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Snow Pea Salad
|Snow peas
|2 Cup (32 tbs)
|Red bell peppers/Green bell peppers
|2 Large, sliced into rings
|1⁄2 Pound, sliced
|Walnut oil
|1 Cup (16 tbs)
|Champagne vinegar/White wine vinegar
|2 Tablespoon
|Granulated sugar
|1 Tablespoon
|Lemon juice
|1 Tablespoon
|Sesame seeds
|1 Tablespoon, toasted
1) In a small container, combine oil, vinegar, sugar and lemon juice and keep the oil mixture aside.
2) Boil the snow peas briefly for 1 to 2 minutes and drain.
3) Wash thoroughly with cold running water to prevent cooking any further.
4) Allow to cool.
5) In a large glass bowl, mix together snow peas, pepper rings and mushrooms.
6) Just before serving, toss vegetable mixture gently with oil mixture.
Drizzle sesame seeds on top and serve.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size
Calories 588 Calories from Fat 502
% Daily Value*
Total Fat 57 g87.6%
Saturated Fat 5.3 g26.5%
Trans Fat 0 g
Cholesterol 0 mg
Sodium 9.1 mg0.4%
Total Carbohydrates 18 g5.8%
Dietary Fiber 4.5 g18%
Sugars 10.7 g
Protein 5 g10.7%
Vitamin A 57.6% Vitamin C 222.1%
Calcium 7.9% Iron 15.7%
*Based on a 2000 Calorie diet


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Lot 104: 2.46CTW Genuine VS2-SI1/G-H Diamond & SOLID 14K White Gold Ring (W/Arthritic Band) *NICE*
Fine Antiques, Art & Celebrity Memorabilia
by GWS Auctions
October 22, 2016
Beverly Hills, CA, USALive Auction
Looking for the realized and estimated price?
Description: 2.46CTW Genuine VS2-SI1/G-H Diamond & SOLID 14K White Gold Ring (W/Arthritic Band) *NICE* - This ring is made of solid 14K white gold and has a unique, arthritic band (a band with an open & close clasp for easy removal). This ring contains: (1) 0.60CT marquise-cut center diamond, (2) 0.40CT each marquise-cut diamonds, (2) 0.28CT each marquise-cut diamonds, (4) 0.10CT each tapered baguette-cut diamond partitions & (2) 0.05CT each tapered baguette-cut diamond partitions. The diamonds range from VS2-SI1, and are all stunningly eye clean and fiery white. Ring size: 6. (6.4 grams)
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subset: commoncrawl


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Womens Air Jordans 3 (III) Fluff Purple Yellow
|Air Jordan
|Womens Air Jordan 3
$150.00 $86.00
|In Stock, shipping in 24 hours. Free Shipping!
It looks like he had been gifted with a range of the Womens Air Jordan 3 for the Christmas, putting him along the short list of people who never live in Eugene that have access to the sneakers.Recently we gave you a preview of what was to return for the Womens Air Jordans 3 (III) Fluff Purple Yellow for facets of 2014 and we noted which your Womens Air Jordans 3 (III) Fluff Purple Yellow would be releasing before long.Premium white moist cloth builds the stunning Air Jordan 3 Low model, while dunkelhrrutige hits their extraordinary rubber tongue, and thus sport blue comes with some color stand out over the tongue logos, midsole detailing and traditional lacelocks.Keep tuned in to Sole Financial institution for further details.
Tags : Air Jordan 3 Low ,


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Top Site - Voting
This project received 8 bids from talented freelancers with an average bid price of $268 USD.Get free quotes for a project like this
Project Budget$30 - $250 USD
Total Bids8
Project Description
I need someone to make me a website like [url removed, login to view] .
Before you apply, look into this website to be sure of all the scripts used there.
I don't really care about design, i will make it by myself, I want all the scripts from that site.
Skills Required
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- Set your budget and the timeframe
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- Forbes
- The New York Times
- Time
- Wall Street Journal
- Times Online


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Wego.com.sg recently found 12 hotels in Lordsburg, United States. Book last minute hotel or find great hotel deals such as Comfort Inn & Suites Lordsburg for S$135 per night. Save more on cheap hotels with rates from as low as S$95 per night.
Filters help you save time when choosing where to stay. Try clicking on a few now...
Lordsburg Hotels 3 Star Hotels Lordsburg Cheap Hotels Lordsburg Cheap Accommodation Lordsburg
Flights to United States Flights from United States United States Airports United States Airlines


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Direct Drive Power Washer. Check out our complete line of Gas Powered Cold Water Pressure and Power Washers including our direct driven power washers that are perfect for applications ranging from consumer and light-duty residential applications to full scale commercial cleaning projects.
Direct Drive Power Washer
Regular Price: $1,831.00
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Need more power than a basic residential pressure washer? Step up to this 4.0 GPM @ 4000 PSI with a 13HP Honda Engine, and a high quality GP Pump.
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< Return to Product Page
Customer Reviews
Mini Moso Charcoal Deodorizers
Overall Rating:
Customer Reviews
Rated 4.8 out of 5Â by 5 reviewers.
Rated 4 out of 5Â by RC62 Seem effective I have a couple pairs of these. But, I have many shoes and don't wear the same pairs all the time, so I don't rely on these too much. But, if you have wet shoes or boots, smelly shoes and teenagers, then get these! They are useful! Who wants to smell shoes that smell like a used bathroom? May 30, 2016
Rated 5 out of 5Â by Like2Shop Charcoal Deodorizers really work! These are the best deodorizers I have ever used. Mini Moso deodorizers absorbs odors better than any product! Sprays leave nasty odors in my shoes; other inserts don work very well. Mini Moso actually works!! You have to remember to put them in the sun for 1 hour once a month as the sun will clean out the impurities that the bamboo charcoal has absorbed. I own 3 pairs. January 3, 2016
Rated 5 out of 5Â by bjewelry Deodorizers for shoes and boots Love these deodorizers, I am getting some for gifts and a pair for me. December 16, 2015
Rated 5 out of 5Â by mamabrown No more stinky shoes I've tried Lysol, powders, baking soda, etc. with little success. Finally found something that actually removes odor from my stinky shoes. At the end of the day, I slip these into each sweaty shoe (there's 2 to a pack) and by morning, they smell fresh. Can't wait to try them on my kid's sport shoes! March 1, 2014
Rated 5 out of 5Â by miho This works! was skeptical at first, but this actually works, only took less than 24hrs to notice the difference! March 24, 2013
1-5 of 5


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Brenda S.'s Event Journal
Sunday June 10, 2007 / 6:00pmMemorials By Moonlight Walking Tour Around the Tidal Basin
2 tickets
General Admittance
How was your experience?
This event was very informative, I really enjoyed the tour and would suggest it to anyone visiting or living here in the Washington DC area. Thanks to the tour guide.


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- 1a story created using the imagination, especially one that is full of action and adventure Dickens' 'A Tale of Two Cities' a fairy/moral/romantic, etc. tale His latest book is a delightful children's tale about talking animals. see also folk tale See related entries: Types of story
- 2an exciting spoken description of an event, which may not be completely true I love listening to his tales of life at sea. I've heard tales of people seeing ghosts in that house. The team's tale of woe continued on Saturday (= they lost another match). Her experiences provide a cautionary tale (= a warning) for us all. see also telltale Word Origin Old English talu 'telling, something told', of Germanic origin; related to Dutch taal
BrE BrE//teɪl//; NAmE NAmE//teɪl//Types of story
to survive a difficult or dangerous experience so that you can tell others what really happened
live, etc. to tell the talejump to other results
(disapproving) an old idea or belief that has been proved not to be scientific
an old wives' talejump to other results
to give some information that is different from what you expect or have been told
tell a different story/talejump to other results
to explain itself, without needing any further explanation or comment Her face told its own story.
tell its own tale/storyjump to other results
to tell somebody about something that another person has done wrong related noun telltale
tell tales (about something/on somebody)jump to other results
Check pronunciation: tale


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Soros Firm Buys Morgan Stanley, Citigroup, JPMorgan ChaseKatherine Burton
Soros Fund Management LLC, the family office that oversees $24 billion, bought Morgan Stanley and added to its holdings of Citigroup Inc. and JPMorgan Chase & Co. in the fourth quarter, according to a regulatory filing.
Billionaire George Soros's firm bought 4.1 million shares of Morgan Stanley, valued at $79 million as of Dec. 31, today's filing with the Securities and Exchange Commission shows. The firm added 6.6 million shares of Citigroup, bringing the value of its stake to $321 million. It added 1.6 million shares of JPMorgan, increasing the position's value to $113 million.
Before it's here, it's on the Bloomberg Terminal. LEARN MORE


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Holy Land Foundation Protest
1100 West Commerce Street
Dallas, Texas 75202
Today, I took this shot of a protest across the street from the federal courts building. Not only was this my first "protest in action" photo but I was also driving by at approximately 30 mph. Now, that's talent! :-)
So why were these people there? See below:
( 2007.08.17 article from WFAA.com) DALLAS - Police are on high alert as the Holy Land Foundation trial continues in downtown Dallas. The trial is centered around terrorism and The Holy Land Foundation, who is accused of funneling money to terrorist organizations. The department has a command post near the federal courthouse to watch for suspicious people.
"We evaluate continually everyday, and will be during the trial," said Deputy Chief Vince Golbeck, Dallas Police Department.
Several recent incidents prompted the Dallas Police Department and federal authorities to beef up security in some areas. Last week, police received information about a suspicious man who appeared to be Middle Eastern and was said to have been spotted checking out a TXU plant near downtown two days in a row. "We are staying alert on those particular properties," Golbeck said.
Also, police said the lead U.S. attorney in the case, Jim Jacks, had his car vandalized at his home in North Dallas. However, there were also other cars in the neighborhood vandalized as well. But as a precaution, the U.S. Marshall's Office has stepped up security at Jacks' home.
Police said many of the tips are not terror related, but are checking everything, leaving nothing to chance. "The majority of these things we are going to be very conservative, very thorough to track down any kind of intel that comes to us like that," Golbeck said.


