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November 02, 2012
Airdrie family deserves fair investigation into dropped sex assault case: Anderson
EDMONTON, AB (November 2, 2012): After having questions about a dropped Airdrie sex assault case during question period heckled by several PC members and being cut off by the PC Speaker, the Wildrose Official Opposition is pleased the government may be changing their tune by promising an investigation into why the case was thrown out.
However, Wildrose MLA for Airdrie Rob Anderson said comments made in the media from the Alberta Justice department appear to be pre-judging the investigation's results. The victim's family was told the case was thrown out due to a lack of resources that caused administrative delays, but a ministry official is today quoted as saying that is "categorically" not the reason. "We're pleased the government has agreed to investigate the matter but it looks as though their mind is already made up," Anderson said. "This victim and her family were already denied their day in court. At the very least, they deserve a fair and unbiased investigation as to why, and the Alberta public is entitled to a full report." The assaults on the victim began when she was nine years old and continued over several years. In 2009, a man was charged with sexual assault in connection with the attacks, but a judge threw the case out last month because it took too long to get to trial. Anderson attempted to ask about the case in the Legislature yesterday but was initially shut down when too many PC MLAs objected to the questions and Speaker Gene Zwozdesky inexplicably cut off Anderson's microphone in response. Anderson was eventually able to ask the questions. The Speaker told Anderson he did not need to meet with him after question period to discuss the incident as the Speaker had stated he would during question period and the PC MLA who interjected during the exchange apologized to Anderson after QP. "I hope we can all co-operate now to help this family get the answers they deserve," Anderson said. "This shouldn't be about politics - the fact is these are tough questions and the Government needs to answer them. If it makes some of them feel uncomfortable - tough! Imagine how the victim and their family must feel."


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What It Does:
Helps prevent build-up of pool or spa chemicals (chlorine, bromine) and hard water minerals (copper, calcium, and magnesium) while providing ultimate conditioning of hair and scalp. Helps prevent green hair caused by copper. Leaves hair silky soft, manageable, full of shine and free of minerals.
Who Needs It:
Swimmers and spa users exposing their hair to pool or spa chemicals and minerals that can attach on to the hair and scalp and cause dryness, lack of body, damage, discoloration, hair loss, flaky, itchy scalp, and the inability for chemical services to properly process.
How You Use It:
Shampoo with Malibu Swimmers' Action Shampoo. Rinse and squeeze excess water from hair. Distribute Malibu Swimmers' Action Conditioner evenly and massage throughout hair. Leave on for at least one minute. The longer the exposure, the greater the benefits. Rinse with cool water and style as usual.
Key Ingredients and their benefits:
- Ascorbic Acid (Vitamin C) helps protect hair against free radical damage caused by exposure to the sun and air.
- EDTA is a chelating agent that helps lift minerals.
- Vitamin E Acetate helps protect hair against free radical damage, moisturizes hair and scalp tissue.
- Panthenol penetrates the hair shaft to protect hair's strength and moisture.
- Botanical extracts provide therapeutic benefit to hair and scalp tissue.

subset: commoncrawl


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Product Spotlight
Featured Research
Quick Stats
You are here
SeaChoice, FCL Forge Sustainable Seafood Partnership
Western Canadian grocery wholesaler Federated Co-operatives, Ltd. (FCL) and SeaChoice, a Canada-wide sustainable seafood program, have joined forces to make it easier for the co-op's shoppers to buy seafood sourced from environmentally friendly fisheries and aquaculture operations.
Western Canadian grocery wholesaler Federated Co-operatives, Ltd. (FCL) and SeaChoice, a Canada-wide sustainable seafood program, have joined forces to make it easier for the co-op's shoppers to buy seafood sourced from environmentally friendly fisheries and aquaculture operations.
Saskatoon, Saskatchewan-based FCL, which is owned by about 260 retail co-ops throughout western Canada, will work with SeaChoice to assess the co-op's seafood supply against sustainability criteria, enhance sustainable seafood procurement, and educate employees and customers on sustainable seafood options.
According to Bill Wareham, SeaChoice representative and marine conservation specialist for the David Suzuki Foundation, the partnership with FCL helps the company "take action that promotes ocean-friendly seafood and minimizes sales of products that harm the marine environment. The future of our oceans depends on initiatives like this that help protect wild fish stocks and the web of life in the marine environments."
Under the new commitment, FCL and SeaChoice will develop and implement a six-point sustainability plan for the procurement and sale of seafood in its stores. The retailer has pledged to offer customers sustainable seafood options and reducing the procurement of unsustainable seafood; provide transparency and traceability information on seafood products sold at its co-op stores, collect and share information on seafood sustainability procurement and sales; educate its employees, suppliers and customers on sustainable seafood; and urge policymakers and seafood producers to improve and develop laws, regulations and certification systems supporting the production of environmentally friendly seafood.
As part of its early efforts to adopt a sustainable seafood policy, FCL will work to identify seafood choices for its customers that meet the SeaChoice "Best Choice" list criteria, which is available online at www.seachoice.org.
Other retailers working with SeaChoice include Langley, British Columbia-based Overwaitea Food Group (OFG), which runs 117 stores in western Canada.
A national initiative operated by the Canadian Parks and Wilderness Society, David Suzuki Foundation, Ecology Action Centre, Living Oceans Society and Sierra Club BC, working in collaboration with the Monterey Bay Aquarium's Seafood Watch program, Vancouver, British Columbia-based SeaChoice provides science-based sustainability assessments of seafood to Canadian consumers, fishermen, chefs and businesses.


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Yellow House Land Company:
An Inventory of Its Records, 1912-1943, at the Southwest Collection/Special Collections Library
Biographical Sketch
This trust estate, an agency of Malcolm Hiram Reed, organized in 1923 to purchase 300,000 acres of land in Hockley and Lamb counties of Texas from the heirs of George W. Littlefield. The land was then resold in small plots to farmers. It is the successor of the Littlefield Lands Company. Like other land developers their company helped to settle the West Texas region, which eventually prospered through the decades.
Scope and Contents
The collection contains legal materials, financial materials, business correspondence, and minutes of the Yellow House Land Company. The bulk of the collection is records of closed sales from property sold in Hockley and Lamb counties.
Access Restrictions
Open for research
Related Material
Administrative Information
Preferred Citation
Yellow House Land Company Records, 1912-1943, Southwest Collection/Special Collections Library, Texas Tech University, Lubbock, Texas
Collection accession #(s):
Gift, 1958-1965
Processed by
Abel Ramirez, April 2004
Additional Sources
Collection Inventory


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Card Name: Mondronen Shaman
Mana Cost: {3}{R}
Colors: Red
Type: Creature - Human Werewolf Shaman
Types: Creature
Sub-Types: Human,Werewolf,Shaman
Rarity: Rare
Artist: Mike Sass
Number: 98a
Power / Toughness: 3/3
Card Text
At the beginning of each upkeep, if no spells were cast last turn, transform Mondronen Shaman.
Sorry, it looks like your browser doesn't have cookies enabled. Dave & Adam's only uses cookies to keep track of your shopping cart while you browse dacardworld.com. Please enable cookies to further enjoy your shopping experience. If you have cookies enabled, please refresh your browser.


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Facebook's New 'Reaction' Buttons are Finally Here! [VIDEO]
Facebook is debuting their new Reactions like buttons.
Facebook is debuting their new Reactions like buttons.
We've been asking for it for years, and now we're finally getting it.
Frustrations over Facebook's lax privacy policies have risen quite high in recent times, which makes the fact that someone was able to hack the social site and make public some of Mark Zuckerberg's private photos all the more satisfying.
This video is hilarious...and a little AWKWARD too... as Mark Zuckerberg meets Jesse Eisenberg for the first time. Video after the jump.


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Here's how you can install Pepper Flash Player 12.0 in Ubuntu 14.04.
1. Open a terminal window.
2. Type in the following commands then hit Enter after each.
sudo apt-get update
sudo apt-get install pepflashplugin-nonfree
sudo update-pepperflashplugin-nonfree --install
Hello Linux Geeksters. As you may know, Pepper Flash is a version of Adobe Flash Player, specially developed for Chromium, which is open-source, and so, can't get the Flash Player API implemented by default, due to proprietary licence issues.More at Linux G
Additional Ubuntu Resources
This article is published as part of the IDG Contributor Network. Want to Join?


