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Jackson Browne gear & memories
Gabriel Currie and Echopark Guitars...
Visit with Jim Campilongo - new album!
Bill Nash's excellent S-63 Strat
Bill Nash Interview
Nash NGDP fuzz
Dr. Z MAZ38
Dr. Z on building exceptional amps
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Lemongrass AVEDA Salons and Spas offer 4 locations in the DFW area, providing bridal hair and makeup services by talented, educated, experienced stylists.
If you are visiting our Hulen or University locations in Fort Worth, be sure to experience our AVEDA spa, which offer waxing and facials, services for brides, the bridal party, and even the groom!
Lemongrass does offer services within our salon locations.
Please contact our special events coordinator, Amber, to coordinate your special day at email@example.com

subset: commoncrawl


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Siren Lace Textured Mesh Longline Underwired Padded Bra A-DD
incl. VAT
Product Details
- Product Shape:Long line
- Fastening:Hook and eye
- Padded
- Wired
- Adjustable straps
- Center bow feature
- 46% polyamide, 37% polyester and 17% elastane,(exclusive of trimmings)
Care Instructions
- Machine washable
Delivery & Returns Information
- FREE Standard Delivery on orders over 45€, 4-6 days.
- FREE Tracked Express delivery on orders over 75€, 2-3 days
You can return any clothing or home furnishing for a refund, provided you have the parcel summary document or other proof of purchase. Items must be returned in a re-saleable condition within 35 days of receipt. After 35 days you may still be entitled to a refund in accordance with your legal rights.
Returns should be processed by your local Post Office. Please obtain a receipt of proof of postage as, we may ask for proof of postage if the item does not reach us. Refunds for any items received as a gift will be credited to the ordering customer's account via the same method of payment.
Read our full Refund Policy for more information.


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Starting a new topic rather than continue derailing the "What type of red wine do I like" one.
Let's drop the cheap and expensive outliers and the old-school exceptions that need to be aged before you drink them and talk about California reds that (1) cost $20-40 bottle retail / $10-18 a glass in restaurants, (2) are technically well made in the sense that the winemakers are producing more or less the kind of wine they intend to, and (3) intended for drinking immediately after purchase.
I'm not unable to find such wines that I like, I just find it very, very difficult. I doubt I like more than one in 50 that I taste. As a noted in that other topic, when I go to places with "locavore" all-California lists, I typically taste all the reds, don't find one I want to drink, and end up drinking white when I would really prefer red.
My palate is not particularly eccentric in this regard. The food-hostility of these wines is one reason so many SF Bay Area restaurants that otherwise focus on local products have wine lists dominated by European imports.
What's going on?
First, most California vineyards are planted with inappropriate varieties. If you're trying to make a moderately priced wine for immediate consumption, nine times out of ten the most suitable grape variety or blend is not one that winemakers in Bordeaux and Burgundy use for their most expensive wines.
Next, the grapes are super-ripe. Traditionally, most grapes for red wine were picked at 21-23 Brix. Thanks to Robert Parker and mass hysteria, the average for Napa Valley Cabernet Sauvignon in 2008 was 25.7. The pendulum may be swinging back, but it has a ways to go.
These super-ripe grapes result in wines that are high in alcohol, which is exacerbated by many winemakers using new yeasts bred so that they would produce higher alcohol levels even at traditional brix levels. In the pre-Parker era, California wines (except for Zinfandel) were typically 12.5%. Today, it's rare to find a red wine under 14%, and 15% is common. That's 12 to 20% more alcohol, a radical change. It's not impossible for a balanced wine to have such a high level of alcohol, but (except for Zinfandel) it's rare.
Riper grapes have different, softer tannins and usually lower acid, which means the wines are more dominated by soft fruit flavors (which can be a plus with grape varieties that are traditionally used to make softer, fruitier wines). Softer tannins and less acid means they typically don't age well, though that's not an issue for wines intended for immediate consumption (unless you buy more than you can drink up while it's at its peak).
Another change from tradition is ubiquitous use of new oak. That used to be too expensive for moderately priced wines, but researchers at UC Davis figured out that you could use alternatives such as chips instead of barrels. Among other things, new oak makes the wine taste sweeter, exacerbating the super-ripe fruit flavors.
To my palate, without the harder tannins and higher acid of less-ripe fruit, the flavors of new oak are clashing and unpleasant. Some people obviously like the combination, but it's hard for me to imagine what food you could pair with such wines.


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Boko Haram Plans To Launch Attacks Using Beggars – Army
The Media Coordinator of the Operation Lafiya Dole in Borno State, Col. Mustapha Anka, said this in a statement in Maiduguri.
"The attention of the Operation Lafiya Dole has been drawn to a terrorists' intention to use beggars to cause mayhem in the state.
"Under the plan, concealed explosives in bundles of firewood are to be placed in crowded and strategic areas of Maiduguri and environs to cause havoc," the News Agency of Nigeria quoted Anka as saying in the statement.


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The Entrepreneur Podcast for Parents – Our Latest Episode
Ready for the Easy Revenue Generation Tool You're Probably Missing?
The Entrepreneur Magazine for Parents – Our Latest Post
Are you having difficulty changing from parent to business owner, and back again quickly?
Join our FREE Facebook community, and create maximum revenue in minimal time NOW with the rest of us Power Parents!


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You hear it all of the time. The problem is that the government is borrowing from China to fund our stupid spending programs, or popular subsidies, or tax cuts. Mitt Romney (remember him?), in a presidential debate, defined his criterion for deciding whether spending is worthwhile thusly: "Is the program so critical it's worth borrowing from China to pay for it?" Big Bird famously didn't meet that test.
Read Full Article »


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A 'Less Judgmental' Crist Reviewing Fla. Gay-Adoption Ban
Independent Florida Governor Charlie Crist (pictured) has an interesting decision to make: Whether the Sunshine State drop a lawsuit that seeks to uphold the state's ban on gay couples adopting children. Click here for the story, from the Orlando Sentinel.
The background goes like this: In early 2008, a judge in Miami threw out the ban in a case involving a gay man who adopted two boys. That decision was appealed by the defendant in the case, the state Department of Children of Families.
A decision on the appeal is pending. But Crist said Tuesday that he is reviewing the appeal. Crist has recently said he doesn't support he ban.
But here's the twist: the plaintiff in the case, Martin Gill, doesn't want . . .


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June 20, 2014
ARVE Error: no id set
We posted the whole "Dwyane Wade Is The Most Mispelled Sports Name" story. Well, Jimmy Kimmel makes a game of the wacky headline by heading to the streets of Hollywood Boulevard and offers random people $100 if they spell Dwayne's name correctly. Hilarity ensues…
CategoriesMiami HeatNews


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As we continue to improve the new Web Branch, we will be moving to publishing weekly release notes with details on the added features, improvements and bug fixes right on the Web Branch log in page. For larger releases, like today's release, we will provide additional details and screenshots as appropriate.
Today's release includes improvements to the Dashboard, a new Services menu item and many other smaller changes. To see the details, visit the Web Branch log in page.
Tuesday, June 30, 2015
Web Branch Release Notes
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Family supports Eat'n Park's Tips for Tots
As Eat'n Park collects funds for youth hospitals across its market region, waitress Taurniece Cromerdie and her daughter are helping McKeesporters understand how Children's Hospital of Pittsburgh touches the lives of everyday people.
Through Dec. 13, Eat'n Park is running its 26th annual Caring for Kids campaign. In McKeesport on Wednesday, servers took part in the Tips for Tots initiative that has helped to raise more than $5 million for children's hospitals over the past quarter century.
Cromerdie, an Eat'n Park waitress for the past 18 years, introduced customers to her daughter Naudia Mya Cromerdie-Grigsby, 11, who was treated at Pittsburgh's UPMC facility when she was diagnosed with a parvo virus infection at age 3.
"I didn't even know that people could get parvo," Cromerdie said of the virus that, in most forms, is deadly to canines.
Naudia was lethargic and simply wasn't acting like herself, her mother recalled. Already familiar with Children's Hospital because of a sickle-cell disease diagnosis at birth, Cromerdie took her daughter straight to the hospital, assuming she soon would have a medical crisis related to her condition.
"Within 20 minutes, Naudia went from the sickle-cell floor to the ICU," Cromerdie said. "The virus attacked her major organs almost instantly."
Naudia was on life support and had undergone a month of treatment when her family was prepared for the worst.
"She was gone," Cromerdie said. "But she's here now, and she's a blessing. When we visit Children's Hospital today, they say it's a miracle that she made it through."
During her recovery, Naudia was on transplant waiting lists for several vital organs; and over time her body has healed. She's now off the transplant lists, and her care consists of regular checkups with UPMC physicians.
"Children's Hospital is a part of my life," Cromerdie said. "Being a waitress at Eat'n Park, I'm proud to be helping out. Tips for Tots is the least we can do to help a hospital that helps so many people."
McKeesport Mayor Michael Cherepko visited the restaurant twice on Wednesday to check in with the Eat'n Park staff and guest servers including Councilman Keith Soles, police Chief Bryan J. Washowich and officer Justin Toth.
"It's an exciting opportunity for community leaders to take part in this program, which donates 100 percent of collected tips to children's hospitals," Cherepko said.
Cherepko served as Naudia's fifth-grade teacher for the first half of the 2011-12 academic year, before taking over the mayor's office.
"She's a wonderful young lady," he said. "She's a good student who is very kind-hearted. She's the type of kid who gets along with everyone."
Cherepko said it's amazing what Children's Hospital was able to do for Naudia's family. He said support from programs such as Tips for Tots helps youth hospitals make a difference in kids' lives on a daily basis.
Jennifer R. Vertullo is a staff writer for Trib Total Media. She can be reached at 412-664-9161, ext. 1956, or email@example.com.


