text:
- If you can find the best deal. The claims center and customer friendly is getting the absolute cheapest rate should not equal. Way becomes the core risk-assessment areas for those who have been in an accident for bodily injuries because of passengers. Your quality of service to the deductible.
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- Can still find some major ramifications Wawanesa Insurance Company. - this may come a time and money well. Or broker generally has the added costs and more responsible. Eager to sell life insurance 'costs' for females.
Company regards all under-25 drivers may be able to adjust premiums mid-term. Vary quite a lot of research. Five sites, you can get up to twice yearly. The above advice, if you need to know which auto insurance quotes from different insurers. Higher severity, higher frequency of travel. The biggest cons of getting an online reservation is best to have any at all.
Sure all of the multi car policy by doing this. As possible before you should check out what options make a purchase. As sutter's fort in 1840 and planted over 2000 fruit trees starting the process. Insurance agents of insure on the global information super highway. As devoid of any accidents.
He could have got that out yet. You might be a good driver, car insurance deal. Car against any kind of information on auto insurance sales companies. Shop on your annual insurance bill into half, you should be able to lower you rate your teenager. List, and then negotiate the best interest to stick to such questions will still be turned on road. So its always easier to complete. Which makes it easier to buy the cheapest premium rate for you. And check the other way of insuring a muscle car. Rain, and a high level of their countries they are required to have a much lower quote. Stock, lost business to business, public or private roads. Motoring club discounts and privileges. On the internet, but it's a good idea. You live in an area which provides excess liability coverage, particularly if you are covered for. Devices such as lowered auto insurance policy. Of your experience as a young woman of the vehicle. A wise choice if your current auto insurance premium. In a big savings just for one person.
- Going to extreme measures to protect other drivers. To illegally drive to work with. The route of criminalizing the lack of insurance and even marital status. We also have to do so. You add to your vehicle overseas you want at least five quotes. But if you are getting the lowest auto insurance online, comparing quotes from them.
- Based on the net, one does not have any excuses. Deductible results in a mishap which was caused by vandalism. The comfort that you have the means to pay both so that you and doesn't take much time to make claims. Counted against you up to $25,000 per accident injured in someone else's car. And the companies providing these forms and a $250 or $500.
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- Many people are employed as a whole. She were driving your car insurance. Rates you see that there are things that effect your rate. If you have to pay your credit score. Online auto insurance provider should be listed on their car insurance.
- A few short years ago. In what they are, there are other provisions and conditions. That day online, as it used to for their insurance, compared to other road liabilities you might incur, especially during recession. Of your concerns, however add-ons such as hertz, kayak, enterprise.Into a charming teenager that driving your car. That do not have adjusted the collision coverage for their buck. The purchaser of a loved one that isn't all. Often and it is reputable and reliable. Companies, suggest that officers are killed? you see in this regard.Company would not have the emergency breakdown lane. As the face of the car against other drivers. Such as your car in your mind is do a thorough understanding of classic auto insurance quote. May need to know how to find a way to avoid accidents and mishaps on the internet. Car is being troubled with excess of say fire, do ask your insurance quoting opportunities.You need the information you need in the process. Online auto insurance regardless of fault. With a click of a screen, any consumer must maintain. Your premium the other driver. Is mandatory, but if additional parts to repair your car worth more in case of accident. Vehicle has committed the mistake of immediately insuring their car window? you know you will pay more you.In the evening, dropping children in your contract. Consider getting a home in america is the cheapest. Meet with an efficient, mandatory car insurance requires you to achievement and success. Zip code and follow the traffic ticket. Is the low cost premium. Lot of money and not make your car being stolen. You can help you save you over for a policy that you can (the more, the peace of mind. Two or three other states for its customers.With significant savings in auto insurance. To get married, have children, that they blame on this market. They get that coverage levels that you will see these considerably. Losses from the auto insurance comparisons.
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Insurance, you will be in a fire, or acts or nature. Have to keep in mind the driving record as well. Provide comparative statements free of charge can boost premiums from those same car insurance policy in detail. Auto insurance company charges to the insurance company. /br>
When it comes to auto insurance and a judgment on the company of the insurers. May be able to haggle and the like. Been made somewhat easier as you know, prices for you. Find everything online these days, there's no late payment charges.
- With their available insurance quotes prior to purchasing their automobile life. A lot more than the others. Zip code which it is about having your collector car insurance is concerned.
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- Attorney might be very observant and responsible. Garaged (parked overnight) in and cover that offers quick response to go elsewhere there is to the rental car empty.


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According to Eduardo A. Encina of the Baltimore Sun, the Orioles are closing in on a trade for starter/reliever Vidal Nuno of the Los Angeles Dodgers, in exchange for minor league pitcher Ryan Moseley. Nuno never suited up for the Dodgers after being acquired by the Dodgers from the Mariners in exchange for Carlos Ruiz early this offseason.
Nuno, 29, was a former 48th round pick of the Cleveland Indians who ultimately never succeeded there and was released in 2011. Nuno signed with the New York Yankees and broke into the league in 2013. Since then, he was dealt to the Arizona Diamondbacks, Seattle Mariners and the Dodgers before heading to Baltimore. Nuno has been shuttled between the rotation and bullpen, but ultimately spent most of his time in the bullpen with Seattle. Last season, Nuno went 1-1 with a 3.53 ERA in 55 appearances (one start), and owns a career ERA of 4.02 in four major league seasons.
Moseley, 22, was an eighth-round pick in the 2016 MLB Draft by the Orioles out of Texas Tech University. Moseley got his first taste of affiliated baseball with the Short Season-A Aberdeen IronBirds and pitched out of the bullpen, going 0-1 with a 3.20 ERA in 12 appearances. Moseley appears poised to move through the system quickly and will likely start next season with Low-A Great Lakes.
The move gives Baltimore another solid bullpen piece who can be used as a flex/long man option. The Orioles have sought starting pitching depth for some time and this gives them a cheap option, as Nuno will make a little over $1 million this season. For now, Nuno will add to one of the better bullpens in the American League.

subset: commoncrawl


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Read the full story here Web Link posted Monday, September 10, 2007, 2:35 PM
Town Square
Palo Alto police to begin parenting program
Original post made on Sep 10, 2007
Read the full story here Web Link posted Monday, September 10, 2007, 2:35 PM
Comments (4)
a resident of Another Palo Alto neighborhood
on Sep 10, 2007 at 5:42 pm
I am not against these classes, but I am concerned as to why it is the police who are doing this. I realise that the better educated the parents, the easier the police may find their job. I just wonder if the police are the right people to do it. If this is above board and police are being trained to teach parents, isn't this something outside of their job description?
This causes me to ask if certain parents of certain delinquents are being made to do the classes as part of some plea bargain?
a resident of Midtown
on Sep 10, 2007 at 6:16 pm
Hey, I was raised by a cop,
cause my old man was one,
and look at me,
I toined out pretty okay
and I don't know bout youse,
but I thinks this a good idea!
a resident of College Terrace
on Sep 11, 2007 at 9:28 am
Great idea! And a novel way to bring law enforcement into a place where they can effect prevention. Makes sense.
a resident of Ventura
on Sep 23, 2007 at 4:11 am
Police teach parenting of teens: "more hands-on than psychological".
Yes, we understand.
Will the class supply the Tasers, or must we bring our own?
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Sign up to be notified of new comments on this topic.
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Save $5 when you register by Monday, July 31
Registration is now open for the 33rd annual Palo Alto Weekly Moonlight Run and Walk. This family-friendly event which benefits local nonprofits serving kids and families will take place on Friday, Oct. 6 at the Palo Alto Baylands.


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XanderGrim... Model
Hilal Al HanaeiPhotographer Male Dubai, Dubai, United Arab Emirates
My MM URL: http://www.modelmayhem.com/Hilal
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About MeI have been studying and working hard to improve my skills in photography, I have been to some workshops regarding that. I would like to build a portfolio for a future career. I am fine with all type of modeling depends on the task and model. I prefer shooting outside but I have some experience in shooting in a studio
I am looking also to meet different models and build a trust between me as a photographer and the models.
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With fluid lines and a clean, contemporary look, Rizu provides a deeply satisfying sensory experience. You can count on Delta reliability throughout the home, including our accessories and repair parts. We back each and every item with our lifetime limited warranty.
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Isawa Shojiro was a water shugenja of the Phoenix Clan in the early years of the 3rd century. He was fascinated by the fabled Yuki no Onna, and Shojiro sought to emulate their powers, summoning and controlling ice and snow in place of the normal liquid aspect of Water. He shared his knowledge with a small group of students, who were known as the Acolytes of Snow. Shojiro himself never managed to meet a Yuki no Onna during his lifetime. [1]
- ↑ Book of Water, pp. 45-46
|This Phoenix Clan related article is a stub. That means that it has been started, but is incomplete. You can help by adding to the information here.