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Nonconventional soil additives
Every year in Iowa, products are touted to producers as being the cure for crop production and economic woes. The old adage states "If it sounds too good to be true, then it probably is." How do you know whether a particular product is a viable fertilizer and supplies crop nutrients, or has some proven effect on soil that will improve productivity? The standard advice is to demand unbiased research results that document all claims, and to discount testimonials. If well-documented research isn't available, then be suspicious of claims.
In Iowa, there are laws and rules administered by the Iowa Department of Agriculture and Land Stewardship (IDALS) for review and register of fertilizer and soil conditioner products. So your first question should be, "Is this product registered for the claims made?" Check with the IDALS Feed and Fertilizer Bureau to find out by contacting Terry L. Jensen at (515) 281-8599 or email@example.com.
According to the Iowa Fertilizer Law (Chapter 200), the definition of fertilizer is "any substance containing one or more recognized plant nutrient which is used for its plant nutrient content and which is designed for use and claimed to have value in promoting plant growth …" Fertilizers have a guaranteed analysis that is "the minimum percentage of plant nutrients claimed and reported as total nitrogen (N), available phosphorus (P) or P2O5 or both, soluble potassium (K) or K2O or both." According to the Fertilizer and Agricultural Lime Rule (Chapter 43), specific additional plant food elements beside N, P, and K can be guaranteed: calcium, magnesium, sulfur, boron, chlorine, cobalt, copper, iron, manganese, molybdenum, sodium, and zinc. There is a minimum analysis (Table 1). For mixed fertilizers, the sum of the guaranteed analysis of total N, available P2O5, and soluble K2O must be 20 percent or greater. Thus, materials below these levels cannot be registered as a fertilizer (this rule does not apply to specialty fertilizers for nonfarm use or materials ordinarily applied directly to plant foliage).
Soil conditioners
According to Chapter 200, the definition of soil conditioner is "any substance which when added to the soil or applied to plants will produce a favorable growth, yield or quality of crop or soil flora or fauna or other soil characteristics, other than a fertilizer, recognized pesticide, unmanipulated animal and vegetable manures or calcium and magnesium carbonate materials used primarily for correcting soil acidity." According to Chapter 43, product claims may be substantiated by one of two methods: 1) efficacy testing or 2) available research data relevant to Iowa crops and soils. Efficacy testing information includes the following:
Fertilizer and soil conditioners submitted for registration may be required to be tested for a minimum of two growing seasons in at least three Iowa crop reporting districts. The results of testing are reviewed by the secretary's pesticide and fertilizer advisory committee.
The bottom line is to ask whether a product being promoted is registered with the state, and if it isn't, then to question why and check with IDALS. Also, registrations can be reviewed to determine that a product meets the claims for which the registration was originally granted, and registrations can be cancelled if evidence exists that fraudulent or deceptive practices have been used.
If you want to access research results on specific products, the North Central Region Experiment Station committee NCR-103, "Non-Traditional Soil Amendments and Growth Stimulants," publishes a compendium and two supplements, titled "Compendium of Research Reports on Use of Non-Traditional Materials for Crop Production." These publications are available from the Iowa State University Iowa State University Extension Distribution Center by calling (515) 294-5247 or via email@example.com, http://www.extension.iastate.edu/pubs, or county extension offices. Also, this committee compiles a listing of "Nonconventional Soil Additives: Products, Companies, Ingredients, and Claims," which is available on the Web at http://www.soils.wisc.edu/extension/hottopics/index.htm. Laws and rules administered by the Feed and Fertilizer Bureau of IDALS can be found at http://www.agriculture.state.ia.us/FertLawsRules.htm.
In summary, be wary of products with claims that seem extraordinary relative to common wisdom and known research. In Iowa, we are blessed with highly productive soils that have many attributes to support good crop productivity. It is the rare soil amendment that might further enhance the natural soil attributes. Supplementing the soil supply of crop nutrients when needed or replacing nutrients removed through crop harvest with guaranteed analysis fertilizer products, manure, or liming materials is a well-documented and research supported activity. Attempting to change soil through soil amendments for common field crops typically is not.
Table 1. Minimum percentage of nutrient content for fertilizers to be offered for sale, sold, or distributed in Iowa.
Adapted from Chapter 200 and 43, IDALS.
This article originally appeared on pages 39-40 of the IC-490 (5) -- April 21, 2003 issue.
Updated 04/20/2003 - 1:00pm


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Israel-News Today
Week in Review
Slowly, the peace moves forward (HA´ARETZ NEWS) By Amos Harel 02/01/05)
Source: http://www.haaretz.com/hasen/spages/534407.html
HA'ARETZ} NEWS SERVICE Articles-Index-Top
At the changing-of-the-guard ceremony for the head of the Central Command in Jerusalem on Sunday, many of the Israel Defense Forces officers in the audience could be seen keeping their eyes somewhat nervously on their pagers. The frequent glances were not the result of an abundance of operational messages, but the very opposite - the pagers were uncustomarily quiet.
Sunday saw just a solitary shooting incident throughout the territories, and those who had become accustomed to an incessant stream of reports since the collapse of the first hudna (cease-fire) in August 2003 are going to need some time to adjust to the fall-off in the extent of the incidents.
The new arrangement in the territories is supposed to stabilize this week: The Palestinian security forces have completed their deployment in the Gaza Strip; Israel and the Palestinian Authority are discussing handing over control of a number of West Bank cities to the Palestinians; and the IDF is already conducting itself in keeping with new game rules, as defined by Chief of Staff Moshe Ya´alon - a halt to offensive operations in the Gaza Strip, and restricting such operations in the West Bank to dealing with "ticking time bombs."
Nevertheless, the localized outburst of violence in the Strip yesterday - the killing of a 10-year-old Palestinian girl, and Hamas´s response of shelling Gush Katif settlements - testifies to the fact that the process is still in its infancy.
A visit at the beginning of the week to two sectors in the territories - Hebron and the northern Gaza Strip - gave a clear picture of the caution with which the IDF is approaching the new situation, and the limited trust it places in the new-old Palestinian partner.
To the letter
Ya´alon´s directives, as far as can be discerned, are being implemented to the letter. Since Saturday security forces have made just one arrest in the territories. Nevertheless, security cooperation between the sides is just kicking off. There has yet to be a return to the pre-conflict days when Israel would pass on names of suspected terror attack planners to the PA, which would then rush to apprehend them.
In the West Bank, at least, the IDF will be the one that deals, when necessary, with the ticking time bombs. And when it comes to Gaza, only specific moves are being coordinated.
A few days ago, the IDF located a Qassam rocket ready for launching in Beit Hanun; the PA picked it up too, and a Palestinian policeman dismantled the launcher. The impression is that the IDF has a counterpart to work with in Gaza: Musa Arafat. In the West Bank, however, it doesn´t - and in both areas, neither the IDF nor the Shin Bet security service has enough trust in the Palestinian partner to provide it with substantial intelligence.
The IDF continues to man positions in the broad corridors its established around the access roads to the Gaza settlements, and soldiers in Hebron are still at posts set up in PA neighborhoods and on hills overlooking settler homes in the city.
On the other hand, the IDF is no longer sending patrols deep into PA territory in Hebron, and there is a definite increase in the presence of Palestinian security forces, albeit unarmed, in the city. The relative quiet of late can be seen as proof that only the Palestinians can do it: The force applied by the IDF during the course of the conflict wasn´t what reined in the terror; it took an internal Palestinian decision.
Ya´alon presented an alternative explanation: At the ceremony on Sunday, he said "our ongoing war on terror led the other side to take a decision to end unilaterally the terror." (© Copyright 2005 Haaretz. 02/01/05)
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I live in canada and require a car that can handle snow and ice, which is another reason some of my favorite cars are a no go for me.
Just if anyone can recommend any cars and broden my scope of cool vehicals that would be cool I'm open to some very new ideas....
I love E30 BMW's but im not sure ill be able to afford one in excellent shape.
These cars aren't prospects as i cannot afford many but just to give you all an idea of the style of cars I like:
Honda/Acura NSX,
BMW E30 325e,
Honda Prelude,
Nissan Skyline,
Mazda rx7
Volvo C70s


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The State Government has a vision for building a stronger South Australia, with manufacturing remaining crucial to the State's economy.
The State Government is committed to supporting automotive component supply chain companies and workers affected by the closures of Holden, Ford, and Toyota.
Through the Automotive Transformation Taskforce, services are available to help workers find new careers and to support businesses to transition into new markets.
- Under the Automotive Workers in Transition Program, eligible workers can access career and training advice, formal recognition of skills, and financial assistance for training, vocational licenses, or travel to get to a training or new work location.
- Spouses/partners of eligible workers can also access career and training advice.
- Under the Automotive Supplier Diversification Program, eligible businesses can access funding for corporate mentoring, diversification strategy and market development, and retooling, for example.
Leadership and expert advice to the Taskforce is provided by the Automotive Transformation Taskforce Board which guides the delivery of programs and services.
For more information contact
Automotive Transformation Taskforce
Department of State Development
Phone 1800 774 010
Email email@example.com
GM Holden
If you are a Holden employee in South Australia, you can email the Holden SA Transition Team email@example.com for more information on the support available.
A Transition Centre is located on site at Elizabeth and is open to GM Holden and automotive component supply chain workers to seek further advice and support on future job pathways.
The State Government is working with GM Holden to ensure the Elizabeth production site remains an important contributor to the state and regional economies.


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This Forum has been archived
Forums: Admin Central Index → Technical Help → Disable image name on hover
Central's forums are a place for the community to help other members.
To contact staff directly or to report bugs, please use Special:Contact.
To contact staff directly or to report bugs, please use Special:Contact.
Note: This topic has been unedited for 423 days. It is considered archived - the discussion is over. Do not add to unless it really needs a response.
I'm making a sort of "puzzle" for my users that I will send out periodically, which they have to solve (each image makes one letter, for an example see the Sandbox that spells "WILL"). However, when you mouse over each image, it reveals the answer. I would like a way to disable this just for one page (I don't want it disabled for the whole wiki). Iggyvolz 23:32, July 5, 2012 (UTC)
- The image names give away the answer. Your only option is to rename them. -- pecoes 12:13, July 06, 2012 (UTC)
- The problem is, that is how the template works. I guess I could take a picture of the whole thing and use that. Iggyvolz 13:30, July 6, 2012 (UTC)
- That's the only solution, I'm afraid. At least here at Wikia. -- pecoes 13:45, July 06, 2012 (UTC)
- Matthew2602 posted on my wall that he could make a solution, and I can't post on Message Walls due to a glitch. If he's seeing this, I would be happy to have a script. Iggyvolz 17:28, July 12, 2012 (UTC)


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Take CableTV.com's Advice and Make DIRECTV Your Next TV Service Provider
Are you looking for the best TV service provider in your city? Independent consumer surveys collected data to help you evaluate the cable companies and other television services available in your area. In fact, this data shows that DIRECTV beat out all the other satellite TV and cable companies in Salisbury, NH and has been ranked as the top choice.
When you sign up for TV service, most major networks are included with your package, regardless of your location. Unfortunately, that isn't always the case with smaller channels. TV packages in Salisbury typically feature different channel offerings, even when compared to the same plan in a different city. So cost isn't the only factor you need to think about when looking for the best cable company in Salisbury, NH—you also need to consider channel options. Choose a premier package and you'll enjoy all the local channels you love, along with major networks and plenty of New England Patriots and coverage. To make your decision easier, we've compiled information on all the networks and special features offered by the highest rated providers available in Salisbury.
CableTV.com can help you find the cable company in Salisbury, NH that satisfies your needs. Our ratings go beyond information about packages and pricing—they also reveal which TV providers have the most reliable service and the fastest, friendliest customer support.