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Bones Brigade: An Autobiography
This documentary follows six teenagers who formed a skateboarding team in the 1980s, revitalizing the sport and becoming its top athletes: Steve Caballero, Tommy Guerrero, Tony Hawk, Mike McGill, Lance Mountain and Rodney Mullen.
Rent DVDs for only $4.99 a month.
Movie Details
- Director
- Stacy Peralta
- This movie is
- Inspiring
- Format
- Ratings
NRNot rated. This movie has not been rated by the MPAA.
14Common Sense rating OK for kids 14+
Widescreen Anamorphic 1.85:1Subtitles
Spanish (Neutral)Closed captioned
YesLanguage and sound
English: Dolby Digital 5.1Other features
Color; interactive menus; scene access.
Bones Brigade: An AutobiographyClose
For Parents
Common Sense Note
Parents need to know that Bones Brigade: An Autobiography is a documentary about a team of famous skateboarders directed by Stacy Peralta (who previously made the best skateboarding documentary to date: Dogtown and Z-Boys). Like that movie, this one has a wealth of old photos and home movie footage to draw from, showing teen boys training hard, developing skills, and goofing off. Language is strong in the present-day interviews, including about a dozen uses of "f--k" and a few uses of "s--t." One of the main characters is shown to have married and had a child while still relatively young (late teens/early twenties). In the older footage, a teen girl is very briefly -- and blurrily -- topless, and teens are very briefly seen drinking beer. In photos, one of the main characters drinks an entire bottle of "mescal." Teen skaters are shown occasionally wiping out and mildly injured.
- Sexual Content
- At an impromptu skating event, a teen girl is possibly topless, though it's very blurry, very briefly, and seen from a distance. One of the main interviewees is shown to have been married and had a baby at a relatively young age, though this isn't discussed in detail.
- Violence
- Some skateboarding wipeouts and mild injuries are shown in some of the flashback footage.
- Language
- About a dozen uses of "f--k," and a few uses of "s--t" during the interviews. Other words include "damn," "hell," "idiot," and "retarded."
- Social Behavior
- Bones Brigade shows that by practicing and developing skills, teens can excel at something they love and perhaps even make a living at it. Though the skateboarding world is sometimes depicted as rebellious and irresponsible, these folks demonstrate that you can succeed without becoming corrupted. It also celebrates the benefit of teamwork.
- Consumerism
- In some of the older footage and photos, logos for Pepsi and Coke can be seen. "Slurpee" is mentioned. Tony Hawk is a brand in and of himself.
- Drugs / Tobacco / Alcohol
- In one sequence depicting an impromptu skating event, teens are (very briefly) glimpsed drinking beer. One of the interviewees mentions this as well. In old photos, one of the main characters is shown drinking an entire bottle of "mescal" (and eating the worm).
- Age appropriate
- Not an issue
- Depends on your kid and your family
- Not appropriate for kids of the age most likely to want to see it


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| This article doesn't have a proper headlining picture.
You can help American Girl Wiki by locating a catalog picture, scanning an image or illustration, or taking a picture and uploading it. Please follow our Good Article Images Guidelines.
The Ruffle Skirt is a My American Girl item released in 2012 in American Girl stores. Retail cost is $12.
Dress Like Your Doll
The matching girls' skirt was available for a cost of $28, it was sold on the website for a discounted price of $14.


text: 697 FXUS63 KAPX 261047 AFDAPX Area Forecast Discussion National Weather Service Gaylord MI 647 AM EDT Wed Jul 26 2017 .NEAR TERM...(Today through Tonight) Issued at 350 AM EDT Wed Jul 26 2017 ...Showers and storms through tonight... High Impact Weather Potential...Periodic thunderstorms will impact the region through tonight. A few stronger storms possible...severe risk low. Pattern Synopsis/Forecast...Overnight surface/upper-air/satellite analysis reveals a compact short wave and attending surface low rolling through central and northern Ontario. Trailing cold front extends down through Minnesota and continues to secondary low pressure in the central plains. Plume of moisture/instability extends northeastward ahead of the front up through the western Great Lakes and just starting to nose it`s way into the northern/western parts of the CWA early this morning. Several clusters of showers/storms continue to develop along the advancing moisture gradient...and have substantially increased in coverage in the last few hours across the northern lakes region and back through northern/central Wisconsin in response to an increasing low level jet leaning into the region. Several rounds of showers/storms are on tap through tonight. Parent short wave and surface low will track into western Quebec through tonight as trailing cold front folds down across the U.P. through the afternoon...then sags down through northern lower Michigan tonight. As mentioned above...clusters of showers/storms have increased substantially across the northern Great Lakes region and back through northern/central Wisconsin just in the last few hours...in response an increasing low level jet acting on the incoming moisture plume. Convection is lining up to move through a good portion of the CWA over the next several hours and thus plan on having fairly high PoPs for most areas. Precip may thin out for a time heading into the afternoon...as the low level jet relaxes. But suspect we see renewed development this afternoon across northern lower Michigan and particularly down toward Saginaw Bay as we warm/destabilize a bit...with showers/storms lingering into tonight as the front sags down through the region. Primary Forecast Concerns...Severe thunderstorm potential through tonight. Respectable 0-6 KM bulk shear values supportive of organized updrafts will be in place today...particularly this morning through midday as a core of 50 knot mid level winds punches across the northern lakes. However...overall instability remains on the modest side...with an axis of elevated MUCAPE (just pushing 1000 J/KG) nosing across the region this morning and MLCAPE values only reaching around 500 J/KG through the day. Certainly could see a few stronger storms owing to the decent shear profiles. But latest SPC DAY ONE outlook has kept us out of the higher severe thunderstorm risk categories today and tonight...which looks reasonable. && .SHORT TERM...(Thursday through Friday) Issued at 350 AM EDT Wed Jul 26 2017 ...Isolate showers Thursday morning improving weather going into the weekend... High Impact Weather Potential...none Pattern Synopsis/Forecast...Models show the main sfc trough running from the sfc low near ILX to Saginaw Bay to the low in Quebec. The ECMWF shows light rain lagging back in N lower, while the GFS is a little further south. However, by 18z/Thu, the rain is out of the forecast area. The sfc high builds into the region and another 500 mb shortwave trough looks to be moving into the Upper Great Lakes overnight. It looks potent enough for rain to come into the region, but not sure. There is little moisture at 850 mb and especially in the 700-500 mb layer. This also has implications for Friday as the models begin to kick off rain showers under the low around 15z/Fri, but the feature is out of the region by 00z. So think that it should remain dry, but this is lower confidence idea. However, it looks like it does dry out by 00z/Saturday and remains so Friday night. Primary Forecast concerns...As mentioned above, the 500 mb low/shortwave trough moving through becomes the main concern. The consensus model doesn`t pick up on it, while the GFS and ECMWF would suggest that there would be low chance or slight chance pops on Friday as the low moves through. Best guess is that since the models don`t have the rain in the same places as its counter part, that blending of the models washes out the pops for the rain. This will need to be watched. However, this would only dampen a few locations around the forecast area. .LONG TERM...(Friday night through Tuesday) Issued at 350 AM EDT Wed Jul 26 2017 ...High pressure looks to dominate the weekend... High Impact Weather Potential...Elevated fire weather conditions possible for Saturday and Sunday but it is a low chance probability. Extended (Saturday through Tuesday)...Saturday and Sunday will continue dry with the high pressure over the region. Sunday night, the GFS shows a cold front moving through with light rain, while the ECMWF is dry with high pressure continuing over the region. The blend of the consensus model brings chance pops on Monday as the front moves through, while the ECWMF continues to be dry with high pressure and that remains through Tuesday. The GFS dries out post frontal. So Tuesday is looking dry for the time being. && .AVIATION...(For the 12Z TAFS through 12Z Thursday morning) Issued at 647 AM EDT Wed Jul 26 2017 Round of showers and some non-severe thunderstorms will roll through northern lower Michigan this morning. Conditions are and will remain largely VFR...but brief periods of MVFR weather is possible with some of the heavier showers/storms. Precip thins out mid to late morning. But expect additional showers and storms to move through the region this afternoon into early evening. Difficult to nail down timing of TSTMs this afternoon/early evening at this juncture...so expect later amendments. && .MARINE... Issued at 350 AM EDT Wed Jul 26 2017 Gusty south-southwesterly winds will impact the region today... particularly the nearshore areas of Lake Michigan. Small craft advisories will remain in place through the day into early evening for northern Lake Michigan. Winds veer northerly and diminish quickly tonight and will remain on the lighter side through Thursday. && .APX WATCHES/WARNINGS/ADVISORIES... MI...BEACH HAZARDS STATEMENT through this evening for MIZ015. LH...NONE. LM...SMALL CRAFT ADVISORY until 8 PM EDT this evening for LMZ323-341- 342-344>346. LS...SMALL CRAFT ADVISORY until noon EDT today for LSZ321. && $$ NEAR TERM...BA SHORT TERM...JL LONG TERM...JL AVIATION...BA MARINE...BA