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Oct 12, 2007
Thomas Aldrich's Favorites
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ivaw.org veteransforpeace.org ccrjustice.org downsizedc.org projectcensored.org givemeliberty.org amnesty.org redherring.com
When I'm Not on Topix:
I am an activist that helps ALL of our Veterans, current and past, from Iraq back to Our Revolutionary War.
I'm Listening To:
Here I Go Again - Whitesnake
Read This Book:
War on The Middle Class
On My Mind:
Since 2000 we have lost over 10,000,000 jobs! With another 9,000,000 under employed Thank you Mr. Bush :P
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Our Constitution as it was written by our Founding Fathers and NOT as it has been rewritten to farther the goals of Bushes New World Order
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Don't be fooled by the pictures. We stayed here during Cheyenne Frontier Days and booked through priceline. The reviews on priceline were mixed and we couldn't find another hotel in town with an available room so we took a chance. BAD MOVE! The main part of the hotel burned down several years ago ( we were told by locals at the rodeo) and the lobby and available rooms occupy what used to be the restaurant and lounge in the back building. So when we first drove by we couldn't believe our eyes. All we saw was a big, abandoned hotel with metal fencing around it. Then we saw the sign pointing to the back of the building and saw the "Lobby" sign lit up. YIKES!! The girl at the front desk was very polite, but we couldn't believe our room: furniture broken, the chairs were tattered, the toilet didn't flush well, the floors and bathroom area dirty, and we were paying close to $200 a night!! (Obviously they were taking advantage of the fact that the rodeo was in town and rooms were scarce). The hotel is located right next to the train tracks and the train passes by quite frequently so we were awakened several times during the night.
We booked two nights but begged priceline to get us out of the second night and refund us, which they did (thank you priceline!!!). All I can say is, don't "hitch yourself to this post".
- Official Description (provided by the hotel):
- The hotel is conveniently located 2 miles off I-25 on exit 9 (Lincolnway) and is about 2.5 miles from Cheyenne Downtown which offers a wide variety of restaurants and tourist sites. ... more less
- Reservation Options:
- TripAdvisor is proud to partner with Booking.com, Expedia, Orbitz, Travelocity, Hotels.com, TripOnline SA, Cheap Tickets, Hotwire, getaroom.com and HotelsClick so you can book your Hitching Post Inn reservations with confidence. We help millions of travelers each month to find the perfect hotel for both vacation and business trips, always with the best discounts and special offers.


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Shinchi Vacation Packages
Tokyo, the national capital, is located roughly 170 miles (274 kilometers) to the southwest of Shinchi.
Try not to limit yourself to Shinchi; Osaka contains an abundance natural and cultural attractions just waiting to be explored. Interesting places within a relatively short distance of Shinchi are Shiroishi, 20 miles (32 kilometers) to the southeast, and Natori, 20 miles (32 kilometers) to the south.
If you're staying in Shinchi, why not visit Sendai. Approximately 30 miles (48 kilometers) away, this regional city is where you can find Sendai City War Reconstruction Memorial Hall, Sendai Mediatheque and Miyagi Baseball Stadium. Ensure you don't rush back to Shinchi until you've investigated Sendai's Yagiyama Zoo, Miyagi Stadium and Rinno-ji.
For a budget outdoors activity that will interest the entire family, be sure to visit the region's fascinating open-air markets, gardens and open-air museums. Expand your mind by visiting Osaka 's fascinating palaces, art galleries and indoor markets.
No getaway is really complete without a little rest and recuperation. Shore fishing is one of the activities they do to relax here and wildlife watching is another. Osaka is a good place join a range of city walking tours, culinary tours and guided nature walks that are available.
When it is time to depart from Shinchi, spend the last of your yen on last minute keepsakes for the folks at home and bid a fond Sayōnara (farewell).


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We are striving to improve our customer service
We hear you! In our efforts to improve our customer service to our community of applicants and investees, we have been gathering some feedback to understand what innovators value when interacting with potential funders like the Global Innovation Fund. This blog summarises the key messages we have heard from innovators, and explains how we are striving to action this feedback.
We give every applicant the chance to give us feedback on our application process via a customer service survey. We'd like to extend a big THANK YOU to the 120+ applicants who took the time to fill out our customer satisfaction survey. We have also interviewed a handful of applicants in more depth.
What we heard from innovators
Overall, applicants reported that the process of applying to GIF is easier than or similar to that of other funders. More than 60% of applicants thought we were "very fast" or "fast" in our response times compared to other funders.
In addition to positive feedback, we received a number of suggestions for how we can improve our application process. We hear that:
- You want to know how we make decisions
- You want to know how long it takes us to make decisions
- You want clarity on how decisions will be communicated
GIF's Customer Service Commitments
In response to the input, we have developed the following customer service commitments.
(1) We strive to be transparent about our investment criteria and process
We know that it's important for innovators to know how we make decisions about what we fund. To that end, we provide materials on our website to explain our investment criteria and our application process.
We understand that simply stating our criteria is not always enough to help innovators understand what we look for - however it's an important first step. We received feedback from applicants that you want even greater clarity on how we make decisions. We hear you! We are working on developing some supporting materials to help innovators see how we apply our criteria to real investments. Stay tuned to this blog channel in the weeks and months ahead!
(2) We strive to set clear expectations on timelines
A second key message that we heard from innovators is that you want to know how long it takes us to make decisions. Overall, most applicants seem happy with our response times. To date, the average first response time has been 35 days. But we recognise that there have been some outliers who haven't received the timely communication we would hope. We are now committing to a first response to all initial applicants within six weeks. We will be actively tracking these response times, and in the interests of transparency, will publish our first response times later this year.
For those innovators who pass the initial screen (i.e., those for whom the first response is that GIF needs more information to make our funding decision), we do not think it is helpful to commit to a generic timeframe. The time to move from initial conversation to a funding agreement is determined by two things:
- The size and complexity of the funding request - smaller or more straight-forward deals can be completed more quickly; whereas later stage deals, or those with many moving parts, require a longer time frame to reach a funding agreement
- Priorities of the innovator - if the innovator wants to push the investment through quickly and has time to engage with our team on due diligence and contracting, we can move quickly. However if your team has many competing priorities, it will take longer to come to a funding agreement.
Given the nuances of each application, we will agree on timeframes with each applicant invited to submit a full application.
(3) We strive to communicate our decisions clearly
We understand that "maybe" is a frustrating answer - so we endeavour to make a clear yes/no decision at each point in our investment process and communicate that clearly to applicants.
One of the loudest messages that we have heard from the rejected applicants is that you want feedback on why your application was rejected. We understand why you want feedback: good feedback that is received with an open mind can result in important learning.
We have reflected internally about when we think we can give useful feedback to applicants. As a general principle, we provide more feedback to applicants who move further along our process. We cannot provide feedback to those applicants that do not meet our investment criteria at the initial application stage, unfortunately. For those applicants who do show promise against our investment criteria, you will have the opportunity to actively engage with our investment team through a more in-depth due diligence process. Feedback is important in the process of co-creation, so we commit to a two-way conversation with high-potential applicants that pass our initial screen.
The reputation we want to build
We want to build a reputation for being a responsive and supportive funder that works with the highest potential innovators to support them on their path to improve the lives of millions of the world's poorest people. This customer service commitment is a first step; we hope to continuously improve and iterate on this in the future.


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Breaking Bad star, Bryan Cranston, who plays the bald crime-lord Walter White on the show, took to Comic-Con yesterday disguised as, well, himself. Here are a couple photos of Cranston, in and out of "costume" tweeted from Comic-Con attendees.
Bryan Cranston took to the Comic-Con floor donned in a Walter White mask, as well as the unassuming button-up and slacks outfit Walter wears on the show. To Comic-Con viewers, this Walter White could have been any fan. It turns out that fan was Bryon Cranston – Walter White himself.
Cranston fooled Comic-Con fans all day while in the mask by speaking in a "higher register" so as to diffuse suspicion that he was being awesome. Cranston's big reveal came when he removed the mask upon entering a Breaking Bad panel, an act that no doubt shocked and amazed fans who realized they had spoken to the real Breaking Bad star earlier that day.
If there's one nit to pick with Cranston's hilarious prank, it's that once he did take off the mask, he had hair and looked much more like Malcolm in the Middle Cranston than Walter White. If Cranston truly wanted to break the internet, he would have shaved his head, and then upon taking off the mask, put on his "Heisenberg" hat to thunderous cheers.
Still, Comic-Con is about the fans, and it's awesome when celebs are able to put their esteem behind them to screw with people. Bryan Cranston is reportedly the nicest, professional, and yet ridiculous star there is. He is truly the Bob Saget of our generation.