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Interim Dean Greg Maggs and the GW Law Alumni Association invite you to join us for a complimentary reception for alumni and friends of the Law School hosted by Gary Rovner, JD '95. Professor Jonathan Turley will also be in attendance to visit with alumni and guests.
Foley & Lardner LLP
321 North Clark Street
Suite 2800
Chicago, IL 60654
RSVP by Monday, April 15th


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I told everyone I'd be writing about this in my last article. You all owe me $5.
In today's time and age, there is one thing that I am frustrated over time and time again. I may write about politics a lot, but it's all normal, just heightened activity. I'm not frustrated by school, or work, or social life.
Ok, a bit of social life.
It's not so much my personal social life, but it's society in general and their reaction to social media. Being a thoughtful, intelligent member of society, I'm often aware of my friends and family members reacting similarly to me, but to different objects.
For instance, I react to political and world news. My mother reacts to world news. My father, to business and economic news. My sisters react to Justin Bieber.
When I'm actually surfing the celebrity sites, which is rare, most of the time the stories posted are on the same celebrities, over and over, almost in a creepy, stalking way. Twitter is the worst, because it's always trending the same #topics. Every day.
Exhibit 1? Charlie Sheen. He's popped into the celebrity news like Trump has in the political area. He's the ultimate example of today's media and population reacting and stalking the person with the most moronic statements. It's what we are about today. A vast majority of my friends follow the media-whoring morons: Sheen, Lohan, Bieber (who isn't a moron but still a media whore).
I feel like the media has really lost touch with humanity. It's skewed our priorities to make money. The media today has taught us to pay attention to the stupid people, the ones coming back for seconds, for thirds, for two-hundreths. It's pushing the things that aren't really important into center stage.
If Michael Jackson had died on the day Tunisia overcame their dictator, what do you think would have been the center story of the day? People today would have cared more about Jackson's death than a country's revolution. It's pathetic. Someone dying usually does not alter the course of the world. A country's revolution always does.
So why does the population react to it so much? Twitter reacts to Lady Gaga, not Libya. I know that because I've tried it out, and found it a waste of my personal time. I can't stand those who think that celebrities are the thing because they are merely human and will never teach us a lesson. Jersey Shore has nothing in terms of material, just a bunch of swearing, abusing morons. America loves it.
I blame the media for the downfall of our integrity, for glorifying the famous stupid people. I blame it for making the population nothing more than a herd of sheep, constantly feeding it things that do not matter.


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Lena Dunham Needs For You To Get Over The Amount of Nudity on 'Girls'
The cast of Girls, which has been renewed for a fourth season, took part in a panel discussion at the HBO portion of the 2014 Winter Television Critics Association tour at the Langham Hotel yesterday in Pasadena, California.
Lena Dunham wasn't all that pleased with one journalist's remark about Dunham's character and her penchant for not wearing clothes.
"I don't get the purpose of all of the nudity on the show … Your character is often naked just at random times for no reason," the reporter pointed out, according to People.
"It's because it's a realistic expression of what it's like to be alive, I think, and I totally get it," Dunham shot back. "If you are not into me, that's your problem, and you're going to have to work that out with professionals."
Producer Judd Apatow quickly came to Dunham's defense, publicly chastising the reporter.
"That was a very clumsily stated question that's offensive on its face, and you should read it and discuss it with other people how you did that," Apatow said from the stage. "It's very offensive."
It didn't stop there. Apatow then asked the reporter if he had a girlfriend, adding "Does she like you?"
Season three of Girls premieres this Sunday on HBO.


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As I look back, I'm convinced that impulsively quitting my job was the single greatest decision I have made in my life so far.
Reckless, stupid, irresponsible, challenging, stressful, frustrating…
…liberating and empowering.
As I look back, I'm convinced that impulsively quitting my job was the single greatest decision I have made in my life so far.
Reckless, stupid, irresponsible, challenging, stressful, frustrating…
…liberating and empowering.
"…it turned out that getting fired from Apple was the best thing that could have ever happened to me. The heaviness of being successful was replaced by the lightness of being a beginner again, less sure about everything. It freed me to enter one of the most creative periods of my life."
I'm not presumptuous enough to actually compare myself to Steve Jobs. I can, though, relate to some of the emotion involved in this change in his life. On January first I chose to leave my job of the last five years and it has been one of the most liberating and empowering experiences of my life. I have no intention of disparaging Apple Retail. I truly believe in the values and vision that were created by Ron Johnson, the mastermind of the Apple Stores' unparalleled success over the last ten years. I have the upmost respect for my peers and all of those who have the courage to be the face of Apple and offer customers a level of service that does justice to the products Apple designs. I miss the opportunity to work closely with friends I've made and the chance to meet customers, to care about their problems, and to share my love for technology and Apple's products. Working as an Apple Store employee has been a defining part of my life, but the time had come for me to move on. Three things have brought me to this. Coincidentally enough, they can each be summed up by a quote from my boss of these years, Steve Jobs.
"I have looked in the mirror every morning and asked myself: 'If today were the last day of my life, would I want to do what I am about to do today?' And whenever the answer has been 'No' for too many days in a row, I know I need to change something."
The first reason I'm moving on is really quite simple. I'm passionate about the products Apple produces, but I'm not passionate about sales transactions, processing returns, answering phones, and various other made-up roles. Again, I have always enjoyed my job; I love the people and the products. Nonetheless, my job as a Specialist was not giving me the types of experiences I would hope to have on "the last day of my life." Apple Retail was always meant to be a temporary solution, a fun way to make a little money—and it has been, but at this point I feel it has been "too many days in a row."
"When you grow up you tend to get told the world is the way it is and you're life is just to live your life inside the world. Try not to bash into the walls too much. Try to have a nice family, have fun, save a little money. That's a very limited life. Life can be much broader once you discover one simple fact: Everything around you that you call life was made up by people that were no smarter than you and you can change it, you can influence it, you can build your own things that other people can use. Once you learn that, you'll never be the same again."
I can't say that my potential can compare in any way to the genius of Steve, but I do believe in the ability of those who are committed enough to really design and create great things. I can relate particularly well to this idea, because this is my passion. Nothing is more satisfying to me than to imagine, design, and create. (As an aside, I don't include maintain on that list, as anyone who has lived with me or seen my bedroom can understand.) The last three weeks I have found purpose to getting out of bed. I have been able to fill my days with interesting, motivating, and productive activities. It's certainly not easy; The significant amount of extra responsibility to manage myself and my own success is quite intimidating and at times overwhelming, but I must say it's worth it. This might be a good time to point out that I am fully aware that my efforts and projects may or may not eventually pay the bills. I hope they do. We will see. I may at some point have to come back to "reality" and return to a "real job," but the most important thing for me at this time—win or loose—is to have these experiences, to feel like I can do more than be part of system that is someone else's vision and design. I can have my own vision. "[I] can build [my] own things that other people can use."
The last concept that I can relate to and is attributed to Jobs is the idea that being a pirate is better than joining the Navy. Shortly after I quit, I used some variation of this as my Facebook status and the responses were quite revealing. Some of the comments that were made include, "Navy has better health care," "In the Navy you get better weapons, better food, and a good retirement," "Pirates make more," and "Pirates all the way! ;)"
How important are better weapons, better food, and a good retirement? Time will tell.
Click here to read about my next thing.


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I have a bear rug that I am doing and I am going to send it off to a professional rugger to get the felt sewn on. I was wondering if anyone has any suggestions on who to send it to. I wan't a high quality professional to do it.
thanks gordon
Return to The Taxidermy Industry Category Menu
There are some great ones all around the country so to save shipping, we might be able to help you better if we knew where you were.
The wonderful ladies name is Betty Jackson. They are located in High Ridge Missouri and I beleive their phone number is 314 677 0012. They are awesome at rugging your hides. Good Luck. Dan Hudzik
I've been around the horn a few times and for my money nobody does a better, or as-good a job as BJ's. They get all my work..PERIOD!
Good luck, JL
I thought Betty had sold that. Still, we don't know where the guy is. There is a fantastic one in PA and Brenda Duval of Wisconsin does a great bear as does Mary (Hilyard)Krueger of Arizona.


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Ever wondered what goes on in the minds of LOTR badficcers, and why they come up with their horrible creations? Here's a handy primer.
Category: Lord of the Rings - Rating: R - Genres: Angst, Horror, Humor, Parody - Characters: Aragorn, Arwen, Bilbo, Elrond, Eomer, Eowyn, Faramir, Frodo, Galadriel, Legolas, Merry, Pippin, Sam - Warnings: [!!!] - Published: 2006-10-10 - Updated: 2006-10-10 - 2480 words - Complete
Site design Copyright 2005, K&D Lynch. All stories contained in this archive are the property of their respective authors, and the owners of this site claim no responsibility for their contents.
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ATLANTA - Members of a civil rights group that has pushed for years to reopen investigations into the decades-old lynching at Moore's Ford Bridge near Monroe said Friday they are pleased that the U.S. Department of Justice has decided to take another look.
"You just feel like the rule of law is going to prevail," said state Rep. Tyrone Brooks, D-Atlanta and the president of the Georgia Association of Black Elected Officials.
The group has pushed since at least 1998 for new investigations into killings of two black couples - George and Mae Murray Dorsey and Roger and Dorothy Malcom - in the summer of 1946.
Mr. Brooks said his group has been aware for more than a year that the FBI was looking into the Moore's Ford case again but kept quiet to avoid interfering.
But at a news conference Tuesday, U.S. Attorney General Alberto Gonzales and FBI Director Robert Mueller announced the Justice Department was reopening investigations of some civil rights-era killings, including Moore's Ford.
The Georgia Bureau of Investigation reopened its investigation several years ago on the orders of then-Gov. Roy Barnes.
Mr. Brooks said local activists have kept up with at least five living suspects named in the original FBI report after the killings.
Mr. Brooks' group and local organizations have also been talking to witnesses who have come forward in recent years.
"We're going to be making an appeal to the suspects to turn themselves in and avoid a long ordeal," Mr. Brooks said. "If we know of five, obviously the law enforcement community could know of many more than we do."
Mr. Brooks also said the group is working to boost its reward fund for information leading to the arrest and conviction of those guilty of the murders, currently at $27,000, to $100,000.
The black officials' association also announced its annual April 4 commemoration of the lynchings to coincide with the anniversary of the assassination of the Rev. Martin Luther King Jr. and its July 25 re-enactment of the killings.
Mr. Brooks said one of the living suspects usually drives his pickup across the bridge during the re-enactments.
"He drives across real slow and he'll look at us, and we'll look back," Mr. Brooks said.
Reach Brandon Larrabee at (678) 977-3709 or email@example.com.
The Georgia Association of Black Elected Officials is asking witnesses or those who have information about the 1946 lynchings at Moore's Ford Bridge to come forward.
The association can be reached by phone at (404) 656-6372 and (404) 372-1894, or through its Web site, www.gabeo.org.