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It only takes a minute to get started. Fill out our uncomplicated sign up form and login to the website.
This step is up to you but will greatly enhance your chances of getting interest from other Christian members on the site. Let people know a bit about yourself, your hobbies and atach your nicest photo of yourself.
Utilise our website's state-of-the-art search to discover members who meet your requirements. Or just look through listings of Christian singles and discover people that you like.
Located someone who is attractive to you? Drop them a line or just a wink and see how you get along. With a little luck you'll be on your way to a lifetime of happiness.
It only takes a few minutes


text: 25 photos $10,750,000 4.05 acres 6 Beds | 14 Baths | 7 ½ Baths | 20620 sqft 1700 South Warson Road, Ladue, MO 63124 - Saint Louis County 4.05 acres Ladue, MO First time on the market: Extraordinary Ladue estate offers approx. 30,000 sq. ft. of living space. The home has stunning formal spaces, with exceptional finish details including limestone and marble floors, 16 fireplaces, main floor master suite with study, separate dressing rooms and "zen" garden; 5 individual second floor guest suites, 2 employee apartments, elevator and 6-car garage. Other features include a sweeping double staircase with balcony overlooking living and main dining rooms, 5,000-bottle wine cellar and adjoining casual dining room with private garden terrace, custom bars, library, fabulous kitchen, breakfast and hearth rooms. Walkout lower level has brasserie party room with 21' bar, billiards and game rooms, 2-lane bowlin


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- "So, say you're gonna do what this guy did, leave S.H.I.E.L.D., go off the grid. What's your move? I'd hide in the wheel well of a 747 flight from Newark to Singapore."
- ―Melinda May
Newark is a city located in the state of New Jersey.
Newark was the point of departure of the 747 flight where Melinda May would hide if she ever wanted to disappear, abandoning S.H.I.E.L.D. and getting "off-grid" just like Richard Lumley did. She chose that flight because it passed over 3,000 miles of land and a dozen countries, so she would be hard to track.[1]


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No public recommendations added yet.
Kirti Stores
Address: 3, Dattani Shoping Centre, Vasanji Lalji Road, Kandivali (west), Mumbai, Maharashtra, 400067
Phone number: 22 - 28017936Post A Recommendation Bookmark this Business Claim this Business
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how can I define this wall boundary condition ?
how can I define the wall boundary condition which the heat flux fluctuates with the time? Must I use the tool of UDF?
Re: how can I define this wall boundary condition
It's the way I'd do it.
It's really not that difficult to write a simple UDF like that. There is a good example in the UDF docs that should be easy to modify.
See section 10.1.2 of the UDF guide, entitled "Transient Velocity Inlet Profile for Flow in a Tube" (udf/node230.htm) in my installation.
|All times are GMT -4. The time now is 09:07.


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Jane Austen Centre
History, tourist information, and nearby accommodation
The Centre is open year round, and special group tours may be arranged. The centre is wheelchair accessible for the ground floor only.
About Jane Austen Centre
Address: 40 Gay Street, Queen Square, Bath, Somerset, England, BA1 2NT
Attraction Type: Museum
Location: On Gay Street, 5 minutes walk from the Roman Baths, well signposted.
Website: Jane Austen Centre
Phone: 01225 443000
Location map
Photo Credit: David Ross and Britain Express
We've 'tagged' this attraction information to help you find related historic attractions and learn more about major time periods mentioned.
Find other attractions tagged with:
Jane Austen (Person) - Regency (Architecture) -
Heritage Rated from 1- 5 (low-exceptional) on historic interest
Bath Assembly Rooms - 0.1 miles (Historic Building)
Fashion Museum, Bath - 0.1 miles (Museum)
Bath Postal Museum - 0.2 miles (Museum)
William Herschel Museum - 0.2 miles (Museum)
Victoria Art Gallery - 0.2 miles (Museum)
Museum of Bath Architecture - 0.2 miles (Museum)
Museum of Bath at Work - 0.2 miles (Museum)
No 1 Royal Crescent - 0.3 miles (Historic Building)
Nearest Accommodation to Jane Austen Centre:
Nearest Self Catering Cottages
Bath, Somerset
Sleeps: 10
Stay from: £1237.00 - 2442.00
Bath, Somerset
Sleeps: 2
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Sleeps: 6
Stay from: £495.00 - 1101.00
More self catering near Jane Austen Centre >>
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Bath, Somerset
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From: £85.00
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From: £139.00
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text:
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The community is for people working in government to share best practice and experiences, discuss problems, and improve skills.
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Anyone who's interested in learning about accessibility or sharing knowledge and experiences can join the community, especially people working as:
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Get involved
Join the accessibility Google group.
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Read these guides to learn more about accessibility:
- Content design: planning, managing and writing content
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- Writing content for everyone
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Accessibility in the Service Manual
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Help us keep the Service Manual up to date by:
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Money Hustle Collage Tee
- 50% cotton, 50% polyester
- Machine wash
- Model wears size Medium
- Imported
While dreams of money showers may dance in your head, how about trying to make it a reality? This jersey knit tee features a printed collage of Benjamin's, flags and "Keep It 100" at the front. One day…one day.


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As momentum builds for the implementation of the Courts Reform (Scotland) Act 2014 and the issue of expenses reform is opened up for debate, two draft bills were also proposed by two Scottish MSPs. The Scottish Government has been busy with proposals for change that will be of significant importance to the insurance sector.
Read the full article on our dedicated website for insurance professionals >>This information is intended as a general discussion surrounding the topics covered and is for guidance purposes only. It does not constitute legal advice and should not be regarded as a substitute for taking legal advice. DWF is not responsible for any activity undertaken based on this information.