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I think we be talking past each other to some extent - terminology on my
part. By "linked", I mean in the general sense - not that they are
close to each other in any physiological sense, only that they are on
the same chromosome. Maybe at opposite ends ;-) I'm not even
slightly familiar with frequencies/probabilities of crossing over &
relationships to distances along chromosome 'legs' (arms?), tho I
vaguely remember reading about studies of such. As I recall, the
farther away, much higher probability of crossing over, which is why I
guess the close ones are considered 'linked'.
> is not the same as factors linked by having
> genes closely aligned physically on same chromosome
This is what I thought you were implying. ;-)
> All my genetics and statistic forensics say these two samples are not
> from same two parents.
Mary Lou, any chance pot/refrigerator baggie labels migrated from one
year to the next?
The one cross I saw the biggest color difference from first year to
second year germination was definitely the same pot (IMM X CSONG). But
there was some overlap, sort of, of colors. Most germinated the first
year - mostly creams, a few whites, one yellow, two rosepurple selfs,
one bluepurple amoena. Second year several whites (including one
t-bearded - so much for first year easy germ of t factor with no
anthycyanin), a few "pink" anthocyanin amoenas, one bluepurple amoena,
no creams, no selfs. Enough overlap of 'types' that I can believe that
color was random in relation to chilling requirements.
Another question - are you saying that crossing two "poor quality"
inbred lines always produces "good quality" offspring? Surely not.
Depends on what other 'junk' is dragged along in the two inbred lines.
If both inbred lines have (back to irises) summer crud susceptible
foliage, then I'd expect the outcross seedlings to have it too. Same
concept with outcrossing two inbred pink lines - if both have early
germination, why would it go away in the offspring? I can't understand
why that trait would suddenly acquire a "normal" distribution.
Linda Mann


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What you need to know about Colorado Springs, CO
Colorado Springs is located in El Paso County, Colorado. The population is 71% White, 5% Black, 3% Asian and 1% Native American or Native Hawaiian. 5% identify as another race or ethnicity, or two or more races. 15% of residents are of Hispanic or Latino origin. The median income is $54,228 and the median home value is $208,600. For more on schools, real estate, business and health care in Colorado Springs, see each of the tabs above. Walkability can be a key driver of the overall livability of a community. Colorado Springs has a Walk Score ® of 33.
Why Colorado Springs is a Great Place to Live
Outdoor Adventures in Colorado Springs, CO
Colleges and Universities in Colorado Springs, CO
Best Trails in Colorado Springs, CO
Up by 8.22%
Up by 8.22%
10 Things to Do in Colorado SpringsLet's put it this way: Plenty of outdoor options bathed in scenic beauty.
Why Colorado Springs is a Great Place to LiveThe country's official Olympic City USA is on an upward trajectory
Nearby Cities
10 Best Cities for VeteransSupport for those who served is strong in these cities
A Change Is Coming to Middle School ClassroomsSee the Change USA works to increase the number of Colorado Springs middle school students pursuing STEM-related careers
Homes Owned 58.79%
58.79% Complete
Homes Rented 41.21%
41.21% Complete
Real Estate
In Colorado Springs, Colorado
Ready to move to Colorado Springs? There are a variety of housing options in the area. Click below to learn about the recent trends, average home values and even a few current real estate listings in Colorado Springs.
Median Home Price
Lower than the state average
Lower than the state average
Outdoor Adventures in Colorado Springs, COColorado Springs is a mecca for outdoor adventures, including plenty of opportunities for hiking, biking, climbing, skiing and water sports.
Innovation Keeps Colorado Springs Businesses on the Cutting EdgeTwo Colorado Springs businesses stay on the cutting edge of their respective industries thanks to their dedication to innovation.
In Colorado Springs, Colorado
The quality of a school district is often a top factor for parents considering where to live. Strong school systems attract new residents and help their cities prosper. See how Colorado Springs rates.
Median Household Income
Higher than the national average
Higher than the national average
In Colorado Springs, Colorado
Being a great place to live and being a great place to work often go hand-in-hand. Here's what the business and employment climate is like in Colorado Springs.
Colleges and Universities in Colorado Springs, COWhether the traditional student pursuing a four-year degree or the displaced worker honing his skills, residents can find a program that to meets their needs in Colorado Springs.
The Military Has a Strong Presence in Colorado SpringsHow strong of a presence? Nearly half of the economy stems from it.
Most residents are
5-17 Years old
5-17 Years old
Best Trails in Colorado Springs, COColorado Springs is blessed with an incredible trail system, taking you up peaks, through parks, or by natural wonders.
High Altitude Gardening in Colorado SpringsTips on how to garden in higher elevations, courtesy of experts at Colorado State University.
In Colorado Springs, Colorado
Access to quality hospitals, doctors and health-care providers is just part of what makes a healthy community. Here's how to get and stay healthy in Colorado Springs.
Bachelor's Degree 36%
36% Complete
Graduated High School 93%
93% Complete


text:
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Lot 569: Battery Operated Line Mar Spanking Bear Toy in Original Box-1950s
Vintage, Antique and Collectible Toys - 500 Lots
by Kraft Auction Service
29 October 2016
Lebanon, IN, USALive Auction
Looking for the realised and estimated price?
Description: Box has wear at corners and edges. Toy has minor wear, with a tear across the top. Toy is clean with minor wear.
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Is Home incorrect or missing key business information? Update your Hotfrog listing today.
Contact details
Are these details correct?
Hotfrog INDIA provides information regarding Home in Mumbai Maharashtra. Home is located at 31/1 A Powai Chowk Mulund Colony, Mumbai Maharashtra 400082 and provides shaving services. Contact them on +91 8976540123 or by visiting them on their website http://www.gorbys.in.
Is Home in 31/1 A Powai Chowk Mulund Colony, Mumbai Maharashtra 400082 your business? Claim your listing and attract more leads by adding more content, photos and other business details.
We have more shaving services in Mumbai Maharashtra available on Hotfrog INDIA. You can update your search for Mumbai shaving by location, keyword or service options.
Best Buliding, 'A' Wing, 5th Floor, SV Road Opposite Andheri Railway Station Mumbai Maharashtra 400058
6th Floor Meadows, Sahar Plaza Complex, J B Nagar Andheri Kurla Road Mumbai Maharashtra 400 059
89/91 Prabhu Shree Ram Mandir Marg , Shop No. G - 9 , 4'th Kumbharwada Mumbai Maharashtra 400004
Off CST Road Kalina Mumbai Maharashtra 400 098
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In her new novel, Susie Boyt introduces 38-year old Harriet Goodman as she visits her psychiatrist for the last time. Having survived a childhood of physical and emotional abuse, Harriet is determined that, after ten years of therapy and a recent large inheritance, she can turn her life around and become a "triumph".
Written in the third person, past tense, the novel immerses us in the nervous whirling of Harriet's consciousness. Her disjointed thought process is dizzying, heightened by rallying mantras and exclamation marks. Eccentric, archaic Harriet even describes her mindset using self- help manual titles ("Putting up with Stuff"). Boyt has a gift for creating loveable protagonists and, despite Harriet's oddities, we're won over by this oversized, sweet-hearted misfit who announces that she will open a school, "a sort of paradise for children". Harriet's school is indeed triumphant. Her rose-coloured vision is complete down to the bird detail in the wallpaper, the softly playing Schubert and the cherry -checked uniforms. Opening day is a success and Harriet is magnificent, wishing that there was "a courtly portraitist with... a chevalier's moustache, to record the fact".
But Harriet's "school of one's own" is more than philanthropy. It's the vehicle by which she can finally win her family's approval and achieve self-worth. The juxtaposition of time-frames means that, by the second chapter, we know that two years later the school is closed amid scandal. We also witness Harriet as a child attacked by her mother, her escape to a "peaceful" mental institution and the estrangement from her brother. Neglected and abused, Harriet in her need to create a microcosm of a happy, caring world becomes all the more poignant.
Boyt has studied Henry James and his stylistic influence is visible, both in the vibrant intensity of Harriet's character and the rich dramatisation of her consciousness. She also uses a Jamesian method of testing moral courage, for while Boyt usually bestows a happy ending, she has resisted one here to great effect.
The shadow of failure and scandal gives the book its emotional depth and ironic humour, and sets a tragic contrast to the sweetness of Harriet's character. However, there are smaller triumphs for our heroine: an acknowledgement from a former pupil that she was "really inspired", an understanding of her brother's cruelty, and a recognition that she has done something wonderful and may do so again.
In My Judy Garland Life, Boyt did what novelists rarely do and revealed the person behind the prose. It's impossible not to draw some similarities between the awkward, humble, witty Boyt in that memoir and Harriet - most strikingly in the optimism that, regardless of obstacle, there is always hope of better. Harriet's ultimate downfall doesn't stop us cheering her on, nor does it diminish her achievement.Reuse content


text:
The battle between the city of Chicago and Uber, a startup seeking to disrupt the taxi and limo industries, heated up this week. Uber has already faced unwanted attention because its pricing model for taxis conflicted with Chicago regulations. Now the city is introducing new regulations for Uber's separate sedan service that the company says could force it out of the Chicago market.
Uber sedans charge based on time and distance traveled. The company's Chicago rates start at $3.50 per mile when the car is moving faster than 11 miles per hour and 85 cents per minute when the car is stuck in traffic.