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April 09, 2013 at 01:23:14 Apr 09, 2013 @ 01:23
No sus shit, the best fighting moves and all that. Really pissed off that i dont know how to fight, and im ready to humiliate a nigga.
Not reading bruh, talking about videos or just tips or some shit.Wait, you think you can learn how to fight by reading about it? And even then, reading about it on a fashion forum?
Chicks barely dancin, glancin every chance they get, like - oh shit, he's so HANSUM
cosign. all you gotta do is chop that nigga on the jugular notch which is right around that lil dip you got in ya throat areaSon in a fight you ain't even got to fight. Just hit that nigga with a quick throat chop & be on with your day.
When a nigga run up on you, just make your hand as stiff as possible & chop that nigga with the swiftness to finish the fight right there. Just in case you might gotta kick that nigga in the skull when he is grasping for air on the ground.
But always remember the number 1 rule, Violence is never the answer.
He's cute? I bet he ain't #Hansum
Im tryna hurt somebody not kill them.fuck that shit , cop a choppa son .
.wa do dem.
be careful what you pretend to be
1:03 to 1:20 had me dying
Instagram: @gillysuits_
pussyIm tryna hurt somebody not kill them.fuck that shit , cop a choppa son .
yeah bruh cus hes gunna throw his brain at themOriginally posted by Inactive UserYou want to humiliate someone by beating them up but you can't fight?
Use your brain.
bein edgy aint all its cracked up to be
"im a very deep minded person" - Tracy Jackson


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Sky Player, BskyB's online TV service, will shortly become available on Humax's range of Freeview HD set-top boxes. It's already confirmed for a number of other devices.
First to get the web-based TV service is the recently-launched Humax HD-FOX T2. Sky Player will also be available on the forthcoming Humax Freeview HD PVR (personal video recorder) with 500GB hard drive, due out later this year.
Sky Player offers Sky customers the opportunity to access a wide range of live TV channels as well as a video-on-demand service online, with content from a range of broadcasters.
Graham North, commercial director of Humax, says: "The addition of Sky Player on our Freeview HD boxes ensures Humax is positioned to offer consumers an exciting way to experience digital TV, with great content and a range of home networking and content sharing features."
Sky says its web TV player is confirmed for launch on Fetch TV, Cello IP-enabled TVs, 3View and now Humax Freeview set-top boxes.


text: About ProAir ® HFA with Dose CounterProAir ® HFA (albuterol sulfate) Inhalation Aerosol with dose counter is indicated in patients 4 years of age and older for the treatment or prevention of bronchospasm with reversible obstructive airway disease and for the prevention of exercise-induced bronchospasm. In March 2012, the U.S. Food and Drug Administration approved the addition of the dose counter, an innovation designed to help patients, as well as their caregivers, keep track of the number of doses remaining in the inhaler. ProAir ® HFA with dose counter supports the FDA's recommendations for such mechanisms, as a means to track the number of doses remaining. Dose counters reduce the risk of utilizing an inhaler that no longer contains the medication required. The safety and efficacy of ProAir ® HFA was determined through clinical trials comparing it to a marketed active comparator albuterol HFA inhaler and placebo HFA aerosol. In clinical trials of adults and children with asthma, ProAir ® HFA produced bronchodilator responses greater than those observed with a matched placebo HFA inhalation aerosol and comparable safety and efficacy to the active comparator albuterol HFA inhaler. Important Safety Information If your symptoms become significantly worse when you use ProAir ® HFA, contact your doctor immediately. This may indicate either a worsening of your asthma or a reaction to the medication, which may rarely occur with the first use of a new canister of ProAir ® HFA. Either of these could be life-threatening. What to tell your doctor before using ProAir ® HFA: If you have a heart, blood, or seizure disorder, high blood pressure, diabetes, or an overactive thyroid, be sure to tell your doctor. Also make sure your doctor knows all medications you are taking, especially heart medications and drugs that treat depression, because some medications may interfere with how well your asthma medications work. Do not exceed the recommended dose. Side effects associated with ProAir ® HFA included headache, rapid heartbeat, pain, dizziness, and irritation of the throat and nose. You are encouraged to report negative side effects of prescription drugs to the FDA. Visit www.fda.gov/medwatch, or call 1-800-FDA-1088. Please see full prescribing information at http://www.proairhfa.com/pdf/ProAirPrescribingInformation.pdf. About Teva Teva Pharmaceutical Industries Ltd. (NYSE: TEVA) is a leading global pharmaceutical company, committed to increasing access to high-quality healthcare by developing, producing and marketing affordable generic drugs as well as specialty pharmaceuticals and active pharmaceutical ingredients. Headquartered in Israel, Teva is a world leading generic drug maker, with a global product portfolio of more than 1,300 molecules and a direct presence in about 60 countries. Teva's branded businesses focus on CNS, oncology, pain, respiratory and women's health therapeutic areas. Teva currently employs approximately 46,000 people around the world and reached $18.3 billion in net revenues in 2011.


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The always modest, always charming Howell Raines, former executive editor of the New York Times, has a new autobiography out, "The One that Got Away," a sequel to his 1993 memoir "Fly Fishing Through the Midlife Crisis."
Dipping into his latest book on his love of fly fishing, we find that Raines is still rising to the conservative-bashing bait.
On page 189, he lets fly with thoughts about liberal bugbear Fox News:
"Fox, by its mere existence, undercuts the argument that the public is starved for 'fair' news, and not just because Fox shills for the Republican Party and panders to the latest of America's periodic religious manias. The key to understanding Fox News is to grasp the anomalous fact that its consumers know its 'news' is made up. It matters not when critics point this out to Foxite consumers because they've understood it from the outset. That's why they're there. Its chief fictioneer, Roger Ailes, had been making up news in plain sight for a half century."
When it comes to "making up news," Fox has nothing on Raines, who, before he was ignominiously drummed out of the Times' top slot editor slot three years ago, constantly
He puts down his fishing rod and picks up his brass knuckles to go after Fox again on page 242:
"Fox Television showed us the future -- outright lies and paranoid opinions packaged as news under the oversight of Rupert [Murdoch], a flagrant pirate, and Roger Ailes, an unprincipled Nixon thug who had assumed a journalistic disguise in much the same way that the intergalactic insect in Men in Black shrugged into the borrowed skin of a hapless hillbilly."
For more examples of New York Times bias, visit TimesWatch.
pushed a ludicrous personal crusade to get women admitted to Augusta National Golf Club (home of the Masters golf tournament) even while the country prepared for war in Iraq.


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Hilf uns: Erscheint dieser Pornostar in diesem Video?
Pornstar Name:
Does anyone know the name of this fantastic lady? I want to see more of her.
Dat Ass!! It's the most perfect asshole I have ever seen. And girl is gorgeous too. A mans dream come true. Her guy is soo lucky (if she got one that is, but I can't imagine this girl being alone for long) but probably got no clue (as usual)
now that's a real woman, none of that tiny butt plug or 4inch dick anal. She's a true anal queen
I will dowhatever it takes to play with your girl! SAY THE WORD! WHAT DO YOU WANT! I WANT YOU! You have a fansastic ass and I am betting your pussy is just as good! Please let me know you will take my little 10 inch dick!
I wanna fuck your loose ass
Beautiful body!!! Love how you hold that dong with your feet and take it up your ass!!! Would love to play in holes for a while!!!
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It's a bit unfair to tell you about the movies that don't open until Tuesday but you may want plan ahead for Christmas afternoon, when you've opened all the presents and are in desperate need of something to do.
The Christmas Day opening most likely to have folks skipping the roast beast is "Les Misérables," the long-awaited movie adaptation of the beloved Broadway musical. It's the story of Jean Valjean (Hugh Jackman), an ex-convict who has jumped parole and forged a new identity, but still has the law in the form of the dogged Inspector Javert (Russell Crowe) breathing down his neck. Valjean encounters the tragic Fantine (Anne Hathaway), and adopts her daughter Cosette as his own, and then watches the adult Cosette (Amanda Seyfried) fall for the idealistic Marius (Eddie Redmayne), one of the young rebels on the barricades during the Paris uprising of 1852. Alas, director Tom Hooper ("The King's Speech") goes for the human drama, with close-ups tighter than the actors' optometrists would go, and misses the grandeur of the piece.
Somewhat better is "Django Unchained," director-writer Quentin Tarantino's free-form homage to spaghetti Westerns and blaxploitation flicks. Jamie Foxx stars as Django, a freed slave who becomes deputy to a chatty bounty hunter (Christoph Waltz, who steals the show) as he tries to free his beloved wife (Kerry Washington) from a nasty Mississippi plantation owner (Leonardo DiCaprio). Tarantino employs his bag of tricks more judiciously than he did in "Inglorious Basterds." The results are just as audacious and violent, but more human.
Lastly, in every sense, is "Parental Guidance," a family comedy that is hijacked by Billy Crystal's rampaging ego. Crystal and Bette Midler play an older couple who visit their grandkids, and bristle when their daughter (Marisa Tomei) shows them her modern parenting techniques. Crystal plays every scene like a Borscht Belt comic in mid-monologue, with creaky gags and an overbearing presence. Anyone who drags their family to this one deserves coal in their stocking next Christmas.