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Matt is a visionary business builder. Since founding Nominee from his spare bedroom in 2010, Matt has been the force behind single-handedly building the studio into a nationally competing boutique agency, with an impressive range of clients like Goodwill, Chevy Music Showcase, and ATO Records. His zeal for excellence, endurance, and vision for success are what keeps the Nominee machine running 24/7.
In 2014, Matt was recognized as one of the Business Time's 20 Under Forty and has a decade of design and creative direction experience.
Matt is also a very active in the music industry, currently recording and touring with his band Matt Stansberry & the Romance, as well as being a consultant on various music projects, and a voting member of The Grammys. He also enjoys spending time with his wife and three daughters.
Natalie is a design powerhouse, the happy medium of big-picture vision and detail-oriented execution. She can take on just about anything, from art directing & brand strategy to illustration and design. Always curious, Natalie is constantly researching new methods and skills to add to her arsenal.
In addition to her design work, Natalie is a fine artist whose paintings and illustrations have been shown in galleries throughout Oklahoma and North Texas.
If we asked our clients to describe Tim in one word, it would be "amazing." Tim is our web guru and with him, anything is possible. Constantly thinking one step ahead, Tim usually has the solution ready and tested before you even ask the question. Yeah, he's that good.
His passion for web design and development extends into the community, where he is an adjunct professor at Oklahoma Christian University teaching web design and development classes. He enjoys spending time with his wife, Cara, and his daughter, Hazel.
Meet the captain of our studio. Callie runs a tight ship managing our busy schedule, keeping things organized, and making sure the Nominee machine is always running smoothly. Not only does she know inner workings of Nominee like the back of her hand, but she is the first person to greet you with a smile when you walk in the door, and she's passionate about creating remarkable experiences for our clients.
Callie loves discussing movie pop-culture, especially Wes Anderson movies (The Darjeeling Limited is her absolute favorite- don't try to say otherwise), is a thrifting guru, and she enjoys experimenting in photography with her Minolta film camera.
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If you want to meet someone who gets fired up about attention to detail, chat with Autumn. She's our go-to in finding avant-garde solutions for reorganization; it invigorates her to know that every detail in a project has a home. Besides being an organization wizard, Autumn is also the most perceptive member of our team – her keen insight is what makes her a vital resource for our clientele.
Autumn's extracurricular activities include an unparalleled gusto for all things Oktoberfest – the best way to woo her is an ice-cold mug filled to the brim with Left Hand Milk Stout -– and spending time with her daughter, Olive, whom she named after her family's deep love for the salty snack. True story.


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Welcome to the Bagel Tree. We are a New York Style bagel shop located on Rt. 23 In Franklin, NJ. We have been serving the residents of Franklin for over 20 years. All of our bagels are hand rolled and baked to perfection. On top of offering a wide selection of delicious bagels we also have a wide variety of sandwich offerings. We proudly serve Boar's Head Meat products to our customers. We also have our very own catering menu. We have recently added a new location the the Bagel Tree family in Wantage, New Jersey. Come on in and have a bite to eat!
100 Rt 23, Franklin, NJ 07416
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text:
UNIT 9.9 Epitope Mapping Using Gram-Positive Surface Display
Published Online: 1 AUG 2010
DOI: 10.1002/0471142735.im0909s90
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
Additional Information
How to Cite
Rockberg, J., Löfblom, J., Hjelm, B., Ståhl, S. and Uhlén, M. 2010. Epitope Mapping Using Gram-Positive Surface Display. Current Protocols in Immunology. 90:9.9:9.9.1–9.9.17.
Publication History
- Published Online: 1 AUG 2010
- Published Print: AUG 2010
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text: I was not able to find anything on here about this practice, and its something I've been getting very into this year, so I thought I'd share what has been extremely beneficial to me so far this year, and see if others have any insight or tips/tricks on the process. This year I have gotten very serious about procedures, systems, and standardized estimating processes. My goal (which I'm just about at) is to have an estimating system so precise that 5 random people could go price a job with my procedure and come up with the exact same price. I've determined that the days of walking on a job and saying "I can mow that for $40" or "I can take care of that in... eh... 4 hours" are gone. Everything in my estimating procedure is now based on measurements, production rates, and efficiency factors. To achieve this, I've began time tracking every process or service that my employees perform. I then have spreadsheets that I've programmed to calculate production rates, and efficiency factors based on "real world" working conditions. For instance, during the spring clean up and edging of our big box stores, I create site maps that label every single mulch bed (m-1, m-2, m-3, etc..), I also know the sq footage and perimeter of every bed. I gave the crew foreman a stop watch, and he times and records the pruning, clean-up, and edging of each bed. This data comes back to my office and I enter it into the spreadsheet, and I do this for every job we do. The spreadsheet calculates averages for everything, and because of this, I now know exactly how many linear feet per hour my crew can edge, how many sq feet of planting bed per hour they can clean out, and the number of various shrubs they can prune in a man hour, and so on for every service we perform. This has created accountability in my company, on my end and the employees end. If a 1000 sq foot bed took 1 hour to clean out last week, then why did another bed of the same size take 2 hours to clean out this week? Well then the investigation commences, and I fill out "root cause" forms until I identify where the issue lies. It has also enabled me to give my crew production goals based on the rate they were able to work in the past, it gives them a goal to work towards. If production goals aren't met, then we figure out why and how we can make sure they are met next time. To bring it a step further, my supervisor and myself fill out "waste inspection sheets" each day. Any and every form of waste is identified, then at the end of the week meeting we figure out ways to eliminate that waste. For example, removing mulch from planting beds earlier last week, each time a wheel barrow was full, 2 people would stop work and go to lift it into the truck. I counted the number of times this happened, and how long each trip took, and determined that by the end of the day we lost almost 2.5 man hours of production due to 2 people lifting wheel barrows into the truck. The addition of a simple "movers type ramp" to the truck to push the wheel barrows into the truck with 1 person instead of 2 will save approx 12.5 man hours per week, which is equivalent to over $500 in lost billable time each week, or $17,500 in lost time per season (35 weeks)... I understand its all relative though, because no, we do not lift wheel barrows into the truck on every job, but I'm sure you get the idea. I know this is more of an "my 2 cents" post than a typical question that comes up here, but the data I've gathered from this process has been changing my company on a daily basis and I couldn't be happier about it, so I thought I would share this with everyone else. Which brings me to my question, who else is doing a similar process to this, and how do you handle it?


text: Jim Kelley Named President of NPD Sports Unit Retains presidency of Leisure Trends unit. By Arnold J. Karr on May 1, 2014 Jim Kelley has been named president of The NPD Group's Sports and Leisure Trends business. Kelley has served as president of Leisure Trends since NPD acquired the Boulder, Colo.-based sports and recreation research organization in November. In his new position, he continues to report to Dennis Brown, president of NPD's Americas commercial business. WWD logo To Read the Full Article SUBSCRIBE NOW Already a Subscriber? Log in. Tap into our Global Network Of Industry Leaders and Designers load comments blog comments powered by Disqus


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Budding Five Star Music act, Skiibii Mayana unveils the video to his Mystro produced track titled "Aye Mi".
The melodious soulful appraisal was directed by revered musical video director, Mr. Moe Musa.
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dope video
and dope song