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New Ridge Route Video, sort of
After finally resolving some software issues, I have finally been able to start editing and posting videos again. My most recent one was taken in November 2015 along the Ridge Route from the Tumble Inn to State 138 at Neenach. It shows some of the issues the road is facing presently regarding erosion and a lack of maintenance.
Highway Videos site
Interesting site with videos and information about a few of the highways in Southern California and Arizona.
Finding Old Highways – A Guide – Part 3 – Bridges
Bridges and culverts can be great indicators of when a highway was built and by whom it was built. In California, there are a few different styles of railing which can be used to put a relative date on a particular bridge when they don't have date stamps on them. Other indicators may be used when the bridge rail has been modified or even replaced by modern rail. Styling elements on the superstructure vary depending on age. Older bridges tend to be more ornate than more modern structures. This page will serve as a guide to these styles and help to increase understanding of what goes into the design of these bridges.
Continue reading Finding Old Highways – A Guide – Part 3 – Bridges
Image of the Week – 2/9/2017
Image of the Week – 2/3/2017
Web forum for old highways
This is a really interesting site that has a discussion forum for old highways throughout the US. Help contribute to the discussions!
Winter Guide to Traversing Mission Valley Roadways
San Diego's Mission Valley can be quite a challenge during the winter. Most of the crossings of the San Diego River are low and not bridged. As a result, when it rains these crossings can be closed rather quickly. This greatly affects the ability to transect the valley along these roadways. The freeways, I-5, State 163, I-805, and I-15 are built with bridges and high enough to not flood. This is a guide to what is normally closed during storms.
From west to east:
Pacific Highway – Bridge – not closed
Morena Blvd – Bridge – not closed
Fashion Valley Road – closed more often than not during storms. This crossing has been rebuilt several times as well.
Avenida Del Rio – closed regularly during storms.
Mission Center Road – low crossing built a little higher than average. Still floods during major storms but is strong enough to avoid being damaged as a result.
Camino Del Este – Still low but built strong. Closes during major storms.
Qualcomm Way – Closes during very large storms. Built a little higher to help keep it open during major storms.
Ward Road – very low crossing and subject to closure during storms.
San Diego Mission Road – low crossing and subject to closure.
Friars Road (east) – Bridge – does not close.
Now, in the event all of these low-level crossings are closed, it is probably best to simply avoid Mission Valley in general. If you have to be there, I would suggest taking the 163 or 15 to cross the San Diego River. It may be a convoluted and circuitous route to use, but it is your only choice. Mission Valley circulation wasn't designed with the river to flood in mind, unfortunately. Some developments, such as Fashion Valley Mall, was at least partly designed for flooding. The southern parking structure was built with the lower floor to be flooded and still allow for use of the rest of the structure. Even MTS built the San Diego Trolley Green Line with the floods in mind. Most of it is elevated through the valley.
Small reorganization and updates
I've been working on the site a bit lately, and have been making some modifications. No pages are going away, so don't think that. New pages, however, are coming and older pages are being moved. I am developing a new page (or series depending on where it goes) on southern California's park lands. In particular, the national, state, and county level parks that help to preserve what this region was before settlement of the lands. This new page (or series) will help to locate these parks and give some information about them.
Due to this new page, I have also decided to consolidate some of the pages on this site into a new "Recreation" page. This will include the kayaking, bicycling, motorcycling, hiking, as well as the new parks page. In time, these pages may expand beyond their current scope to cover more areas. For now, they will remain mostly the same. The menu has already been updated to reflect these new changes, so keep that in mind while you explore this site.
With these particular subjects in mind (kayaking, bicycling, motorcycling, hiking, and parks), let us know if you would like to see any new areas covered by those pages or are looking for more detailed information about what is already covered.
Future Website Ideas
So far, it looks like there is interest in something for US 395 and US 466. As it stands, US 395 at least has something for San Diego County (still in progress) and the current signed route as far as its intersection with State 14 near Inyokern. US 466 is still pending. I am interested in working on a tour, of sorts, of the highway from Bakersfield to Barstow. What I lack to create the tour is photos. I have traversed the route many times but need photos of the route in Barstow, Hinkley, Kramer Junction, Boron, Tehachapi, Keene, Edison, and approaches to Bakersfield. I'm still looking for additional ideas for the site, if there are any. The pages I have talked about thus far will take a while to produce. There is still some research to be done on those roadways.


text:
After bug 700639, each time the debugger gets to a paused state the editor contents are refreshed, even though no change has occurred. The reason is that the XUL menulist we are now using needs to generate a change event every time a new script is loaded, which triggers the editor refresh. In the HTML select element case we could add options without triggering it, but I don't think that's possible now.
But even if it were, the problem stems from the fact that when entering a paused state the debugger client sends a "scripts" protocol request to the server to retrieve any scripts the server is aware of. This is currently redundant, since the client also handles "newScript" notifications from the server each time a new script appears. The better fix would arguably be to remove the "scripts" request, as well as the "scriptsadded" and "scriptscleared" handlers from the debugger client.
The one time when a "scripts" request would be required is when opening the debugger UI. The debugger server may have already received a list of scripts in the page before the debugger UI was ready to handle such notifications, so it would be necessary to query once.
Created attachment 592710 [details] [diff] [review]
Working patch
Simple fix. I didn't include a test, because it looks complicated to write a proper one and it doesn't seem to make much sense for a one-liner, to be honest.


text:
Archives |
Draught for Drought
Draught for Drought exceeded its expectations and had a full house at the fundraiser/awareness campaign put on by Keep Indianapolis Beautiful September 14th.
By Brittany Cravens
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text:
Gift Premium Subscription To ScunionBrewCo
Join Date:09-28-2011
Last Activity:02-12-2012 8:07 PM
5 gal. Apple/Cranberry/Blackberry Cider
5 gal. Apfelwein
- 5 gal. Apfelwein
- Single
- Georgia
Posted in thread: How many gallons of EdWort's Apfelwein have been made? on 12-29-2011 at 05:29 AM
Just bottled my second round, damn the headaches...21,784.5 + 5 = 21,789.5
Posted in thread: 100 year old fermentaion chamber on 12-27-2011 at 03:25 AM
So I am a new cider maker, and do not ferment beer at all. I have been exploring cheap
fermentation chambers to expand my product to the 20 gallon per batch step. I have looked at
plastic conical indu...
Posted in thread: Carboy!!!!!!!!! on 12-25-2011 at 11:07 AM
Good to know. Go big or go home then!
Posted in thread: Carboy!!!!!!!!! on 12-25-2011 at 10:25 AM
Why? Just use 6 gallon. I brew 5 gallon batches in my 30 gallon primary if my others are busy.
Well not anymore I have a ton of fermenters now but I used too.25 gallons worth of headspace
volume oxyge...
Posted in thread: How many gallons of EdWort's Apfelwein have been made? on 12-23-2011 at 10:40 AM
I have developed an obsession with this and am compelled to constantly have a primary full.
21731.5 + 10 = 21741.5
Posted in thread: Cider With Blackberries on 12-15-2011 at 04:48 AM
Looks amazing UpstateMike, and I currently have a clone of this in primary fermentation right
now. Quick question, do you worry about bacteria on the berries at all? I get that wild yeast
might be pre...


text:
--- In email@example.com, Deb Cranford <estrail1rider@...> wrote:
The 4 for today that I will be working on are attached. Anyoneknowing right off....all help appreciated. I am learning!The first two are of the same iris. The small flowered on is 29" tall...is it still a TB?The white is 44" tall and the flower is 7" tall!! I have neverseen such a huge bloom!
> Deb
I don't know who it is.... but I just discovered your #3 for today in a bed of irises planted by my husbands grandmother. That probably makes it at least 60 years old.