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|Publication number
|Publication type
|Application number
|Publication date
|Oct 23, 1979
|Filing date
|May 19, 1977
|Priority date
|May 21, 1976
|Also published as
|Publication number
|Edward De Maeyer, Jaqueline De Maeyer nee Guignard
|Original Assignee
|Agence Nationale De Valorisation De La Recherche (Anvar)
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (7), Non-Patent Citations (2), Referenced by (36), Classifications (10)
|External Links: USPTO, USPTO Assignment, Espacenet
The invention is concerned with a process for the purification of preparations having a therapeutic activity, and in particular an interferon-type activity.
It also embraces the corresponding purified preparations and their use as the active ingredient of medicines.
It is known that interferon is a therapeutic product of great value, and specifically by reason of its remarkable anti-viral and immuno-depressive properties.
According to the usual methods of synthesis in vitro, this product is obtained by the action of virus or of chemical substances upon tissue cultures. The resultant preparations however also contain contaminating substances. Specifically, such preparations contain proteins which have a pyrogenous action and besides display inconvenient ability to induce a specific sensitisation in the recipient.
Consequently the crude interferon preparations cannot be used directly, in particular for the applications envisaged hereinabove, and must be purified.
Various methods have been suggested, specifically methods for purification by filtration through gel or through ion exchangers.
The use of these methods for the purification of interferon preparations however runs up against difficulties specifically due to very small yields.
Methods of affinity chromatography have also been suggested.
As is known these methods are based upon a selective affinity between an adsorbing phase constituted by a ligand fixed in covalent manner upon a solid support and some given product. The passage of a preparation containing the product through the adsorbing phase enables the product to be retained in a selective manner upon the ligand and thus enables it to be separated from the preparation containing it. The recovery of the product thus fixed can then be achieved with the help of an appropriate eluant.
The use of these particular techniques for the purification of interferon preparations however runs up against difficulties, and the attempts carried out up to the present time have not led to satisfactory results.
Thus the methods of affinity chromatography which, for the purification of the interferon, use anti-interferon specific antibodies have not yet been developed upon a large scale due to the difficulties of manufacturing anti-interferon anti-sera.
The object of the present invention is to remedy these inconveniences and provide a process which will make it possible to obtain, in an extremely simple manner and moreover with excellent yields, interferon preparations which possess a high content of products with interferon-type activity, to a major extent freed from contaminating proteins, and consequently capable of use in therapy.
Another object is to provide purified preparations in aqueous solution and thereby directly capable of use in therapy.
For that purpose, according to the process of the invention a crude preparation of interferon is put in contact with an adsorbent equilibrated with a buffer whose concentration in salts is isomolar or hypomolar relative to that of blood, the said adsorbent comprising on the one hand a gel forming a solid support, the size of the pores in which is such that they permit the passage of macro-molecules, and on the other hand a compound fixed upon the said support, playing the role of ligand vis-a-vis the products which it is desired selectively to separate, and which possesses a polycyclic structure of the kind of the blue chromophore, sold under the Trade Mark BLUE CIBACRON F3GA, of the formula ##STR1## or an equivalent product, whose configuration permits the interferon to be retained in a selective manner, the amount of adsorbent employed and the period of contact between the ligand and the preparation to be purified being sufficient to enable the desired fixation of the products with interferon-type activity from the above-indicated preparations, and that, for recovery of the products thus fixed, one elutes with the help of a buffer whose concentration in salts is hypermolar by comparison with that of blood by contacting the products fixed on the adsorbent with said buffer.
By these means it is possible to separate interferon with remarkable selectivity from preparations containing it, these preparations being either of human or of animal origin and constituted specifically by the supernatent liquids of tissue cultures, either freshly transplanted (primary or secondary cultures) or continuously grown, where the cells have been induced to form interferon, advantageously in a culture medium freed from serum, by a virus or by a synthetic nucleic acid such as poly I.C.
The separated interferon can easily be recovered, in practice in an almost total manner and without it being denatured, by carrying out an elution under the appropriate conditions indicated above. Furthermore, the advantage of this process is increased by the fact that the adsorption capacity of the adsorbent employed is practically undamaged. Consequently, this adsorbent can be reused for numerous purification operations, which is extremely advantageous from the economic view point.
In one embodiment of the invention the gel forming the solid support for the adsorbent comprises a polysaccharide such as agarose or a derivative thereof, specifically a modified agarose in which the polysaccharide chains are cross-linked in a three-dimensional sieve, in particular that sold under Trade Mark SEPHAROSE or again a polyacrylamide or an acrylic resin.
The ligand coupled to the solid support is advantageously provided by CIBACRON BLUE F3GA itself, or the product sold under the Trade Mark CIBACRON BLUE 3 GA (which differs essentially from the previous one by the presence of a hydrogen atom in place of the --SO2 ONa upon the phenyl radical at the extremity of the chain), or again by some equivalent product. By "equivalent product" there is meant, in both the description and the claims, a product which even if it differs from the blue chromophores indicated above in the kind and/or the position of its substitutions nevertheless makes it possible to secure the desired selective fixation of interferon.
The cyclic compound employed as ligand can without inconvenience be attached to a polysaccharide, specifically to a polysaccharide of high molecular weight, of the order of 2 million, designated DEXTRAN 2000. In this case it is the polysaccharide which is attached to the solid support.
A preferred compound of this kind, that is to say constituted by a cyclic compound attached to DEXTRAN 2000, is the product known under the trade mark BLUE DEXTRAN 2000, which corresponds to the aforesaid CIBACRON BLUE F3GA attached to DEXTRAN 2000. This product is advantegeously attached via the intermediary of the DEXTRAN 2000 upon a solid support constituted by agarose, and particular SEPHAROSE, which corresponds to the product sold under the trade mark BLUE DEXTRAN SEPHAROSE.
Although the reasons are not understood, it is a fact of great use from the practical view point that the aforesaid polycyclic compound of the CIBACRON BLUE F3GA kind, or its equivalent, displays a great affinity for products with interferon-type activity. Adhering to the conditions of iso- or hypo-molarity set out above, created advantageously by means of a buffer having a molarity of about 0.01 to 0.15 M, upon simple contact the said products are selectively adsorbed, while the major part of the undesirable substances present in the preparations are not retained.
It is particularly convenient to bring the adsorbent and the preparation to be purified into contact within a chromatography column, by making the preparation to be purified pass through this column. The column packed with the adsorbent is equilibrated with the aforesaid buffer whose pH is of the order of 6 to 8, preferably around neutrality.
Crude preparations of interferon are used which have interferon titres of about 104 to 106 International Units per 0.1 to 0.2 g of proteins, which gives a specific activity (International Units of Interferon, or U.I.F., per mg of protein) lying between 105 and 107. These preparations can be employed as such for passage through the column, or they can first be subjected to a preliminary dialysis against the buffer which served to equilibrate the said column. Operating under conditions isomolar with the tissue culture medium, the yield of interferon is about 80% to 90%, while it approaches 100% under hypomolar conditions. In the latter event, a particularly suitable buffer is an aqueous solution of Tris-HCl, pH 7.5 containing 10 mM of Tris-HCl ("Tris" is the abbreviation currently adopted to denote tris-[hydroxy-methylaminomethane]).
The stage of separating the products with interferon-type activity from the crude preparations which contain them is of course also within the scope of the invention.
Moreover from another aspect the invention provides, as new industrial products, the products with interferon-type activity attached upon the adsorbent phases indicated hereinabove. One preferred group of these products is constituted by those fixed upon BLUE DEXTRAN SEPHAROSE complex.
The fixed products can be recovered by increasing the concentration of salts in the buffer--specifically by the addition of acetates, phosphates, chlorides or other salts--in such a manner as to create a hypermolar concentration of salts relative to that of the blood, and at least 0.5 M and preferably 1 M.
This recovery of the products with interferon-type activity is advantageously carried out by elution of the column.
It will be noted that said recovery may entirely be achieved in an aqueous medium. The works done by the inventors have in fact evidenced that the products having an interferon type activity fixed on the ligand could be recovered almost entirely without having to resort to organic solvents and this by using a hypermolar buffer, such as indicated hereabove, in an aqueous solution.
The results obtained are equally satisfactory, whichever be the origin of the interferons, they may particularly originate either from fibroblastes or from leucocytes.
Upon recovery of the eluted fractions, one has purified interferon preparations with an increased content of interferon and practically wholly freed from contaminating proteins, and these indeed in a proportion of at least 90% as compared to the proteins initially present.
Besides, the conditions employed for the elution make it possible to recover the major part of the products with interferon-type activity in a fraction of the eluate of small volume, which has the advantage of making available preparations having an increased concentration in interferon as compared with the concentration in the crude preparations.
Thus, even starting from the better crude preparations of interferon described hereinabove, having an already high specific activity of the order of 107 International Units per mg of protein, by using the process of this invention it is possible in a single stage and with an almost total yield to obtain interferon preparations 30 to 40 times purer as compared with the crude preparations, and which have a specific activity lying between 1 and 5�108 International Units of Interferon. This specific activity is amongst the highest which has been obtained by other, extremely complex and costly methods, which moreover have the disadvantage of giving mediocre yields.
The scope of the invention also includes purified preparations. More particularly the invention concerns the preparations obtained by purification of crude fibroblast or leucocyte interferon preparations.
Said purified preparations have highly appreciable therapeutical properties and more especially a most interesting antiviral action, as evidenced by the various tests carried out.
It will particularly be referred to their action against the hepatitis B virus, which has been evidenced by administering to patients suffering from chronical hepatitis B, preparations, purified as per the invention, of human fibroblast or leucocyte interferon. The tests have been carried out according to the techniques described by Desmyter et al. in The Lancet, p. 645-647, 1976 and Greenberg et al. in Med. 295 p. 517-522, 1976. A considerable improvement of the hepatic functions could be observed in these patients, due to the replacement of the infected cells by non infected cells protected by the interferon.
These interferon preparations have also proved effective against herpes simplex and particularly for preventing from the recurrence of herpetic keratitis. The tests carried out, resorting to human interferon preparations, purified as per the invention according to the techniques described by kaufman and al. in J. Infect. Dis. 133 (suppt) A 165-168, 1976 have demonstrated the protective action of such preparations against infections caused by the virus and their inhibitive action against the recurrence of hepatic keratitis.
Moreover, said preparations reduce the frequency and the duration of viral complications more especially the ones resulting from herpes Zoster, as evidenced when carrying out the tests according to Jordan et al. in J. Infect. Dis 130, 56-62, 1974. On the other, they seem to increase the life duration of patients suffering from osteosarcome. Such properties may be evidenced by carrying out tests according to Strander et al. Acto Orthop. Scand. 45 p. 958-959, 1974.
All above tests have clearly demonstrated the perfect innocuousness of the products having interferon-type activity according to the invention, as well as the absence of noxious secondary effects resulting from their administration.
Moreover, considering the high degree of purity of the preparations purified according to the invention, the latter will advantageously utilized as active principles of drugs (medicines).
The interferon being specific of the species, it is then necessary for therapeutic applications for example in man to have interferon originating from humain cells.
The medicaments of the invention, which contain said purified preparations as active ingredients, can be administered, more especially by the intramuscular or sub-cutaneous route, in the form of an aqueous solution. A unit posological dose of 106 to 108 can be employed.
The treatment of chronical hepatitis is indicated as an example: patients are administered a 107 units dosis of fibroblaste interferon at alternate days, during two weeks.
Osteosarcomes may be treated by administering 3�106 units dosis of leucocyte interferon three times a week for periods which may range from 6 months to 1 year.
In order that the invention and its advantages may be well understood it will now be described in further detail, though only by way of illustration, in the following Examples:
The process for manufacture of BLUE DEXTRAN SEPHAROSE gels has been described by L. D. Ryan and C. S. Vestling (Arch. Biochem. Biophys., [1974], 160, 279-284). Since that publication, it has become no longer necessary to undertake the activation of the SEPHAROSE by cyanogen bromide according to Cuatrecasas (P. Cuatrecasas, J. Biol. Chem., [1970], 3059-3065) because there are ready lyophilised commercial preparations for the employment of already activated SEPHAROSE.
One employs SEPHAROSE 4B activated with cyanogen bromide and BLUE DEXTRAN 2000, such as those sold by Pharmacia (Uppsala, Sweden).
(a) Preparation of the SEPHAROSE gel (CNBr SEPHAROSE 4B).
In order to swell the gel and to eliminate the bactericidal agents present in the lyophilised preparation, the desired quantity of CNBr-SEPHAROSE is washed on a ground glass filter with a 1 mM solution of HCl, for about a quarter of an hour. The desired quantity of CNBr-SEPHAROSE is worked out upon the basis that one gram of dry product gives about 3.5 ml of gel. For washing one uses 200 ml of HCl 1 mM per gram of dry weight of CNBr-SEPHAROSE. The gel thus formed is immediately coupled with BLUE DEXTRAN 2000.
(b) Coupling of the SEPHAROSE gel with BLUE DEXTRAN 2000.
The quantity of BLUE DEXTRAN necessary for the coupling is worked out upon the basis that one gram of dry weight of SEPHAROSE is capable of fixing from 80 to 100 mg of BLUE DEXTRAN.
This quantity is dissolved in an alkaline buffer having a pH of the order of 8 to 10, especially a 0.4 M carbonate buffer of pH 10 (20 mg of BLUE DEXTRAN 2000 while easily dissolved in one ml of carbonate buffer).
The solution of BLUE DEXTRAN is poured onto the SEPHAROSE gel previously rinsed with 3 or 4 times its volume of 0.4 M carbonate buffer pH 10. The mixture of the gel and the BLUE DEXTRAN is poured into a deep glass flask and subjected to slight agitation, for example with a revolving paddle wheel, for about 18 to 24 hours, at a temperature of the order of +4� C., in order to promote the coupling.
After the coupling stage, the BLUE DEXTRAN SEPHAROSE product obtained is rinsed, on a ground glass filter, with an excess of 0.4 M carbonate buffer pH 10, in order to eliminate any non-fixed BLUE DEXTRAN, until the buffer remains colourless (optical density equal to or less than 0.02).
In order to eliminate the active sites which have not been fixed to the BLUE DEXTRAN, the product of the coupling is then put in contact with a solution of an alkanolamine, particularly 1 M ethanolamine at pH 8, for about two hours at a temperature of +4� C. For this operation, the product of the coupling is preferably transferred from the flask into another container.
To make the gel resistant to variations in pH, in molarity or in anything else, it is then subjected to three rinsing cycles, each cycle consisting of alternate rinsing firstly with an acid solution consisting of a 0.1 M acetate buffer at pH 4.0 to which 1 M NaCl has been added, and secondly with an alkaline solution which consists of a 0.1 M borate buffer at pH 8.5, to which 1 M NaCl has been added.
After these rinsing cycles, the gel is equilibrated with a 10 mM Tris-HCl buffer pH 7.5 to which a bactericidal and anti-septic agent has been added, in particular 0.02% sodium azide.
The BLUE DEXTRAN SEPHAROSE is poured into a chromatography column of the appropriate dimensions (it is estimated that one ml is capable of fixing about 8 million International Units of Interferon).
Before the very first use of the column, it is rinsed with 20 times its empty volume of the 10 mM Tris buffer pH 7.5 to which 0.02% of sodium azide has been added.
To eliminate the molecules of BLUE DEXTRAN which might tend to detach themselves, the elution buffer, consisting of the 10 mM Tris-HCl pH 7.5 to which 1 M NaCl has been added, is passed through until the optical density at 254 nm remains at 0.
(In the case of columns of large dimensions, it is recommended to undertake several successive rinsing cycles with the Tris buffer with and without NaCl in order to strengthen the adaptation of the column to variations in pH, in molarity or other variations).
A BLUE DEXTRAN SEPHAROSE column can be used for several months and for numerous cycles of interferon purification provided that it does not become contaminated by bacteria. It is therefore desirable to keep it, outside periods of use, under sodium azide and in the cold, at about +4� C.
Since however sodium azide is toxic for cells, it is then necessary before carrying out fixation of interferon to pass through the column at least three times its empty volume of the Tris-HCl buffer without azide in order to eliminate any trace of that antiseptic.
The interferon preparation to be purified consists of 32 ml of a supernatent liquid from a freshly-transplanted culture of human fibroblasts, induced to produce interferon by poly I.C. The titre is 6250 International Units of Interferon per ml, the content of proteins is 1 mg per ml (the high content of proteins in this preparation is due to the fact that it has been enriched with human albumin for its preservation). The specific activity of the said preparation is thus 6.2�104 International Units of Interferon per mg of protein.
The preparation is dialysed, at a temperature of about +4� C., for fifteen hours, against the equilibration buffer for the column, in this particular case the 0.01 M Tris-HCl pH 7.5.
32 ml of the preparation are passed, at a rate of 1 ml per minute, through a column 1.6 cm in diameter, containing a BLUE DEXTRAN SEPHAROSE gel according to Example 1 up to a height of 10.5 cm. The column is rinsed as a preliminary matter with at least three times its empty volume of buffer without azide. The sorption phase is monitored on a recorder of optical densities or OD at 280 nm.
As soon as all the preparation has permeated into the gel, it is washed by passing through the column from five to eight times the empty volume of the 10 mM Tris-HCl buffer of pH 7.5 in aqueous solution so as to ensure the elimination of non-adsorbed contaminating substances.
A fall in OD is observed, which finally comes to rest at zero.
To recover the interferon, an aqueous solution of 10 mM Tris-HCl of pH 7.5, to which this time there has been added 1 M of NaCl, is made to pass through the column, with an output of 0.5 to 0.7 ml/mn.
Fractions of 6 ml are collected in tubes, whose OD is measured to monitor the adsorption phase. The results obtained with the fractions in tubes numbered 8 to 11 are given hereinafter, in terms of their interferon titre and their content of proteins.
______________________________________ International Units Proteins SpecificTube No. of interferon/ml (μg/ml) activity______________________________________ 8 6 250 250 9 61 900 278 2.2 � 10610 830 16011 660 80______________________________________
It is found that the products with interferon-type activity are mainly recovered in tube No. 9. The interferon titre (U.I.F/ml) is ten times greater than that found in the starting material, while the content of proteins is about 25% of that of the crude preparation; the specific activity has been increased by a factor of 36, or other words the preparation which emerges from the BLUE DEXTRAN SEPHAROSE column has been purified 36 times as compared with the crude preparation.
It is found that the recovery of the chromatographed interferon carried on in aqueous medium is complete. In fact, in tube No. 9 some 6 ml were recovered of an interferon whose titre is 61900 units per ml, thus in total 371,400 units of interferon. This compares with the starting material consisting of 32 ml of a preparation containing 6250 units per ml, thus 200,000 units in total.
This example moreover only seeks to show the effectiveness of the method even in the case of a crude preparation of human interferon which was mediocre at the outset, as much because of its slight content of interferon as also because of its artifial contamination with albumin.
After the desorption phase of the products with interferon-type activity, the column is regenerated by causing about ten times its empty volume of the 10 mM Tris-HCl buffer pH 7.5 to which sodium azide has been added at a rate of 0.02% to pass through the column. In this way the column is made ready for re-use and can meanwhile be preserved at low temperature.
After several cycles of use, the gel can be regenerated to remove any traces of proteins which have become fixed thereon, and to do this the column is rinsed with five times its empty volume of 3 M KCl.
The effectiveness of the method for purification of interferon upon BLUE DEXTRAN SEPHAROSE gel is properly illustrated by Example 3, where chromatography is carried out upon a preparation of mouse interferon not previously enriched artificially with further proteins.
The interferon preparation to be purified consists of the supernatent liquid of a tissue culture (cells C 243) induced to produce interferon by the virus of Newcastle disease. After inactivation of the virus at pH 2, the supernatent liquid is dialysed against 0.01 M Tris-HCl pH 7.5, for about 14 hours, at a temperature of about +4� C.
The interferon titre of the preparation is initially 1.5�106 International Units of interferon per ml, its content in proteins is 0.1 mg/ml and its specific activity is 1.5�107 Interferon Units/mg of proteins.
100 ml of the preparation containing the interferon is made to pass through a BLUE DEXTRAN SEPHAROSE column according to Example 1, of 1.6 cm diameter, and containing the gel to a height of 10. cm, operating as in Example 2. After the column has been permeated by the preparation and rinsed, desorption is carried out with an aqueous solution of Tris-HCl, 0.01 M enriched with 1 M NaCl, recovering 6 ml fractions. The interferon is principally recovered in tubes 15 and 16 as shown by the results set out in the table below.
______________________________________ Specific activity (units of inter- Proteins feron per mg ofTube No. U.I.F./ml (μg/ml) proteins)______________________________________14 2.4 � 105 7015 2.3 � 107 111 2.1 � 10816 5.8 � 106 12 4.8 � 10817 2.4 � 105 1118 1.2 � 105 10______________________________________
As indicated above, the products with interferon-type activity are recovered principally in two of the tubes, mainly tubes No. 15 and No. 16.
As compared with the crude preparation, the increase in the specific activity, that is to say the degree of purification, is respectively 15 times for tube No. 15 and 35 times for tube No. 16. The specific activity of tube 16 in particular (4.8�108 units of interferon per mg of proteins) is up amongst the highest which have ever been published up to the present time.
Furthermore, the recovery of the interferon present in the crude preparation is complete.
In the 100 ml of the starting preparation, there were in toto some 1.5�106 �100, and thus 1.5�108, Interferon Units.
Now, having recovered all the products with interferon-type activity in the two 6 ml fractions each will be, respectively:
for tube No. 15: 2.3�107 �6 totalling 13.8�107 units
for tube No. 16: 5.8�106 �6 totalling 3.4�107 units
which is to say, on adding together the values for interferon activity observed in tubes 15 and 16, there are in toto 1.8�108 units.
The remarkable selectivity of the process of the invention should again be emphasised, since it makes it possible simultaneously to enrich the interferon activity while eliminating the major part of the contaminating proteins.
|Cited Patent
|Filing date
|Publication date
|Aug 2, 1963
|Aug 9, 1966
|Glaxo Lab Ltd
|Process of purifying interferon with alumino-silicate adsorbents
|Nov 30, 1964
|Dec 3, 1968
|Glaxo Lab Ltd
|Purification of interferon-containing material
|Dec 24, 1970
|Nov 20, 1973
|M Ho
|Interferon production
|Oct 2, 1974
|Aug 17, 1976
|G. D. Searle & Co.
|Interferon purification
|Apr 3, 1975
|Sep 21, 1976
|Stichting Rega V.Z.W.
|Stabilization of interferon
|Aug 4, 1975
|Apr 5, 1977
|Research Corporation
|Selective removal of albumin from blood fluid and compositions therefore
|Jul 22, 1974
|Aug 9, 1977
|Charles Chany
|Pharmaceutical formulation of interferon insolubilized by fixation on a support
|Chem. Abstract, vol. 77, 1972, 84911y, Boehme et al.
|Proc. Soc. Exptl. Biol. & Med., vol. 153, No. 3, Dec., 1976, Cesario et al.
|Citing Patent
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|Publication date
|Oct 12, 1979
|Jul 14, 1981
|E. I. Du Pont De Nemours And Company
|Purification of interferon
|Aug 6, 1980
|Feb 9, 1982
|The Green Cross Corporation
|Process for recovering interferon
|Jul 6, 1981
|Aug 10, 1982
|The Green Cross Corporation
|Process for recovering interferon
|Mar 24, 1981
|Dec 7, 1982
|Skurkovich Simon V
|Method and apparatus for the treatment of autoimmune and allergic diseases
|Jun 7, 1982
|Aug 14, 1984
|Mochida Pharmaceutical Co., Ltd.
|Method for purifying interferon
|Feb 7, 1984
|Feb 12, 1985
|Interferon Sciences, Inc.
|Method for purifying gamma-interferon
|Dec 3, 1982
|Apr 8, 1986
|Skurkovich Simon V
|Method of immonosuppression after transplantation of cells, tissues and organs
|Dec 3, 1982
|Aug 12, 1986
|Simon V. Skurkovich
|Methods for the treatment of pathological conditions by removing interferon from the organism
|Feb 14, 1984
|Apr 14, 1987
|Health Research, Inc. (Roswell Park Division)
|Method for the large scale purification of human fibroblast interferon
|Jan 23, 1986
|Dec 29, 1987
|Hoffmann-La Roche Inc.
|Adsorbent for protein purification
|Jul 9, 1986
|Feb 7, 1989
|E. I. Du Pont De Nemours And Company
|Preparation of a protein fraction exhibiting cell growth-inhibiting activity
|Mar 25, 1986
|Apr 25, 1989
|S.V.S. Laboratories, Inc.
|Method for treating AIDS and other immune deficiencies and immune disorders
|Dec 19, 1991
|Sep 1, 1992
|Ube Industries, Ltd.
|Body fluid purifying material and method for purifying body fluid by use thereof
|May 27, 1992
|Jul 18, 1995
|Viragen Inc.
|Method for modulating specific activity of inteferon alpha
|May 25, 1995
|Sep 12, 2000
|Schering Corporation
|Highly purified species of human leukocyte interferon
|Jul 10, 1998
|Sep 11, 2001
|Immunocept, L.L.C.
|Hemofiltration systems, methods, and devices used to treat inflammatory mediator related disease
|May 23, 1995
|Jun 25, 2002
|Schering Corporation
|Process for purifying human leukocyte interferon
|Jul 25, 2001
|May 4, 2004
|Immunocept, L.L.C.
|Hemofiltration systems, methods and devices used to treat inflammatory mediator related disease
|Mar 23, 2001
|May 18, 2004
|Immunocept, L.L.C.
|Method and system for providing therapeutic agents with hemofiltration for reducing inflammatory mediator related diseases
|May 15, 2001
|Sep 7, 2004
|Immunocept, L.L.C.
|Method and system for colloid exchange therapy
|May 12, 2004
|Nov 6, 2007
|Immunocept, L.L.C.
|Hemofiltration methods for treatment of diseases in a mammal
|Mar 23, 2006
|Apr 21, 2009
|Imunocept, L.L.C.
|Method and system for colloid exchange therapy
|Mar 8, 2004
|Apr 28, 2009
|Immunocept, L.L.C.
|Method and system for colloid exchange therapy
|Nov 5, 2007
|Jul 20, 2010
|Immunocept, L.L.C.
|Hemofiltration systems, methods and devices for treatment of chronic and acute diseases
|Mar 8, 2004
|Sep 9, 2004
|Immunocept, L.L.C.
|Method and system for colloid exchange therapy
|Apr 16, 2004
|Oct 7, 2004
|Matson James R
|Hemofiltration systems, methods and devices used to treat inflammatory mediator related disease
|May 12, 2004
|Feb 10, 2005
|Immunocept, L.L.C.
|Hemofiltration systems, methods and devices for treatment of chronic and acute diseases
|Aug 11, 2003
|Jun 15, 2006
|Jacek Rozga
|Selective plasma exchange therapy
|Mar 23, 2006
|Jul 27, 2006
|David Radunsky
|Method and system for colloid exchange therapy
|Nov 5, 2007
|May 15, 2008
|Matson James R
|Hemofiltration systems, methods and devices for treatment of chronic and acute diseases
|Jun 21, 1982
|Feb 17, 1983
|Mochida Pharm Co Ltd
|Verfahren zur reinigung von interferon
|Mar 18, 1982
|Apr 26, 1990
|Simon Rockville Md. Us Skurkovich
|Title not available
|Oct 10, 1980
|Apr 22, 1981
|E.I. Du Pont De Nemours And Company
|Purification of interferon
|Oct 10, 1980
|Nov 25, 1981
|E.I. Du Pont De Nemours And Company
|Purification of interferon
|Feb 26, 2002
|Jul 22, 2009
|Maxygen Aps
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|U.S. Classification
|530/351, 530/413, 530/417, 530/415, 424/85.4
|International Classification
|Cooperative Classification
|European Classification