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Updated 04/01/2014Your profile? Update now
Nick Vaughn, Attorney
444 Enterprise Dr.Somerset, KY 42501
Phones: (606) 677-0597
Fax: 606-677-0598
Vaughn, Nicholas C.A. (Attorney)
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444 Enterprise Dr.Somerset, KY 42501
(606) 677-0597
When viewing a listing, consider the state advertising restrictions to which lawyers and law firms must adhere, as well as our West Legal Directory disclaimers. Some lawyers publish comparative information regarding the services that they provide which may be subject to specific comparative communications restrictions.
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VT Edition Interview: Oliver Goodenough on Internet legalities
12/28/07 1:33PM By Jane Lindholm
| Download MP3
The internet has been described as the wild west of the fast expanding public communications frontier. A libel suit filed just about a month ago against a Brattleboro website could help define the shape of law and order in this new legal territory. The site itself, is a new idea: an online community news website that describes its contributors as "citizen journalists."
Someone made some allegedly libelous comments on the site, about a co-worker's personal life. Now the woman who believes her reputation has been damaged is suing - not only the person who made the online comments but the operators of the website for allowing the comments to be posted.
Oliver Goodenough is a Professor at Vermont Law School and an expert in communications and libel law. He spoke with VPR's Jane Lindholm about the case.


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U.S. Department of Transportation
Federal Highway Administration
1200 New Jersey Avenue, SE
Washington, DC 20590
Federal Highway Administration Research and Technology
Coordinating, Developing, and Delivering Highway Transportation Innovations
The focus of the Long-Term Bridge Performance (LTBP) program is to better understand bridge performance. In order to define and identify performance and performance issues, a study was initiated to synthesize research activities relevant to bridge performance both in the United States and abroad. The study also focused on heuristics (experienced-based), expert opinions, assumptions, and generalizations to provide a clear and comprehensive perspective on the concept of bridge performance.
This study resulted in a draft report titled "Performance Primer," which provides a deeper understanding of the meaning of bridge performance and how it can be reliably measured to improve the performance of the highway transportation system. The primer helped shape the direction of the LTBP program and provided a platform for the types of data needed to measure performance. This report will be available in August 2010.
The Federal Highway Administration (FHWA) LTBP recently hosted a geotechnical workshop called "Identifying Bridge Substructure Performance Issues." Participants represented State highway agencies, FHWA, academia, and consulting. The transportation leaders also included a cross section of structural, geotechnical, and hydraulics engineers.
The purpose of the workshop was to consider overall bridge performance and identify geotechnical performance metrics or indicators that correspond to good and poor performance. As a follow-up to the focus group findings, participants were asked to fill in the gaps since very little information was provided concerning foundations, substructure, and hydraulics. There were three sets of breakout sessions: "Bridge Performance Issues," "Data Needs and Data Gaps," and "Tools, Technology Development and Monitoring." Each session was designed to generate creative thought and advanced solutions for holistic bridge performance. As a result, the LTBP program was provided with recommendations that were collected on how to identify, prioritize, and address substructure and foundation performance issues. A final report on the workshop will be available by August 2010.
The LTBP program aims at a better understanding of the principles underlying performance issues of bridge structures, an understanding that ultimately will lead to a better way of managing the bridge infrastructure by minimizing life-cycle costs and improving current asset management strategies. In order to achieve this goal, during the LTBP program large amounts of data will be collected from different sources including legacy data, detailed visual inspection reports, environmental information, and monitoring or instrumentation data. In order to efficiently manage, organize, and utilize this vast amount of data, the LTBP program is developing an open, scalable, and extensive data management and analysis infrastructure. This data infrastructure integrates LTBP data with other data sets such as NBI, PONTIS, weather, and traffic, thereby providing a single source of information for researchers, bridge owners, and other stakeholders. Once integrated, the data infrastructure should provide ways for querying and mining this vast amount of information to help in better understanding deterioration processes and defining better life cycle cost models.
Users will be able access and query the data warehouse anytime from anywhere via a state-of-the-art Web-based application, the Bridge Portal, to better understand deterioration processes and define life cycle cost models. Users can search for bridge structures based on cross-domain criteria, such as bridge type or age, weather statistics, traffic information, geographic area, and condition ratings from the past 20 years. Advanced visualization tools will provide a map-based user interface and dynamic, graphical presentations of query results. Built in statistical applications will allow users to compare a specific bridge to the average condition ratings of structures of a similar type, age, or location.
The LTBP Bridge Portal is currently being beta-tested. It is anticipated that it will be made public in spring 2011.
In order to have consistency in data collection for the program, another LTBP software tool, the Bridge Assessment Decision Tool (BADT), was created to query data collection protocols. In the initial phase of the LTBP program, commonly available and proven bridge health monitoring, nondestructive evaluation (NDE), and inspection techniques and data collection protocols that will be used throughout the program have been identified and developed. Data collection protocols will be tested, verified and refined in the pilot study phase of the LTBP program. The focus of these protocols has been on high priority data collection methods, which are techniques common to all the pilot bridges.
The BADT houses the data collection protocols in a database and makes connections between the specific measurement and evaluation techniques, general types of techniques (i.e., visual inspection, NDE, monitoring), data types, types of deterioration, and bridge components. A Web-based application then allows users to run queries using these connections as search criteria. A query returns the protocol sheet for a given measurement technique, including a detailed description, equipment required, technique procedure, suggested frequency of use, and output data.
The BADT was created to ensure consistency in the data collection throughout the LTBP program. Initially, it will be utilized by the LTBP contractor. However, it is anticipated that the BADT will be made public at the end of the pilot study in 2012.
A LTBP pilot study phase of work got underway in September 2009, to validate and refine the methods, protocols, and guidelines—developed in the first phase of the program—for gathering reliable information and to ensure consistency in the data that will be collected in the long-term phase of the program. The experiences and results from inspecting, instrumenting, testing, and monitoring the pilot study bridges will be a major determinant in the final design of the various LTBP experimental studies.
In addition to assessing the effectiveness and usefulness of the chosen data protocols, the pilot studies will provide critical knowledge on: difficulties in accessing components of the bridge, maintaining safe and satisfactory traffic flow during onsite activities, realistic costs associated with all aspects of the field testing, key factors in coordinating with the bridge owners, issues related to durability and reliability of instrumentation, and data transmission methods.
The seven pilot bridges are being chosen to represent a cross-section of the most common superstructure types and environmental conditions seen throughout the United States. These bridges are located in California, Florida, Minnesota, New Jersey, New York, Utah, and Virginia, creating a broad geographic distribution. Selection criteria are also established to ensure that as many variables as possible governing bridge performance can be examined during the pilot phase. Thus far, detailed inspection of pilot bridges in New Jersey, California, Utah, and Virginia has been completed. All are highway overpasses typical of the bridges that the LTBP program will focus on. The detailed inspection of the three remaining pilot bridges in Minnesota, New York and Florida is expected to get underway in May 2010. The pilot study phase is expected to continue through 2011.
The first issue of the LTBP Newsletter will be available in spring of 2010. There will be four issues annually. To receive a copy, please send an e-mail along with your affiliation to www.fhwa.dot.gov/publications to Hamid Ghasemi: email@example.com
The LTBP program and its current activities will be featured at the following conferences. [More]
|Office of Infrastructure R&D
|Infrastructure R&D Program
»Long-Term Bridge
Performance Program
»Long-Term Pavement
Performance Program
|Infrastructure R&D Experts
|Infrastructure R&D Laboratories
|Infrastructure R&D Projects
|Infrastructure R&D Publications
|Infrastructure R&D Topics
|FHWA Launches Flagship Initiative to Collect Nationwide Data on Highway Bridges
|FHWA Deploys LTBP Program
|National Bridge Inventory
|AASHTO Bridges Subcommittee
|Strategic Highway Research Program 2
Robert Zobel
LTBP Program Coordinator
E-mail email@example.com
Yamayra Rodriguez-Otero
LTBP Development and Outreach Engineer
E-mail email@example.com
Turner-Fairbank Highway Research Center
6300 Georgetown Pike
McLean, VA 22101
|Driving Directions/Map Click here for directions and a map to theTurner-Fairbank Highway Research Center facility.


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Boletellus russellii Russell's Bolete
The cap surface tends to break up into patches. The pores are large, yellow and angular. Deep flutes run down the stalk giving it a shaggy appearance.
Caps are 3-9 cm across, dry, felty to scaly, convex, light-brown to ochre, and sometimes breaking up into patches. Flesh is yellowish, not staining. Tubes are cream to greenish yellow, and not staining. Pores are large and angular. Stalks are up to 20 cm tall by 2.5 cm wide, red-brown, and with deep longitudinal flutes forming a shaggy, raised network. Spore prints are dark brown. Widespread and not uncommon, it fruits under hardwoods especially oaks. Edible.