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When you are running an e-commerce website selling merchandise or service, you will be wondering: must i obtain a merchant services)? Gurus myself the same question for a few years before I "took the plunge" too.
Obviously you know, there are popular online payment methods such as Paypal, Google Checkout, or Amazon Payment which accept bank cards in the customers. If you use these payment methods, you'll be able to say that you accept debit card payment, indirectly. These internet payment services all charge similar fees close to $0.30 per transaction plus a amount of a couple of.5-3% of the transaction amount. Kind you'll want to accept charge card directly?
Obviously, accepting charge card directly gives your customers more options. After customer adds item(s) to cart, he/she is lead to a page to spend. On the page, he/she will find a different accessibility of paying by way of a credit-based card. Should you be the buyer, which method can you would rather use? Before we even start to guess, I'll explain to you an investment log collected from one of individuals websites:
The transaction log shows 40 most up-to-date consecutive orders from the site with payment methods. You can see that we accept Paypal, Google Checkout, and direct bank card (gateway_cc inside log).
One of several 40 orders, 13 transactions were paid by Paypal, 5 by Google Checkout, and the rest by direct bank card - a whopping 55% in the customers thought i would pay by direct plastic card! Our experience established that by accepting credit card payment directly, our sales increased by about 50%. OF course this percentage is a little unique of the proportion of transactions paid by plastic card because some customers has been "forced" to select from Paypal and Google Checkout, while some might simply leave when they actually don't could do with Paypal or Google Checkout.
Why would some individuals prefer using direct credit card? Listed below are my guesses:
1, A great deal of plastic card gives 1% in which about 5% money back or some type of equivalent for instance airline mileage.
2, Undergoing Paypal or Google Checkout means your own personal information explains a different party. If you are intending to repay by credit-based card anyway, why opt using an additional Alternative party? This is true specifically customer does not curently have a Paypal or Google Checkout account. Don't you have an adequate amount of "accounts" already?
3, More (privacy) sensitive web users could be concerned that Google or Paypal is collecting their preferences and data.
4, A few of them probably have bad bad knowledge of Paypal or Google Checkout.
Merchant card account fees can be like Paypal and Google checkout fees. You can obtain better rate as soon as your sales volume is larger.
Other than giving your visitors more choices to pay, there may be another reason that a merchant services is critical. Paypal and Google checkout use a number of restrictions around the belongings you sell. For example, both paypal and Google checkout prohibit weapons including knives as well as paintball items, tobacco related products, etc. For information on their restrictions, you have to carefully read their TOS. That has a merchant credit card, you don't get of the people restrictions.
A very good processing account supplier (we have used) are available here. Act and acquire business relationship make use of you are prepared of your e-commerce business!
Related posts:
Should you Accept Debit card for ones e-Commerce Site?
When you Accept Debit card for the e-Commerce Site?


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Last week, the S&P Dow Jones Indices - via data from the S&P Healthcare Economic Composite Index - reported the healthcare sector's annual growth decelerated in March, resulting in the lowest rate since January 2005. With many industries within the sector seeing historically low growth rates, we wanted to see which healthcare companies investors felt were most likely to exceed expectations. We ran a screen with this in mind, focusing specifically on insider and institutional investor behavior within the healthcare sector.
To start, we began with a universe of companies experiencing insider buying with net purchases over the last six months representing more than 2% of share float. Since insiders are employed at the company and have nonpublic information, their stock buying suggests they are bullish about the firm's future and believe current share price is undervalued. We then selected the healthcare stocks from that list.
Two percent may seem minimal, but considering these are often individual investors putting enough money on the line to collectively acquire that percentage of float is significant.
And why six months? A 1968 study by Professors James H. Lorie and Victor Niederhoffer showed that when number of insider buyers is at least two more than the number of insider sellers "the stock can be expected to outperform the market during the next six months." Furthermore, "insiders tend to buy more often than usual before large price increases and to sell more than usual before price decreases."
Next, we screened for stocks with bullish sentiment from institutional investors, with significant net institutional purchases over the last quarter representing at least 5% of share float. This indicates that institutional investors such as hedge fund managers and mutual fund managers expect these names to outperform into the future.
The List
For an interactive version of this chart, click on the image below. Analyst ratings sourced from Zacks Investment Research
Do you think these stocks will soon realize their potential? Use this list as a starting point for your own analysis.
1. Pacific Biosciences of California, Inc. (NASDAQ:PACB): Develops, manufactures, and markets an integrated platform for genetic analysis.
- Market cap at $160.49M, most recent closing price at $2.42.
- Over the last six months, insiders were net buyers of 1400000 shares, which represents about 3.73% of the company's 37.55M share float.
- Net institutional purchases in the current quarter at 2.7M shares, which represents about 7.19% of the company's float of 37.55M shares. The 2 top holders of the stock are Deerfield Management and Maverick Capital Ltd.
During the first quarter of 2013, Pacific Biosciences of California, Inc. reported $5.6 million in revenue, which is a 44% decrease from the same period in 2012. The company sold 3 DNA sequencing PacBio RS systems in the quarter, which contributed $1.9 million to total revenue. It also saw increased usage of the RS systems and received 4 new orders, bringing its systems backlog up to 6. April marked the launch of the PacBio RS II, which provides the longest sequences available today. The company gave existing RS systems customers the opportunity to upgrade to the RS II for $20,000 prior to launch, and at the end of March, Pacific Biosciences received 31 upgrade offers, representing over 40% of its customers.
As Seeking Alpha contributor Craig Keolanui notes, Pacific Biosciences' success depends on proving its products' value to members of the scientific community. The company faces stiff competition from peers Illumina (NASDAQ:ILMN) and Life Technologies (NASDAQ:LIFE). GenomeWeb reports the latter recently received approval from Health Canada for diagnostic use of its 3500 series Sanger sequencers, which were also approved by the USDA back in February. Keeping this in mind, investors should keep an eye on how Pacific Biosciences fares with 100k Pathogen Genome Project where the company is using its Single Molecule, Real-Time (NASDAQ:SMRT) technology to sequence the genomes from 1,000 foodborne pathogen samples. In the earnings call, CEO Michael Hunkapiller stated that the company expects to see more revenue from that project in the second quarter.
2. Infinity Pharmaceuticals, Inc. (NASDAQ:INFI): Engages in the discovery and development of medicines for the treatment of cancer and other conditions in the United States.
- Market cap at $1.23B, most recent closing price at $24.91.
- Over the last six months, insiders were net buyers of 5380195 shares, which represents about 13.09% of the company's 41.11M share float.
- Net institutional purchases in the current quarter at 3.8M shares, which represents about 9.24% of the company's float of 41.11M shares. The 2 top holders of the stock are FMR, LLC and Orbimed Advisors LLC.
Infinity Pharmaceuticals suffered a blow last week after J.P. Morgan expressed concern over the drug maker's presentation of its IPI-145 drug at the upcoming ASCO conference. Insider Monkey reports that J.P. Morgan and investors - shares fell by more than 20% following the comments - are wary of the company's ability to penetrate a crowded market of leukemia drugs. The Motley Fool notes that competitors Roche and Gilead Sciences (NASDAQ:GILD) had positive findings from studies for the GA101 drug and idelalisib drugs, respectively. Both drugs have exhibited an ability to shrink tumors in patients.
Infinity Pharmaceuticals didn't record any revenue during the first quarter of 2013. The $25.2 million revenue generated during the same period last year came from the company's now-terminated strategic relationship with Purdue/Mundipharma. In the company's earning call, CFO Lawrence Bloch expressed optimism for future projects and the drug maker's $300 million in cash investments and worldwide commercial rights for its programs. The company's lung cancer drug, the Hsp90 inhibitor retaspimycin hydrochloride, is currently in trials, with results coming forth later this year. While this market is less congested than the aforementioned leukemia one, Reuters reports that Roche received FDA approval on its drug Tarceva last week for testing in non-small cell lung cancer patients with specific mutations of the EGFR gene.
3. Aegerion Pharmaceuticals, Inc. (AEGR): Engages in the development and commercialization of novel therapeutics to treat severe lipid disorders.
- Market cap at $1.77B, most recent closing price at $61.57.
- Over the last six months, insiders were net buyers of 624767 shares, which represents about 2.61% of the company's 23.97M share float.
- Net institutional purchases in the current quarter at 5.9M shares, which represents about 24.61% of the company's float of 23.97M shares. The 2 top holders of the stock are FMR LLC and Perceptive Advisors LLC.
Last quarter Aegerion commercially launched Juxtapid, its orphan drug developed to lower cholesterol in those suffering from the rare genetic disease homozygous familial hypercholesterolemia (HoFH), which prevents cholesterol from being cleared from the body. The biotech company has since seen 185 global prescriptions written for the medication, which places the company well on track to reach its goal of 250-300 patients by the end of the year, as reported by The Street. In an earnings call at the end of April, CEO Marc Beer noted that the number of Juxtapid prescriptions issued each week has been steadily increasing since the launch and, as of April 30, was at 12 a week. Furthermore, Juxtapid's steep pricing - one year of therapy costs $250,00 - is expected to help Aegerion generate $15 - $25 million in revenue this year.
The company does face immediate and long-term competition on the HoFH front. Isis Pharmarceuticals launched its HoFH orphan drug Kynamro last quarter, though the company didn't provide any numbers during last week's earnings call. At $176,000 a year, Kynamro is a cheaper alternative to Juxtapid, which Decision Resources cites as a potentially significant factor depending on how much insurance carriers are willing to cover given the presence of two similar options on the market. Additionally, at the end of April, Forbes's Matthew Herper wrote that Amgen, a PCSK9 inhibitor that helps lower cholesterol, may pose a long term threat to Juxtapid, especially with its appealing $20,000. However, the drug's efficacy on HoFH patients remains to be seen; in European preliminary trials, Amgen lowered cholesterol by 15 - 49% for 8 patients compared to Juxtapid and Kynamro's 50%.
*Insider ownership information sourced from Yahoo! Finance. Institutional data sourced from Fidelity. All other data sourced from Finviz.
Disclosure: I have no positions in any stocks mentioned, and no plans to initiate any positions within the next 72 hours.
Business relationship disclosure: Business relationship disclosure: Kapitall is a team of analysts. This article was written by Mary-Lynn Cesar, one of our writers. We did not receive compensation for this article (other than from Seeking Alpha), and we have no business relationship with any company whose stock is mentioned in this article.


