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Grand Theft Wiki:Staff
This page documents official Policy on Grand Theft Wiki. It concerns standards that all users should follow. When editing this page, please ensure that your revision reflects consensus. When in doubt, discuss first on the talk page. |
Staff are users on Grand Theft Wiki with the responsibility to keep the wiki running smoothly. The main job of all Staff is to help users, so don't hesitate to ask a member of Staff if you have any problems or questions.
Certain Staff also have various rights, administrative tools and authority over the wiki. There are three levels of Staff on Grand Theft Wiki.
No users other than the ones mentioned here have any administrative or decision-making authority. Trusted Users are not staff members, but simply regular users trusted with some restricted tools.
List of Staff
Position | Logo | People | Description | Rights |
Executive Manager |
Executive Managers are the owners of Grand Theft Wiki, and have the highest level of administrative control. Some rights are reserved for Executives only. |
| ||
Manager | Managers (Bureaucrats) are the leaders of the staff team. They supervise the other admins, and make sure everything possible is being done to keep this wiki running smoothly |
| ||
Administrator | Administrators (Admins or SysOps) are members of Staff with tools and authority to solve disputes and keep the wiki running smoothly. The main part of their job is to help other users; but they enforce the policy of Grand Theft Wiki, and occasionally need to step in and use their additional tools to solve problems. |
Full details of rights for each user group can be seen on Special:ListGroupRights
Other user groups
Several other user groups and rights exist on the wiki, but are not staff positions.
These include: Trusted users, Autoconfirmed, Bots, Checkuser, Oversight, Patroller
Inactive Staff
These users are previous or inactive staff.
- Thai420 (Bureaucrat)
- Thorpe (Sysop)
- Chief of Staff (Sysop)
- GTW-Chris (Sysop)
- GTW-George (Patroller)
- GTW-47 (Patroller)
- Blakegripling ph (Patroller)
- Eganio (Administrator)
- MattyDienhoff (Moderator)
- BloodyGTA (Moderator)
- GTA4PC (Moderator)
- Hardrock182 (Administrator)
- Paperworktemple (formerly Masterpogihaha) (Administrator)
- GuildKnight (Administrator/Manager)
- Gta-mysteries (Administrator)
- Chimpso (Administrator)
- MrLanceVanceDance (Inspector)
- Silver Infernus (Administrator)
- User:JFletcher (Administrator/Manager)
Promotion to Staff
See Grand Theft Wiki:Promotion to see the process for nominating staff for promotion, and to provide nominations and opinions.


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Welcome to Tacoma World!
You are currently viewing as a guest! To get full-access, you need to register for a FREE account.
As a registered member, you'll be able to:- Participate in all Tacoma discussion topics
- Communicate privately with other Tacoma owners from around the world
- Post your own photos in our Members Gallery
- Access all special features of the site
Hi lift jack?
Discussion in '2nd Gen. Tacomas (2005-2015)' started by oneshot808, Nov 10, 2011.

subset: commoncrawl


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Subscribe to Sport SchedulesAn Internet Calendar Subscription is a calendar that you download and view in either iCal or Outlook. However, unlike a schedule download for calender applications, Internet Calendar Subscriptions are updated automatically. For Outlook:
The Internet Calendar opens in side-by-side view in the Outlook Calendar and is added to the Navigation Pane in Calendar view under Other Calendars. Please note that send/receive errors will be present in Outlook. While Outlook is able to recognize the start and stop time of events for all functional purposes, all times are interpreted as "floating" start times, causing error (0x00040023). For iCal:
Online Calendar ApplicationsYou can subscribe to sport calendars using internet calendar applications by such providers as Google, and Yahoo. To add a subscription to your calendar, copy/paste your desired sport link below into your internet calendar application of choice when adding a subscription via URL. Please note that game start times are not displayed correctly within Google Calendar, as they are displayed in GMT time.
For more detailed instructions for Google and Yahoo calendar applications, click below:
Download Sport Schedules


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iMomoko is offering a 10% off student discount. To claim this offer you'll first need to sign up with Student Beans. Here you can verify your student status by providing an official university email. Once verified, you will gain access to an exclusive 10% off student discount code from iMomoko. This offer cannot be combined with any other promotion.
If you have additional questions about iMomoko student discount and eligibility requirements you can contact them here.
Other Ways to Save at iMomoko:
- Check for iMomoko coupons - BrokeScholar tracks all iMomoko coupon codes and discounts on our iMomoko coupon codes page. Today's top iMomoko deal: 15% Off Your Total with code DIGIDIP15
- Follow iMomoko on social media – Often iMomoko releases special deals to their followers on their Facebook and Twitter pages.
Similar Stores with Student Discounts:
Student Discount FAQs
What types of businesses offer student discounts?
Students will be happy to know that nearly every type of business offers student discounts. According to BrokeScholars database of student discounts, fashion and beauty retailers lead the way for online student discounts. For in-person offers, restaurants and events are most common.
How do I prove I'm a student?
How you verify your student status will depend on whether the student discount is for in-store or online use. For use at a physical location, you will simply need to show a valid form of student ID. For use online, retailers will require you to sign up with an official university email address to confirm eligibility. This can vary company to company, so check with the issuing business directly.
I was unable to verify my student status online, what should I do?
If you are a valid student but unable to verify your account, do not panic. If you attend a smaller university your school simply may not be listed in the verification system. Contact the company offering the discount directly. They will ask for other forms of verification, such as a government issued ID and a transcript or acceptance letter.
How many times can I use a student discount?
This will depend on the specific discount you wish to use. Based on our data, it seems most student discounts at restaurants and physical stores will offer students a discount on every purchase. However, student discounts offered by online retailers tend to only work once, or during a specific time of year. Read the fine print of the student discount you intend to use for the best answer.
Can I combine my student discount with other promotions?
The answer to this depends on the company offering the student discount. For online retailers, often you will be able to combine your student discount with ongoing sales and free shipping offers, but not with other promo codes. In-store offers seem to be more flexible, allowing you to apply print coupons and your student discount together.