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Radically Ordinary
The ordinary Christian life is not the opposite of the radical Christian life. The ordinary Christian life is a radical life. The ordinary Christian life is a life of daily trusting Christ; daily repenting of our sins; daily abiding in …Read More
Every Jot and Tittle
Although we don't like to admit it, the reason many of us don't read the Bible regularly is because we are afraid of it. We are afraid of the Bible because we are ignorant of the Bible. Many …Read More
In Christ
Repetitio mater studiorum est. "Repetition is the mother of all learning." The Apostle Paul understood this. Under the inspiration and superintendence of the Holy Spirit, Paul constantly repeated the foundational truths of biblical doctrine, and he did so not only …Read More
Church Growth and the Sovereignty of God
It seems that every time I meet a pastor from another church, he asks me the common, unsolicited, ecclesiastical question of the twenty-first-century: "How big is your church?" Most pastors are usually a bit confounded when I respond …Read More
Uncontrollable Anxiety
In the middle of writing my column this month I deleted what I wrote and have started over because I just received word from one of my closest friends that his wife, pregnant with their long-awaited second child, might be …Read More
Ten Years Later
The world has changed. We are not the same people we were on September 10, 2001. The events of September 11, 2001, and the events that followed in ensuing years have not only changed America but nations and peoples throughout …Read More
Living Life CORAM DEO, Before The Face Of God
As a publication of Ligonier Ministries, Tabletalk exists to equip Christians to articulate what they believe and why they believe it. It is our foremost desire to awaken as many people as possible to the holiness of God by proclaiming …Read More
Proclaiming Life to Captives
I am writing this on the twenty-fifth anniversary of President Ronald Reagan's release of the "Personhood Proclamation." On January 14, 1988, Reagan released the following declaration: "America has given a great gift to the world, a gift that drew …Read More
Telling the Truth
Nearly forty years ago, in his book The Church at the End of the 20th Century, Francis A. Schaeffer penned the following: "Does the church have a future in our generation? …I believe the church is in real danger. It …Read More
A Life of Faith and Forgiveness
If you travel to Wittenberg, Germany, the birthplace of the Protestant Reformation, you may find yourself scratching your head wondering how Martin Luther managed to nail his 95 theses to the solid-bronze door of the 500-year-old castle church. It wouldn …Read More
The Wisdom of Listening
Wise men are difficult to find. We must go to great lengths - we must search high and low in order to find a man who is truly wise. I grieve for my children and my children's children as I …Read More
Preserved by God
Winston Churchill is often quoted as saying, "Sure I am of this, that you have only to endure to conquer. You have only to persevere to save yourselves." Considering what he accomplished in his life, such a statement is …Read More
Relevant, Old Paths
My dad was fifty-two years old when I was born. When I was thirteen, he asked me if I was embarrassed that he was so much older than my friends' dads. I told him I wasn't embarrassed but that …Read More
The Heart of Words
Words are powerful. They transform lives and make history. They birth nations and topple empires. They make peace and fuel wars. They make covenants in marriage and wound those we most cherish. They change hearts and give news of eternal …Read More
Does Jesus Got You?
In 2003, the International Dairy Foods Association celebrated its tenth anniversary of the "Got Milk?" advertising campaign. During the past ten years, just about everyone from Michael Jordan to the Cookie Monster has been featured wearing a milk mustache …Read More
Burk Parsons
Dr. Burk Parsons is copastor of Saint Andrew's Chapel in Sanford, Fla., a Ligonier Ministries teaching fellow, vice president of publishing for Ligonier Ministries, and editor of Tabletalk magazine. He is an ordained minister in the Presbyterian Church in America and director of the Church Planting Fellowship. He is on Twitter @BurkParsons.


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Get Pet Questions Answered by Pet Experts
My name is XXXXX XXXXX I am a Vet tech who works with all times of Small animals and Wildlife. I do need to ask a couple of questions.
Do you know if Chi Chi ingested any part of the tube?
Was the tube metal or plastic?
Did she just chew through the tube or do you think she ate a large amount of the Ointment?
What the formula the type with pain killer or without?
Hi Joan,
She may have ingested part of the tube. I did find small pieces of it on my counter but it's hard to tell if she actually ate any of it.
The tube is plastic on the outside but is silver on the inside. I'm not sure if that means that it's metal.
It looks like she ingested some of the ointment. There is a good amount gone from where she was chewing through the tube. Possibly up to a teaspoon.
It was the original ointment.
Hi Pamela,
The Neosporin itself is not toxic and it is used for minor wounds. It will cause gastrointestinal upset and diarrhea if too much was ingested. In a Squirrel that young it can get serious because of dehydration. I would be prepared with some Pedialyte and fluids if needed. As far as the tube if she chewed it and swallowed pieces of the tube, that can also be a problem, because depending on the size of the pieces they could cause an intestinal blockage or perforation passing through the intestinal tract. I do suggest contacting a Vet for supportive care at this point. My concern is not toxicity, but a possible blockage from pieces of the tube and diarrhea leading to dehydration. The Vet may want to give you something for diarrhea and shhot an X ray to see if the pieces of tube will pass. In the mean time watch the stools carefully for any signs of blood in the stool from the pieces of the tube and monitor for the onset of diarrhea. It may not hurt to start Chi Chi on some Bene Bac which is a Pro biotic to help with gut flora and please make sure you giving a good calcium supplement and getting Chi Chi into the sun for UVB rays. Please let me know if you have further questions or need help. My goal is to assist you in helping Chi Chi. Joan
My regular veterinarian won't be in his office until tomorrow morning at 8:00 am. Do you think it's ok to wait until then? So far, she seems ok. I'd rather not take her to an emergency veterinarian, as most of them don't deal with squirrels. My vet is a wildlife rehabilitator and very good with all kinds of animals, especially exotics. I have pedialyte on hand from when we first rescued her, so I can give it to her, if needed. How long after ingestion do you think the diarrhea will start? Is it possible that she may not even get diarrhea?
She should be ok until tomorrow as long as she does not start with the diarrhea or appear to have an gastrointestinal pain from pieces of the tube. She may not have ingested enough to cause problems, but I need to make you aware of the worst case scenario and hope it does not come to that. I am hoping that she did not ingest too much, since it does not taste good. In any case vigilance will be important with her and if she does seem in distress, I would try to reach your Vet as most Vets that do rehab will come in to the office for an emergency. If you cannot reach him then the ER Vet would be needed. For now monitor her and you can even offer some Pedialyte to help flush her system. Joan


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Ready for another community mind-teaser? How do seemingly different concepts – namely, preservation and change – intertwine seamlessly at the birthplace of Orange County?
Luckily, you don't have to stretch your imagination for this one. The answer is right before your eyes every day along El Camino Real. With consistent support from many volunteers and others dedicated to preserving our historic landmark, Mission San Juan Capistrano is redefining and re-creating visible magic while maintaining its historical purpose.
California history comes alive and preservation efforts continue to progress, as the new gatehouse opening symbolizes. Additional traditions continue throughout the year. Looking ahead, the summer will see the always-anticipated return of the Music Under the Stars concert series. Even this seasonal musical celebration provides funding for preservation of our city's treasure.
Coming this month are more enhancements to a visit to the Jewel of the Missions as the famed swallows return to Capistrano:
• March 19: St. Joseph's Day and Return of the Swallows Celebration: 10 a.m. to 2 p.m. on the mission grounds, including ringing of the historic bells, live music, community presentations, performances, history of St. Joseph's Day and the swallows legend. Lecture from 2 to 3 p.m. by Charles Brown, professor of biological sciences at the University of Tulsa, Okla., who is considered the preeminent expert on swallows and their migration patterns.
• March 23: Mariachis at the Mission: 2-4 p.m., after the Swallows Day Parade downtown.
Find time to walk the mission grounds, feel the special aura and the melding of old and new. But while you're soaking in the wonders, give some thought to the old adage, the more things change, the more they stay the same. It's not just another oxymoron.
More information: www.missionsjc.com or 949-234-1321.
Robert Manniello is a medical doctor who is a longtime contributor to the Capistrano Valley News. You can contact him at firstname.lastname@example.com.


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Lost in spice – easy tomato and egg brunch
This looks like your standard eggs and tomato dish right? But there are lots of flavours going on in here and the end result is a super tasty, incredibly easy brunch or even supper. We have been travelling quite a lot during these school holidays and every time we come home this is the kind of food I crave, easy to put together, simple and homely. I add lots of chili and a squeeze of lemon juice to mine, and if it's handy, half an avocado too.
2 tbsp olive oil
1 brown onion, finely sliced
3 cloves garlic, finely chopped
1 tsp caraway seeds
1/2 tsp ground coriander
1/2 tsp chilli flakes
1 can Ardmona Duo tomatoes
1 cup ripe cherry tomatoes
1 egg per person
Parsley, finely chopped
Sourdough bread
Bring a frying pan to medium heat, add the olive oil and cook the onions for about five minutes, or until soft. Add the garlic and spices and cook for another minute or two. Pour in the tomatoes, tinned and fresh, and cook for about 10 minutes on a gentle simmer. In another pan, fry the eggs sunny-side-up. Serve each person a good spoonful of the tomatoes, topped with the egg and place a big plate of bread slices with good butter in the centre of the table.