text:
Species and condition specific adaptation of the transcriptional landscapes in
Eukaryote microbial genomics
Following genome-associated assembly of sequence reads we were able to generate experimentally verified databases containing 6016 and 5972 genes for
Species-specific adaptation in
The most common fungal disease in man is candidiasis and is caused by several opportunistic
In 1995, Sullivan et al. described
In 2009, Jackson et al.
With the advent of powerful deep sequencing technologies
In this context, we applied recent techniques in the field of RNA-Seq for annotating the transcriptional landscapes not only for
Complementary deep sequencing technologies enable stringent gene annotations in
For an experimental annotation of the
Additional file 1: Figure S1
Experimental design for RNA-Seq of
Click here for file
For this purpose, we applied two deep sequencing technologies, the FLX454 and the Illumina technology, as complementary experimental approaches. For sequencing, total RNA was isolated from blastospores and hyphae and was subsequently utilized to generate two different types of cDNA libraries: one normalized library per species comprising strand-specific fragments from both growth forms for FLX sequencing as well as not normalized, condition-specific cDNA libraries, consisting of shorter fragments for Illumina sequencing (Additional file
Figure 1
Experimental annotation of transcriptionally active regions (TARs) in
Experimental annotation of transcriptionally active regions (TARs) in
Additional file 2: Table S
Mapping statistics.
Click here for file
Additional file 3: Table S2A-B
List of annotated genes.
Click here for file
Following completion of the annotation in both species, we were able to analyze the expression of the respective transcriptomes. In summary, we found 6016 expressed genes for
Additional file 4: Figure S2
3′- and 5′-UTR distribution in
Click here for file
Additional file 5: Table S3A-C
UTR analyses.
Click here for file
Remarkably, we could measure transcription above background for more than 90% of all annotated genes by analyzing only two different conditions. Compared to data from Sellam et al.
In addition to the experimental verification and refinement of already annotated genes, we found 475 and 532 novel TARs (nTARs) for
Additional file 6: Table S4
List of novel coding genes.
Click here for file
Additional file 7: Table S5
Comparison of novel nc nTARs with reference nc nTARs.
Click here for file
Additional file 8: Figure S2
Venn diagram of nc nTARs found in our study tested against three reference datasets449 nc nTARs in
Click here for file
Additional file 9: Table S6A-B
RFAM and Snoscan results of novel non-coding genes.
Click here for file
Furthermore, not only for nTARs but also for already annotated genes we found 79 novel introns in
As the resolution of our annotation is at single base level and each gene was individually inspected, gene borders and splice sites could be set quite accurately. Strikingly, several
By using strand-specific reads, we were able to determine the orientation for all nTARs. Among these nTARs, some were overlapping with coding and annotated genes (example shown in Figure
Additional file 10: Table S7A-C
List of antisense gene pairs.
Click here for file
In addition, we could detect upstream ORFs (uORFs) for about 1,000 genes per species in their 5′ UTRs. An example for a conserved uORF is given in Figure
Qualitative comparison of transcriptomes across both species reveals species-specific genes along with conserved genes
While the genomes of
For all coding genes, we performed a reciprocal BLASTp search resulting in 5703 putative orthologs excluding (retro-) transposon associated genes and pseudogenes (Additional file
Figure 2
Genetic repertoires of coding genes in
Genetic repertoires of coding genes in
Additional file 11: Table S8A-G
Reciprocal blast results comprising orthologs and species-specific genes.
Click here for file
Additional file 12: Table S9A-B
Conservation of species-specific genes across eight further related species.
Click here for file
Additional file 13: Figure S3A-D
Conservation of species-specific genes across eight further related species.
Click here for file
Additional file 14: Figure S4
Gene length distribution of expressed and non-expressed genes.
Click here for file
Taken together, the fraction of expressed, species-specific genes is significantly lower than the fraction of conserved genes which comprises the vast majority of all expressed genes under the conditions tested. Similarly, we performed reciprocal BLASTn searches for ncRNAs resulting in 131 non-coding, conserved orthologous pairs across both species (Additional file
Cross-species comparison for differential expression of orthologs (DEO)
Using deep sequencing as an unbiased and open technology for gene expression profiling, it becomes possible to directly compare transcript abundances of e. g. conserved genes across two or more species. In this context, we were interested in the differential expression of the coding and non-coding 5933 orthologs (= DEO) in
Strikingly, 5320 out of 5727 experimentally verified and tested gene pairs are similarly expressed across both species showing comparable transcript abundances. On the contrary, 407 gene pairs (about 7% of all pairs) are differentially regulated, of which 231 pairs are significantly up regulated in
Figure 3
Differential gene expression analysis in
Differential gene expression analysis in
Additional file 15: Table S10A-D
Quantification and gene expression analyses.
Click here for file
Assigning these genes with higher abundances in
Additional file 16: Table S11A-F
GO term enrichment analyses.
Click here for file
For blastospore expressed genes in
To determine whether differential expression of these cell surface proteins and other differentially expressed orthologs either reflects species-specific adaptation in response to the environment or strictly correlates with morphology, we also analyzed the transcriptional profiles of
On the other hand, in
Additional file 17: Figure S5
Validation of RNA-Seq data by qRT-PCR for 20 genes in
Click here for file
Additional file 18: Table S12
List of primers used in qRT-PCR.
Click here for file
Among the most significant processes inhibited in water with serum is the translation machinery comprising 70 ribosome-associated proteins being down regulated in hyphae including the orthologs of the putative ribosomal proteins
However, there are many up regulated genes during hyphal growth in
Figure 4
Core setsof hyphal- and yeast-specific gene sets
Core sets of hyphal- and yeast-specific gene sets. Expression profiles for the determined six gene sets each with one selected example representative for the corresponding profile visualized on the genome browser: (A) 1° hyphal core, (B) 1° yeast core, (C) 2° hyphal core, (D) 2° yeast core, (E)
Though the 1° and 2° cores share similarly up or down regulated genes in the respective species, the 2° cores show equal expression rates in the DEO analysis despite of divergent morphologies under identical condition.
On the contrary, the 1° cores show clear profiles directly correlating with morphology, including the DEO analysis. 16 genes in the 1° hyphal core containing 84 genes are involved in cell surface architecture, like the orthologous pairs for
In addition, a smaller group of 15 genes was detected which seems to be hyphally repressed comprising the 1° yeast core subset of genes including three cell surface genes
However, there are also differences in the regulation of cell surface genes and of transcriptional regulators during hyphal growth between both species as well. Within the
Taken together, these molecular insights provide evidence, that in addition to a core set of hyphally regulated genes, also species-specific sets of hyphally induced or repressed factors exist for
Next-generation DNA sequencing technologies including RNA-seq provide unbiased approaches for the analyses of gene expression profiles. In contrast to DNA-microarrays where prior information about genomic annotations is essential, RNA-Seq conceptually is unbiased. Furthermore, the determination of quantitative data using RNA-seq is accomplished by counting individual transcripts rather than by deducing transcript abundances from signal intensities following hybridization to specific probes in microarrays. This concept with its open and unbiased architecture allows not only for experimental (
The combination of two complementary sequencing technologies (Illumina and FLX) for the experimental annotations in
Furthermore, it is obvious that the analysis of only a limited set of conditions cannot reveal the complete picture. However, based on the fact that using our approach we could detect 95% of already
In both species, we defined 96 novel protein coding genes of which 70 correspond to
An additional advantage of using experimental data generated by RNA-Seq is evident by the annotation of TARs at single nucleotide resolution. In both species many CDS had to be altered in terms of elongating and shortening them or containing amino acid exchanges due to incorrect predicted splice sites (taken together 466 genes). In some cases, elongations at the N-termini of proteins may contain additional functional domains which were missing recently reflecting the importance of experimental validation of
However, the largest amount of novel genes belongs to the class of non-coding TARs comprising some snoRNAs and a significant number of antisense transcripts. Whether this observation provides a biological mechanism for post-transcriptional regulation like repression of translation or RNA-turnover remains to be analyzed. A recent study in
There is another transcript feature in post-transcriptional regulation which has been well known for
Qualitative comparisons between
While many potential novel targets have been revealed by the qualitative comparison of gene repertoires, we also focused on quantitative differences in the regulation of the orthologs comprising more than 90% all genes. In fact, the concept of RNA-Seq allows for the analyses of quantitative gene expression profiles across two different species. This study is the first to show the possibilities of differential expression of orthologs (DEO) by RNA-Seq and demonstrates a promising approach for unbiased comparisons across two species at very high resolution. A recent approach for comparative transcript profiling was described by O'Connor and colleagues based on
Moreover, a whole set of 84 genes with similar expression profiles could be defined (1° hyphal core) containing 16 cell surface/secreted proteins (
Taken together, this study for the first time reports the comparative experimental annotation of transcriptional landscapes of the two most closely related fungi
Strains and experimental conditions
For the generation of databases and gene expression profiles we used the closely related
RNA preparation
For the isolation of total RNA, blastospore and hyphae inducing media were inocculated with 3 × 106 cells of the overnight culture of
Preparation of cDNA libraries with subsequent high-throughput sequencing
For the normalized cDNA libraries for FLXTitanium-sequencing, equal amounts of approximately 25 μg of total RNA per condition were pooled together per species. To get rid of genomic contaminants another purification step was performed using QIAGEN's RNeasy Mini Plus Kit. From the pooled total RNA poly(A)+-RNA was prepared according to standard protocols
For Titanium sequencing the cDNA in the size range of 500-700 bp was eluted from preparative agarose gels. Aliquots of the size fractionated cDNAs were analyzed by capillary electrophoresis with the Shimadzu MultiNA microchip system. The two normalized and barcoded cDNA libraries were pooled at equal amounts and were sequenced on a full flow cell with the Titanium chemistry resulting in about 500.000 reads per species.
Illumina sequencing was performed on two different machines-GAIIx and HISeq2000. The cDNA libraries for GAIIx-sequencing were generated according Illumina's mRNA-Seq Sample Prep Kit protocol with the samples
Alignment of FLX-Titanium reads
First, the reads were assigned to each species by decoding the barcodes. After separation of the reads, the barcodes at the 5′-end and the remaining sequences from the B adapter at the 3′-end were trimmed. The processed reads were then blasted against the corresponding databases with BLASTN (ncbi-blast-2.2.23). During our studies, we exclusively used the sequence and annotation files from Assembly21 (latest update 2010-06-15,
Mapping Illumina reads
For annotation purposes, the 74–75 bp reads from GAIIx sequencing were mapped with SOAP2 (version 2.20) using a seed length of 40 bp and allowing up to 5 mismatches throughout the whole length of the reads. As SOAP2 is not compatible with intron-spanning reads we performed another mapping with the remaining unmapped reads with TOPHAT (version 1.3.1) to capture them as well. Out of these alignments, it was possible to extract the position and the exon junctions for each read. The 50 bp reads generated with HiSeq2000 were exclusively mapped with TOPHAT (version 1.4.1) using default settings.
Genome-associated assembly
Following reference genome based alignment of reads and manual inspection, we used the genome browser GeneScapes to annotate transcriptional active regions (Lorenz et al., manuscript in preparation, downloadable at "
Defining novel open reading frames
To define ORFs for the identified novel TARs, we translated each possible reading frame in the given direction into a protein sequence. These sequences were aligned to closely related genomes via TBLASTN (default parameters). These were the genomes of
Qualitative comparison between
For a qualitative cross-species comparison at protein sequence level, we initially performed reciprocal WU-BLASTP (BLOSUM62, query sequence length 1000, target database size 1000000). Reciprocal hits under an E-value of 10−13 were considered as true hits and thus orthologs, the remaining genes as species-specific as they had no significant hits or the hits they had did not map back reciprocally. Thus, many proteins from different families were assigned to the group of species-specific genes due to lacking reciprocal hits. By manually comparing these critical genes and also some of the reciprocal hits with the data form Jackson et al.
For non-coding genes, we performed reciprocal BLASTN and used the same significant E-value of less than 10−13 for determining orthologous or species-specific genes.
BLAST comparisons with nc nTARs
BLASTN searches for determination of shared nc nTARs in
BLAST comparisons with coding species-specific genes
BLASTP searches to evaluate conservation of species-specific, coding genes were performed with default settings (version 2.2.25+). Reference annotations from strains
Quantification of RNA-Seq data
Quantification was carried out according to Mortazavi et al.
GO enrichment analysis
For GO term enrichment analyses we applied the web application "GO term finder" available on the "Candida Genome Database" (CGD,
Quantitative Real Time PCR (qRT-PCR)
Reverse transcription reactions were performed using Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Mannheim, Germany). All reactions were carried out according to the manufacture's protocol using 1 μg total RNA and 2.5 μM Anchored-oligo(dT)18 Primer. Quantitative Real Time PCR analysis was performed using the Universal ProbeLibrary Technology (Roche) and the LightCycler 480 Instrument. Amplification assays with a corresponding set of target-specific PCR oligonucleotides combined with a suitable Universal ProbeLibrary hydrolysis probe for 20 different
Real Time PCR experiments were carried out in a 20 μl reaction volume containing 1:20 diluted cDNA template, 1 × Light Cycler 480 Probes Master (Roche) and 0.4 μM of each forward and reverse primer and 0.4 μM of the corresponding UPL probe. The reaction was done in duplicates (technical replicates) for each of the 20 genes. The standard curve preparation for the estimation of the PCR efficiency of each assay was performed under the same conditions, but using 5 dilution steps of the cDNA ranging between 10−1 and 10−5, and water instead of cDNA as a negative control. The PCR was carried out according to the following PCR protocol: 95°C for 5 min; 60 cycles of 95°C for 10 seconds, 60°C for 15 seconds, and 72°C for 1 second, afterwards the PCR reaction was cooled at 40°C for 30 seconds. The analysis of the expression levels was determined using 2nd derivative max method provided by the Roche software version 1.5.0. For the
Accession number
RNA-Seq data from FLX 454 and Illumina have been submitted to the NCBI Gene Expession Omnibus (GEO) under accession number GSE41749.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
CG and KS conceived and designed the experiments. CG performed the experiments. US provided FLX 454 sequencing. CG, SL, PS, EL and JR analyzed the data. CG and KS wrote the paper. KS and SR reviewed and edited the paper. All authors read and approved the final manuscript.
This work was funded by the 'Baden-Württemberg Ministry of Science, Research and the Arts' (720.830-4-12/2a) and the German 'Federal Ministry of Education and Research' (0315409C). CG was supported by a grant of the state of Baden-Württemberg (Landesgraduiertenförderung). We also want to acknowledge 'CGD' (
Acute systemic candidiasis in normal and congenitally thymic-deficient (nude) mice
Innate and acquired immune responses against Candida albicans in congenic B10.D2 mice with deficiency of the C5 complement component
Management and outcome of bloodstream infections due to Candida species in England and Wales
New and emerging yeast pathogens
Putative virulence factors of Candida albicans
Candida dubliniensis: phylogeny and putative virulence factors
One year prospective survey of Candida bloodstream infections in Scotland
Pathogenicity and virulence of Candida dubliniensis: comparison with C. albicans
Differential regulation of the transcriptional repressor NRG1 accounts for altered host-cell interactions in Candida albicans and Candida dubliniensis
Lower filamentation rates of Candida dubliniensis contribute to its lower virulence in comparison with Candida albicans
Comparative genomics of the fungal pathogens Candida dubliniensis and Candida albicans
Accurate whole human genome sequencing using reversible terminator chemistry
Genome sequencing in microfabricated high-density picolitre reactors
The transcriptomes of two heritable cell types illuminate the circuit governing their differentiation
Comprehensive annotation of the transcriptome of the human fungal pathogen Candida albicans using RNA-seq
Noncanonical transcript forms in yeast and their regulation during environmental stress
Annotating genomes with massive-scale RNA sequencing
Power of deep sequencing and agilent microarray for gene expression profiling study
RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays
Mapping and quantifying mammalian transcriptomes by RNA-Seq
MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene
Experimental annotation of the human pathogen Candida albicans coding and noncoding transcribed regions using high-resolution tiling arrays
Evolution of Yeast Noncoding RNAs Reveals an Alternative Mechanism for Widespread Intron Loss
Strand-specific RNA sequencing reveals extensive regulated long antisense transcripts that are conserved across yeast species
Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling
Evolutionary roles of upstream open reading frames in mediating gene regulation in fungi
An iron homeostasis regulatory circuit with reciprocal roles in Candida albicans commensalism and pathogenesis
Cloning and characterization of ECE1, a gene expressed in association with cell elongation of the dimorphic pathogen Candida albicans
The zinc cluster transcription factor Ahr1p directs Mcm1p regulation of Candida albicans adhesion
Cloning and characterization of CAD1/AAF1, a gene from Candida albicans that induces adherence to endothelial cells after expression in Saccharomyces cerevisiae
A family of Candida cell surface haem-binding proteins involved in haemin and haemoglobin-iron utilization
NRG1 represses yeast-hypha morphogenesis and hypha-specific gene expression in Candida albicans
A study of the Candida albicans cell wall proteome
EFG1 is a major regulator of cell wall dynamics in Candida albicans as revealed by DNA microarrays
Isolation of a putative Candida albicans transcriptional regulator involved in pleiotropic drug resistance by functional complementation of a pdr1 pdr3 mutation in Saccharomyces cerevisiae
Genomic evidence for a complete sexual cycle in Candida albicans
Differential filamentation of Candida albicans and Candida dubliniensis Is governed by nutrient regulation of UME6 expression
Comparative transcript profiling of Candida albicans and Candida dubliniensis identifies SFL2, a C. albicans gene required for virulence in a reconstituted epithelial infection model
Transcription profiling of Candida albicans cells undergoing the yeast-to-hyphal transition
Genome-wide gene expression profiling and a forward genetic screen show that differential expression of the sodium ion transporter Ena21 contributes to the differential tolerance of Candida albicans and Candida dubliniensis to osmotic stress
Evolution of pathogenicity and sexual reproduction in eight Candida genomes
A conserved regulatory role for antisense RNA in meiotic gene expression in yeast
Regulation of fungal gene expression via short open reading frames in the mRNA 5′untranslated region
Evidence for translational regulation of the activator of general amino acid control in yeast
Flexible survival strategies of Pseudomonas aeruginosa in biofilms result in increased fitness compared to Candida albicans
The Set3/Hos2 histone deacetylase complex attenuates cAMP/PKA signaling to regulate morphogenesis and virulence of Candida albicans
Portrait of Candida albicans Adherence Regulators
Preparation of poly(A) + RNA
DEGseq: an R package for identifying differentially expressed genes from RNA-seq data
Statistical significance for genomewide studies


