running at 70-100 per cent of your maximum heart rate for 15 minutes. You can take a resting heart-rate test and personalise your heart-rate training zones. You don't have to use the Jabra Sport app, although it does include some really useful features.īefore beginning any activity tracking you can do one of four tests so that the app has a good benchmark on which to base its calculations. If you already use a different app - like Strava - for running, you can still use your phone to track your run and just use the earphones as the heart-rate monitor if you'd like. While the Bragi Dash Pro was improved in this regard over its first generation attempt, the Jabra Elite Sport is still the more consistent product. It hasn't mattered if the earphones were tracking a run with an iPhone in a pouch around our waist, or if we were simply listening to music at our desk in the office, they've stayed consistently connected the entire time. The Bluetooth connectivity performance has been superb in all of our testing.
Instead of describing yourself as a " Carthage award-winning graphic artist ," you might write that you are a " multi-talented graphic artist who won the 2019 Carthage Award for artistic excellence and innovation." Even if your reader is not familiar with your specific industry's awards, they will likely respond well to positive keywords like "innovation." However, you will need to describe them in a way that any reader can understand. Writing a short bio is similar to writing a resume in that you will want to highlight your professional accomplishments. Read more: Core Values: 84 Powerful Examples for the Workplace 4. Your philosophy should give your reader an accurate idea of what matters the most to you. Your professional philosophy could relate to expanding your expertise, contributing to society or learning new skills. This should answer the question, "Why do you do what you do?" Your personal philosophy may involve serving others, providing for your family or making the most of your opportunities. State your philosophyĪfter introducing yourself, you may want to relate your personal or professional mission statement. Related: How To Write a Professional Bio (With Examples) 3. ![]() This might look something like " My name is John Grayson and I am a recent college graduate with a Bachelor's Degree in Web Design and Development." For example, your bio might begin with " Sarah Hayes is the Senior Marketing Director for Blooming Street Creative." If you are in the process of finding a job, you can list your most recent title or your college major. In the first sentence of your short bio, you will need to give your name and your current job title. Related: How To Write a Personal Bio That Draws Attention 2. Your voice should accurately represent who you are as a person, so you might also adjust your voice to be more professional, witty, friendly or reserved, depending on your personality. However, if you are adding a bio to your company page and all your coworkers used third-person in their paragraphs, you should certainly follow suit. Writing in the first person means using the words "I" and "me", and writing in the third person means using your name.įor instance, if you are writing a slightly informal bio for your Twitter profile, first-person would probably be the best fit. For our purposes, choosing a voice involves deciding whether you are writing in the first or third person. The first step in writing a short bio is deciding on a voice. Here are some steps you can follow to help you write a successful short bio: 1. Writing a short bio can be a deceptively challenging task. Read more: 5 Steps for Great Business Writing How to write a short bio ![]() Short bios are brief by definition so it is important to determine which of these topics is most important to make the best impression on your readers. Your company name or personal brand statement Your short bio is often the first (and perhaps only) thing a potential employer, client or contact will read before deciding to contact you, so it is important that your bio be accurate, informative and memorable. ![]() Short bios are typically used to provide a summary of an individual's accomplishments, an overview of their career history and a description of their professional goals. You can often find short bios on social media profiles, personal websites and company team directories. Short bios are concise, biographical paragraphs that professionals use to introduce themselves. ![]() HP reserves the right to change or cancel this offer at any time. Requires acceptance of Terms and conditions of Sales and Service. Offer will end 11.59pm Thursday 30th November 2023 or while stocks last. Maximum spend of £4500 including VAT per customer during the event. ![]() Purchases are for personal use only and not for resale. Code promotions cannot be used in conjunction with any other code and are single use per customer. For promo code discounts, enter respective code to redeem the advertised product discount. *BLACK FRIDAY EVENT - Terms and Conditions: HP Ink, Toner & Paper are excluded from this promotion.Offer will end 5pm Thursday 23rd November 2023 or while stocks last. Promo code promotions cannot be used in conjunction with any other code and are single use per customer. *BLACK FRIDAY EVENT - Terms and Conditions: For promo code discounts, enter respective code to redeem the advertised product discount.It’s a very handy feature that will certainly appeal to a number of people. This meant that after setting up Windows 10 for the first time, we were able to access the internet immediately without having to connect to a Wi-Fi network and enter in a password. Windows 10 does a great job of integrating LTE connectivity, and our review sample came with a Vodafone SIM card already installed. HP also claims this is the world’s first gigabit class LTE Intel-based laptop, offering speeds comparable (and even above) home broadband connections. While a growing number of laptops come with LTE connectivity, it’s still relatively rare, and certainly helps the HP Folio Spectre stand out among its competitors. Not only does this mean you can access the internet from almost anywhere, as long as there is network coverage, but it also means you don’t have to rely on public Wi-Fi hotspots, which can often come with security implications. This allows you to insert a SIM card into the laptop and connect to the internet via a cellular connection. Windows 10’s robust support for stylus input must be commended here as well, as it’s put to good use by the Folio Spectre.