text: Fingers Crossed Emoticons X= 1.5 (4 Votes) A hand with crossed fingers; may be used to wish someone good luck. More Info Direction Vertical Category Expressions Less Info (yn) 1.0 (2 Votes) Inserts a picture of a hand with fingers crossed in MSN/Live Messenger. Represents a hopeful gesture, but may also mean "not really" when saying you are going to do something. A "secret" emoticon not officially published by Microsoft. More Info Direction N/A (text) Category Actions Shortcuts Live MessengerWindows 8Windows Phone Less Info Last Updated: September 22, 2012


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Paparazzi Accessories by SheDiva
Bracelets, Handmade Jewelry, Bridal Jewelry
What is Paparazzi Accessories?
Paparazzi is a Direct Sales and Party Plan Company, which specializes in affordable, fashionable accessories. We sell earrings, necklaces, lanyards, bracelets, rings, hair clips and headbands - all for $5.00 each! Also have little girl items for $1.00 each! Two sisters and their husbands opened Paparazzi in December 2010 in Utah. Their goal and mission is to bring self-confidence, empowerment, and independence to women and families all over the world. They seek to fulfill this mission through the following avenues: First, they seek to offer women an affordable option to be fashionable and feel great. Second, they offer a real home-based business opportunity - independent of "recruiting" friends & neighbors. Consultants are able to carry inventory, host parties, invite associates, have fun, and sell their inventory directly to the customer. Plus, they are able to sell as much and as often as they would like. Third, they offer future independence through the Paparazzi Compensation Plan by Consultants training others to be successful. This gives each Consultant the ability to build a long-term residual income, giving them a potential for real independence.
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Audi R8 Crash Site on Bandra Worli Sealink 2015
Notice the Audi A4 which hit the R8 from behind @0:21 This was the accident on the Bandra Worli Sealink involving an Audi R8 an Audi A4 and a Hyundai i10 when the Parx Supercar Rally had just begun. The following video was shot after the accident. www.facebook.com/supercoolcars instagram.com/rich1409
Parx Supercar show 2013 & Audi R8 Fire in Mumbai [HD]
Bugatti Veyron in Hyderabad (India) Part 1
Spotted this W16 behemoth on the roads of Hyderabad. Enjoy !!
Audi R8 Accident 2013 - Burning of Audi R8
Audi R8 Accident - Burning of Audi R8


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This review is from: ZyXEL NBG5715 Simultaneous Dual-Band Wireless N900 Media Router IEEE 802.11a/b/g/n
Pros: stable 2.4Ghz connection
Cons: 5Ghz SSID works for a while and then disappears until manual router restart. This is an expensive dual-band router, if 5G is not working then what the point?
Customer support responds relatively quickly first, but then forgets about you.
Keeping it 2 eggs because it works at least in 2.4Ghz mode.
This review is from: EDIMAX EW-7722UTn N300 USB 2.0 Mini-size Wireless Adapter
Pros: Did not have an opportunity to enjoy any
Cons: Does not work in N mode with TP-Link TL-WR1043ND (both 2.4Ghz).
G works but not N. Even with security turned off.
I also have 3 other adapters working fine with router above, so something is wrong with this adapter.
This review is from: GIGABYTE GA-MA785GM-US2H AM3/AM2+/AM2 AMD 785G HDMI Micro ATX AMD Motherboard
Pros: Inexpensive.
Most of functionality does work, seems to be stable, but I have two problems (see Cons)
Cons: 1) HDMI output does not suport 1080p@60Hz. Only 20Hz.
Support is totally not helpful, they're suggesting doing some weird stuff, like "make a screenshot" of this problem %-\. My LCDTV works with other PCs at this frequency, but not with this one. Have to use DVI instead as a workaround. (x3 more wires, unfortunately)
2) Does not wake up correctly using S3 mode. Had to switch to S1.
Other Thoughts: Definitely, would not recommend.READ FULL REVIEW


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Do tunes every get stuck in your ears? I call sticky tunes ear worms. Here are the worms traveling through air, time, and the culture right now that are pleasing to my ear. What are your ear worms? Hum (A Prius for Everyone)(Do you know who sings this?)Well, it's a nice idea. I'd like to think everyone could afford a Prius. But they are giving away this tune for free and it's stuck in my head via a Prius worm FireworkKaty PerryWhat a great song from the pop p … [Read more...]
Ear Worms: the sound track in my head
Cardinal Dolan & Stephen Colbert meet Fordham c/o Jim Martin, SJ to evangelize anew
I was not there last night, where new kinds of Bronx cheers must have been heard when Cardinal Dolan of New York and Stephen Colbert of Comedy Central's "Colbert Report" met at Fordham University. Their host was Rev. James Martin, SJ, a lover of good humor and author of "Between Heaven and Mirth."Here are a couple of paragraphs from Jim's post at "America" magazine this morning and a link to the whole thing. Jim also explains why there was no television … [Read more...]
October Baby on DVD today
Earlier this year I reviewed "October Baby" for On Faith and Media:"October Baby" tells the fictional story of Hannah (newcomer Rachel Hendrix), a 19-year old college student who collapses during the performance of a play. In an effort to find out why she is having medical issues and nightmares her parents tell her she is adopted. This overwhelming news sends her and her best friend, Jason, on a road trip to find her real mother.Along the way Rachel must sort thro … [Read more...]
For Greater Glory on DVD Sept 11
In 1917 the Mexican Constitutional Congress adopted a new constitution. It confirmed the separation of church and state first decreed in the 1857 constitution, returned subsoil rights to the government from ownership and control by foreign corporations, established the basis for secular education, and provided for land reforms. Five articles restricted the power and liberty of the Catholic church. These forbade public worship outside of churches, restricted the church's right to own property, c … [Read more...]
American Bible Challenge
When I first heard of the Game Show Network's idea for a new game show, I wondered: How is it no one ever thought of it before? There is so much material in the scriptures that if the producers play it right, they could have many seasons. It could be a way to bring Christians together around the Word.On the set of Game Show Network's "The American Bible Challenge" with host Jeff Foxworthy. ((c) GSN)This hourlong show that tests Bible knowledge is hosted by comedian Jeff Foxworthy ("Are You S … [Read more...]
Bows, arrows and a sword: Girls rock this year on film
2012 is turning out to be a banner year for films with significant female lead characters: warriors, weapons and all.With an estimated budget of $185 million, the new Pixar/Disney 3-D animated feature, "Brave," opened June 22 and grossed $66 million. Made on half that budget, "The Hunger Games" opened March 25 with a box office of $152.5 million and has brought $400 million in North America and nearly $650 million worldwide. (The DVD is due out Aug. 18.) "Snow White and the Huntsman," made … [Read more...]
BRAVE and nine more reviews at "Sister Rose Goes to the Movies"
For reviews of BRAVE, Madagascar 3, MEN IN BLACK 3, The Avengers, CHIMPANZEE, Dark Shadows, SNOW WHITE AND THE HUNTSMAN, Battleship, and THE LUCKY ONE click on Sister Rose Goes to the Movies. … [Read more...]


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Seven Layer Dean Dip
|Canned black beans
|15 Ounce, drained, rinsed (1 Can)
|Chopped fresh cilantro
|3 Teaspoon
|Chili powder
|1 Teaspoon
|1 Small
|Lemon juice
|2 Teaspoon
|Chopped jalapeno peppers
|2 Tablespoon
|1 Medium, seeded and finely chopped
|Shredded monterey jack cheese
|1⁄2 Cup (8 tbs) (Nonfat Or Reduced-Fat)
|Thinly sliced scallions
|1⁄4 Cup (4 tbs)
|Chopped black olives
|1⁄4 Cup (4 tbs)
1. Place the beans, cilantro, and chili powder in a food processor, and process until smooth. Spread the mixture in a 9-inch deep dish pie pan or a shallow serving dish.
2. Peel and finely chop the avocado. Toss with the lemon juice, and set aside.
3. Spread the jalapeno peppers over the bean mixture. Spread the avocados over the jalapenos. Create layers of the remaining ingredients in the order listed.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 813 Calories from Fat 332
% Daily Value*
Total Fat 39 g60.4%
Saturated Fat 12.4 g62.2%
Trans Fat 0 g
Cholesterol 49.8 mg16.6%
Sodium 2232.7 mg93%
Total Carbohydrates 83 g27.7%
Dietary Fiber 30 g120.1%
Sugars 9.9 g
Protein 38 g76.6%
Vitamin A 95.5% Vitamin C 93.9%
Calcium 100.2% Iron 33.6%
*Based on a 2000 Calorie diet


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YOU CAN provide hope and help to people with ALS.
You can help fund urgently needed research into finding an effective treatment and a cure for ALS and support services to those with ALS. Please consider providing a planned gift to ALS Canada.
We can have a conversation with you and your advisors to determine if a planned gift fits your desires, financial objectives, family needs, tax situation and our funding objectives. All discussions are confidential and will not obligate you to make a gift. Some of the many benefits of making a planned gift to ALS Canada include:
There are a variety of ways to give a gift, they include:
To learn more, please request our Planned Giving Fact Sheets and Will Planning Guide by contacting Jill Bridgman, Vice President, Fund Development toll free at 1-800-267-4257 ext 216 or by email at jbridgman (at) als (dot) ca.
ALS Canada strongly recommends that you seek professional advice to ensure your financial goals are considered, your tax situation reviewed, and your planned gift is tailored to your circumstances.
A properly drafted Will is the only way to provide an orderly and timely plan for the settlement of your life.