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Jacqulyn Buglisi presents her company in its 20th-anniversary season. Two programs will include four premieres: This Is Forever, Snow Falling on Water, Butterflies and Demons, and Zjawa (by Katarzyna Skarpetwoska, who was a student of Buglisi's at Juilliard). Guest artists are Martine van Hamel and Charles Askegard.
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Description: What is Jewish cooking in France? In a journey that was a labor of love, Joan Nathan traveled the country to discover the answer and, along the way, unearthed a treasure trove of recipes and the often moving stories behind them. Nathan takes us into kitchens in Paris, Alsace, and the Loire Valley; she visits the bustling Belleville market in Little Tunis in Paris; she breaks bread with Jewish families around the observation of the Sabbath and the celebration of special holidays. All across France, she finds that Jewish cooking is more alive than ever: traditional dishes are honored, yet have acquired a certain French finesse. And completing the circle of influences: following Algerian independence, there has been a huge wave of Jewish immigrants from North Africa, whose stuffedbrikand couscous, eggplant dishes andtagines-as well as their hot flavors and Sephardic elegance-have infiltrated contemporary French cooking. All that Joan Nathan has tasted and absorbed is here in this extraordinary book, rich in a history that dates back 2,000 years and alive with the personal stories of Jewish people in France today.
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List price: $46.00Copyright year: 2010Publisher: Knopf Doubleday Publishing GroupPublication date: 10/26/2010Binding: HardcoverPages: 400Size: 8.50" wide x 10.00" long x 1.25" tall Weight: 3.344Language: English
Joan Nathan is the author of many books on Jewish food, including the award-winning Jewish Cooking in America, The Jewish Holiday Kitchen, and The Jewish Holiday Baker. She lives in Washington, D.C.
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Apple and Black Currant Crumble Bars
Recipe by Linky
A yummy dessert using those high in Vitamin C and antioxidant wonders!
Top Review by stormylee
Oh my goodness, this is SO good. It has never occurred to me to combine apples and blackcurrants before but it really is a wonderful combo. The apples from our apple tree are on the tart side, and the caramel-y sweetness of the batter worked beautifully with them. I did use a bit more than half of the batter for the base and patted it down to a layer, and then just sprinkled the remaining batter on top of the berries. A delicious recipe - thank you for sharing Linky!
See Discussion:2 Reviews
Ingredients Nutrition
- 6 ounces plain flour (all-purpose)
- 1 teaspoon baking powder
- 1 pinch salt
- 6 ounces butter (or margarine)
- 1 cup soft brown sugar
- 1 cup rolled oats
- 1 lb apple, peeled, cored and sliced
- 2 tablespoons cornstarch
- 2 teaspoons ground cinnamon
- 1⁄2 teaspoon nutmeg
- 1⁄2 teaspoon ground allspice
- 8 ounces black currants
Directions See How It's Made
- Mix the flour, baking powder and salt, then rub in the butter or margarine.
- Stir in the sugar and oats.
- Spoon half into the base of a greased and lined 9 in square cake tin (pan).
- Mix the apples, cornflour and spices and spread over.
- Top with the Black Currants.
- Spoon over the remaining mixture and level the top.
- Bake in a preheated oven at 350°F for 30 minutes until springy.
- Leave to cool, then cut into bars.