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Do you want to shop from Shoeinc.com? Are you considering placing order? Wondering if the store is genuine? Read this review to know everything about the online store.
Shoeinc.com: Is This Website Legit?
The Shoeinc online store other wise known as Shoeinc.com is an online store that sells Shoe like Sneakers, Laces, etc. at suspicious discount price. Unfortunately, We research and found out some drawbacks about the store. Below are the red flags attached to the store-
5 Reasons Why Shoeinc Is Not A Genuine Website:
Poor Customer Support:
The means of contact on Shoeinc.com online website as email@example.com is fake. Mails sent to the address are not replied and the phone number+49 2151 3873740 is not functioning.. How then can customers contact the store after placing order? This is suspicious!
No Business Address:
Shoeinc failed to provide physical business addresses on the Contact Us Page of the website. How then can customers locate the store in case of refund/return of goods?
Lacks Security:
There is no trust seal from Norton, McAfee etc. As a result of this, Customers financial and personal details are not secured and are vulnerable to online hackers. Beware!
Discount Offer:
Shoeinc de offers discount for all its products which looks too good to be true. Remember the popular saying that if it looks too good to be true it is definitely not true. Offering of low rice and high discount is a bait used by online scammers to lure buyers. Don't be deceived!
No Social Media Presence:
Could it be that the people behind Shoeinc online store are afraid of being exposed on social media plat form? There is no social media Icon provided on the website. It does not lead to its official page. We searched and could not find anywhere social media page belonging to the store store.
Other Reasons Why You Not Should Shop From This Store Are As Follows:
Trust Score:
Shoeinc.de has a very poor trust score. This red flag should not be over looked and buyers are advised to take caution and avoid getting scammed.
It is quite obvious that the items on display on this store are stolen from other websites.
Should You Buy From Shoeinc?
No! With the above red flags, Buyers are advised not to buy from this store because after placing order, There is high tendency of not receiving any product and or you will receive a low quality goods. Online scammers are known to be using cheap discount prices to get customers. This is simply an example of bait and switch scam.
How Can You Check If An Online Store Is Legit?
- Customers Reviews
- Domain Registration Details
- Security Of Website
- Social Media Presence
- Discount Offer
- Business Address/contact
Things To Note While Shopping Online:
Receiving Of Low Quality/Inferior Product:
While shopping online, you should be careful not to fall into the hands of these fraud star.
Not Receiving Product:
It is no news that online scammers tends to lure buyers by offering low prices and high discount rate and after order has been made, customers will end up not receiving goods they have paid for.
Hacking Of Credit Card:
Credit card details are prone to being hacked by these fraudulent online stores. Sometimes you pay double the amount of the goods or you get debited for a product you did you order.
Scam VIP Membership:
Have you heard of VIP membership scam? This is one of the ways online scammers are defrauding customers. Here when an order has been made, You are automatically subscribed as to VIP membership club without your consent and get charged for it. Xemsy is an example of scam online store with such format.
Does Payment With PayPal On Shoeinc Store Guarantee Safety?
The answer is no! These days paying with PayPal does not necessarily mean that you will get refund immediately. Scam online stores uses fake tracking number which might have been shipped to your although not related to you. And this fake tracking number is been inserted into your PayPal transaction. Unfortunately for you PayPal sees this as complete transaction and makes it almost impossible to get your money back.
How Do You Get Your Money Back?
Are you an online shopper and have been scammed severally by these fake online stores? Have you order items online and after payment you can't reach the vendor? Read here to get more information on what to do.
From our findings, Shoeinc.com, just like Rewindppd, Sketchsteps, online stores we have reviewed are not trustworthy online stores. . As a result of this, We cannot vouch for its authenticity. Hence, customers are advise to tread with caution. Don't forget Federal Trade Commission (FTC) at https://reportfraud.ftc.gov/ and the Better Business Bureau (BBB) at https://www.bbb.org/scamtracker/. Reach out to them and report the website.
According to FTC, fraudulent online stores is one of the most reported fraud categories in 2022. Reports online show that more than one thousand persons fall prey to this kind of scam monthly. This is as a result of unchecked advertising on social media, cheap prices, and unsecured payment systems.
If you've fallen prey to this kind of scam, kindly contact your bank for a chargeback.
Meanwhile, have you shopped from this online store? Was your experience pleasant or sour? Please let us know in the comment section.
For more online store scams reviews, click here!


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21" Clip In Remy Hair Extensions: Dark Brown/ Monroe Blonde No. P4-613
Dark Brown No 4 and Monroe Blonde 613 is a perfect mix to deepen your naturally dark hair and give it just the right shades of light blonde highlights. Your hair is already beautiful, so why not show it off a bit to the world? A solid and silky set of these with add volume, shine, and depth to your hair. Did we mention that you can wash and style our extensions just like your own natural hair? Our exclusive clip in extensions are made of the highest quality, hand-picked 100% Remy Human Hair. Treat yourself to a set of our Dark Brown and Platinum Blonde Mix Clip In Hair Extensions and indulge yourself in full and longer
The 21" hair extensions, our best seller, will open your eyes to a new world of long luscious hair. They are meant to turn eyes and bring out your sex appeal. You will not go wrong with a set of these extensions. Length and volume here we come!
- 100% Remy Human Hair Extensions
- 7 clip in pieces
- 140-150 Grams of hair.
Add instant length and volume to your hair with Celebrity Strands Red Carpet Worthy Hair. Make all heads turn as you walk with confidence into any room. Our 21″ clip in hair extensions have been a top seller across the country. Top quality 100% Remy human hair with clips that will stay in place as you engage in any activity. These natural looking hair extensions can be straightened, dyed, and curled as if it was your own hair.
Shipping and Return Policy
When will I get my hair extensions?
We at Celebrity Strands pride ourselves in having a fast and easy shipping experience for our customers. If your order is placed before noon we will try our best to ship your order out the same day. It can take between 5-7 business days (US only) for your order to arrive (this includes processing time). All our orders are sent out Monday-Friday, this excluded weekends and holidays.
Shipping times varies depending on your location. Orders outside of the US can take between 5-15 business days to arrive.
- If we notice any irregularities with your order shipping times may increase. Please provide us with
- contact information if we need to reach you about your order.
- Didn't pick the right color the first time? As long as your set is unopened,
- we will gladly exchange your Celebrity Strands Hair Extension set for the right color you're looking for. Our hair extension quality is very high and we rarely have issued returns, but we understand the issue might come up for a refund, and will process your order based on our policy below.
- Human Hair Products like our Celebrity Strands 100% Remy Clip In Extensions cannot be exchanged if they are opened, worn, or tampered with in any way. They are a hygienic product and we stand behind our policy of hygiene and public safety. This safety and health policy is followed by the majority of hair extensions suppliers nationwide. We take this matter very seriously and will not accept any returns or exchanges if we notice tampering or any sort of usage.
How long do I have to return/exchange my hair extensions?
- We make returns and exchanges easy and smooth.
- First, contact us at
- within our exchange period thirty (30) days from your invoice date.
- Second, we will respond and issue you an RMA* (Return Merchandise Authorization)
- Third, we will then provide you with a return address to send your set of extensions to. After we receive your Celebrity Strands Hair Extension set, we will inspect each item to make sure they qualify for an exchange or return. To qualify, all items must be unopened, in the original packaging with no tampering, and include a copy of your order invoice. Products that have been opened, worn, or tampered with will not be exchanged or returned.
- *Any product(s) returned to us without an RMA will be denied and returned to sender.
Do I have to pay shipping for my return/exchange?
- You are responsible for all shipping costs and must include a tracking number with your shipment to make sure the package arrives. You also must write "Return, No Duties" on your package before shipping it back to us. We will not be responsible for any lost items.
When will I receive my exchange/refund?
- After we have received, inspected and qualified your returned hair extensions, we will process your request and send out your exchange or issue you a refund (MUST be within 30 DAYS from your invoice date) A full refund will be issued to the original payment method used when ordering. Remember, no returns or refunds will be issued if hair extensions have been opened, or taken out of the cardboard backing they are shipped in.
How to Care for your Hair
It is important to take great care of your hair extensions because the better you care for them the better and longer they will care of you!
- Make sure to always comb out your hair extensions starting from the tips. Do not tug or pull the hair too hard in order to prevent hair from falling out of the weft.
- We recommend that you wash your Celebrity Strands Clip-In Extensions after every 6-10 uses or as needed.
- Don't forget that even though your hair extensions are made from 100% human hair you still need to be gentle with them. Do not over-process or over-dye your hair.
- Celebrity Strands Clip In Hair Extensions last from six months to two years depending on how well they are cared for and how often they are worn. Since they're made from 100% real Remy human hair, they can change with you and your hair style needs.
Want this color in a different length?