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Some Common Page Plus Problems
From HowardForums Wiki
- If you use the web form to port your number, often nothing will happen. Follow up the next day by calling and asking for the 'porting department'. Better still, port by calling PP rather than using the web page.
- The $12 startup credit is no longer provided for porting. For new numbers, check your account page and if you don't see the $12 credit, call. You can also port after you activate in order to get the credit.
- Make sure you are happy with your number BEFORE you activate Unlimited Talk n Text. You can't change your number in the middle of a cycle without cancelling the plan early and losing what you prepaid.
- Applying the appropriate card should activate the corresponding plan (e.g. $39.95 card activates Unlimited Talk n Text) but sometimes it doesn't work. If so, you may be charged for calls/texts you thought were included in your plan. Check your account page, and if there's a problem, call.
- Some (many?) phones will show a roaming icon outside your home area, but this does not mean you are roaming. The only indication of 'real roaming' is that when you make an outgoing call, you are asked to dial again . This is a pain when driving, and wouldn't it be better if they just said "You are roaming"---no, that would make too much sense. For incoming calls, there is NO indication; you'll find out later that you were charged 59 cents/min.
- Occasionally you call will not go through, you will get a low balance or zero balance message that is clearly false. Just dial again. There has been one report that incoming callers have heard this message, and the call has not gone through. This is a more serious problem.
- PP's call log does not show total number of texts or total call minutes (useful for those on plans with a quota). The only way they'll tell you that you have exceeded your quota is that they'll start charging you extra, or calls will stop going through if you have zero balance. (As of March '11, the "My Account Summary" page on the user's o/l account showed data, text, and minutes remaining. Likely, all quota accounts now do.)
- Data may not be activated until you call and ask. If you have any difficulty getting your phone to work with data, this is the first thing you should suspect. (They may say "every line is automatically activated", but it's not true; insist that they check.)
- All newer Verizon phones should work, but technical assistance is limited. Even if you are paying for a plan that includes data, it is your responsibility to get the phone working with data. If that's not something you feel comfortable doing, buy your phone from PP. PDAs, eg. phones running Windows Mobile or Palm OS, should need no setup.
- PP's call log does not show how much data you have used (useful for those on plans with a quota). The only way they'll tell you that you have exceeded your quota is that they'll start charging you extra, or data will stop working if you have zero balance.
- The PP call log has delayed updates. This is potentially a problem for people testing data usage. Allow enough time for the log to update before assuming you are not being charged.
Source(s): phones solutions


text: 11/22/16 11/23/12 POPSUGAR Celebrity Celebrity Couples George Clooney and Stacy Keibler's Thankgiving in Cabo Stacy Keibler and Cindy Crawford Wear Bikinis For a Cabo Turkey Day With George and Randy November 23, 2012 by Allie Merriam 0 Shares Chat with us on Facebook Messenger. Learn what's trending across POPSUGAR. View on One Page Photo 2 of 37 Start Slideshow ADVERTISEMENT () Share This Link Copy What's Your Reaction? Thanks for your reaction Don't forget to share this with your friends! ← Use Arrows Keys → 2 Previous Next Celebrity CouplesRande GerberCelebrity VacationsThanksgivingStacy KeiblerGeorge ClooneyBikini


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In one photo of Boston bombing victim Martin Richard, 8, he is dressed in all white and holding a First Communion banner.
His name is glued jauntily, and a bit crookedly, in white felt letters, at the top. His mother must have let him do most of the work because all of the felt symbols included in the banner kit are used: a brown dove, pink heart, white host, red alpha and omega, mustard-colored chalice, grapes, chaffs of wheat and even the loaf of bread are glued on the banner.
Another mom would have advised restraint, selecting only a certain color scheme, vetoing her child if he wanted to cram everything in there. Another mom would have done most of the work. God bless Denise Richard for letting her son decorate his First Communion banner the way he wanted.
Wonder Boy and I worked on his banner a week before the bombings. I spread out the same felt pieces Martin Richard used in his banner, helped him find the white letters of his name, and hot glued his chosen design. Wonder Boy chose the brown cross, gold chalice, and host for his smaller banner. He carefully drew a small cross on the host with gold glitter. He wanted to include a bunch of grapes and the loaf of bread but it wouldn't fit.
In the end, my son was happy with his banner, which will mark our place in the church pews on May 4, when he receives his First Holy Communion. Later, it will be framed and have pride of place in the room he shares with his brother.
I will take a similar picture on May 4 that the Martins did early this year, of their own boy, now, unbelievably, gone. I will have my 8-year-old son hold his banner before him, and I will tell him to smile. I will make sure everything is perfect before I press the shutter. Then I will give him a hug and hurry him along. There will be lunch waiting, and guests ready to greet him on his special day.
I will pause to admire this banner, a bond between two boys, one of whom will stay forever 8. If I move the banner just so in the sunlight, the gold glitter in the cross glistens just so.
Columnist Anissa Rivera can be reached at firstname.lastname@example.com.


text:
GROVES-L ArchivesArchiver > GROVES > 2001-02 > 0981934782
From: Pat Berry <>
Subject: Groves
Date: Sun, 11 Feb 2001 15:39:42 -0800
Posted on: GROVES Queries
Reply Here: http://cgi.rootsweb.com/~genbbs/genbbs.cgi/surnames/g/r/GROVES/queries/7
Surname: Groves
I am a descendant of Joe and Stephen Groves. I know they lived in Adams
Co., Illinois, and Joe and Elizabeth (Witt) eventually moved to Missouri.
In your large database, do yu have a Joe or Stephen? I have no idea where
Stephen relocated to Illinois from. Thanks
This thread:
|Groves by Pat Berry <>


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by Staff Writers
Beijing (AFP) June 18, 2013
Foreign direct investment (FDI) into China rose during the first five months of this year, the government announced Tuesday, paced by strong increases from the European Union and United States.
Incoming FDI, which excludes financial sectors, rose 1.0 percent to $47.6 billion from January through May, the Commerce Ministry announced. It was also up 0.3 percent in May to $9.26 billion from the same month last year, the ministry said.
"Investment from the EU and the US continued to increase rather rapidly" in the first five months, ministry spokesman Shen Danyang told reporters.
EU investment into the world's second-largest economy rose 24.1 percent $3.45 billion over that period. Investment from the United States increased 22.6 percent to $1.58 billion.
The rise in EU investments come as Brussels and Beijing are involved in a series of disputes covering products ranging from steel pipes to solar panels that have sparked fears of a trade war between the economic giants.
China said earlier this month it will deal "appropriately" with the EU's decision to challenge it at the World Trade Organisation after Beijing slapped duties on some steel products.
China has launched an anti-dumping probe into imports of EU wine and chemicals while the EU has slapped tariffs on Chinese solar panels and threatened an investigation into the country's key telecom equipment firms.
The vast majority of the FDI investment into China comes from a group of 10 Asian countries and regions including Hong Kong, Taiwan, Japan and Singapore. Investment inflows from those locales gained 0.57 percent on-year to $40.9 billion over the five-month period.
Hong Kong accounted for the majority of that figure at $30.5 billion.
"The FDI situation has been relatively stable and good," Shen said, calling it an endorsement of "China's competitiveness and the acknowledgement of China's investment environment by global investors."
FDI into China fell in 2012 for the first time in three years owing to global economic uncertainties led by Europe's debt crisis, an economic slowdown at home and regional political tensions.
China's economy grew at its slowest pace in 13 years in 2012 as gross domestic product expanded 7.8 percent in the face of weakness at home and in key overseas markets.
China's overseas investment in non-financial sectors in January-May, meanwhile, rose 20 percent year-on-year to $34.3 billion, the commerce ministry said.
China's investment into Hong Kong, the Association of Southeast Asian Nations, the European Union, Australia, the US, Russia and Japan totalled $26.9 billion, up 16 percent on-year and accounting for 78 percent of the total in the first 5 months, according to the ministry.
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text: Emily is a young composer of contemporary music, native of Singapore. Recent commissions include those from the Singapore Lyric Opera Children's Choir, Philharmonic Orchestra Society and Ding Yi Music Company. A recipient of a doctoral fellowship from Brandeis University, she will be pursuing her Ph.D. in Composition and Music Theory there with David Rakowski, Eric Chasalow and Yu-Hui Chang next fall. She holds Masters degrees in composition and music theory pedagogy from the Peabody Conservatory, Johns Hopkins University, graduating with the Otto Ortman award in composition and Anonymous Award in music theory. Emily also graduated from the Yong Siew Toh Conservatory of Music, National University of Singapore, as valedictorian of her class, and studied composition with Ho Chee Kong and double bass with Guennadi Mouzyka. Emily had also been conferred the Associate and Licentiate diplomas of the Trinity College London in Music Theory, Criticism and Literature, and also holds a diploma in double bass performance from the Associated Board of the Royal Schools of Music (London).


text:
A prestigious address, along the prime stretch of Sheikh Zayed Road, in the heart of the executive business district. Minutes from the World trade Center exhibition halls and the emerging DIFC, and with direct access to the Dubai Media and Internet cities and Jebel Ali Free Zone. Also a central location to reach major shopping malls, beaches, golf courses and all that's happening in the cosmopolitan city of Dubai. Steps away from DIFC metro station. 5 minutes drive to the Dubai Mall and Burj Khalifa and Down town.
Number One Tower Suites Dubai
in a larger map
©2013 Number One Tower Suites All Rights Reserved
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text:
Shelburne County teachers add their voices to province-wide walkout
SHELBURNE COUNTY, N.S. – For the first time in its 122-year history, the NSTU held a teachers' strike on Friday, Feb. 17.
By Greg Bennett
The Coast Guard
Although some local municipal councils are looking for a pledge from Nova Scotia's opposition leaders that they would turn back changes to Shelburne County's electoral boundaries, there appear to be few takers.
The Coast Guard contacted representatives of both parties for comment on the issue. Liberal leader Stephen McNeil did not return an answer by press time but was expected to provide an answer by the end of the week.
Progressive Conservative leader Jamie Baillie did reply but made no promises on the issue other than to not interfere with the work of the electoral boundaries commission.
"I share the concerns of the people of Shelburne County, who are outraged by the political interference of one, majority party, in the independent process to redraw electoral boundaries. I am disheartened by the turmoil your community has been put through during this process but I believe any politician who promises to redraw the lines him or herself, is committing the same sin the NDP did," wrote Baillie. "I believe that someone who wants to be premier needs to be honest about this and other issues.
Baillie's only commitment was that a PC government would restore the power of a "truly independent boundary commission, which must work with the people to determine fair representation. I respect and honour the independence of the commission and believe they will make the right decisions in the future."
In December a bill splitting Shelburne County's provincial political boundaries passed the third and final reading in the legislature by a 26-22 vote.
As a result, Barrington and the rest of the western part of the county will be added to the formerly protected Acadian riding of Argyle while the rest of Shelburne County east of the Clyde will amalgamate with Queens County to form a new Queens-Shelburne constituency.


