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text:
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text:
Pushing aside instincts.
I hear Gabby's thoughts, they're almost shouting at me. She thinks of Alex with fondness, and me with resentment. I sigh. She's trying to hide her tears, but I can still see them, glistening in the sunlight. We stay there, unaffected by the light, the silence growing colder between us. The voice in the back of my head, the one that tells me to be less harsh, is berating me for my words. I scowl at the floor, trying to ignore it. But how can you ignore the parental instinct? Regardless of how long you push it aside, it never goes away.
'Sit down, Gabby.' I say quietly. She hesitates as I get up to let her sit by the window. Well, massive hole. Reluctantly she sits down and I perch on the edge of the seat next to her. What she says next puts everything I was about to say to the back of my mind.
'How are you sitting in the sun? I thought vampires were supposed to burn...'
'It takes about two hours for a vampire to become badly hurt by the sun, Gabby. After about an hour they start to burn, but it's only as bad as human's sunburn.' I shrug, looking away as she wipes her eyes. 'Otherwise we wouldn't last long as a species, would we?' I smile slightly. I look up at Konrad again. He's managed to survive however many hundreds of years like this. So why is it so hard for the rest of us? She remains silent, but I can still hear her thoughts.
'What about silver...? What does that do to vampires?' she asks after a while.
'Silver...' I mutter, wondering how I can explain that one. 'Real silver – not that crap that humans think is silver – has properties that can be used by all manner of creatures. Vampires, demons, angels, shape shifters. Et cetera. As for what it can do? It has thousands of different uses, really. For the most part it depends on how it's been created and what the creator wants to use it for. It can be used to restrain a creature, or to enhance the powers they already have.' I pause and look at her. 'It's a hard one to explain, that one. I would usually say ask Konrad, but he's one of the Elders I warned you about earlier... I'm sure there's someone around other than him that can explain it better than me.'
Her thoughts are still swirling. 'Is it something I could learn to use?' she asks.
'No. I've spent the last seventy years or so trying to get it to work for me, and had only minimal success.' I tell her with a laugh as I remember my last attempt to use silver. I ended up branding myself with it. I trace the burn marks on my arm through my jumper, not needing to see what pattern they formed, though my right hand stays wedged into my pocket, fingers protectively wrapped around the blob.


text: AUBURN HILLS, Mich., Jan. 4, 2013 /PRNewswire/ -- As the 2013 FIRST (For Inspiration and Recognition of Science and Technology) robotics season starts on Jan. 5, The Chrysler Foundation awarded grants totaling $147,000 to 25 elementary, middle and high school teams in Alabama, Arizona, Indiana, Michigan and Ontario, Canada. The grants will help the teams offset costs associated with items such as registration fees, parts and materials and team apparel. The Chrysler Foundation will award $29,000 in additional funds to boost the championship runs of teams qualifying to compete at the FIRST Championship, April 24-27, in St. Louis, Mo. In 2012, nine teams received additional funds. This year, The Chrysler Foundation has committed a total of $181,000 to FIRST teams and programs. "The FIRST program is an ideal platform for inspiring the next generation of science and technology leaders that are vital to our industry as well as many others," said Mark Chernoby, Head of Vehicle Engineering and Vice President - Executive Coordinator, Chrysler Group LLC. "We hope that by engaging students at an early age, we can provide the spark that inspires them to become tomorrow's engineers." The Chrysler Foundation has awarded grants to the following schools and their respective teams: FIRST Robotics - $6,400 grant (per team) Arizona Kingman High School Indiana Western High School ( Russiaville) Michigan Andover & Lahser High Schools ( Bloomfield Hills) Ann Arbor Huron High School Birmingham Seaholm & Birmingham Groves High Schools Bishop Foley Catholic High School ( Madison Heights) Chelsea High Schools Dundee High School Goodrich High School* Grand Blanc High School Hartland High School Lake Orion High School Macomb Academy of Arts and Sciences (Armada)Notre Dame Preparatory ( Auburn Hills) Oakland Tech Northeast ( Pontiac) OSM Tech Academy at Clarkston High School Oxford Community Schools Pontiac High School Rochester Adams and Stoney Creek High Schools ( Rochester Hills) Walled Lake Schools Warren Consolidated Schools *Two teams are based at this school


text:
Defense Official: Russia Has Short Range Missiles in South Ossetia
"Anything such as that, or any other military equipment that was moved in would be in violation of this cease-fire and should be removed immediately," Pentagon Spokesman Bryan Whitman said. "The only forces that are permitted to remain under the cease fire agreement are the forces that were in there at the Aug. 6 time frame.
Webmaster's Commentary:
And what can the US and NATO do, precisely, to counter this?
Not one heck of a lot, unless they feel the uncontrollable urge to go to war against Russia in behalf of Georgia.