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Prices, specials, features and availability subject to change.
Contact for details
IT'S ALL RIGHT HERE! Everything you could wish for in an apartment home is at Jackson Green. Experience Ocean County's most luxurious address! Upon entering this beautifully landscaped community, you will discover brand new apartments offering every comfort for a unique lifestyle with exceptional amenities. At Jackson Green, you're close to everything yet surrounded by the quiet ambiance of natural, open spaces with trees, farmlands, parks and streams. Visit the spectacular new Nature Center or the Manasquan Reservoir, or treat yourself to a day at Six Flags. When you're ready to make your next move, take a step up to Jackson Green.
Lease Terms: 12 month or 24 month lease. Security deposit 1 and one half months rent.
Breed Restriction. Cats Allowed. Dogs Allowed. Call for service animal policy.
128 units
Check Availabilityor call
They take great care of the place. The maintenance is amazing. Their customer service is excellent.
May 22, 2014
Residents are not friendly. It seems that they rent to anyone just to fill the apartmens. You can hear every Read More move and noise that the residents above you do. People slam doors on the way out or into their apartment. When you complain about it at front desk they just brush it off. Applicances are bottom of the line. When I moved out, I cleaned the apartment thoroughly, and was told it wasn't clean. I should have taken pictures before I left. The rent is definately too high.
May 22, 2014
i love the area and my place, clean quiet, and everyone here is very nice.
May 20, 2014
Monday-Friday: 9:00am-5:00pm
Saturday-Sunday: Closed
KRE Group
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Greetings mortals! This is Neo making a blog out of thin air just because I can. I won't promise it's better than anything you read on other blogs on this wiki nowadays (because I'm just as out of ideas as everyone else) but maybe it's something you haven't thought of yet.
Oda is a sexist! Well, duh, I suppose you knew that already. But apart from his depiction of women did you ever notice how most females got DF powers that are associated with femininity or archaic images of women?
- Woshu woshu no Mi (Tsuru) - Doing the washing is one of the chores traditionally done by women. (Probably because leaving the washing to a man can only end in a disaster...)
- Sube sube no Mi (Alvida), Kilo kilo no Mi (Miss Valentine), Bonney's unnamed DF - Women care a lot about their skin/weight/age.
- Mero mero no Mi (Hancock), Hebi hebi no Mi (Sandoria/Marigold) - Gorgon theme. Nuff said. It's actually quite fascinating how close Oda stuck to the original: There were three Gorgons, two immortal and one mortal. The two immortal gorgons were ugly from the start. Medusa, who was mortal, was said to be very beautiful before she got turned into the same kind of monster as her sisters by a jealous goddess. Later she got her head chopped off by Perseus. (if memory serves there is a japanese/east asian saying about beating someone's head off their shoulders, meaning crushing someone's overconfidence - which could be applied to Hancock and Luffy)
- Ori ori no Mi (Hina), Awa awa no Mi (Kalifa) - That's just sexual harassment...
- Hana Hana no Mi (Robin) - The name and the whole makeup have definitely a feminine touch.
The only DFs consumed by girls excluded from this are Toge toge no Mi (Paula), Mogu mogu no Mi (Miss Merry Christmas) and Horo horo no Mi (Perona). Though, one could argue that the spike fruit belongs into the same category as Hina's and Kalifa's DF (if you are into that kind of thing...).
Captain Obvious out. Now you can make your sexist jokes.
PS: It's super unfair there are no female Logia users. :/