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- Description
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Grimm's Toolbox
We love this Toolbox!
It's just the right size for little one's to carry around with them, doing jobs, fixing things - it's a very natural way for them to imitate grown-ups & for role playing.
Ideal for every child - just add imagination!
Hand-crafted in Europe from sustainably sourced wood using completely non-toxic natural oils.
Approx 14.5cm in length, with 3 individual pieces.
Advised for ages 1 years +.
Hand-crafted in Europe from alder wood using completely non-toxic water-based dyes.
Grimm's wooden rainbow is approx 38 cm in length and 18 cm in height.
Advised for ages 2 years +.
We know you'll want your order as quickly as possible, which is why we endeavour to dispatch orders within 24 hours of receipt. In fact, we dispatch most orders the same day they're placed.
Need your order really quickly? No problem! Just select the next day shipping option at the checkout. Just so long as you place your order before midday, we'll get it to you by 1pm the next working day.
If for any reason there will be a delay with your order, we will contact you as soon as possible. If you've any questions, please don't hesitate to contact us on 0845 519 6212 or at firstname.lastname@example.com.
FREE UK Delivery - Standard delivery is free for all UK orders for delivery to all UK addresses. Delivery is normally within 1-3 days. Monday to Saturday, excluding public holidays.
UK Next Day Delivery - guaranteed next working day delivery - £6.95. Monday to Friday, excluding public holidays. Order by 12 noon.
EUROPE: We dispatch items to the EU using a Europe Priority Airmail Signed-For Service, transit time 3-5 days - £8.95.
IRELAND: We dispatch items to Ireland using Fastway Couriers - £2.95. Monday to Friday, excluding public holidays.
INTERNATIONAL: We dispatch items outside the EU using either DHL or an International Priority Airmail Signed-For Service, transit time 5-7 days - £15.95 *(Excluding USA and Canada).


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2.5.1 Conditional compilation symbols
The conditional compilation functionality provided by the
#else, and
#endif directives is controlled through pre-processing expressions (Section 2.5.1) and conditional compilation symbols.
- conditional-symbol:
- Any identifier-or-keyword except true or false
A conditional compilation symbol has two possible states: defined or undefined. At the beginning of the lexical processing of a source file, a conditional compilation symbol is undefined unless it has been explicitly defined by an external mechanism (such as a command-line compiler option). When a
#define directive is processed, the conditional compilation symbol named in that directive becomes defined in that source file. The symbol remains defined until an
#undef directive for that same symbol is processed, or until the end of the source file is reached. An implication of this is that
#define and
#undef directives in one source file have no effect on other source files in the same program.
When referenced in a pre-processing expression, a defined conditional compilation symbol has the Boolean value
true, and an undefined conditional compilation symbol has the Boolean value
false. There is no requirement that conditional compilation symbols be explicitly declared before they are referenced in pre-processing expressions. Instead, undeclared symbols are simply undefined and thus have the value
The name space for conditional compilation symbols is distinct and separate from all other named entities in a C# program. Conditional compilation symbols can only be referenced in
#define and
#undef directives and in pre-processing expressions.