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Call us 7 days a week
Item #: ISWB2052053
Usually ships in 20 to 23 days Estimated lead times do not include weekends or holidays. Transit time is determined by shipping method and destination. Orders requiring credit information might take additional time.
0 reviews | Write a review
with FREE shipping today! (within the contiguous US)
Frequently Purchased Together
Product Specifications


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A table in a relational database is much like a table on paper: It consists of rows and columns. The number and order of the columns is fixed, and each column has a name. The number of rows is variable — it reflects how much data is stored at a given moment. SQL does not make any guarantees about the order of the rows in a table. When a table is read, the rows will appear in an unspecified order, unless sorting is explicitly requested. This is covered in Chapter 7. Furthermore, SQL does not assign unique identifiers to rows, so it is possible to have several completely identical rows in a table. This is a consequence of the mathematical model that underlies SQL but is usually not desirable. Later in this chapter we will see how to deal with this issue.
Each column has a data type. The data type constrains the set of possible values that can be assigned to a column and assigns semantics to the data stored in the column so that it can be used for computations. For instance, a column declared to be of a numerical type will not accept arbitrary text strings, and the data stored in such a column can be used for mathematical computations. By contrast, a column declared to be of a character string type will accept almost any kind of data but it does not lend itself to mathematical calculations, although other operations such as string concatenation are available.
PostgreSQL includes a sizable set of built-in data types that fit many applications. Users can also define their own data types. Most built-in data types have obvious names and semantics, so we defer a detailed explanation to Chapter 8. Some of the frequently used data types are integer
for whole numbers, numeric
for possibly fractional numbers, text
for character strings, date
for dates, time
for time-of-day values, and timestamp
for values containing both date and time.
To create a table, you use the aptly named CREATE TABLE command. In this command you specify at least a name for the new table, the names of the columns and the data type of each column. For example:
CREATE TABLE my_first_table ( first_column text, second_column integer );
This creates a table named my_first_table
with two columns. The first column is named first_column
and has a data type of text
; the second column has the name second_column
and the type integer
. The table and column names follow the identifier syntax explained in Section 4.1.1. The type names are usually also identifiers, but there are some exceptions. Note that the column list is comma-separated and surrounded by parentheses.
Of course, the previous example was heavily contrived. Normally, you would give names to your tables and columns that convey what kind of data they store. So let's look at a more realistic example:
CREATE TABLE products ( product_no integer, name text, price numeric );
(The numeric
type can store fractional components, as would be typical of monetary amounts.)
When you create many interrelated tables it is wise to choose a consistent naming pattern for the tables and columns. For instance, there is a choice of using singular or plural nouns for table names, both of which are favored by some theorist or other.
There is a limit on how many columns a table can contain. Depending on the column types, it is between 250 and 1600. However, defining a table with anywhere near this many columns is highly unusual and often a questionable design.
If you no longer need a table, you can remove it using the DROP TABLE command. For example:
DROP TABLE my_first_table; DROP TABLE products;
Attempting to drop a table that does not exist is an error. Nevertheless, it is common in SQL script files to unconditionally try to drop each table before creating it, ignoring any error messages, so that the script works whether or not the table exists. (If you like, you can use the DROP TABLE IF EXISTS
variant to avoid the error messages, but this is not standard SQL.)
If you need to modify a table that already exists, see Section 5.6 later in this chapter.
With the tools discussed so far you can create fully functional tables. The remainder of this chapter is concerned with adding features to the table definition to ensure data integrity, security, or convenience. If you are eager to fill your tables with data now you can skip ahead to Chapter 6 and read the rest of this chapter later.
If you see anything in the documentation that is not correct, does not match your experience with the particular feature or requires further clarification, please use this form to report a documentation issue.


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Need to print a silhouette patterned paper to wrap a gift or complete a craft?
This free pastel blue silhouette small star outline pattern paper may be just the thing you need!
You'll find that this printable star outline theme scrapbook paper will print in vertical on a piece of 8.5 x 11 inch sheet of computer paper.
This patterned paper is great for wrapping paper, origami, card-making, scrapbooking, and other paper crafts.
I have also posted this exact patterned paper in different colors too. To find them, simply search "color you want + star outline pattern" on the site.
TOU for the Free Pastel Blue Silhouette Small Star Outline Pattern Paper
You may use these files for your own personal, non-commercial use only. You may not distribute or sell the files or claim them as your own. Files may be printed and distributed in educational environments.
If you would like to share the printables with a friend, please send them directly to this page so that they can download a copy for themselves. Please do not link directly to the download file.
Thank you for complying with the terms of use!
Please also see the disclaimer at the footer of the site regarding copyright and character use.
Downloading and Printing
These files are in PDF format. You will need a PDF reader like the free Adobe Reader in order to open the file.
If you don't have Adobe Reader, you can download it for free at Adobe.com.


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Gatto Innamorato
A cartoon cat variant of 😍 Smiling Face With Heart-Eyes. Depicted as yellow on major platforms.
Gatto Innamorato è stata approvata come parte di Unicode 6.0 nel 2010 ed è stata aggiunta a Emoji 1.0 nel 2015.
Emoji PlaygroundNuovo
Mostra altroProssimi eventi
Notizie Recenti
Mostra altroEmoji Mashup Bot's New Home on Emojipedia
First launched just one day before World Emoji Day, the original Emoji Mashup Bot has not been operational for over a year. Today, Emojipedia has debu...
New Earliest Emoji Sets From 1988 & 1990 Uncovered
In 2019 Emojipedia detailed a historic revelation: Docomo's i-mode emojis from 20 years prior were not the first to exist. Now, in 2024, further digit...
iOS 17.4 Emoji Changelog
Today Apple has released its latest emoji update, introducing 118 new emojis including a phoenix, a lime, several gender-neutral family designs, and v...