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All Hilton London Metropole hotel rooms offer highspeed internet access.


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|Skin in Health and Disease
Contact Dermatitis
Contact dermatitis is a common occupational disease and occurs at home to cosmetics, skin care products and to other chemicals. Outdoors plants are common causes of contact dermatitis; poison ivy is prototype of a skin irritant that produces dramatic inflammation on contact. Textile dyes in clothing are a common source of skin problems.
Our strategy of managing skin diseases is to treat them as ecological problems. You get better results if you assume that several factors interact to produce skin disease than if you assume that each disease has a single cause and single cure. The skin is a meeting place for disease- causing agents coming from the outside and the inside at the same time. Skin irritants and allergens combine forces to produce skin chronic skin disease. Food allergy and contact allergy may be two expressions of the same or similar antigens. Food allergy may start eczema, and the inflamed skin, in turn, becomes more vulnerable to irritants, allergens and infections.
Contact dermatitis has been divided into two categories:
1. allergic contact dermatitis 2. and irritant contact dermatitis
Both allergic contact dermatitis and irritant contact dermatitis have similar presentations. Acute skin lesions may be red, swollen and weeping; crusting lesions may appear after a few days. In chronic cases skin reddening, scaling and fissuring are common. The distribution of the lesions may provide clues to the irritant or allergen trigger. Identifying and removing the contact solves the problem.
Allergic Contact Dermatitis
Allergic contact dermatitis is a delayed, type IV allergic reaction. Latex rubber is a well recognized source of contact reactions. Healthcare personnel. now wear latex gloves routinely in all patient encounters and sensitization estimates go as high as 40%. Patients who have been sensitized to skin-contact allergens may develop generalized eczematous inflammation if these allergens or chemically related substances are ingested. For example, a patient with a history of nickel allergy may get a widespread involvement after eating foods rich in nickel. Patients sensitized to latex may cross-react to bananas and kiwifruit. Patients sensitized to topical ethylenediamine may develop generalized inflammation following treatment with aminophylline.
Contact urticaria is a local immediate or delayed hive-like reaction reaction at the site of contact . The skin contact may also cause a generalized allergic reaction with rhinitis, asthma, or anaphylaxis. Natural rubber latex, for example can cause contact urticaria. Symptoms vary from mild itching to asthma and anaphylaxis. Ethylene oxide, isocyanates, chloramine-T, epoxy resins and nickel sulphate have caused IgE-mediated skin reactions.
Ahmed and Richardson described patients with hand dermatitis; their occupations were in healthcare, followed by laborers, service workers, and machinists/mechanics. The most commonly positive patch tests were: formaldehyde & releasers, other preservatives, fragrances, rubber, nickel, and neomycin/bacitracin. Bacitracin is an antibiotic that is used in several types of consumer products, including cosmetics and ophthalmic and skin ointments. Bacitracin is often combined with two other antibiotics, polymyxin and neomycin, in first aid antibiotic products (Polysporin). Increasing reports of allergic contact dermatitis reactions and near fatal anaphylaxis concerns the North American Contact Dermatitis Group.
Allergic contact dermatitis of the feet may be caused by rubber and leather
components of shoes; substances identified by patch testing included
chromated leather, p-tert butylphenol formaldehyde resin,
dithiodimorpholine, 2- mercaptobenthiazole, thiurams and isocyanates. Textile contact dermatitis often presents itching and red patches with or
without fine skin peeling. Huntley suggests: "Usually these lesions develop at
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of knees, buttocks, waistband area, and anterior and posterior axillary folds."
Skin in Health and Disease
is a book about skin health, skin care that offers solutions to specific skin disorders through diet revision an improved nutrition. The good news for many suffers some chronic skin disorders is that complete diet revision will often improve and sometimes resolve skin symptoms. This book focuses on the most common skin disorders that can be controlled by changes in skin care, diet and the environment. Eczema, for example, is sometimes an expression of food allergy, or allergy to contact materials. Dermatologist may deny the food allergy causes, so that their patients often have to make independent decisions. In all aspects of medicine, we advocate responsible self-management and offer this book as primer and reference for self-managers with skin problems.
The book, Skin in Heath and Disease is intended to be used with the Alpha Nutrition Program. This text provides background knowledge, helpful in understanding skin disorders and the relevance of diet revision. Often a food holiday on Alpha ENF is the best way to begin recovery and the Alpha Nutrition program is an ideal way to design a safer, healthier long-term diet. If diet revision is appropriate to solve your Skin Disorders, you can order the the Starter Pack Option.
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Alpha Nutrition is a trademark and a division of Environmed Research
Inc., founded in 1984 at Vancouver, BC, Canada. Online Since 1995.