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Prediction of paclitaxel sensitivity by CDK1 and CDK2 activity in human breast cancer cells
DOI: 10.1186/bcr2231
© Nakayama et al.; licensee BioMed Central Ltd. 2009
Received: 4 August 2008
Accepted: 24 February 2009
Published: 24 February 2009
Paclitaxel is used widely in the treatment of breast cancer. Not all tumors respond to this drug, however, and the characteristics that distinguish resistant tumors from sensitive tumors are not well defined. Activation of the spindle assembly checkpoint is required for paclitaxel-induced cell death. We hypothesized that cyclin-dependent kinase (CDK) 1 activity and CDK2 activity in cancer cells, which reflect the activation state of the spindle assembly checkpoint and the growth state, respectively, predict sensitivity to paclitaxel.
Cell viability assays and DNA and chromatin morphology analyses were performed in human breast cancer cell lines to evaluate sensitivity to paclitaxel and the cell cycle response to paclitaxel. We then examined the specific activities of CDK1 and CDK2 in these cell lines and in xenograft models of human breast cancer before and after paclitaxel treatment. Protein expression and kinase activity of CDKs and cyclins were analyzed using a newly developed assay system.
In the cell lines, biological response to paclitaxel in vitro did not accurately predict sensitivity to paclitaxel in vivo. Among the breast cancer xenograft tumors, however, tumors with significantly increased CDK1 specific activity after paclitaxel treatment were sensitive to paclitaxel in vivo, whereas tumors without such an increase were resistant to paclitaxel in vivo. Baseline CDK2 specific activity was higher in tumors that were sensitive to paclitaxel than in tumors that were resistant to paclitaxel.
The change in CDK1 specific activity of xenograft tumors after paclitaxel treatment and the CDK2 specific activity before paclitaxel treatment are both associated with the drug sensitivity in vivo. Analysis of cyclin-dependent kinase activity in the clinical setting could be a powerful approach for predicting paclitaxel sensitivity.
Paclitaxel is used widely in the treatment of breast cancer and several other solid tumors [1–6]. Paclitaxel is not effective in all tumors, however, and the characteristics that distinguish resistant tumors from sensitive tumors are not well defined. Identifying the tumor molecular characteristics associated with resistance to or sensitivity to paclitaxel would help determine which patients are most likely to benefit from paclitaxel therapy.
Paclitaxel resistance has been attributed to a variety of mechanisms, including upregulation of antiapoptotic Bcl-2 family members such as Bcl-2 and Bcl-XL [7, 8]; upregulation of membrane transporters such as mdr1, which increases drug efflux [9]; point mutations in β-tubulin residues or altered expression of tubulin isotypes, which impair drug–tubulin binding [10, 11]; upregulation of ErbB2 (HER-2) by inhibition of cyclin-dependent kinase (CDK) 1, which causes delayed mitosis [12]; and high expression of microtubule-associated protein tau mRNA, which decreases paclitaxel binding to microtubules and microtubule polymerization [13].
We recently reported that activation of the spindle assembly checkpoint is required for paclitaxel-induced cell death and that inactivation of this checkpoint was correlated with suppression of CDK1 activity [14]. CDK1, in combination with other mitotic cyclins, is a universal master kinase and is required for the regulation of mitosis [15]. Previous studies that used CDK inhibitors or dominant-negative CDK1 constructs showed that CDK1 plays a critical role in paclitaxel-induced cell death [16–18]. CDK1 activity may therefore be a predictor of paclitaxel sensitivity.
Several studies reported that rapidly proliferating tumors have a higher response rate to chemotherapy [19, 20]. Expression levels of cyclin A and cyclin E are suggested to correlate with the proliferation state of cancer cells [21, 22]. These molecules are considered to drive the cell cycle by activating CDK2 [23]. Moreover, a correlation between the effectiveness of paclitaxel and the tumor growth rate has previously been reported [24]. We therefore speculated that high CDK2 activity is required for increased paclitaxel sensitivity.
In the present study we measured both the kinase activity and expression level of CDK1 and CDK2 before and after paclitaxel treatment in human breast cancer cell lines and in xenograft models of human breast cancer. We defined the specific activity of CDK as a kinase activity (unit) of its protein expression (1 μg). We found that changes in CDK1 specific activity after paclitaxel treatment predicted the paclitaxel sensitivity of breast cancer cells and xenograft tumors. Baseline CDK2 specific activity was higher in tumors that were sensitive to paclitaxel than in tumors that were resistant to paclitaxel.
Materials and methods
Cell cultures
MDA-MB-231 and MDA-MB-468 human breast cancer cells were cultured in DMEM–Ham's F12. T47D and MCF-7 human breast cancer ells were cultured in RPMI 1640 medium. Both media were purchased from Sigma (St Louis, MO, USA) and were supplemented with 10% heat-inactivated FBS (Hyclone Laboratories, Logan, UT, USA) and 1% antibiotic–antimycotic solution (Gibco Invitrogen, Carlsbad, CA, USA). All cell lines were maintained at 37°C in a humidified incubator in a 5% carbon dioxide atmosphere.
Preparation of cell lysates
Cultured cells were treated with 100 nM paclitaxel (Calbiochem, Darmstadt, Germany) for 24, 48, or 72 hours, and then these cells were frozen immediately at -80°C. At the time of CDK analysis, the frozen cells were thawed, treated with lysis buffer (0.1% NP-40, 50 mM Tris–Cl (pH 7.4), 50 mM NaF, 5 mM ethylenediamine tetraacetic acid, 1 mM NaVO3, and 0.2% proteinase inhibitor cocktail; Sigma), and centrifuged at 10,000 × g for 5 minutes at 4°C. The obtained lysates were assayed for CDK activity and expression as described below.
Cell viability assay
For cell viability assays, cells were seeded at a concentration of 5 × 103 cells/well in 100 μl culture medium into 96-well culture plates and were incubated for 24 hours. Cells were washed, and fresh culture medium containing various concentrations of paclitaxel (0.1 pM to 100 μM) was added. After 72 hours of treatment, 10 μl WST-1/ECS solution (Chemicon International, Inc., Temecula, CA, USA) was added to each well, and the plates were incubated at 37°C for 4 hours. Absorbance was measured at 440 nm on a microplate reader (Molecular Devices, Sunnyvale, CA, USA).
DNA analysis
For DNA analysis, cells were treated with 100 nM paclitaxel for 24, 48, or 72 hours, after which nonadherent cells were collected from the flask and then the adherent cells removed by trypsinization. The cells were stained with 50 μg/ml propidium iodide in PBS-glucose containing ribonuclease A (2 kU/ml; Sigma) for 1 hour. The DNA content of the cells was measured using a FACSCalibur flow cytometry system and was analyzed with ModFit software (BD Biosciences, San Jose, CA, USA). The DNA content was analyzed at the time corresponding to the doubling time of each cell line (that is, 24 hours after treatment for the MDA-MB-468 and MDA-MB-231 cells, and 48 hours after treatment for the T47D and MCF-7 cells).
Chromatin morphology analysis
For chromatin morphology analysis, cells were treated with 100 nM paclitaxel for 24, 48, or 72 hours, after which nonadherent cells were collected from the flask and then the adherent cells removed by trypsinization. Both cell fractions were mixed, washed with PBS, smeared onto glass slides by cytospin centrifugation at 1,000 × g for 5 minutes, and then stained with aceto-orcein solution (Muto Pure Chemicals Ltd, Tokyo, Japan) to visualize the chromatin. The morphology was analyzed under a light microscope at 20× magnification. The DNA and chromatin morphology was analyzed at the time corresponding to the doubling time of each cell line after paclitaxel treatment (that is, 24 hours after treatment for MDA-MB-468 and MDA-MB-231 cells, and 48 hours after treatment for T47D and MCF-7 cells). The MDA-MB-468 and MDA-MB-231 cells could not be analyzed 48 hours after treatment because of low viability (<10%) at that time.
Analysis of cyclin-dependent kinase and cyclin protein expression
For analysis of CDK and cyclin protein expression, the crude cell lysates containing 2.5 μg total protein were added to the wells of a newly developed dot-blot device (Sysmex Co., Kobe, Japan). The analysis procedure in detail was described in our previous report [25].
Analysis of cyclin-dependent kinase activity
CDK molecules were selectively precipitated from 50 μg total protein with 2 μg corresponding anti-CDK1 or anti-CDK2 antibody and 20 μl protein A sepharose beads (Amersham Pharmacia, Uppsala, Sweden) for 1 hour at 4°C. Fifty microliters of the substrate mixture (containing 10 μg protein substrate (histone H1; Upstate Biotechnology, Lake Placid, NY, USA), 5 mM adenosine 5'-O-(3-thiotriphosphate; Sigma), 20 mM Tris–Cl (pH 7.4), and 0.1% Triton X-100) was added, and the beads were incubated under continuous shaking at 37°C for 30 minutes. The substrate mixture was then collected, and the monothiophosphates introduced as the substrate were labeled further by incubation with 10 mM iodoacetyl biotin (Pierce, Rockford, IL, USA) in coupling buffer (100 mM Tris–Cl (pH 8.5) and 1 mM ethylenediamine tetraacetic acid) for 90 minutes in the dark at room temperature. The reaction was quenched with β-mercaptoethanol, and 0.4 μg thiophosphorylated substrate was applied to the wells of the Sysmex dot-blot device. The wells were blocked with 1% bovine serum albumin for 30 minutes at room temperature, incubated with fluorescein-labeled streptavidin (Vector Laboratories, Burlingame, CA, USA) for 1 hour at 37°C, and washed with Tris-buffered saline. After that, fluorescent images of the dot-blot device membrane were evaluated with a Molecular Imager FX image analyzer (Bio-Rad, Hercules, CA, USA), and the fluorescence intensity of the dots was quantified with the Quantity One program (Bio-Rad).
Cyclin-dependent kinase specific activity
We defined a unit as equivalent to the kinase activity of 1 μg total protein from CDK1 or CDK2.
Xenograft models of breast cancer
MDA-MB-231 and MDA-MB-468 cells were suspended in PBS, and T47D and MCF-7 cells were suspended in a solution of Matrigel (BD, Franklin Lakes, NJ, USA) 50% v/v in PBS. The suspended cells (5 × 106 cells/mouse) were inoculated subcutaneously into the mammary fat pads of 6-week-old female BALB/c nu/nu mice (CLEA Japan Inc., Tokyo, Japan). Because T47D and MCF-7 cells require estrogen for growth, 17β-estradiol pellets (0.72 mg, 60-day release; Innovative Research of America, Sarasota, FL, USA) were implanted subcutaneously in the shoulder region of each mouse before inoculation with the cells. When the tumor masses reached 50 to 80 mm3 (about 10 days after the inoculation), the mice were given daily intraperitoneal injections of paclitaxel (20 mg/kg per day, or one-half of the 50% of lethal dose 40 mg/kg, n = 7) or 15% ethanol solution containing 0.9% NaCl as a vehicle control (n = 8) for 5 days. Two-dimensional tumor measurements were made daily until 12 days after the first dose. The tumor volume was calculated according to the formula: volume = π (short diameter2) × (long diameter)/6.
Unpaired two-sample t tests were used to test differences in tumor size between the control group and the drug-treated groups. This was approved by the animal ethic committee.
The in vivo CDK specific activity was measured in the tumor tissues resected from the mice as follows. Tumor-bearing mice were given a single 20-mg/kg dose of paclitaxel, and 24 hours later they were killed by cervical dislocation. Tumor tissues were resected and lysed in lysis buffer with a homogenizer (HM-100; Sysmex Co.). Welch's t test was used for comparison.
Sensitivity of breast cancer cell lines to paclitaxel
First, we examined the sensitivity of four human breast cancer cell lines to paclitaxel in vitro. The 50% inhibitory concentration (IC50) values determined by cell viability assay were as follows: MDA-MB-468, 1.8 nM; MDA-MB-231, 2.4 nM; T47D, 4.4 nM; and MCF-7 cells, 7.2 nM.
Results of DNA and chromatin analysis of breast cancer cells after paclitaxel treatment in vitro
We next examined the cell cycle response to paclitaxel using DNA analysis and morphologic analyses of chromatin in each of the four breast cancer cell lines.
In the morphologic analysis of chromatin after paclitaxel treatment, MDA-MB-468, MDA-MB-231, and MCF-7 cells showed ring-like staining (Figure 1b, arrows), indicating that paclitaxel induced perinuclear microtubule bundles [26]. T47D cells, in contrast, did not show bundles but did show chromatin condensation, which is indicative of cells arrested in mitosis (Figure 1b, arrowheads).
These results indicated that paclitaxel sensitivity in vitro and the cell biological response were not correlated in the four cell lines.
Specific activities of CDK1 and CDK2 in breast cancer cells after paclitaxel treatment in vitro
We measured the specific activities of CDK1 and CDK2 in the four breast cancer cell lines after treatment with 100 nM paclitaxel. This concentration was chosen to ensure that cells were exposed to levels two log units higher than the IC50 but within the physiologically tolerable range.
Similarly, CDK2 specific activity was increased after paclitaxel treatment in MDA-MB-468 and T47D cells (Figure 2a, right). The increment of CDK2 specific activity after the treatment was 2.1 times in MDA-MB-468 cells and 2.9 times in T47D cells, respectively. The magnitude of the increases in CDK2 specific activity after paclitaxel treatment, however, did not correlate significantly with the IC50 values of paclitaxel in the tested cell lines (R2 = 0.004; data not shown). Cyclin E expression was not related to CDK2 specific activity after paclitaxel treatment in any of the four cell lines (Figure 2a,c).
Sensitivity of tumor xenografts to paclitaxel
Specific activities of CDK1 and CDK2 in breast cancer xenografts after paclitaxel treatment in vivo
CDK2 specific activity was increased after paclitaxel treatment only in the MDA-MB-468 tumors (Figure 4b), even though CDK2 specific activity in vitro was increased after paclitaxel treatment in both MDA-MB-468 and T47D cells (Figure 2a, right). Interestingly, the baseline (before paclitaxel treatment) CDK2 activity was significantly higher in the MDA-MB-468 and MDA-MB-231 tumors than in the T47D and MCF-7 tumors (Figure 4b). The mean value of CDK2 specific activity before the treatment is 20.3 units/ng in MDA-MB-468 cells, 11.2 units/ng in MDA-MB-231 cells, 2.7 in T47D cells, and 6.5 units/ng in MCF-7 cells, respectively (P < 0.01 for each).
We found that an increase in CDK1 specific activity after paclitaxel treatment correlates with sensitivity of the xenografts to paclitaxel, and a lack of change in CDK1 specific activity correlates with a lack of sensitivity of the xenografts to paclitaxel. These findings indicate that analysis of CDK1 activity could be a powerful approach for predicting paclitaxel sensitivity. In our in vivo experiment, the highest CDK1 specific activity value observed after paclitaxel treatment in xenografts of the paclitaxel-resistant T47D and MCF-7 cells was 0.15 units/ng. If this value was used as the cutoff value to distinguish between paclitaxel-sensitive and paclitaxel-resistant xenografts, the CDK1 specific activity would have a positive predictive value of 100% for the determination of paclitaxel sensitivity in our studies. Several reports have shown that conventional in vitro drug sensitivity tests such as the histoculture drug response assay are about 80% accurate in predicting sensitivity [27, 28]; however, this assay is time consuming and tedious. In addition, we observed that the variation in IC50 values for paclitaxel determined by cell viability assay between the four cell lines was less than one order of magnitude (from 1.8 nM to 7.2 nM), which suggests that the conventional assay is difficult to apply in clinical practice. The results of our current study suggest that our newly developed system for measuring CDK activity in vivo would allow more accurate prediction of paclitaxel sensitivity than the conventional assay.
We reported recently that activation of the spindle assembly checkpoint is required for paclitaxel-induced cell death [14]. Assessing the function of the checkpoint in human cancer by analyzing mutation of genes or protein expression, however, would be impractical. Actually, the sensitivity of paclitaxel in vitro was not consistent with the cell biological response in four cell lines. MDA-MB-468 and MDA-MB-231 cells treated with paclitaxel were induced to M-phase arrest and showed ring-like staining, which was followed by apoptosis, suggesting that both cell lines possessed functional spindle assembly checkpoints. In contrast, MCF-7 cells did not show any increase in the sub-G1 fraction after paclitaxel treatment although they showed ring-like staining and increased in the G2/M fraction, indicating that the spindle assembly checkpoint – and consequent induction of apoptosis – was impaired. This response may be related to the known defect in MCF-7 cells in caspase 3 [29], which is necessary for paclitaxel-induced apoptosis [30]. This finding demonstrates that M-phase arrest in response to paclitaxel treatment does not always reflect the induction of apoptosis. In the case of T47D cells, in which no typical ring-like staining was observed, we suspect that there were point mutations in β-tubulins – including important residues for drug–tubulin binding or altered expression of tubulin isotypes. The induction of apoptosis in T47D cells upon treatment with higher concentrations of paclitaxel than the IC50 values may therefore depend on other checkpoints instead of, or in addition to, the spindle assembly checkpoint.
Notably, in the absence of paclitaxel treatment, the paclitaxel-sensitive MDA-MB-468 and MDA-MB-231 xenograft tumors showed much more rapid growth than the resistant T47D and MCF-7 tumors (Figure 3). Several studies reported recently that rapid proliferating tumors have a higher response rate to chemotherapy [19, 20]. Actually, we found that the rapidly growing tumors showed significantly higher CDK2 specific activity without paclitaxel treatment than did the slowly growing tumors (Figure 4b). Moreover, a correlation between the effectiveness of paclitaxel and the tumor growth rate has previously been reported [24]. CDK2 specific activity in breast tumors would therefore be another indicator of paclitaxel sensitivity.
Consequently, accurate prediction could be expected from the combination assay of CDK1 and CDK2 activities. A clinical study is needed to validate our concept of predicting sensitivity to paclitaxel by analyzing the CDK activity in tumor tissues from patients.
The increase in CDK1 specific activity after paclitaxel treatment indicates that a tumor is sensitive to paclitaxel, and a lack of change in CDK1 indicates that a tumor is resistant to paclitaxel. The level of CDK2 specific activity before paclitaxel treatment was shown to correlate with paclitaxel sensitivity in vivo. Consequently, accurate prediction of paclitaxel sensitivity could be realized by a combination assay of CDK1 and CDK2 activities. Validation of our concept with a clinical sample will be needed in future studies.
cyclin-dependent kinase
Dulbecco's modified Eagle's medium
fetal bovine serum
50% inhibitory concentration
phosphate-buffered saline.
The authors thank Dr Tameo Iwasaki and Mr Kaoru Asano of the Central Research Laboratories, Sysmex Corporation, for intensive discussions and for supporting this project. They thank Christine F Wogan, MS, ELS, Department of Scientific Publications, Texas MD Anderson Cancer Center, for editorial review.
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Catherine Zeta-Jones is showing off her newly svelte body in a racy photo spread.
RELATED: Check out Catherine's Allure photo shoot
The Welsh beauty, famed for her curvy figure, has been sporting a slimmer frame recently after losing weight during her grueling run in the Broadway show "A Little Night Music."
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RELATED: Famous Naked Celeb Moments
The spread features Zeta-Jones laying on a bed, exposing her bare behind and staring sexily at the camera.
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She says, "That's when your dancing days and being in theater pay off. When you're doing a quick change, you don't give a s--- who sees you."