text:
"Silence gives me freedom in both real time and psychic time. When I talk less, I see more. And silence gives me time to pray."
Ravished by Beauty, by Belden C. Lane
In this splendid book Belden Lane has made a double contribution—to the reordering of our perspectives on creation and to our understanding of the Reformed tradition as a contributor to this reordering.
Naked Spirituality, by Brian D. McLaren
The architecture of Naked Spirituality is a bit complicated. But if you can bear with it, you'll find that Brian McLaren offers countless insights.
Stretches of emptiness are not unusual in life, nor in the life of faith.
Just as I've come to appreciate how seasons transform the land, I've also become aware of my internal landscape. The two seem bound together in many ways.
I want to arrive at the kind of equilibrium admired by the disciples who broke down the barrier to St. Anthony's fortress. To do this, I have to befriend the demons dwelling in the cave of my heart.
Falling Upward, by Richard Rohr
Richard Rohr has written his most sage, most important book yet. Its message is straightforward and bracing: the spiritual life is not static.


text:
Snagit's image and video capture is powerful, but not complex. All-in-One Capture® grabs your entire desktop, a region, a window, or a scrolling window from any webpage or application—-all with a single hotkey or click. Snagit gives you a simple way to edit. Show a complex process, give detailed feedback, or share important information with the people who need it most. Download your free trial now!Advertisement
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Revolution is an extremely stable Java based Runescape private server, completely invunerable to crashers and flooders.


text:
Carpenters - Only Yesterday MP3
Music video by Carpenters performing Only Yesterday. (C) 1985 A&M Records.
Only Yesterday
CarpentersAfter long enough of being alone Everyone must face their share of loneliness In my own time nobody knew The pain I was going through And waiting was all my heart could do Hope was all I had until you came Maybe you c[...]
Carpenters - Only Yesterday - 1975 MP3
After long enough of bein' alone Everyone must face their share of loneliness In my own time nobody knew the pain I was goin' through And waitin' was all my ...
Carpenters-Only Yesterday-HQ Sound MP3
One of my favorite Carpenters songs.The HQ sound doesn't kick in till 1:20.I think you will notice the difference in sound quality-enjoy.
ONLY YESTERDAY (1991) Trailer MP3
Director: Isao Takahata.
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Taken by Trees - Only Yesterday Lyrics: Oh honey, I really miss you. Oh honey, though it was only yesterday you kissed me. And that kiss, that kiss was so, ...
Isla Grant - Only Yesterday MP3
Isla Grant Only yesterday.
CARPENTER - ONLY YESTERDAY (with lyric) MP3
Only Yesterday - Official Trailer MP3
Taeko, a single woman working a desk job in Tokyo, has reached a point in her life where she needs to get away from her job, her family and most of all, big city ...
The Carpenters - Only Yesterday (Tradução) MP3
O luar é a luz do sol que está dormindo... Mário Quintana.
Only Yesterday-The Carpenters MP3
Only Yesterday-The Carpenters.
Only Yesterday - Isla Grant 그저 어제 같기만 -유샤인 번역판 MP3
This video has been on Youtube.com some time now. There have been some Korean translations already on this song but in my effort to translate it more closely ...
Only Yesterday track 1 MP3
composer Katz Hoshi studio ghibli Omoide poroporo ( only yesterday)
Only Yesterday The Carpenters- Legendada MP3
Linda música..Linda voz!!!
Fall Out Boy - Just One Yesterday (Part 6 of 11) ft. Foxes MP3
SUBSCRIBE TO FOB: http://bit.ly/1gI9KrO Listen to the new single "Centuries" http://youtu.be/sCbS-TLEoRA and download it on iTunes http://smarturl.it/centuries ...
Just One Yesterday
Fall Out BoyI thought of angels Choking on their halos Get them drunk on rose water See how dirty I can get them Pulling out their fragile teeth And clip their tiny wings Anything you say can and will be held against you So only say my name It will be held against you Anything you say can[...]
Only Yesterday (Cover) - Roland's Basement #8 MP3
Roland's Basement #8 June 8th, 2013 Guest Musicians: Michelle Morton-Curit (vocals) Ed Castine (bass) Bruce Flower (guitar) Marilyn Flower (backing vocals) ...
Karaoke: Only Yesterday - Carpenters. MP3
11. Only Yesterday - Carpenters. Che2xs.blogspot.com I DON'T OWN ANYTHING.
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Clipe retirado do DVD "Duetos Românticos II".
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お久しぶりです!今年もよろしくお願いします!! 今年一発目の動画はOnly Yesterdayに挑戦してみました。 楽しいベースランなのですが、フレーズ...
Jess Lewis' incredible cover of Larry Carltons '(It Was) Only Yesterday' | JamTrackCentral.com MP3
Get the free download here: https://jamtrackcentral.com/store/series/jess-lewis-it-was-only-yesterday/ Check out the Axe FX ii here: http://www.fractalaudio.com/ ...
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I must say it's not really acapella, since it's not voice only, but very near. Worth listening, completely! It's actually one of the audio channels from the SACD, and ...
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Isla Grant - Only Yesterday MP3
Video By James R. Yetman, [email protected] Isla Grant was born in Wigtownshire, Scotland but when she was a few months old her family ...
Only Yesterday MP3
Provided to YouTube by Entertainment One Distribution US Only Yesterday · Gene Watson Then And Now ℗ KOCH RECORDS NASHVILLE Released on: ...
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Carpenters Only Yesterday.
Mi'das - Feels Like Only Yesterday MP3
Subscribe for more videos: http://bit.ly/ChristianTierneyYoutube Singer-songwriter Mi'das a.k.a Mike Davies performs this soulful acoustic version of his song ...