text:
Home › Forums › Gear Forums › Gear (General) › ULA Conduit VS MLD Exodus
Mar 23, 2009 at 2:20 pm #1235020
Ok, I'm trying to decide between these two packs so please let me hear your thoughts on which you would prefer. Both have the same capacity of 3200 cu in and both have the same average max load. The difference is the Conduit weighs 17 oz and the Exodus weighs 13.8 oz. Do the extra 3.2 ounces on the conduit make it a more comfortable and durable pack? Let me know. Thank you in advance for your input.
-SidMar 23, 2009 at 2:51 pm #1488190
Oh and also feel free to give me suggestions of other similar backpacks if you prefer it over the two I listed.
-SidMar 23, 2009 at 3:05 pm #1488194
Craig BurtonMember
@missingutahLocale: Smoky Mountains
How much weight do you plan on carrying?
If you can't fill these packs, then I don't recommend either. To me, the design of each requires a full load to carry comfortably.
As far as durability and comfort in relation to weight.. well, I find the opposite to be true; at least for the 2008 models. I would suspect the Exodus to be more durable, as I've explained in a previous post somewhere if you search.
The fixed hipbelt pockets of the Conduit is a luxury that is tough to pass up though.
Be aware that the Conduit is designed to ride a little high — which means, as a skinny guy, it requires a little bit of adjusting as I walk – especially with very light loads.
Though that all sounds favorable for the Exodus; but I'm not lying when I say I favor the Conduit for trips – mainly because of the convenient hipbelt pockets.
You can't go wrong with either though, so long as you are using them for their appropriate purposes and loads.Mar 23, 2009 at 3:28 pm #1488200
Well, my base weight will be under 8 lbs. I am in the process of adjusting my gear list, however, and I may be able to bring it down to under 7 or possibly 6 lbs. I am also looking at Zpacks and the MLD Revelations, and while I do love the idea of the weight savings, I am not too sure about the durability aspect and I would rather not have to buy a $100 pack every few weeks.Mar 23, 2009 at 4:35 pm #1488222
Joe WestingMember
Don't forget about the MLD Prophet! 2900 cubes w/ the extension collar.Mar 23, 2009 at 5:02 pm #1488230
Craig, being that you own the Exodus, you can probably answer this. I plan to use the GG Torso length pad with this pack. Is there a sleeve in the exodus to insert the pad? If not, how do you use the pad to get back support from it? Thanks.Mar 23, 2009 at 5:23 pm #1488246
Chris MorganBPL Member
@chrismorganLocale: Southern Oregon
A GG torso nightlite might be a bit small to use in the Exodus. I have used a 3/8 thinlite coiled twice inside for winter use and this has worked quite well as a frame. For the spring, I plan on buying a 3/4 length nightlite, cutting just just beyond torso length so that the pad encircles the entire inside of the pack.
Don't forget the Exodus has two volume reducer loops on the bottom to shrink the pack.Mar 23, 2009 at 5:39 pm #1488249
Mark BishopBPL Member
@mark_bLocale: Northwest (WA)
The 2009 version of the Conduit has modular (optional) hip belt pockets. I think this is how ULA is able to show an overall weight reduction for the 2009 Conduit. (17 oz. vs. 20 oz last year) I know it's a personal choice but I wouldn't do without the hip belt pockets.Mar 23, 2009 at 8:45 pm #1488302
Ok so what I understand then is the Exodus does NOT have a sleeve to put the pad in, whereas the Conduit has straps to keep a pad in place against the back? Meaning the Exodus would do better with a 3/4 Nightlite that goes all the way around while the Conduit would do better with a Torso nightlight that would be up against the back with the straps? If this is correct, which configurations is more stable?Mar 24, 2009 at 9:52 am #1488407
Kyle PurcellMember
@dufus934Locale: North Texas
I just returned from a short overnight trip with my conduit. The base weight was a little more than yours, but it carries very well. I have used a full length 3/8" ccf pad coiled up. I get extra padding for my back by putting the area with the most pad overlapping against my back in the pack. Then I stuff bag and clothes in first (take up room) then everyting else on top. This is the best way to pack the bag (I think), becasue if you need more room, just push and the bag/clothes compress in the pack.
I love the conduit, and cannot over state this pack's comfort.
You must be logged in to reply to this topic.