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Dream Big Goat ComfortWash/ComfortColor T-Shirt (S-4XL) - Multiple Colors!
- FREE US SHIPPING On all orders over $100
- In stock, ready to ship
ComfortWash & ComfortColor t-shirts are everyone's favorite! Pending color and inventory, brand between Comfort Wash & Comfort Colors can be interchangeable. But no worries, they are the exact same quality tee!
Made to order and printed Direct to Garment in Texas! Items are made to order and will ship in 10-15 business days. (Monday - Friday)
These are unisex and run true to size. We recommend staying with your true t-shirt size when ordering.
Machine wash, cold. Hang dry or tumble dry, low. Do not iron decoration.
If your color is on backorder, you will be contacted to exchange colors or sizes.
Items are made to order and will ship in 10-15 business days.
We hope you love your items! If you need to return an item, we offer store credit or exchange. NO refunds. You can read more about our policy at the bottom of our homepage. WMB can edit the brand of t-shirt to one similar with no notice. Colors may vary from screen to screen and we have done our best to represent the truest color form. These displayed are MOCKUPS. We print direct to garment using high quality inks and printers.
If you have any questions, please email firstname.lastname@example.com
We want to thank you for shopping at Wandering Maverick Boutique.It is our goal at Wandering Maverick Boutique that our customers have a great experience shopping with us, and we take great pride in the unique experience we provide.
The best way to get a hold of us is through our email email@example.com.
- No refunds. Only exchange or store credit.
- All items must be unworn and unwashed.
- Customers are responsible for shipping if they want to return.
- Once WMB delivers to the post office, the package is no longer WMB's responsibility. Please contact the local post office or sign up for text alerts for your package.
- Items that were purchased with 20% off or more are FINAL SALE.
- All grab bags are FINAL SALE - no exception.
- All items purchased in person at shows are FINAL SALE - no exception.
- Canceled orders will be charged a 15% restocking fee.
- If your order is returned to the WMB warehouse due to an insufficient address, you will be required to pay the shipping back and provide a new address to send your order to.
- If you ordered items that contain a PREORDER, the entire order will not ship until it is ready. If you want to receive your other items before that, we can invoice you a separate shipping charge.
- Wandering Maverick is not responsible for additional taxes, fees, postage, customs charges, etc. on international orders.
I ordered the wrong size, how do I exchange it?
Please email firstname.lastname@example.com. Once you email us with your order number and size you want to exchange for, we will give you the correct return address. Please note, the shirt must be returned within 1 week of notifying us. The customer is responsible for shipping back to the warehouse. We encourage you to have tracking information and send it over to us via email when you send your package. Please include your name and order number in the package.
I received a different size than what I ordered, how do I exchange it?
Please email firstname.lastname@example.com within 3 business days of your package being delivered. We will email you a prepaid return label and as soon as it is dropped off in the mail, we will process the correct item to be shipped.
My T-shirt is damaged/incorrect. What should I do?
Please send pictures and your order number to email@example.com so we can determine what the next step will be. Rest assured you will receive exactly what you ordered!
Can you add a discount to my order? I forgot it at checkout.
We cannot go back and discount orders. Please ensure that all codes are put in before hitting submit order.
Can I cancel my order?
If your order has not been sent into production, you may cancel with a 15% restocking fee. This covers fees that we have as a business.
USPS is sending my order back to you, can you please mail it back?
Yes, we will mail your order back to you. The customer will be responsible for providing a new shipping address. Customers will also be responsible to pay for the package to ship back to them if they originally had free shipping.
I want to cancel/pause/edit my Monthly Tee/Crew Club membership.
Head to www.shopwmb.com and log in using the email address associated with your account. From there you can manage your subscription. If you have any troubles, please email us!
Customs is charging me more to get my package, will you refund me or pay the additional?
We will not pay any additional fees/postage at customs. Additionally, we will not refund for what you have to pay. On most international orders, we had to pay more for shipping than what you paid in the first place.
- $6.95 flat-rate US shipping or FREE on US orders over $100!
- Shipping based on weight for all international orders.
- Turnaround time for custom orders is 5-15 BUSINESS DAYS
- Returns on unworn, unwashed items with tags still attached are accepted within 14 days of receipt of item.
- Returns are eligible for store credit only.
- Sale/clearance items are NOT returnable, they are final sale.
- Shipping charges (original and return) are the customer's responsibility.
Subscription Boxes
- Our Monthly Tee box Ships out once a month on the day of purchase you will be billed on a recurring basis monthly on the same day unless you pause or cancel your subscription.
Buy now, pay later.
Split your entire online purchase into 4 interest-free payments, over 6 weeks with no impact to your credit.
2 weeks
4 weeks
6 weeks
Here's how to Sezzle
Shop and add items to your cart as normal!
Choose Sezzle at Checkout! You'll be redirected to Sezzle to Sign Up or Log In to complete your order.
Sezzle it
Your order will be shipped out right away* and your payments will be split up over 6 weeks.
*shipping times subject to merchant shipping policy
Download the App
Shop directory. Reschedule payments. Plus more!


text:
What is this product?
This is a Solution Manual For Systems Analysis and Design, 10th Edition. This is not the textbook. A Test Bank is a collection of exam questions with solutions based on the textbook. However it is not from the textbook. Test Banks or exam guides are useful in test preparation, revisions and self-study. A Solution Manual is a book guide. It has detailed solutions to end of the chapter questions and projects or case studies may be included as well in some guides. These solutions can assist with assignments, homework and book-based learning.
Who can benefit from it?Anyone looking for supplementary material for college textbooks can make use of this product, as long as they're aware of the academic policies in place.
How to make sure it's what you need?Check the full chapter sample to see the type and number of questions that are answered, and if you're satisfied, proceed to add to cart or checkout. If you have any questions or concerns, reach out to the live chat support.
How do I download Solution Manual For Systems Analysis and Design, 10th Edition?After completing the payment, you'll be directed to a page with the download link, which will also be sent to your email for future reference. If you encounter any download issues, reach out to the 24/7 live chat support for assistance.


text:
- 25% Off On New Register
- +91-86170-38699
- firstname.lastname@example.com
A vast majority of the app marketers mainly concentrate on the post-launch app marketing techniques and measures while completely missing on the pre-launch campaign. This prevents the app to create buzz and hype just around the time when the app is launched. As and when you launch the app, already a considerable number of people should expectantly look forward to your app and this requires long-drawn marketing efforts leading up to the app launch event. To create pre-launch buzz and hype about the app a mobile app development company has an array of marketing options like social media campaign, search engine ads, video ads, email campaigns, etc. Apart from online options, you can also reach out to the wider audience with traditional marketing options like outdoor ads, print ads, media ads, and promotional events.
Onum Company
Web Designer, Illustrator
Media Group
Photographer, UI Designer
Torex Company
Motion Designer, Illustrator
Nimkus 2000
PHP Developer, Photographer
Hard Skills
Just as a retail business in real life is remembered not just for its product offerings but also because of its services, support, and customer-friendliness, an app that offers a helpful customer support system for its valued users enjoy more traction and engagement than other apps. Great brands all over the globe enjoy appreciation and popularity because of their customer-friendly support and services.


text:
The Editorial Staff who decide to use of this option will receive the SJIF Value within 7 days.
Register journal - (Free Service)
in a scored and prestigious database of scientific journals.
Manage journal - (Free Service)
This section allows you to place information about the journal, editors, and publisher, etc.
Full list of journals from database of SJIFactor.com. It contains currently over 24000 journals from all over the world.
A proof of being indexed in SJIF Journal Rank List.
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WebTorrent Desktop Download for PC Windows 11. FREE
WebTorrent Desktop
by WebTorrent - Freeware
WebTorrent Desktop (2024) Download for Windows 11 PC/Laptop
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Bryan Shaw never expected to write a research paper about a rare eye cancer.
He's a chemist who works on how metals and proteins interact. But life has a funny way of interrupting the best-laid plans, and now Shaw may be on to a powerful new way to detect retinoblastoma in newborns. Such early detection could mean children with the disease would have a better chance of keeping their eyes and staying alive.
Shaw's scientific odyssey begins in May 2008, when he and his wife, Elizabeth, had a baby boy, Noah. Bryan was a postdoc at Harvard University at the time.
When Noah was 3 months old, Elizabeth noticed that sometimes when she took a flash picture of Noah with their digital camera, she would see a white reflection come back from his eyes instead of the usual red dot. She had read in a parenting magazine that this could be an early sign of retinoblastoma, so naturally as a first-time parent she assumed the worst. As husbands are wont to do, Bryan assured her she was crazy.
But on their next visit to the pediatrician, the doctor dutifully shined a penlight into Noah's eyes, since a white reflection from this simple eye exam can also detect the retinoblastoma tumor. The pediatrician didn't like what she saw, so she sent the Shaws to an ophthalmologist, who confirmed the worst: Noah had tumors in both eyes.
There followed months of chemotherapy, radiation, and ultimately surgery to remove Noah's right eye — and eventually years of checkups.
At first, Shaw says, he was too distraught to think about his son's cancer in scientific terms. But one day when Noah was 2, Bryan was in the waiting room while Noah was getting therapy, and he started thinking about the baby pictures that first revealed the tumor. He wondered if there was a way to track the disease back to the day the tumors first appeared.
He started small, but ultimately he would end up digitizing thousands of pictures he and Elizabeth had taken of Noah, searching for the first sign of leukocoria, as the white-eye condition is technically known.
"It first showed up in a picture taken when Noah was 12 days old," says Shaw. It didn't appear in all the pictures taken around that time. The tumor was small then, and the camera angle had to be just right to get the white reflection. But by 3 months there were days when nearly all the pictures showed signs of leukocoria.
Shaw is now on the faculty of Baylor University in Waco, Texas. He collaborated with the ophthalmologist who treated Noah at the Massachusetts Eye and Ear Infirmary and Noah's oncologists at Dana Farber Cancer Institute to produce a paper published in PLOS ONE.
In addition to pictures of Noah, the paper analyzes family photos from eight other children after they'd been diagnosed with retinoblastoma. The leukocoria is present in all of them.
Using the more than 7,000 pictures Bryan and Elizabeth have taken of Noah, the paper also shows that the amount of leukocoria that the digital cameras can detect increases with the size of his tumor.
Bryan is hoping to get more pictures from families with a child with retinoblastoma. He's also looking for a collaborator who could write software that would automate the process of scanning photos for leukocoria.
The disease is rare — fewer than 12 cases per million children aged 0-4. That works out to about 1 case for every 15,000 live births. Still, Bryan thinks a screening test would help with early detection of the cancer, and might save the vision of kids like Noah.
And speaking of Noah, he's a hoot. His health is good, and the cancers seem to have given up. He likes to draw and sing, and he's anything but shy. He has a prosthesis where his right eye used to be, which makes him look pretty much like any kid his age. And now he has a baby brother Samuel who, happily, doesn't share his disease.
Copyright 2021 NPR. To see more, visit https://www.npr.org.