Īnother feature we’re pleased to see in the HP Folio Spectre is a SIM port for mobile LTE data. It’s got a nice heft to it when in use, and the screen reacts quickly and smoothly to drawing and writing with it. Unlike some premium devices (we’re looking at you, Surface Pro), the Spectre Folio includes a stylus, which makes the price a little more palatable. We’ve been spoiled by thin bezels in premium laptops like the Dell XPS 13, so they feel a little cheap and old fashioned here. The screen is impressively thin, though the bezels, especially at the top, which houses a webcam, and at the bottom, which displays the HP logo, are quite thick. The WLED backlit screen is impressively vibrant, and the IPS panel offers wide viewing angles, which is useful when adjusting the screen to various modes. The brown leather feels pleasant to the touch and helps keep the device in one place on otherwise slippery surfaces, like glass, marble or granite. It certainly feels like a premium device. The hinge is also strong enough for the screen to be angled when in laptop mode without the whole device tipping over. It’s also here where you see how HP has fully integrated the leather into the design. Banish any thoughts of flimsy 2-in-1 laptops with material covers, as the Folio really feels like a robust and well made bit of kit. When adjusting the Spectre Folio into these different modes, the laptop feels sturdy and solid. It’s a pricey proposition, then, with fierce competition, which means HP needs to pull out all the stops to make the Spectre Folio worth the considerable sums of money it is asking for. It’s also comparable in price with the Surface Pro if you buy one with the keyboard cover included. These prices pit the HP Spectre Folio against flagship laptops like the 12-inch MacBook and the Asus ZenBook 3. It’s a bit of a shame that this fully specced-out version isn’t available to buy worldwide, at least for the moment. This costs $1,499 (£1,499).Īustralia doesn’t get that model, but it does get a unique Spectre Folio with the Core i7-8500Y processor, along with 16GB of LPDDR3 RAM and a 512GB M.2 SSD for AU$3,399. There is also a version with a Core i7-8500Y processor with an LTE modem attached for wireless connectivity anywhere, plus the same amount of RAM and storage as the version above. While this version is available in the US and Australia, it's not available in the UK. The screen is a 13.4-inch 1080p (1,920 x 1,080) WLED touchscreen that’s protected by Corning Gorilla Glass. ![]() Proceed to the next step to complete the assembly. A pop-up window will be displayed where one can visually inspect the structure (Fig. Click the "view structure" link next to the Component PDB file link. ![]() If this step is not working the instructor will provide pre-calculated files.ĬHARMM-GUI will display the list of prepared files (Fig. Proceed to the next step to combine membrane components. This could be a problem that will not be resolved by minimization and the system has to be regenerated. Proceed to the next step.ĬHARMM-GUI reports on possible lipid penetrations of rings (not shown). Select CaCl 2 as the salt of choice (Fig. As for the ions, we will use calcium ions for system neutralization and additional concentration. We will use the default Replacement method. The system size summary will be displayed (not shown) as well as two choices for protein insertion into the membrane (Fig. Click again the "Show the system info" button. Observe that summary now has list of lipids (Fig. Remark: for E.coli outer membrane composition composition see e.g., Lugtenberg EJJ & Peters R, BBA, 441, 38–47, 1976. Click the "Show the system info" button to update. Hover over the links "Image" in the row of the chosen lipids and lipid structure will be shown. As a reminder, PE is a zwitterionic (formally neutral) while PG is charged lipid. Enter "90" in the upper and lower leaflet boxes for POPE, and "10" in the upper and lower leaflet boxes for POPG. 6): Build a membrane consisting of phosphatidylethanolamine (POPE) 90% and phosphatidylglycerol (POPG) 10%, which will be our model for the outer membrane of E. Select lipids under the clickable triangles (Fig. We can keep the default value of the water thickness at 17.5 Å. The results of the estimate will be shown in the table (Fig. 5, left) and click the "Show the system info" button. Enter 50 Å as the initial estimate for the size of the cell (Fig. Cross sectional density will be displayed (not shown) as well as system size options (Fig. Proceed to the next step.ġ.2 CHARMM-GUI: select lipids and water thickness. 4) – this assumes the Z-axis is aligned with the principal axis of inertia of the protein. Keep the PDB orientation as a choice (Fig. Open the CHARMM-GUI website () and go to Input Generator –> Membrane Builder –> Bilayer Builder.ġ.1 CHARMM-GUI: load, analyze, and position PDBĬHARMM-GUI provides choice for protein orientation in the membrane. Small modifications might be needed when used on other computers. This tutorial can be completed on iMacs in the SCS VizLab (151 Noyes Lab) or with personal computers with Windows or Mac OS. MobaXTerm for a Windows machine), and CHARMM-GUI web server. Personal computer: Terminal (Mac OSX) or xserver (e.g. Step 4: when job is completed, analyze the results. Step 3: submit the system to the local computer cluster Step 2: download the system to local desktop computer ![]() Step 1: prepare membrane-OmpLA structure using CHARMM-GUI web-server For info on the cluster see SCS Clusters webpage. Outline: we will use the SCS computer cluster to model the systems. CHARMM-GUI is developed by the Im Lab at the Lehigh University ( ).Įstimated time to complete this tutorial is 1.5 hr. Note: NAMD and VMD are developed at the NIH Center for Macromolecular Modeling hosted at the University of Illinois at Urbana-Champaign ( ). In a recent experimental study, the Fleming Lab (Johns Hopkins) used spontaneous folding of OmpLA into the membrane to design a hydrophobicity scale (Moon PC and Fleming KG, PNAS 108, 10174, 2011). The system of interest will be the outer membrane phospholipase A from E. This tutorial is designed to introduce the preparation, modeling, and analysis of membrane-protein system with molecular dynamics (MD) using NAMD, VMD, and CHARMM-GUI. School of Chemical Sciences, University of Illinois Urbana-ChampaignĪugust 17, 2016, Edition 1.1. Basics of membrane-protein interactions using molecular dynamics with NAMD, VMD, and CHARMM-GUI |
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