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Pen and paper - or - movie cam or other type cam. Freeze Plus P spray might me good to have in console of vehicle.
This is a discussion on angry guy wants to fight within the Carry & Defensive Scenarios forums, part of the Defensive Carry Discussions category; We have four...with cars parked in their driveways...perhaps a local contact may help. We've had a few speeders in the summer...usually when groups of us ...
We have four...with cars parked in their driveways...perhaps a local contact may help.
We've had a few speeders in the summer...usually when groups of us are sitting together in someone's driveway...
Last time a construction worker went speeding by, the group all yelled to slow down...the guy came back to a crowd of about 7-8 people...he nodded and waved...and was driving much slower.
We like our 'hood...
In your situation, I would just get his plate number and report it...
When he drove backwards to speak to you, I would have walked away...you're no chicken...your armed, and you need to back down or walk away from loud mouths like him...he'll get his, and it's spinning out in the cosmos right now...just waiting to strike!
Stay armed...use common sense...stay safe!
Proverbs 27:12 says: The prudent see danger and take refuge, but the simple keep going and suffer for it.
Certified Glock Armorer
NRA Life Member
Pen and paper - or - movie cam or other type cam. Freeze Plus P spray might me good to have in console of vehicle.
For God, Family and Country!
"I did not want to look like a chicken."
Man, if you are carrying a concealed weapon you better look like, walk like and cluck like a bloody chicken.
If this had escalated in to a shooting situation a lawyer would have a "field day" with that statement. What is it they say about pride before a fall?
Even if I don't have a duty to retreat, when on my own time, I will if I safely can. In my house is another story, but on the street E&E is what I do best.
I have been in an "Armed Encounter" and between you and me, there is nothing fun, glamorous or heroic about it.
Last edited by BikerRN; April 12th, 2007 at 03:43 AM.
I have problems off and on with a business across the parking lot from mine. A large body shop that employs a large number of "gangsta's". It's boom cars, drag racing in the parking lot, drug deals you name it, we've had it on occasion. I just make it a point to stop in and have a talk with my local PD whenever stupidity reaches intolerable levels or the drugs start up again. Then we suddenly have patrol cars cruising the parking lot several times a day. Amazing how peaceful all those "gangsta's" get when that starts.
Don't be afraid to get LE in on it now. Description, plate number and report his dangerous behaviour every time he tears up the street. Chances are he's had contact with them or soon will. They will just keep their eyes open more and that will certainly help you. As one cop told me when I reported one of the neighboring bad boys statement of " Ain no cop bad enuf to mess wid me", "Oh, we just love those guys! They're a lot of fun!"
If you stand up and be counted, from time to time you may get yourself knocked down. But remember this: A man flattened by an opponent can get up again. A man flattened by conformity stays down for good. ~ Thomas J. Watson, Jr.
Better you decided not to fight.Fighting is against the law.Period.Both sides.No such thing as self defense in a fistfight if you decide to fight back because you are then as guilty as they are of assault,unless you are just outright jumped on and getting beatup and not putting up any defense or are overpowered by their size or skills.Legally,retreat is what you are supposed to do in a hand to hand situation.No matter how skilled or what size you are or who is at fault.There is such a thing in the law as 'equitable' defense.You have to use the same amount of skill or power against an opponent,as they use on you,in hand to hand combat.This is in the public view.Not someone breaking into your home.We know the rules in that situation.But,most people do not know the rules out in public when comes to hand to hand combat because we have been under educated since we were in the 5th grade and having that fight with a kid on the playground.We've been led to believe our whole lives thats its a 'mans' thing to do or we are cowards or yellow if you dont fight back and you run away your a wussie.Well,the law says different, and if you are brought before a judge you'll find out real quick what the law really says about it.Google it for yourselves and study it.I know.Ive been there.Ive studied and continue to study now martial arts and boxing,almost my entire life,and I'm 5'10" by 250lbs.Still.I would walk away no matter.Go to the house quick as you can.Close the door.Call non emergency 911 and report the speeder and aggressor.He comes to your door,ignore him.He comes in the door,unauthorized,then you got the right to protect your self by any means necessary.Out in the street, or the driveway,another story.You arent being a coward.You are being smart.And law abiding.
Last edited by nofearfactor; April 10th, 2007 at 04:14 AM.
Your best weapon is your brain. Don't leave home without it.
Thoughts: Justifiable self defense (A.O.J.).
Explain: How does disarming victims reduce the number of victims?
Reason over Force: Why the Gun is Civilization (Marko Kloos).
Carrying means that we have to walk away from such a situation. If you wish to fight, do not carry a gun.
Call the police and report the incident NOW! I know, you feel silly reporting it this much after the fact. Doesn't matter. Do it anyway.
You need to get it on the record that he threatened you (which is exactly what he did). If this blowhard ever tries to make good on his threats then having your complaint on record ahead of time may very well make the difference between you being charged and not charged. And believe me, no matter how innocent you are, if you get charged it is going to cost you a TON of money!
Battle Plan (n) - a list of things that aren't going to happen if you are attacked.
Blame it on Sixto - now that is a viable plan.
Learning to shoot again : Starting Over
Be a Rattlesnake.
Distance is your friend and Avoid and slither way at all cost. Only after this has failed and your backed into a corner make a loud verbal warning for others to hear, and strike only in fear of being devoured.
Its not about guns...Its about Freedom!
Assistant Instructor @ http://www.green-ops.com/ Located in the Washington D.C. Metropolitan Area "Why should your training be any less special?"
A few folks have already covered it- pride disappears. I would also add to the repertoire: get OC, practice with it; get a koppo or fist-load, practice.
Its all well and good to talk avoidence (I've stayed happy in life with this policy ), but it won't necessarily be a possibility. Have more tools in the box than just a hammer.
He got out, started the verbal "exchange"(aka, "assault"). Your wife should have been on the cell to 911, and you would be perfectly entitled to have your spray (concealed) in hand. One verbal, "Hey, I'm not looking for trouble, we're just walking...", he comes closer, he gets some spice.
We go thru this same thing in our neighborhood with speeders.My kids skateboard and play frisbee and basketball and they know better than to be in the street anyways,but still,you're gonna be at some point and these kids literally have to run for their lives many times.Not much can be done other than reporting maybe.
Just stay out of their way I suppose is the best thing to do.People just want to drop and fight anymore these days over just about anything.My wife is constantly ******* someone off in traffic by screaming at them or whatever for cutting her off or whatever and never thinking of the possibility that someone is going to stop someday and want to rumble with her,or worse.And she couldn't fight her way out of a wet bag with holes in it.A Sunday school teacher with not a violent bone in her body but a bad attitude out on the roads.Me,I'm calm and cool and collected and just wave at em when they cut me off or swerve at me or whatever.Doesnt bother me in the least.I will let anyone in front of me when waiting.People step on my foot and I apologize to them for my foot being there.I'm big and naturally muscular and bulky looking and still have a good frame at 38 from years as a bodybuilder and boxer and athlete when I was younger.But I'm as non offensive as I can be to anybody.I just let it all roll off of me.Lifes too short to get into it with every one I meet.Maybe I'm just getting older and wiser.When I was younger I thought I could whip anybody with my hands and feet.i didnt carry pistol back then,for one,it wasnt legal in Oklahoma.Now,I just carry my pistol wherever I go(legally anyways), keep my mouth shut,my eyes open,look over my shoulder wherever I am, and hope to hell no one ever messes with me or breaks into my home or car or messes with any of my family.Hopefully I can get thru this life without getting killed or worse yet,having to kill someone.
Yep, this is definitely a downside to carrying a gun. You have to look like a wuss sometimes and walk away from a fight. Just remember though, you are doing them (the blowhards) a favor by not getting involved in a fight where they might wind up dead. By reigning in your temper and pride and backing out of a fight, you are literally granting them another day to be an a-hole, which makes you the bigger man.
To address your situation, you probably could have called the police on that guy. Tell them he was speeding through your neighborhood, stopped and confronted you when he saw you watching him, and I guarantee you the police will not take kindly to his behavior. Police will respond to calls about irresponsible or erratic driving, so they would definitely respond to a call like this.