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Gang-Related News Articles
Welcome to the Gang-Related News Articles page. This page provides a list of articles pertaining to gangs and gang-related activities from various U.S. and Canadian news sources. A link to the source of each article is provided. The list of articles can be refined by date and limited to a specific state or Canadian province or territory using the control below.
Wolf and McDonnell promise more action to stop gang violence (Virginia): Congressman Frank Wolf (R-10th) joined Republican nominee for Governor Bob McDonnell on Thursday to unveil a comprehensive plan to reduce gain activity in Virginia.
Source: Loudoun Independent Date: May 28, 2009
Gang violence won't be tolerated in Utah (Salt Lake City, UT): Utah's next governor vowed Thursday to combat gang crime by continuing the work of a state task force established by the current governor.
Source: The Salt Lake Tribune Date: May 28, 2009
Gang with Pasadena ties targeted in huge Maryland sweep (Baltimore, MD): ATF officers and city police raided dozens of locations throughout Baltimore early Thursday, seizing drugs and weapons and arresting people on federal gang racketeering and drug conspiracy charges.
Source: The Baltimore Sun Date: May 28, 2009
New Oklahoma City center helps keep at-risk youth engaged (Oklahoma City, OK): Selling drugs and drinking alcohol were recurring items on Arnelious Kessee's to-do list. He was also accused of committing a burglary last year.
Source: The Oklahoman Date: May 28, 2009
Neb. governor signs bill aimed at gang violence (Lincoln, NE): Nebraska Gov. Dave Heineman has signed a bill that promises stiffer penalties meant to reduce gang violence.
Source: KTIV NewsChannel 4 Date: May 27, 2009
Tucson gang members arrested (Douglas, AZ): Douglas Police and The Arizona State Gang Task Force (GIITEM) arrested 6 criminal street gang members from Tucson for assaulting two male subjects at the 15th St. Park and for various other crimes including participating in a criminal street gang.
Source: Douglas Dispatch Date: May 27, 2009
Festival stabbing appears 'gang-related' (Fruitport, MI): A stabbing at the Fruitport Old Fashioned Days festival Monday appears to be "gang-related," according to Fruitport Township's police chief.
Source: Muskegon Chronicle Date: May 27, 2009
Plans in works to curb gangs in Tacoma (Tacoma, WA): Gangs are continuing to grow in Tacoma, prompting police to rework and bolster their plan for fighting them. "What we are doing isn't working," assistant police chief Bob Sheehan said recently.
Source: The News Tribune Date: May 26, 2009
Federal grants to benefit Pierce County (Pierce County, WA): A $1 million federal grant will help pay for three gang-fighting efforts in Pierce County. Safe Streets recently received the money, which will be spent:
Source: The News Tribune Date: May 26, 2009
Boy, 13, shot to death in East Oakland (Oakland, CA): Josue Lopez-Gil was 13. He had five hamsters, cared for wounded animals and set out bowls of milk for a neighborhood cat.
Source: The Mercury News Date: May 26, 2009
Conference today to address growing problem in Wichita: child sex trafficking (Wichita, KS): Police and social workers are concerned about the growing problem of child sex trafficking in Wichita, so they are holding a conference today called, "Community Action to End Domestic Sexual Exploitation."
Source: Examiner Date: May 26, 2009
Second year of city gang data shows slight decline in activity (Aurora, CO): This year's annual report on gang activity in the city contains a key element that last year's report lacked: comparisons. Now in its second full year, the report tracks the yearly progress Aurora Police Department's Gang Tracking Initiative, an effort started in 2006 that was designed to provide hard data and clear information about the activities of criminal street gangs in the city.
Source: Aurora Sentinel Date: May 26, 2009
Gang rivalry sparks shooting, Hemet police say (Hemet, CA): Police believe two Hemet men getting a pizza on Friday afternoon opened fire on a man who was an associate of a rival gang and was trying to raise money for his friend's funeral.
Source: The Press-Enterprise Date: May 26, 2009
Man shot by Long Beach police is gang member, but name is withheld (Long Beach, CA): Authorities on Tuesday withheld the name of a man killed during an officer involved shooting over the weekend, saying his family has yet to be formally notified of the incident.
Source: Press-Telegram Date: May 26, 2009
Anti-gang group making progress (Orangeburg, SC): The first year's report card on gang intervention in Orangeburg shows progress, but officials say more work needs to be done.
Source: The Times and Democrat Date: May 25, 2009
Two gang assaults reported in Monticello (Monticello, NY): Police in Monticello are investigating two serious gang assault incidents, both of which occurred last Friday.
Source: Mid-Hudson News Date: May 25, 2009
El Paso county attorney seeking civil injunctions against alleged gang members (El Paso, TX): Two fundamental forces will collide in the court case of 24 people whom the county attorney has publicly labeled as members of the Bloods street gang.
Source: Fort Worth Star-Telegram Date: May 25, 2009
Felony gang charge not used a lot (Columbus, OH): Three tattooed dots on a man's hand that officers noted six years ago during a routine booking for a gun charge tipped off Columbus police to another potential investigation.
Source: The Columbus Dispatch Date: May 24, 2009
More Latin Kings plead out in Lubbock court (Lubbock, TX): More members of the Latin Kings street gang pleaded guilty Friday in Lubbock federal court. That's the same street gang accused of a drive by murder in Big Spring last year.
Source: KCBD NewsChannel 11 Date: May 24, 2009
A gang dispute sparks a Portland drive-by shooting, police say (Portand, OR): A dispute between rival Latino gangs led to this week's drive-by shooting in Northeast Portland that wounded the mother of the intended target, according to court papers filed Friday.
Source: The Oregonian Date: May 23, 2009
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