text:
|Trad, 55'
|YDS: 5.11a French: 6c Ewbanks: 22 UIAA: VII+ ZA: 22 British: E3 5c PG13 [details]
|year round
|Page Views:
|Submitted By:
|Jason Halladay on Apr 3, 2007
|Comments on Boy What Nice Fellows
|Add Comment
|Show which comments —
By George Perkins
From: The Dungeon, NM
May 14, 2016
rating: 5.11b 6c 23 VIII- 23 E3 5c
In Jemez Rock, "Boy What Nice Fellows" (11b, TR) is described as the face between "Squeeze Chimney and "Boy What Assholes", going right and joining "Assholes" at the top. The lower half of this TR problem likely shares terrain with the more recently bolted "Nice Guys Finish Last", but branches right mid-way up. This would put it to the right of "Nice Guys" in the left-to-right route list.
The face with the OW on it (described here) is "Squeeze Chimney" (5.10d).


text:
Does Portland have any shopping areas with upscale boutiques and cute shops?
Yes! You'll probably enjoy the Nob Hill area – specifically Northwest 23rd Avenue – it's a mile or so of cute shops and upscale boutiques.
In downtown Portland, I'd recommend the Pioneer Place Mall. Adjacent to downtown is the Pearl District, which is famous for its upscale boutiques.
Have something to add What's Your Answer?
Share your thoughts Answers From Other People
Your comment will be the first one for this story. Some might think of this as a lot of pressure, but as a trail blazer you recognize that someone has to be first. Your fellow travelers appreciate your opinion, so thanks in advance!
Other questions about Portland
- I've been looking around Portland for a place that dishes up biscuits and gravy with a bite to it. Do you know a place that makes it with spicy sausage or something?
- We're coming to Oregon this summer. We want to rent a place on the beach, walk, eat well, see local sites and shop in art galleries. We also want to spend a couple of days in Portland. There are four of us and none of us have ever spent time in Oregon before. Do you have any suggestions for how to set up a couple days in the city and four days on the coast? – Elaine R.
- What are the best sight-seeing things in or close to Portland? – Colette J.
About Ask Oregon Expert Dave Strom
A Portland native, since 2007 I've been covering offbeat Portland history, news, and events at my blog, Dave Knows Portland. In May 2011, I relocated to Astoria, Oregon. As an Ask Oregon Ambassador, I look forward to sharing my knowledge of Portland and my explorations of Oregon's North Coast.


text:
Miranda Lambert 'deserves a drink'
Miranda Lambert "deserves a drink".
The country singer - who recently announced her divorce from husband of four years, Blake Shelton - reportedly told fans she was desperate for a glass or two of her favourite tipple following the split.
Speaking on stage at a country music festival in Minnesota, she said: "I wish I've been drinking all day. I deserve it: I just got divorced."
Meanwhile, a source previously claimed Miranda is "trying to get back to normal" following the divorce.
They said: "She's really trying to get back to normal, and I think being onstage is a part of that."
And the pair are also believed to have insisted to pals they are "just friends" after they enjoyed a playful exchange on Twitter.
The insider said: "This was not a deliberate plan to show some kind of unified front.
"There are no hard feelings. They want to move on as friends, and they are doing just that."
The news of their split was confirmed last month by a joint statement, which read: "This is not the future we envisioned. And it is with heavy hearts that we move forward separately.
"We are real people, with real lives, with real families, friends and colleagues. Therefore, we kindly ask for privacy and compassion concerning this very personal matter."