text:
7147 CLARK POINT, Winneconne WI 54986-0000 $239,900
Features of 7147 CLARK POINT Winneconne WI 54986-0000
- Lots for Sale
- Status: Active
- Mailing City: Winneconne
- County: Winnebago
- Lot Size: 58x192
- School Dist: Winneconne Community Sch Dist
- Property Taxes: $2,316
- Property Tax Year: 2016
- Access Type: Lake
- Listing Company: First Weber Group
Property Description for 7147 CLARK POINT Winneconne WI 54986-0000
7147 CLARK POINT Winneconne WI 54986-0000 - Extremely hard to find waterfront, buildable vacant lot with walk out potential is now available and open to all builders. There are lots of mature trees and the perfect setting for viewing the most beautiful sunsets. 58` of improved shoreline and all the archeological studies are completed so you can start building quickly.
Land Features
- Waterfront/Access: Y
|MLS Number
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text:
February 21st- The 2009 Protagonists Diary
February 21st! Where has the new year gone! That New Year's Baby is toddling around on two legs and taking off!
Diary, I am sad to report I have been AWOL from my writing. :( My jobs have been keeping me very busy! I have a case conference coming up and have to prepare all of the documents fro that, plus all of the documents for another conference I'm not even going to. Luckily, grading the papers of my students is easy. It takes longer to enter the grades into the gradebook than to grade them. And lesson planning is almost done FOR me.
But enough about work. This is the WEEKEND! I don't want to think about work.
The kids are playing various video games, the husband is asking me to clean the house (which I promised I would) so I have to put down my pen and climb out of my chair and get to it!
I've already cut his hair, so I'm making progress on my chore list even this early in the morning! :)
So sorry so short, but I'm still busy!