text:
Potter by Norah Gaughan
Welcome! You are browsing as a guestWould you like to see 25 projects made from this pattern and much more? create a free account What am I missing?
© Berroco Inc.
© Berroco Inc.
This cardigan features crisp ribbing and a polished double breasted button band reminiscent of a pea coat. Straight sleeves flair slightly and leave plenty of room for layering.
For more information, see: http://www.berroco.com/ng5/ng5_potter_pv.html


text:
This section is to serve as an introduction on AAV to colleagues who are relatively new to the field, but are interested in getting more information about it. As could be imagined, this field is quite complex and rapidly changing. After going through hundreds of literatures, we extracted the key pieces of relevant information, and put them together in a way that is relatively easy to follow, for colleagues who don¿t have a Ph.D. degree on AAV.
We will start with the biology of AAV, and the general principle and methods for rAAV production, and we will end with a note on the myth about DNA persistence and relatively long-term AAV transgene expression. There are some important notes that will help you understand further some aspects of AAV better, but it will be overwhelming to follow if we put all of them here, especially for those we are newcomers to AAV. We put those various notes in a separate file (called Supplementary Notes), for those who are interested in reading further.
What is AAV?
The first human adeno-associated virus (AAV) was discovered in 1965, as a contaminant of adenovirus (Ad) preparations (this is probably how it got its name). It is one of the smallest viruses with a non-enveloped capsid of approximately 22 nm. Although 80-90% of adults are sero-positive with AAV2, infection has not been associated with any symptoms or disease.
Because a co-infecting helper virus, such as adenovirus or herpes simplex virus, or induction of cellular stress is usually required for a productive infection to occur, AAV serotypes are ascribed to a separate genus in the Parvoviridae family designated Dependovirus.
AAV has a linear single-stranded DNA (ssDNA) genome of approximately 4.7-kilobases (kb), with two 145 nucleotide-long inverted terminal repeats (ITR) at the termini. The virus does not encode a polymerase and therefore relies on cellular polymerases for genome replication. The ITRs flank the two viral genes - rep (replication) and cap (capsid), encoding non-structural and structural proteins, respectively.
The rep gene, through the use of two promoters and alternative splicing, encodes four regulatory proteins that are dubbed Rep78, Rep68, Rep52 and Rep40. These proteins are involved in AAV genome replication. The cap gene, through alternative splicing and initiation of translation, gives rise to three capsid proteins, VP1 (virion protein 1), VP2 and VP3, with molecular weight of 87, 72 and 62 kDa, respectively. These capsid proteins assemble into a near-spherical protein shell of 60 subunits.
Immunofluorescence results suggest that capsid assembly is confined to the nucleoli of infected cells. Fully assembled AAV capsids enter the nucleoplasm in an AAV Rep-dependent manner. Selective AAV DNA encapsidation is presumably directed by protein-protein interactions between two complexes: (1) the pre-formed empty capsids and (2), complex of Rep78 or Rep68 with the virus genome. Next, the Rep52 and Rep40 proteins are responsible for transferring the AAV genome DNA into empty particles through the pores. It should be noted that, this is just one of the many working hypotheses for AAV replication.
AAV2 attachment is primary mediated by heparan sulphate proteoglycans, while internalization is aided by the co-receptors, such as avb5 and FGFR1 etc. The use of ubiquitous heparan sulphate proteoglycans as docking sites explains in part the well-known broad tropism of this virus. However, the events and processes that regulate the trafficking of AAV particles into the nucleus are still not fully understood.
After entry into the host cell nucleus, AAV can follow either one of two distinct and interchangeable pathways of its life cycle: the lytic and the lysogenic. The former develops in cells infected with a helper virus such as adenovirus or herpes simplex virus (HSV), whereas the latter is established in host cells in the absence of a helper virus.
When AAV infects a human cell alone, its gene expression program is auto-repressed and latency is ensued by preferential integration of the virus genome into a region of roughly 2-kb on the long arm (19q13.3-qter) of human chromosome 19, designated AAVS1. This site-specific integration involves the AAV ITRs and Rep proteins (Rep78, Rep68).
When a latently infected cell is super-infected with a helper virus, such adenovirus or herpes simplex virus, the AAV gene expression program is activated leading to the AAV Rep-mediated rescue (i.e., excision) of the provirus DNA from the host cell chromosome, followed by replication and packaging of the viral genome.
Upon helper virus-induced cell lysis, the newly assembled virions are released. The induction of the lytic phase of the AAV life cycle from a stably integrated provirus can also occur in the absence of a helper virus, though with a lower efficiency, when the host cell is subjected to metabolic inhibitors, DNA damaging agents or genotoxic compounds.
AAV2 is probably the most widely used AAV serotype for in vitro and in vivo gene delivery. It should be noticed that many other serotypes of AAV have been isolated, and the every year there are some new serotypes being discovered. Among them, AAV1, AAV2, AAV5, as well as AAV7, 8 and 9 etc, have been used for gene delivery studies. The question is: among these serotypes of AAV, which serotype(s) is appropriate or best for a particular study model.
Though there are numerous studies trying to compare the efficacy of gene delivery in various models, the data are quite often controversial, if not conflicting. The exact reason for this is not clear. It is generally believed that difference in animal species involved, study protocols, end-point readout, tissue or organs, as well as the procedures used to prepare those recombinant viruses etc, could contribute to this. If possible, it is a good idea to compare the efficacy of gene delivery by several rAAVs carrying a reporter gene, such as GFP or LacZ, to decide which serotype and procedure is the appropriate for your particular study.
Given the fact that the AAV ITRs contain all cis-acting elements involved in genome rescue, replication and packaging, and the fact that the AAV ITRs are segregated from the viral encoding regions, i.e., Rep and Cap gene regions, recombinant AAV vector design can follow the whole gene- removal or "gutless" vector design rationale, as in the retrovirus system. In other words, the cis-acting viral DNA elements involved in genome amplification and packaging are in linkage with the heterologous sequences of interest, whereas the region(s) encoding trans-acting viral factors involved in genome replication and virion assembly are provided in trans. Typically, rAAV particles are generated by transfecting producer cells with a plasmid (AAV cis-plasmid) containing a cloned recombinant AAV genome composed of foreign DNA flanked by the 145 nucleotide-long AAV ITRs, and a separate construct expressing in trans the viral rep and cap genes. The adenovirus helper factors, such as E1A, E1B, E2A, E4ORF6 and VA RNAs, would be provided by either adenovirus infection or transfecting into production cells a third plasmid that provides these adenovirus helper factors. Given that HEK293 cells, a commonly used AAV production cells, already contains the E1A/E1b gene, so the helper factors need to be provided are E2A, E4ORF6 and VA RNAs.
Significant progresses have been regarding the production of recombinant AAV, particularly recombinant AAV2. The production method for other serotypes are derived from method(s) developed for recombinant AAV2. Of numerous approaches to improve AAV vector production, two strategies differing in principles are now widely used, depending on how the adenovirus helper factors are provided.
One method relies on wild-type adenovirus infection into the cell lines that stably harbor AAV rep/cap genes, as well as the AAV vector DNA. The other method, the helper-free method, is based on the adenovirus-free transient transfection of all elements that are required for AAV production in host cells such as HEK293 cells. It generally involved the co-transfection of AAV production cells with 3 plasmids: (1) an AAV2 ITR-containing plasmid carrying the gene of interest; (2), a plasmid that carries the AAV2 Rep-Cap; and (3), a plasmid that provide the helper genes isolated from adenovirus. Although the method using wild-type adenovirus-inducible AAV producer cell lines can be easily scaled up in cultures and produce AAV vectors with very high titers, it is very challenging to completely get rid of the adenovirus from AAV product, and contamination of wild-type adenovirus is highly undesirable in view of vector safety and specificity. On the other hand, although the transient-transfection method generates high-titer AAV vectors that are free of adenovirus, the process is very labor labor-intensive and expensive.
Besides these 2 commonly used procedures, new techniques for producing recombinant AAV have also been developed, though they are not that commonly used as the two we discuss here.
Though AAV2 is the most extensively studied serotype, and significant processes have been made about its production, for some tissues and cell types, it has low transduction efficiency. Therefore, an important area in the development of AAV as a vector concerns the engineering of altered cell tropisms to narrow or broaden rAAV2-mediated gene delivery and to increase infection efficiency. Between various AAV serotypes, the difference in transducing efficiencies could be caused by difference of their respective receptor content on target cells. A strategy to alter rAAV tropism exploits the natural capsid diversity of other serotypes, by packaging recombinant AAV2 genomes into capsids derived from other AAV isolates.
The commonly used approach employs hybrid trans-complementing constructs that encode rep from AAV2 whereas cap is derived from the other serotype displaying the cell tropism of choice. For instance, if it is the rAAV2/5 that is to be produced, and if the production method is based on the helper-free, transient transfection method discussed above, the 1st plasmid and the 3rd plasmid (the adeno helper plasmid) will be the same as discussed for rAAV2 production. However, the 2nd plasmid, the pRepCap will be different. In this plasmid, called pRep2/Cap5, the Rep gene is still derived from AAV2, while the Cap gene is derived from AAV5. The production scheme is the same as the above-mentioned approach for AAV2 production. The resulted rAAV is called rAAV2/5, in which the genome is based on recombinant AAV2, while the capsid is based on AAV5. It is assumed the cell or tissue-tropism displayed by this AAV2/5 hybrid virus should be the same as that of AAV5.
It should be noticed, though the production of these hybrid rAAV vectors are the same as that for rAAV2, the purification procedure could be quite different, due to the differences in capsid properties.
Recombinant AAV gives prolonged and stable expression in numerous animal models without notable toxicity. Before transgene expression can occur, the single stranded rAAV DNA needs to be converted into a transcriptionally functional double-stranded template. It is important to note that this stability does not arise due to foreign DNA insertion into the parental virus pre-integration site (AAVSI), since the absence of rep gene products prevents DNA targeting to the AAVS1 locus.
Now the question is: how is the long-term transgene expression achieved? Recently, it was proposed that, after the single- to double-stranded DNA conversion, a duplex rAAV genome is formed, through either intra- or inter-molecular recombination at the ITRs. This duplex rAAV could either exist in a circular form or linear concatemers. It is the circular forms that are thought to be responsible for vector persistence and long-term transgene expression.
Vector BioLabs offers many adeno-associated virus services to fit your research needs, please inquire for details.