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Contact Website Administrators about golo
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Why must I answer this question?
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Although spammers can rent or buy existing e-mail lists, many opt to use software known as 'e-mail harvesters' (often referred to as 'spam bots') that extract e-mail addresses from web pages. These e-mail harvesters work very much the same way search engine spiders do and try to collect every e-mail adress they encounter on the web. However, the brute force algorithms they use, are not able to answer the simple question above.


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799 Showgirl by Morris Costumes Final Sale
799 Showgirl by Morris Costumes Final Sale
Hair Fiber | Synthetic |
Cap Size | Average |
Additional Information
To get shipping cost estimates, add this item to your shopping cart and enter your country/state/zip code information.
Shipping Method: Order Processing Time: Time in Transit: Total Delivery Time: USPS Priority 1-3 business days 1-3 business days 5-7 business days USPS Priority Express 1-3 business days 1-2 business days 3-5 business days UPS Ground 1-3 business days 2-8 business days 5-10 business days UPS 2nd Day Air 1-3 business days 2 business days 4-6 business days UPS Next Day Air 1-3 business days 1 business day 3-5 business days USPS Priority Mail International 1-3 business days 7-10 business days 8-14 business days USPS Priority Mail International Express 1-3 business days 5-7 business days 6-10 business days UPS Worldwide Express 1-3 business days 5-7 business days 6-9 business days For more details, read our Shipping Disclaimer. -
If for any reason you are unhappy with your purchase, send it back! You have several options.
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- Items must be sent back within 14 business days of receipt in new condition. Items cannot be worn or tried on, must be in the original packaging with all tags attached, and be odor-free. Bellami, Bosley, Henry Margu, Louis Ferre, Milano Collection, costume wigs, costume accessories, wig accessories, clearance sale items, and price matched items are FINAL SALE.*
- The refund amount will be calculated based on return item cost only. For your convenience, return labels are included for domestic orders. A $14 deduction will be made from any refund to the return when used. We are not responsible for lost or damaged packages.
- Promotional items of the product being returned must also be sent back; otherwise retail value will be charged.
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- Items that do not meet these criteria will be subject to a 30% fee; the remaining amount will be refunded to the original method of payment.
Return Process:
All the items we sell are in new and unworn condition. Therefore, we have 2 refund options in place:
- Refund as store credit. Store credit does not expire and you may apply it towards future purchases at any time. Only a registered account is required. Our Hair Gurus can assist you in setting up an account if you don't have one. Restocking fee waived.*
- Refund to the original payment method with a 15% restocking fee. Every item returned is carefully received, examined, and shipped back to our vendors with significant time and costs involved.
At Voguewigs.com, customer satisfaction is our top priority. If for any reason you are unhappy with your purchase, we will gladly exchange or refund the item. Please ship the item back with the return/exchange form included on the backside of your invoice within 10 business days of receipt. See the full details here.


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Welcome to our website! As we have the ability to list over one million items on our website (our selection changes all of the time), it is not feasible for a company our size to record and playback the descriptions on every item on our website. However, if you have a disability we are here to help you. Please call our disability services phone line at 208-664-2131 during regular business hours and one of our kind and friendly personal shoppers will help you navigate through our website, help conduct advanced searches, help you choose the item you are looking for with the specifications you are seeking, read you the specifications of any item and consult with you about the products themselves. There is no charge for the help of this personal shopper for anyone with a disability. Finally, your personal shopper will explain our Privacy Policy and Terms of Service, and help you place an order if you so desire.
Runge Furniture Company in Coeur dAlene, ID is an authorized dealer of Hooker Furniture Products. Hooker Furniture today is a complete home furnishings resource offering the world's greatest selection of quality home entertainment, home office, accent, dining and bedroom furniture, as well as leather and custom seating. During over 85 years of operation, They've sought to display integrity in their relationships with employees, customers, sales representatives, suppliers and the local communities in which they live and work. Product integrity also is paramount.
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text:
September 6, 2022
about costola
At home near the sea. Ever-focused on beauty and the thrilling art of creation; found in the moments that leave me breathless and in awe.
After 15 years of capturing couples and families during the most precious times of their lives, she can truly say that this is exactly where she is meant to be. When she isn't photographing weddings, Andie enjoys the outdoors alongside her husband, Tony and daughters, Ava & Audrey.
Portrait Sessions can be stressful, but if you arrive with a few tricks in your bag, you'll come away with portraits that perfectly capture what family is all about.


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Matthew 26-28: Jesus' life-giving death by Amy Welborn offers a close look at the passion, death, and resurrection of Jesus Christ in Matthew's Gospel.
It is a part of Loyola Press' Six Weeks With the Bible series, which provides individuals or groups plans for concise but thorough 90-minute sessions to learn about and discuss the pertinent Scriptural passages. General guides for how to effectively lead an adult education session are also included. The series is available in paperback and also in Kindle versions.


text:
Air Cylinder, CG1-Z SERIES | SMC Corporation CDG1KWLN20-150Z
Availability: Out of Stock
What is SnapMagic Search?
SnapMagic Search is a free library of symbols & footprints for the CDG1KWLN20-150Z by SMC and for millions of electronic components.
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Part Name | CDG1KWLN20-150Z |
Manufacturer | SMC |
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text:
Please share your impressions, remarks, suggestions to help us improve the new Southplanet website. Don't hesitate to signal any bugs that you happen to come across !
Add your information (With the self-presentation files)
Convention with Wikiafrica / Wikipedia : Southplanet's contents are shared for free according to the licence Creative commons .
© Some artistic content (photos, videos, sound recordings, written texts) is subject to copyright. Copyright is mentioned when and where it applies. Copyrighted material cannot be reproduced.


text:
A small victory but we'll take it.
via the Associated Mess:
A federal judge has ruled a property management company owned by the founder of Domino's Pizza doesn't have to immediately implement mandatory contraception coverage in the health care law.
U.S. District Judge Lawrence Zatkoff ruled Sunday in favor of Tom Monaghan and his Domino's Farms Corp., near Ann Arbor. Monaghan, a devout Roman Catholic, says contraception isn't health care but a "gravely immoral" practice.
Zatkoff granted Monaghan's emergency motion for a temporary restraining order until a final decision is made in the case. The mandate would have taken effect Tuesday.
The government says the contraception mandate benefits women's health and removes financial barriers. There are about a dozen similar lawsuits pending nationwide.
Monaghan sold most of his controlling stake in Domino's Pizza in 1998 to private equity company Bain Capital.