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Italian Hotel Classification is sponsored by the Italian Government. Accommodation facilities are classified through the star system that awards stars according to the structural, service and functional standards of the facility itself.
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The NH Caltagirone Villa San Mauro hotel enjoys a panoramic location, about 5 minutes driving from the city center of Caltagirone in south-eastern Sicily. NH Caltagirone Villa San Mauroprovides a total of 91 air-conditioned guest rooms. Rooms are spacious and tastefully decorated with traditional c...
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Theater setting
Swimming Pool


text:
Happy Birthday Brad Pitt!
Can you believe that this stud of a man, Brad Pitt, is turning the big 5-0 TODAY!
Over 10-year ago the hunky Brad Pitt was featured on the cover of Vanity Fair magazine where he said, "I'm not in a hurry. Listen, if it's not right, I don't want it. I'm telling you straight: it's a damned good time of my life."
Since then the actor's career has only accelerated and now has 6 kids with long time girlfriend, Angelina Jolie.
But we gotta say, the man looks good for 50.


text:
What was all the commotion about in North Surrey on Tuesday morning?
For one hour only, between 8:30 and 9:30 a.m., the Shell gas station at 148 Street and 108 Avenue sold gas for just 10 cents a litre.
"I got 42.5 (litres) and it cost $4.25. It's ridiculous and I love it though. I'm broke and on disability so it really helps."
That is customer Mark Bogdanoff.
It was promotion for a new gasoline.
The lineup was blocks long and got so bad, the intersection was closed and police were called out.
"Police are here to insure that everybody acts like adults and doesn't cut in line and just does their fair share of following the rules,"
says RCMP Staff Sgt. Dale Carr.
Shell was set to hold the deal again later in the morning, at another location.


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Industry transformation — driven by forces as varied as evolving care delivery models, increasing competition and accelerating consolidation — underpins key issues facing hospital executives today
Learn more about:
For a complete listing of Willis Towers Watson services for the hospital industry, download our brochure, Transformation in the Hospital Industry.
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Best Getaways for Under £200!
Yes really, bargain breaks all under £200! They all include flights and some are even all-inclusive so you can eat and drink to your hearts content! Prague Let's kick things off with the beautiful city of Prague. This offer offers the perfect base for exploring the beautiful city of Prague, you can explore the famous historical sights … Continue reading Best Getaways for Under £200!