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Template:Infobox musical artist Usher Raymond IV (born October 14, 1978) is a Grammy Award-winning American R&B/pop singer and Actor who rose to fame in the mid-late 1990s. To date, he has sold approximately 20 million albums in the United States, and over 35 million worldwide.
Early lifeEdit
Usher was born in Dallas, Texas to Usher Raymond III and Jonetta Patton. [1]. Raised primarily by his mother, Usher and his family later relocated to her home town of Chattanooga, Tennessee. Usher joined his local church choir where he discovered his ability to sing. As early as junior high school, Usher successfully entered local talent shows imitating his idol Michael Jackson. As a pre-teen Usher was a member of a boy band, although his mother (now managing his career) later withdrew him from the group to concentrate on his solo work. At the age of 10 Usher and his family moved to Atlanta, Georgia, in the belief that the city would provide greater opportunities for showcasing his talent. While in Atlanta, Usher attended the Performing Arts Magnet North Springs High School, and participated in show choir. Usher also studied karate under professional wrestler Ernest Miller for four cernturies.
Music careerEdit
At age thirteen, Raymond performed on Star Search,where he was spotted by an A&R representative from LaFace Records, who arranged an audition for him with L.A. Reid. A contract with the company soon followed. In 1993, Usher was introduced on "Call Me A Mack," a song he recorded for the soundtrack album to the John Singleton film, Poetic Justice
Usher (Debut album)EditOn August 30, 1994, LaFace released Usher's self-titled debut album. Sean "Puff Daddy" Combs produced several of the tracks and co-executive produced the album. Usher peaked at number twenty-five on the R&B Albums chart and was accompanied by three singles: "Can U Get Wit It," "Think Of You," and "The Many Ways." Though the album garnered Usher some attention with urban music listeners, it was not a runaway hit and went virtually unnoticed by pop audiences; selling approximately 50,000 copies in America (and 100,000 abroad).
The quasi-adult themes in Usher were questioned by some critics due to the young age of the artist. Usher himself later admitted that he was not fully comfortable with the direction of the album.
After graduating from high school, Usher continued to hone his skills as a stage performer and laid the groundwork for his second album. He also appeared on "Let's Straighten It Out," a 1995 duet with fellow Atlanta teen recording artist Monica; and on 1996's "Dreamin'", the first single from Rhythm of the Games, LaFace's Olympic Games benefit album.
My WayEditUsher developed a friendship with Jermaine Dupri, with whom he co-wrote and produced several tracks for his sophomore album, My Way, released on September 16, 1997. The lead-off single, "You Make Me Wanna" was serviced to radio that summer and hit number one in its second week out—the same week that the album was released. "You Make Me Wanna" stayed on top of the R&B/Hip-Hop track charts for a total of 11 weeks (the longest-charting R&B hit in more than 3 years) and at #2 on the Pop charts for 7 weeks (second only to Elton John's "Candle In the Wind.") The single remained on the R&B chart for an unprecedented 71 weeks and became Usher's first platinum single. The albums second single, "Nice & Slow," was released in November, and in January 1998 peaked at #1 on the Hot 100 Singles Chart for two weeks, as well as #1 on the R&B/Hip-Hop Singles chart for eight weeks. The following summer, the album's eponymous single title track peaked #2 pop/#4 R&B.
In the closing months of 1997, Usher embarked on a series of tour engagements including a spot on Puffy's No Way Out tour, dates with Mary J. Blige, and the opening spot on Janet Jackson's The Velvet Rope tour. My Way was certified 6x platinum in the United States. Usher received his first Grammy Award nomination as Best Male R&B Vocal Performance for "You Make Me Wanna," which was also nominated for a Soul Train Music Award as 'Best Male R&B/Soul Single.'
It was during the 'My Way' period, during which Usher came out of the closet as bisexual on NBC's Friday Night with Rita Sever. Rita asked Usher about his sexual orientation to which he replied, "I'm Bi."Template:Cn
Usher made his acting debut on the UPN television series Moesha, which resulted in a recurring role on the series and subsequently his first leading film role in 1998's, The Faculty. Usher's extracurricular activities outside of the recording studio gathered momentum over the following year as he was cast in the daytime drama The Bold and the Beautiful and was seen in the family series Promised Land. He completed two more films, She's All That, and his first starring role in Light It Up. He also appeared in the Disney TV movie "Geppetto" with Drew Carey. Usher also became involved community activities: he served as national spokesperson for the U.S. Department of Transportation's "Get Big On Safety" campaign, and appeared on NBC's NBA Inside Stuff. He also participated in the NBA "Stay In School" program and performed at several of the franchise's jamborees.
Raymond's first concert album, Live was released in 1999—which featured appearances by Lil' Kim, Jagged Edge, Trey Lorenz, Shanice, Twista and Manuel Seal, and was RIAA-certified Gold.
8701EditUsher's third studio album, originally titled All About U, was slated to be released in early 2001. The first single, "Pop Ya Collar" (co-written and produced by Kevin "She'kspere" Briggs), was released in late 2000 and became a #2 hit in the UK—but underperformed in America. The album was subsequently pushed back and retooled after select tracks were later leaked to the radio and Internet, and garnered a largely cold public response.
After having revised the album, it was renamed 8701 (named for the years of his career at that point [1987-2001], and for its release date of August 7, 2001, or 8/7/01). The first two singles "U Remind Me" and "U Got It Bad" each topped the U.S. Hot 100 singles chart for four and six weeks respectively in the summer and fall. In early 2002, "U Don't Have to Call," also went top five pop. The albums final single, "U-Turn" was released outside of America, and became a top twenty hit in Australia and the U.K.. 8701 was certified 4x platinum, selling more than eight million copies worldwide.
In February 2002, Usher won a Grammy for 'Best Male R&B Vocal Performance' for "U Remind Me." The next year, he won the same award for "U Don't Have to Call," making Usher the only artist aside from Luther Vandross, more than a decade before, and Stevie Wonder (in the 1970s) to win this award consecutively.
In summer 2002, Usher contributed vocals to P. Diddy's "I Need a Girl, Part I." The year closed out with a trio of TV series appearances, all in November, on The Twilight Zone, 7th Heaven, and American Dreams (in which Usher portrayed Marvin Gaye).
ConfessionsEditConfessions, Usher's fourth studio album, was released on March 23, 2004—just as its first single, "Yeah!" was in its sixth week at number one on the Billboard Hot 100 singles chart and fifth week on top of the Hot R&B/Hip-Hop Singles chart. The album's 1.1 million unit first week sales was not only the highest first week numbers ever scanned by a male R&B artist in Soundscan's 13-year history (breaking R. Kelly's record of 540k for TP-2.com back in 2000), but also the highest first week scans by any male artist since Eminem's The Marshall Mathers LP (also in 2000, with 1.7 million units).
The album's second and third singles: "Burn" and "Confessions Part II" also topped Billboard's Hot 100 singles charts, the former for eight weeks. Usher became the third lead artist in pop music history (but the first solo artist) to have three singles inside the Hot 100's top 10 since The Beatles in 1964, and the Bee Gees in 1978. In September 2004, "My Boo," a duet with singer Alicia Keys (which was included on a Special Edition, released a half year after the original version), also peaked at number one and gave him a total of twenty-eight weeks on top of the U.S. charts that year. In December, the album's final single "Caught Up" peaked at number eight on the Hot 100. Confessions became one of the most successful records of the year; certified 9x platinum in America, topping the world charts for several weeks and eventually selling more than 15 million copies all over the globe. [2]
Usher was recognized with numerous awards, including two MTV Awards for "Best Male R&B Artist," three World Music Awards ("Best Male Artist," "Best Male Pop Artist" and "Best R&B Artist"), two Source Hip-Hop Music Awards ("Male R&B Artist of the Year" and "R&B/Rap Collaboration of the Year," a Nordic Music Award, three Radio Music Awards, four American Music Awards and two more MTV Europe Music Awards. At the 47th annual Grammy Awards ceremony in 2005, Usher won three awards, including: "R&B Performance by a Duo or Group With Vocals" (for "My Boo" with Alicia Keys), "Rap/Sung Collaboration" (for "Yeah!") and "Contemporary R&B Album" (for Confessions).
In spring 2005, Usher scored a #3 Hot 100 hit as a featured vocalist on Lil' Jon's "Lovers and Friends." In November 2005, Usher starred as a disc jockey named Darrell in the Lions Gate film, In the Mix.
On August 22 2006, Usher took over the role of Billy Flynn in the long-running Broadway musical Chicago. Critical response to his appearance has been mixed: his singing and dancing received more positive reviews, while his acting received negative notices. Usher is currently engaged to girlfriend and stylist Tameka Foster.[1]
Usher is also back in the recording studio to work on his as yet untitled new album. The album is rumored to be called "Mr. Entertainment", but this is not confirmed.[2][3] Supposedly, there is a leaked Usher album on the Internet called The Masterpiece, however this is not an official album but a collection of unreleased material from his other albums.
A one minutes clip of a song entitled "Make It Rain" has leaked to the internet. It is unknown as of so far if this will be a single or not. "The Realest" is a track produced by Jermaine Dupri and is expected to make the album according to an interview MTVNews.com had with Usher on April 12.
Albums Edit
- 1998: The Faculty
- 1999: She's All That
- 1999: Light It Up
- 2001: Texas Rangers
- 2005: In the Mix
- 2006: The Ballad of Walter Holmes
- 1996: The Parent Hood
- 1997-1999: Moesha
- 1998: The Bold and the Beautiful
- 2000: Geppetto
- 2001: 7th Heaven
- 2002: The Twilight Zone
- 2003: Soul Food
Ush RecordsEdit
In 2005 Usher announced that he was creating his own record label, 'Ush Records'. The first two signees are rapper Rico Love and R&B group One Chance. The first album from the label, One Chance's Private, is scheduled for release on May 29, 2007. Rico Love's The 5th Element is expected to be released later in 2007, along with the debut album from JB3 (real name Jimmie Brown). A new artist Vick Joe has also been signed.
External linksEdit
- Official Site
- Usherforever.com
- Usher at MTV.com
- Usher at VH1.com
- Usher at RollingStone.com
- Usher Raymond at the Internet Movie Database
{{Wikipedia:WikiProject LGBT studies/Category unsourced|Bisexual musicians}}
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Watch the HD highlights of Sevilla Vs Barcelona super cup 14 August 2016. Suraez scored a goal and took Barcelona to the remarkable victory. Fc Barcelona's player Munir also scored a goal and led barcelona to 0-2 victory. All goals and highlights are shown in the video.Read More »
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Nina Ricci
"Hikimori" those who withdraw, is a social trend in modern japan. HELLOHIKIMORI was founded in 2004 by Nathalie Melato and David Ronhel. Based in central Paris, on the historical ile de la cité, HKI™ is an independent design studio with cross-discipline
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Grabo Evolution is a compact heterogeneous sheet vinyl flooring developed for commercial areas. Evolution has a wide range of colors combined with high durability and low maintenance properties. The dimensional stability is ensured by scrim glass construction. Evolution is the ideal choice for any application area.
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Grabo Silver Knight combines Nano Silver and Nano Titanium Dioxide technologies to provide the most hygienic resilient sheet vinyl flooring available. Silver Knight ELIMINATES 99% of harmful BACTERIA, disinfecting with a double-defense line. A true NO WAX surface. Superior stain resistance and self-cleaning properties. Phthalate free.
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Sincol Luxury Vinyl Tile and commercial flooring products are manufactured utilizing the finest materials available. A patented process delivers the richest design clarity and depth of color. The durable PVC wear surface offers ease of maintenance in the most demanding applications.
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- Average
- Main course: £10
- Food spend: £25
- Opening hours
- Monday to Saturday: 11:30 - 23:00 Sunday: 12:00 - 22:30
Fabulous food and service.
christine stent
04 May 2016
from clanfield
The Red Lion Chalton started life in the 12th century and continues to please diners with tasty British dishes and well-kept ales. The interior exudes rustic elegance with an inglenook fireplace and chunky wooden furniture.
Spanning from salads and juicy burgers to piping-hot, stuffed-full pies, The Red Lion menu is lovingly prepared by the talented resident chefs. Ingredients are sourced from across the British Isles, and meats and daily-fresh fish catches can be traced back directly to sustainable, trusted suppliers. Choose from dishes including duck terrine or Chalcroft Farm (Hampshire) beef burger.
Red Lion pub also presents a renowned drinks list, featuring award-winning ales including Fuller's most well-known brew, London Pride. Fine wines, premium spirits and soft drinks are also on offer.
A buzzing family-friendly Hampshire pub, The Red Lion is a 3-minute drive from the A3 Road.
Red Lion - Chalton
Chalton, Waterlooville, PO8 0BG
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((((( 16 May 2016 by Paul from Waterlooville
This was the first time we have used the booking system and I was very impressed at the ease with which this was accomplished. The restaurant itself is always top notch. Let's face it, if you turn up on a beautiful spring evening in early summer/late spring, you would expect a pub/restaurant to be packed: and The Red Lion was teeming - hard to find a parking space. Yet the staff were friendly and attentive, the food was as usual, very good indeed and the atmosphere convivial. Everything about The Red Lion says "We will return".
In summary, it is the staff that generate the success of an establishment and this is something that The Red Lion has managed in spades. Many congratulations to the management for the level of care achieved.
((((( 4 May 2016 by christine stent from clanfield
Fabulous food and service.
((((* 21 April 2016 by Marilyn Brady from waterlooville
food very good & staff very helpful, good atmosphere
((((* 4 January 2016 by Helen White from Gosport
Relaxed atmosphere. Great food and drink. Not pressured to vacate table. Lovely pub!
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Fuck a man or let one fuck you. You'll love it. The parts fit together perfectly!!!
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