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Apache HTTP Server Version 2.4
Description: | User-specific directories |
Status: | Base |
Module Identifier: | userdir_module |
Source File: | mod_userdir.c |
This module allows user-specific directories to be accessed using the
Description: | Location of the user-specific directories |
Syntax: | UserDir directory-filename [directory-filename] ...
Context: | server config, virtual host |
Status: | Base |
Module: | mod_userdir |
The UserDir
directive sets the real
directory in a user's home directory to use when a request for a
document for a user is received. Directory-filename is
one of the following:
. This turns off
all username-to-directory translations except those
explicitly named with the enabled
keyword (see
followed by a
space-delimited list of usernames. Usernames that appear in
such a list will never have directory translation
performed, even if they appear in an enabled
followed by a
space-delimited list of usernames. These usernames will have
directory translation performed even if a global disable is
in effect, but not if they also appear in a
clause.If neither the enabled
nor the
keywords appear in the
directive, the argument is treated as a
filename pattern, and is used to turn the name into a directory
specification. A request for
will be
translated to:
UserDir directive used | Translated path |
UserDir public_html | ~bob/public_html/one/two.html |
UserDir /usr/web | /usr/web/bob/one/two.html |
UserDir /home/*/www | /home/bob/www/one/two.html |
The following directives will send redirects to the client:
UserDir directive used | Translated path |
UserDir http://www.example.com/users | http://www.example.com/users/bob/one/two.html |
UserDir http://www.example.com/*/usr | http://www.example.com/bob/usr/one/two.html |
UserDir http://www.example.com/~*/ | http://www.example.com/~bob/one/two.html |
"UserDir ./"
would map "/~root"
- which is probably undesirable. It is strongly
recommended that your configuration include a "UserDir
disabled root
" declaration. See also the Directory
directive and the Security Tips page for
more information.
Additional examples:
To allow a few users to have UserDir
directories, but
not anyone else, use the following:
UserDir disabled UserDir enabled user1 user2 user3
To allow most users to have UserDir
directories, but
deny this to a few, use the following:
UserDir disabled user4 user5 user6
It is also possible to specify alternative user directories. If you use a command like:
UserDir "public_html" "/usr/web" "http://www.example.com/"
With a request for
, will try to
find the page at ~bob/public_html/one/two.html
first, then
, and finally it will send a
redirect to http://www.example.com/bob/one/two.html
If you add a redirect, it must be the last alternative in the list. Apache httpd cannot determine if the redirect succeeded or not, so if you have the redirect earlier in the list, that will always be the alternative that is used.
User directory substitution is not active by default in versions
2.1.4 and later. In earlier versions, UserDir public_html
was assumed if no UserDir
directive was present.
Lists of specific enabled and disabled users are replaced, not merged, from global to virtual host scope


text:
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Answers to your FedEx Authorized ShipCenter pickup and dropoff questions


text:
Visits to page
Now online: 1
Today: 28
This week: 280
Overall: 386371 Book links
Comments about our book "No Oil in the Lamp Fuel, Faith and the Energy Crisis"
'The biblical call to be stewards of creation requires us to use our God-given resources with care. I highly recommend this book to all Christians who are concerned with the biggest issue of our day – the future of the planet.'Matthew Sleeth, Executive Director, Blessed Earth
'I found this a really helpful read. It's scientifically well-informed and nicely Biblically-based, but also full of practical information for those of us who want to try to do something about this really important issue. We don't know what our immediate future holds or what we and our children and grandchildren will be having to deal with in the coming years. We do know, though, that we must live our lives seeking justice and shalom through all we do. This book will help us take some steps towards that.'
Ruth Valerio author of "'L' is for Lifestyle: Christian Living That Doesn't Cost the Earth"
'No Oil in the Lamp is a great starting point for Christians who have heard about peak oil but never quite looked into it. (If you're wondering about the title, see Matthew 25) But it's also one of the best books I've read on oil depletion generally.'
Jeremy Williams at "Make Wealth history"
"this is a well-researched and comprehensive guide to where we are energy-wise and how Christians can respond practically both in their own homes, and via their churches to a growing crisis."
Catherine von Ruhland at "Third Way" magazine
"...there is so much in what is a fact- and idea-filled boolc that it should be commended to a wide readership to study closely, for its understanding and advice about things that can be, and are being, done by families, community groups and churches."
"New Directions" magazine
"The book will be an excellent if demanding primer for
eco-congregations. It could form a valuable start for discussions in eco-congregations around issues that are hanging all the time. It does not answer all the questions but it can start discussions on a range of issues."Eco-congregation Scotland
"Much of the material in No Oil in the Lamp is difficult to bear and so I highly recommend this book for a shared reading. These are some of the biggest issues of our time and deserve collective lament, reflection, and action. In summary, I came away from the book with these simple insights about how my own imagination needs to be transformed:"
Resilience, not Sustainability.
Hope, not Fear.
Transition, not Panic.
Generosity, not Hoarding."
" a brilliant book worth checking out."
Geoff Maddock
Englewood Review of Books
"In this efficient, authoritative introduction to the peak oil problem, Mellen and Hollow summarise the problem, and describe a range of possible near-Futures ranging from the utopian to the apocalyptic."
"Life and Work"
"...it is an important read for all of us, both as Christians and as citizens"
"The Reader Magazine"
"The fact is that I read the whole book in just one and a bit readings, I could simply not put it down. Easy to read, full of facts, some quite frightening, very well referenced indeed, and quite inspirational. As far as Peak Oil is concerned the world is sleep-walking into a bleak future unless facts are faced. The authors of this book face those facts in easy to read
detail and show that there really is a way forward if action is taken.""Green Christian Magazine"
The authors have a profound vision of the church as salt and light in the preparations for an energy-constrained future, and the book provides the information and inspiration needed to get started.
"Prism Magazine"
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Our Friends
Category Archives: air pollution
We need to talk about woodburners
Woodburners come up in the news a lot at the moment. And not in a good way. They are much maligned as a source of air pollution. Confession time I have a woodburner which I love, so it was with … Continue reading
particulate pollution
Last week I wrote a short blog on what I thought the three most pressing environmental problems were. For all three I gave a brief assessment on the will to deal with them. In no particular order I decided to … Continue reading
Three pressing environmental problems
Hearing an article on the news on plastics the other day got me thinking about I thought were the three pressing environmental problems. I came up with plastic pollution, Climate change (obviously) and particulate pollution. I'm going to have a … Continue reading
Does nature have inherent value?
Does nature have inherent value? On the news over the last week there has been a lot of talk over the loss of songbirds in Europe, mainly France where the survey was done. The same is true here though. Last … Continue reading
let there be light
Let there be light is not only God's command in Genesis 1v3 but also a TEARFUND campaign to get the UK government to fund small scale renewables and batteries in the developing world. Its worth reminding ourselves as we read … Continue reading
air pollution
I've read a recent bit of research on air pollution from China. The research looked at particulate (PM10) levels across the major cities in China and the effect on mortality. The findings were unsurprising in many ways but also showed … Continue reading