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British Travel Awards Vote now
Famous for its stunning landscapes, crystal clear waters and white sands, Alghero lies on the North West coast of the island and is a historic walled town with a pretty harbour and fabulous coastline. The 'Centro Storico' or 'Old Town' is the hub of Alghero and is called Barcelonetta or 'Little Barcelona' with streets named in both Catalan and Italian. In fact, you'll often hear the locals shouting out to their neighbours in the local Catalan dialect. Surrounded by the ancient walls or 'Bastioni' overlooking the sea, Alghero's narrow cobbled streets are the perfect place to explore and unwind. The magnificent Santa Maria Cathedral and the churches of San Francesco and San Michele are an architectural delight, whilst the many piazzas offer the chance to simply sit and sip a cappuccino and watch the world go by. Shop till you drop in the boutique filled streets and then take a bite to eat in one of the many restaurants followed by an ice cream from one of the gelaterias and then a stroll along the seafront.
Most restaurants in Sardinia will serve fresh and quality produce, along with Italian wine. Seafood is always a good option, especially when by the coast - ask to see your fish before it is cooked for you. Also, whilst in Alghero, try the Seadas - fritters stuffed with cheese and lemon peel and topped with Sardinian honey.
The nightlife in Alghero is quite relaxed and relatively low-key, but you will find an array of good bars and restaurants to spend your evenings over a quality meal and a few drinks. You will find somewhere to spend the evening to cater for all tastes, from an Irish pub, friendly local Italian places, relaxed lounge bars and stylish cocktails bars with live music. Many places will cater for you to sit outside, so you can enjoy the warmth of the summer evenings and the sunset over the harbour.Information updated 17 Mar 2014
Speciality of the area
An exclusive type of red coral, found only off the coast of Sardinia can be found in the towns jewellery shops.
General Services
Food Shopping
High Street
The details displayed are for general information purposes only and were correct as at the last updated date. Please note that details (including opening and closing times) may change. In particular, some facilities may only be available during high season. Attractions are not connected to James Villa Holidays and we do not endorse or recommend any supplier or attraction.
There are a few winerys in the area of Alghero, however the closest being Santa Maria la Palma. A family-run winery with Sardinian vineyards, the estate produces wine and grappas for you to sample and purchase.
The Alghero marina is a large natural port, situated just along from the cathedral and city walls. From here you can take a boat trip to Neptune's Grotto, a series of marine caves located on the imposing cliff of Capo Caccia.
Le Bombarde amusement park for families is close to the beach of Le Bombarde, which offers different play areas for children, and a cafe for mums and dads.
Spend an afternoon meandering around the winding streets of the historical old town and its Piazza Civica, an open square lined with boutique shops an restaurants. Follow on with a stroll along the city walls, originally built to protect against maritime threats. From the height of the walls you can enjoy stunning views out towards Capo Caccia and Neptune's Grotto.
The five chapels of Cattedrale di Santa Maria, dedicated to the virgin Mary, can be found in all it's gothic glory at Piazza del Duomo. It was founded in the 14th century and has recently been refurbished. If feeling fit, you can wind your way up the bell tower!
Porto Conte National Park and Capo Caccia extends from the north of Alghero and round the coastline, until it is almost opposite the town. The Capo Caccia is a marine protected area of limestone cliffs, that juts out of the land with views looking back onto Alghero. The national park is a perfect place to walk with the family, exploring the flora and fauna, bird watching and taking in the beautiful scenery.
Take in the medieval Catalan influenced architecture, found all around the old town.
Rock climbing
Ten pin bowling, Bike hire, Cycle route, Fishing, Bird watching
Sail boats, Scuba diving, Snorkelling
The details displayed are for general information purposes only and were correct as at the last updated date. Please note that details (including opening and closing times) may change. In particular, some facilities may only be available during high season. Attractions are not connected to James Villa Holidays and we do not endorse or recommend any supplier or attraction.
New Years Day
Festival Alguer - The Anfit...
Calvalcata Sarda - Sassari
Alghero's Cup - Alghero
Alghero Challenge Cup - Alg...
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Our Holiday Season 2016
Can you even believe that Christmas Eve is tomorrow? We are so ready to finish work for the day and start our own joyous celebrations! Bridget gets out early for Finals (yay!) & Casey is finishing up a few things at work and then we are both officially on WINTER BREAK!!
But before we take the weekend off to celebrate Christmas, we wanted to share with you our latest seasonal video.
Like our summer video and fall video, we captured all of the activities we did during the holiday season. From our Christmas card photo shoot, to cookie day, to days outside in the snow and adventures in downtown Chicago…our 2016 Holiday video has it all! We hope you guys enjoy seeing a glimpse behind-the-scenes around here just as much as we love capturing these memories and sharing them with you.
Holiday 2016 Video
We still can't believe Christmas is this weekend…. where does the time go?!
We hope you have a wonderful Christmas! We'll be back here on Monday with our December Happy Hour post, and then we'll officially be out of the office until we get back from our trip to Mexico (more on those vacay details and our posting schedule next week).
And most importantly, thanks for all of you have done to make this holiday season a magical one here on the DIY Playbook!! We feel so blessed to have you in our lives and beyond grateful for your unwavering support.From the bottom of our hearts, Happiest Holidays from our homes to yours.
Catch up on some of our past videos here:
Leave a Comment
10 thoughts on "Our Holiday Season 2016"
Kerri Hansen
Love ❤ the video! I am so happy I have discovered your blog this year, I can't wait to see what you guys have lined up for 2017!
I want to wish you and your families a very Merry Christmas and Happy Holiday season. Enjoy Mexico, have some guacamole for me! Hugs from North Vancouver, Canada 🇨🇦!↓
DIY Playbook
We are so happy to have you Kerri! Merry Christmas to you & your family!
And yes….there will be plenty of guac happening. haha!
Merry Christmas to you both!↓
DIY Playbook
You too Trang! 🙂
Kara D
It's been a whirlwind holiday season for me as well! Enjoy your winter break, wear comfy clothes, sip some bubbly and eat some donuts! May it be drama-free and bright! Merry Christmas and Happy New Year!↓
DIY Playbook
Bubbly & Donuts….you know us too well Kara! Merry Christmas to you!
shawnna griffin
hey girls -love the video! Merry Christmas!↓
DIY Playbook
Merry Christmas to you Shawnna!
Colleen Bansley
Merry Christmas Casey & Bridget and to your families too!! This video is so cute!! Can't wait to see what is in store for the DIY Playbook in 2017!!!↓
DIY Playbook
Thanks so much Colleen! And thank you for the adorable holiday card. Cutest family ever! 🙂