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Azusa Care and Control Defense
Defending Against A Care and Control Charge in Azusa
DUI offenders charged with impaired driving face severe and harsh punishments in Azusa. In such cases, it is always advisable to contact an experienced and reliable DUI Lawyer.
As per section 23152 of the California Vehicle Code; everyone commits an offense who operates a motor vehicle or vessel or operates or assists in the operation of an aircraft or of railway equipment or has the care and control of a motor vehicle, vessel, aircraft or railway equipment, whether it is in motion or not; while the person's ability to operate the vehicle, vessel, aircraft or railway equipment is impaired by alcohol or by a drug or having consumed alcohol in such a quantity that the concentration in the person's blood exceeds eighty milligrams of alcohol in one hundred milliliters of blood.
If you are facing a care and control charge, it is necessary for you to hire a reputable DUI lawyer who will fight to protect you from punishment arising from the case. Our firm offers you reliable DUI lawyers in Azusa who specialize in dealing with DUI offenses and who will strive to protect you from a conviction in an impaired care and control case.
What Constitutes A Care and Control Over 80 mg DUI Charge in Azusa, CA
In order to constitute a DUI offense as well as for you to be convicted of a DUI, you do not have to actually be driving a vehicle when you're impaired by alcohol or drugs. Whether the engine of the vehicle is running or not, if you are able to set the vehicle in motion, it is enough to put you in a situation when you can be convicted of a DUI. The reason for this is that you have the ability to be in care and control of the vehicle and, csonequently, it will be presumed that you did so while your ability was impaired.
How to Avoid a Azusa Care and Control Charge Legally?
If you have been found drunk while sitting in the drivers seat of a vehicle, you will be attracting far more charges than when you are sitting in any other position of the vehicle. After investigation, the police will conclude that you were in impaired care and control of a moving vehicle and will argue the case based on the fact that you were fully capable of setting the vehicle in motion, unless there is evidence to show otherwise. The burden will then be upon you to refute this and convince the court that you had no intention or capability to set the vehicle in motion. In such a case, it is always advisable to stay out of the vehicle or hire a sober driver. By doing so you will be sure to avoid a care and control charge.
Getting in Touch With A Reliable Azusa DUI Lawyer To Fight Your Care and Control Charge
After assessing your case and preparing it thoroughly for representation in court, our lawyer will be able to mobilize the various ways to avoid the care and control charge. A high-quality and experienced DUI lawyer will fight to convince the court that you had no intention or capability to set the vehicle in motion.
Do Not Hesitate To Contact Us If You Have Received A DUI Offense in Azusa
Types Of DUI Charges:
Consequences Of A DUI:
Azusa Laws Regarding Care and Control Over 80mg and Impaired Care and Control Charges
As per Section 23152(e) and Section 23152(f) of the California Vehicle Code, you are said to be committing a DUI offense and being in care and control of a motor vehicle while you are driving under the influence of drugs or alcohol or driving while being influenced by a combination of both. Impaired care and control does not necessarily mean driving a vehicle. In fact, a police officer can prosecute you if he has concrete reasons to form an opinion that your ability is impaired due to the influence of alcohol or any other drug irrespective of whether you were sitting on the driver's seat or the other seats, even if the vehicle is not in motion.
Section 23152(b) of the California Vehicle Code explains and defines Over 80 as an extreme offense. As per this section, it is an offense to operate or be in care and control of a motor vehicle with over 80 mg of alcohol in 100 milliliters of blood. Getting charged with care and control over 80 mg means, that you were in care and control of a motor vehicle and your blood alcohol concentration being more than 0.08 which is a criminal offense and has very serious consequences.
Breaking the Azusa Care and Control Criminal Code and Its Consequences
The care and control criminal code consequences are very serious in Azusa and California. Notably, as per the California Vehicle code DUI offenses are considered as a crime and its consequences are very serious with suspension of license, a treatment or education program, fine or probable jail time and most of all, a criminal record. A good DUI lawyer is your only option to protect you from a DUI conviction. We offer you one of the most reputable DUI lawyers in Azusa, California and can provide you protection from a DUI conviction and its consequences. We always make it our duty to provide the best services and guidance in DUI cases to all Azusa clients.


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Foil Stamping
Foil stamping is an instant attention grabber. From business cards and letterhead to wedding invitations and thank-you notes, foil stamped text and graphics provide an accent of sophistication and create a lasting impression. We are experts in the foil stamping process and can help you use it to the best effect. So contact us today and add a touch of class to your next document.