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The Shack — based on the best-selling novel about a grieving father who spends a weekend with the three Persons of the Trinity — comes to theatres next week, and the studio behind the film has released a few new clips to whet our appetites.
Watch: Four new clips from The Shack tap into the movie's theological themes
Newsbites: The Jesus movies past and future edition!
A few quick items about three recent and upcoming movies about Jesus.
Watch: The trailer for Season 2 of CNN's Finding Jesus
I must confess, I kind of missed the first season of Finding Jesus: Faith, Fact, Forgery when it aired on CNN two years ago. At the time, I was writing quite extensively about National Geographic's Killing Jesus and NBC's A.D. The Bible Continues, and there just wasn't time to follow a third program as well, as intriguing as it seemed.
Newsbites: The religious TV shows in development issue!
Three quick items about TV shows with a religious hook that are now in development.
Win a DVD copy of Hillsong: Let Hope Rise!
Last Friday, the Aussie worship band Hillsong United released a new music video for a song of theirs that will appear on the soundtrack for The Shack. As it happens, Hillsong had a movie of their own last year, and I have a DVD copy to give away. The disc includes the entire 103-minute documentary, called Hillsong: Let Hope Rise, as well as four bonus songs and almost 20 minutes of extra behind-the-scenes footage — and if you'd like to have it, you can send your name and mailing address to firstname.lastname@example.com by this Sunday, February 19. I'll pick a winner at random next week. (The disc is coded for Region 1, so it will work on DVD players in the U.S. and Canada.)
Watch: Hillsong United's new music video for The Shack
The powers that be have released another song from the soundtrack to The Shack, and with it another music video containing new bits of footage from the film. The studio has also released three new TV spots. Check 'em all out below the jump.


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Dominus Estate 2004
Winemaker Notes
The 2004 vintage is deep red in color, with an intense nose of vanilla, black currants and cedar. The elegant integration of the tannins and fruit is noticed immediately. Flavors of warm mulled spices on the mid-palate complement the balanced and complex structure. The long, aromatic finish exhibits notes of... View More
Critical Acclaim
This very generous and well-ripened offering starts out with an involving display of cherries, cassis, herbs, sweet oak and loam in its deep and decidedly complex aromas, and it follows with equally wide-ranging flavors that show a real sense of layering and plenty of depth. Ripe, but always in balance and firmed by a good dose of youthful tannins, this latest Dominus bottling is among the winery's very best efforts ever, and it is certain to get better yet over the next decade.
The elegant, dark plum/purple/garnet-colored 2004 Dominus exhibits a big, sweet, cedary, roasted herb, mocha, black cherry liqueur, black currant, and earth-scented perfume. Medium to full-bodied with silky tannin as well as a luscious mouthfeel and finish, it should be more accessible in its youth than many of its predecessors, but easily last for two decades
DominusView all wine
His love of Napa Valley lingered and in 1981, he discovered the historic Napanook vineyard, a... View More


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Grey Jays
Expand Messages
- Greetings! I run a small organic farm in Elk, and as we have a very active bird population I have started on the path of becoming a birder. My father in law is coming to visit this week, from Carmel. He is serious about his list. I saw that there are Grey Jays in Van Damme, and would love to go with him to see them. Never have seen one here. Any suggestions as to where to start?
Many thanks,
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Nova by SinglenautView Demo Store
Nova is a sleek theme designed by our friends at Singlenaut, and it sets the stage by displaying your beautiful photos front and center, while keeping the rest of the design concise and out of the way.
- Responsive and optimized for any type of mobile or desktop device
- Image slideshow on the Home page
- Set the number of featured products on the Home page
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- Display your store's logo in the header
- Display a repeating background image on every page
- Display product inventory bars on Product pages
- Show a search field in the header
- Customizable welcome message on the Home page
- Choose either a stacked product layout with large images, or switch to a simple grid view of product thumbnails
- Recommended Home page slideshow image size is 1200 x 800 pixels
- Use the Home page welcome message to advertise promotions, tell your company's story, or simply say hi to your customers
- Use the Home page slideshow to advertise discounts or show off detailed product shots
If you're on a paid plan and don't mind getting your hands dirty in the code, then here are a few common advanced customization questions we get for this theme.
How can I remove the store name from the top left corner of the page?
Head to Customize Design > Advanced > Layout and remove the following code from the header section of the page:

{{ store.name }}

Since your store name serves as a link back to your Home page, you may want to add your Home link to your logo/header. To do that, head to Customize Design > Advanced > Layout and look for the following code:

And change it to:

How can I remove the products on the Home page?
To remove the featured products from the Home page, head to Customize Design > Advanced > Home and remove the following code:
{% else %}
{% if products != blank %}

{% for product in products %}

{% for image in product.images %}

{% endfor %}

{{ product.name | truncate: 75 }}

{% case product.status %}
{% when 'active' %}

{% if product.on_sale %}On sale!{% endif %} {{ product.default_price | money_with_sign }}

{% when 'sold-out' %}

Sold out

{% when 'coming-soon' %}

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{% endcase %}
{% endfor %}
All Products
{% else %}

No products found.

{% endif %}


text: Sony sort of pre-empted the rush to wrist-based technology, updating its original offering to the SmartWatch 2 late last year, along with the introduction of the Sony Smartband at CES. But it is in no rush to sign up with Android Wear.Speaking to CNET, Ravi Nookala, head of Sony Mobile's US business, stated that it plans to use its own technology for the foreseeable future: 'We've already invested time and resources on this platform, and we will continue in that direction.'Sony wasn't on the original list of Android Wear partners and as such, the statement comes as no surprise. But it isn't quite the end of the road. Sony followed up the comments with a statement that keeps the door open just a little.'We're excited about the potential of Android Wear to extend the mobile OS experience into wearable devices,' it said. 'While we are currently focused on our in-market wearable offering, including SmartWatch 2, we continue to work closely with Google as a key partner and continue to evaluate opportunities across a number of areas as we extend our SmartWear Experience.'So perhaps it's a case of seeing where both SmartWatch goes and where Android Wear goes before making a final decision. Sony has its own platform, apps and customer base. It isn't going to give that up until someone demonstrates some big numbers on a rival platform.Source: CNET


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Let us know when your receive great advice so we can thank our lawyers and point others with similar questions in the right direction.Got it!
I am an owner of a wedding photography business and I am inquiring about a wedding contract issue. Our contract states that a 50% non-refundable down payment is required to secure a wedding date. A client is wanting to cancel her booking and she wants to get her payment back. We told her at our initial meeting that the deposit is non-refundable, and we went over the contract with the client, she filled out her information on the contract and gave us the deposit, but she forgot to sign it. I need advice on the validity of the contract, and I am inquiring to find out if legal representation is needed in this case.
Depending on the size of the contract, you should probably refund the money. Without her signature, she would have a good argument that she didn't agree to the nonrefundable deposit. You would have an argument that she agreed and the written contract plus payment is evidence of her behaving in accordance with the contract terms. The question is: do you want to spend a few thousand dollars on attorney fees and damage your reputation?Ask a similar question
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Apprenticeship program is really bright aspect of any professionalism industry for applicants/ apprentices and for employers both. If you are an aspiring learner looking forward to media apprenticeship, travel apprenticeship or may be for electrician apprenticeships qld and even actually in any industry for building your professional career in that particular field, the options aren't less. Else you may be a company owner who is looking forward to enroll the apprentice program in your company but little non-informative about it. Both sort of people – a wannabe apprentice and employer needs to acknowledge their respective benefits which they will get from an apprenticeship program.
Benefits of Apprenticeship to an apprentice
Qualification of a person counted by its degree or certificates which comes along heavy tuition fees but qualification essential in any industry to work the practically requires to learn those skills. Some sort of apprenticeship may need or benefited with some pre-knowledge or course but rest of vocational based industries can professionally acquired by you as starting your career with apprentice program. You should know these benefits or advantage which will push you towards apprentice program.
- Apprenticeship program are structurized training where you will be qualified as learning the particular practical or vocational skills towards achieving the qualification in that industry for further building career in it. You will be gaining those knowledge and skill set as literally working there as apprentice
- Apprenticeship offers an initial entry in your interested career or industry along with your study or may even before studying of it. You will start a good pay or may lucky to get smart amount in very earlier stag e of your career
- It is experience, dealing with real problems and issues rather than reading it theoretically. You will develop more qualified skills and confidence in that industry.
- There will no tuition or heavy admission fees, no education loans and no debt. You will earn with earning which is the best part of it.
- Apprentice has immense and wide area of choice as there are more than 500 types of apprentices. Nearly every industry with all kind of jobs has apprentice position.
Benefits of Apprenticeship to an employer/Company
More and more companies are determining the importance of apprenticeship program and ending up including it in their company. As company owner, you should look for these benefits and surely get an apprentice program for betterment of your company
- The apprentice programs are generators of skilled workers for the company in coming future. It is sort of investment where they are training the skills of apprentice according to the company's process or functionality. They can be future leaders of your company.
- The skill gaps and process lapses in your company can be filled by these apprentices where minor or non-primary but essential works are efficiently done by them. The apprenticeship program is source of bright employees which will be very well established with company already.
- It improves productivity, tackles and resolves skill shortage, cultivates fresh ideas and perspectives and also boosts the company's overall quality of production.


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1 work Add another?
Analysis of nonhomogeneous polar-orthotropic circular disks of varying thickness1 edition - first published in 1962
Links (outside Open Library)
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History Created April 1, 2008 · 2 revisions
|September 9, 2008
|Edited by RenameBot
|fix author name
|April 1, 2008
|Created by an anonymous user
|initial import


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Crime Scene Forensic Photography Job Description Salary Education Forensic means "having to do with law
Use Photography to support evidence
Photograph evidence ,accidents and crime scenes
Different Techniques to accurately record evidence
On call
create a complete and accurate picture of the crime scene.
Used for trials Usually work for:
Police department
Forensic Services company
Coroner's office
Insurance companies Depends on;
$50,000 annually
good pension and benefits Strong background and qualifications in photography
Most start as Crime Scene Investigators or Scene of Crime Officers
General Training in CSP
Specialist Training in
Experience in digital imaging
National Occupational Standard for photo imaging
Science( bio/ chem) at least one A level or higher
and math Criminal Justice Strayer University Manansas Virgina University of Phoenix Phoenix , Arizona Criminal Justice University of Nevada Natural Sciences Las Vegas Prerequisites No Set Standards
higher Education Higher pay
Depends who you work for
Police Traning
No Formal Training Criminology ,Forensics Degrees Experience Infrared and ultraviolet Cameras, computers, Image production By: Olivia Nevarez
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Alex Nevarezon 14 May 2013
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