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Greetings! We are Sharma and Sharma Advocates, a distinguished firm renowned for delivering exceptional legal services in Bijnor. Our company offers top-notch Legal Services in Bijnor. Our company is dedicated to delivering exceptional legal services and personalized attention to our valued clients in Bijnor. Our team of experienced lawyers is dedicated to securing positive results for our clients across a diverse range of legal issues in Bijnor. Our business offers expert legal assistance for any matter, who will handle your case with precision and diligence in Bijnor.
With a team of highly skilled and experienced lawyers, we provide comprehensive representation in various areas of law in Bijnor. Sharma and Sharma Advocates is a prominent Law Firm in Bijnor renowned for its exceptional legal services. Our firm is dedicated to delivering personalized attention and achieving favorable outcomes for our clients in Bijnor. Whether you require assistance with any legal matter, we have the expertise to guide you through the legal process in Bijnor.
Our firm is honored to employ some of the nation's most qualified and reputable lawyers in Bijnor. If you need the Best Lawyers in Bijnor, you need to look no further than our firm. With their extensive knowledge of Indian laws and regulations, our top lawyers are able to manage complex legal cases and provide clients with effective strategies to achieve favorable outcomes in Bijnor. Our team consists of renowned legal professionals specializing in various areas of the law in Bijnor.
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'Round-the-horn site-directed mutagenesis
back to protocols | ||
'Round the Horn Site-directed mutagenesis
Submitted by Sean Moore
I was in graduate school and had to make several codon substitutions at the same position in a cloned gene. I came up with this protocol and called it "Around-the-Horn", a phrase used in baseball meaning to throw the ball through all of the base positions in a circle. The phrase originally referred to "Rounding Cape Horn of South America" a very difficult, but rewarding journey ("Round-the-horn" is not difficult). Subsequently, a group published a paper describing the process, but I was first, take full credit, and have used it in ways not described by others.
I was used to doing "Quickchange" (Stratagene) wherein two complementary oligos containing the desired mutation are used to prime extensions all the way around the plasmid. Quickchange is a linear amplification (not PCR) and each round of extensions is performed on the plasmid template. There are several problems and limitations with Quickchange:
- The size of the mutation is limited to a few base pairs.
- Each primer has to be long to provide sufficient annealing on both sides of the mutation (money).
- Non-optimal extension conditions will result in "end-filling" wherein the polymerase fills the recessed 3' end of the product instead of extending the primer on the template (this is especially a problem as the reaction progresses and is why only 12 or so cycles are recommended). The blunt-end products won't give you any transformants and also mislead the researcher into thinking their reaction went great because they see a product on a gel, but the product is a dead end.
- For each different mutation at a given position, a new primer pair has to be ordered (mo' money).
'Round-the-horn is a PCR-based mutagenesis. The mutations are contained in one or both of the primers. The primers are phosphorylated so that the PCR product can be ligated into a circle and used to transform cells. The procedure has the advantages that the primers are smaller, a visible product on a gel means your reaction worked and must contain the encoded mutations, only one primer needs to be changed to change the mutation at the same site. If you are concerned with mutations elsewhere in your plasmid, you should sub-clone the segment of interest into a "fresh" vector (this is the case for any site-directed protocol). Finally, as opposed to standard site-directed mutagenesis, this procedure exponentially amplifies the plasmid, leading to increased yields, increased transformation efficiencies, and the potential to create a large library with randomized residues.
This protocol can be cited directly OWW, but I get frequently asked for the first paper to use it. When I was nearing the end of my graduate work, I made the first mutations in the P22 scaffolding protein; found in this reference:
However, if my shoddy memory serves correctly, there is another publication that came out shortly before mine, also from my graduate lab, the technique was used to introduce point mutations in HIV CA. I recall advising our technician, but I may be wrong. It is simply listed as "PCR mutagenesis".
Primer design
In this example, the Forward primer (primer A in the image above) has intended mutation at its 5' end. Design such that the annealing portion has Tm ~60-63º.
The reverse primer in this example resembles the complementary strand and abuts the site of mutation.
- If the change needed is large, both primers can have mutant 5' ends. Alternately, two rounds of PCR can be performed to build the desired PCR product.
- Primers without mutations at their 5' ends can be spaced apart on the template with a gap between them to make deletions of any desired size.
- You can amplify a low-copy plasmid with primers that introduce cloning sites for receiving an insert. The PCR product becomes the "vector" component of the ligation reaction. In this case, you don't need to phosphorylate the primers because the restriction digest of the ends will leave 5' phosphates. An advantage using PCR to prepare your vector is that background "vector re-ligation" reactions are practically eliminated.
- Remember to transform bacteria that have no restriction system as the plasmid will be unmodified. This is not essential, but really helps.
Primer Phosphorylation
- Primer stock should be at 100 μM in water or TE.
- For each primer, mix:
- 37 μL water
- 5 μL 10 Kinase reaction buffer (comes with PNK)
- 1 μL 50 mM MgSO4 (comes with most polymerases)
- 5 μL primer (10 μM final)
- 1 μL 100 mM ATP
- 1 μL PNK
- Incubate at 37°C for 30-60 minutes.
- Heat each reaction to kill the PNK (65°C for 20 minutes or 95°C for 5 min).
The Mixture
Set up the following 50μL mixture on ice. After mixing, put into pre-heated PCR block. Keep in mind that your particular volumes may be different, some newer fusion polymerases (recommended) use 5X and 2X buffers.
- 40 μL water
- 5 μL 10X polymerase buffer
- 1.5 μL forward primer (0.3 μM final)
- 1.5 μL reverse primer
- 0.5 μL dNTPs (20 mM each)
- 1 μL template (mini-prep'd plasmid)
- 0.5 μL polymerase (not Taq, use Vent, Deep Vent, 9°N, Pfu, or a faster fusion enzyme)
PCR program
- 55-58°C 30" (~5°C below Tm, higher for fusion enzymes)
- 72°C 2' per kb being amplified (much less if using a fusion enzyme)
- Goto (2) 25-27 more times
- cool to 18°C for a minute or so (there's no need to hold at 4, you just cooked the sample for an hour!)
- end
Product Preparation
Run 5 μL on a gel. I put EtBr in the gel so I can check to see if there is a product within a few minutes. If you see the band, digest the template in the PCR reactions by adding 1 μL of DpnI, mix, and incubate for at least 60 minutes at 37º.
Run a preparative gel of the PCR product, excise the band, and use a kit to extract the product.
Product ligation
- 2.3 μL water*
- 2 μL PCR product
- 0.5 μL 10X ligation buffer
- 0.2 μL ligase
- 5 μL
If there is not much product, replace the water fraction with more PCR product.
Incubate overnight at room temp. These blunt ligations can be pretty slow. Some enzymes (e.g. 9° North and Taq) leave crap on the 3' ends. This greatly reduces the efficiency of the blunt-end ligations.
Works best with electroporation or fresh TSS cells. Transform 4 μL into 50 μL cells. Plate all of it. You should get 20 to 1000 colonies. Many times, I have forgotten to phosphorylate the primers before doing the PCR. You can successfully phosphorylate the purified PCR product before ligation, but the ligation/transformation will be less efficient. PNK prefers exposed 5' ends.
We have been using NEB's Q5 enzyme almost exlusively now. A rough estimate of 0.5 ng of template, 300 nM primers, and 2-3 degrees below the NEB recommended anneal. Also, use NEB's GC enhancer and 30 sec/kb.
I have had reactions that just wouldn't produce product (for whatever reason). When doing PCR, it is difficult to predict the yield or quality of a product. The longer the product is supposed to be, the greater the chance of competing reactions taking over. Usually, people will adjust the Mg++ concentration to influence polymerase behavior or adjust the annealing temp to influence primer annealing and polymerase extension.
Keep in mind that when the sample temperature is raised from the annealing temperature (say 55) to the optimal extension temperature (say 72), the oligo would normally melt off of the template. The reason primer extension works is the polymerase slowly extends the oligo during annealing and while the temperature is rising so that when the 72 degree mark is hit, the extended product is long enough to keep the primer attached and the polymerase happy. Therefore, oligo composition, the type of polymerase, the annealing temp, and ramp time can have significant effects on the reaction.
Take the following example: Each reaction had 1 μL of the same mini-prep, and all reactions were done with the same program at the same time. Three different primer pairs were used and three different enzymes. I have found that the surest way to get a successful 'Horn reaction is to set up the reactions with different enzymes (historically Deep Vent and Turbo Pfu, but now an array of fusion enzymes) rather than to set up arrays of Mg++ concentration or a gradient of annealing temps. The desired product is marked with an arrow. Note that the different enzymes gave varied performance simply depending on the primer pair used. Consider the cost of your time repeating steps relative to the cost of the enzyme aliquot. We now use NEB's Q5 enzyme with excellent results, even on >10 kb plasmids.


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Men's Shirt 12 (40)
Please read all instructions before you purchase items.
1. Follow the link in the purchase button in the box to the right.
2. Add item to your cart and then proceed to the check-out or add additional items if you wish to then proceed to the check-out.
3. Login to your Amazon account or create an account.
4. Add a new address. (Note: Our Ongole Homes have a different address than our Hyderabad Homes, if you are buying items for children in both locations you will need to make two separate purchases.) Send this gift to the address below:
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Military Colony 3rd Line
Mongamuru Road
Ongole, Andra Pradesh 523001 India
Phone Number: 889-772-1039
5. Continue to Checkout, select delivery speed (keep in mind that you should plan for all gifts to arrive prior to Dec. 20 to ensure they arrive in time for Christmas.)
6. Add payment and confirm purchase.
7. IMPORTANT: After you complete your purchase, be sure to fill out the Christmas Gift Purchase Form below to let us know you purchased items, otherwise we will not know that an item has been purchased until it shows up on our doorstep.
8. Once your receive a confirmation email or shipping emails please forward them to email@example.com so we can keep track of all packages headed our way and so we can thank you!
Men's Shirt 12 (40)
Sarah's Covenant Homes
Military Colony 3rd Line
Mongamuru Road
Ongole, Andra Pradesh 523001 India
Phone Number: 889-772-1039


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Tips and Tricks for Overheating |
To cool down an engine on the verge of overheating, turn OFF the air conditioner and turn ON the heater full blast. This will help dissipate the heat from the engine. Stop the car if this doesn't cool down the engine. This is not a permanent fix. Visit our shop and have the problem permanently fixed. |
Avoid overheating by releasing load on your engine. If you are climbing a long hill, turn off the air conditioner or shift into a lower gear. This will allow you to get over the hill without additionally straining your engine. |


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