text:
Tax Power of Attorney South Dakota Form – Adobe PDF
License / Price: Free
Version: Adobe PDF (.pdf) Fillable
File size: 289 Kb
3,843 Downloads
Use this tax power of attorney form (POA) to assign and authorize your tax powers to your agent. In most cases this will be your CPA (Certified Public Accountant) and the document will be attached to your tax filing. The form must be signed in the presence of a notary public in order for it to be accepted by the South Dakota Department of Revenue.
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A knowledge-sharing seminar was organised in Cedefop's premises in Thessaloniki on 22 May. European Training Foundation (ETF) experts presented their work on organising qualifications systems in the ETF partner countries as well as on skills and migration.
What is vocational education and training (VET) like in the smallest European Union Member State? Watch our video to get an overview of Malta's VET system in a nutshell.
A new online toolkit for tackling early leaving from education and training was launched at Cedefop's policy learning forum on vocational education and training (VET) as a solution to leaving education early (16-17/5) in Thessaloniki.
At an international conference organised by Italy's Regione Umbria on 10 May in Brussels, Cedefop Director James Calleja said that 'recognition, reputation and employability are the key indicators of quality in adult education and learning.'
The May 2017 issue of Skillset and match, Cedefop's magazine promoting learning for work, is now available to read and download.
A community of practitioners to promote quality in vocational education and training (VET) delivery has been set up by Cedefop in close cooperation with the European associations of VET providers active in the cooperative network Vet4EU2.


text:
Status: Open (Call business to confirm holiday hours)
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John E Patricelli, MD provides surgery in ELMER, NJ. The most common types of surgeons are: General, Cardiovascular, Otolaryngology, Neurological, Reconstructive, Neonatal and Orthopedic.
Please call John E Patricelli, MD at (609) 358-7330 to schedule an appointment in ELMER, NJ or to get more information.
Be the first NJ.com user to add photos or videos of John E Patricelli Md
Patricelli John E MD is located at 350 Front St, Elmer, NJ. This business specializes in Doctors & Clinics.
Patricelli John E MD can be found at Front St 350. The following is offered: Doctors & Clinics. The entry is present with us since Sep 8, 2010 and was last updated on Nov 14, 2013. In Elmer there are 20 other Doctors & Clinics. An overview can be found here.


text: Starburst Botanical Ring Regular Price: $275 Special Price $137.50 Qty: Size Guide Add to Bag Add to Wishlist Stunning Art Deco details embolden this conversation-sparking cocktail ring, featuring pavé and statement Swarovski crystal stones, including a colorful cushion-cut centerpiece. Set in genuine gold-plated pewter, it adds an opulent touch to your everyday bands, and shines as a standalone accent to simple-and-sleek looks. • 2" long x 1" wide • Pewter / crystal / brass • Nickel-free / lead-free / cadmium-free • Made in USA • Free ground shipping & returns on full-price U.S. orders Style# R17J414X Share Contact us


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text:
I pulled my scarf around my neck so I could hide my gills. It was going to be a pain I could tell but I was going to have to live with it for the time being. I couldn't do anything about my hair, which was blue, but hopefully that wouldn't be too out of place.
I looked at the clock on the wall. That can't be right, half past! I'm late!
'Bye mum!' I yelled over my shoulder as I flew out our front door. Oh God, oh God, oh God! I couldn't run fast enough, I was much better at swimming. I saw the docks, everyone was already on board but the teacher is still waiting next to the boat.
'Late Nina? That's out of character for you.'
'I know I'm sorry miss.' I hurry up the gangplank and onto the boat. My class looked so different with all their animal attributes covered up but I managed to spot my best friend, Harper and made my way over to the seat next to him.
'Nice scarf.'
'Nice hat.' I pulled it off his head and his rabbit ears came out.
'Don't do that, you know I can't let people see them.' He grabbed the hat back from me and stuffed his ears back inside it.
I smiled, it was so easy to wind him up and it felt so satisfying. Luckily he knew I was joking so we got on well.
'Ready to go Harper?'
'Are you?' He raised one eyebrow in that quizzical look he does.
'Yes I am.'


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The Art and Science of Synaesthesia
The Institute for Art and Olfaction
Tuesday, April 23, 2013 from 7:00 PM to 9:00 PM (PDT)
Event Details
Smell psychologist Dr. Avery Gilbert leads an interactive exploration of synaesthesia, the multisensory phenomena that connect smell, sight and sound.
Once thought to be rare, synaesthesia has inspired artists, musicians, poets and perfumers. Dr. Gilbert contends that most people experience some form of synaesthetic perception: Join us to learn how the brain creates these associations, and how they have been used in art and performance from the ancient Greeks to Smell-O-Vision.
Dr. Gilbert, a scientific pioneer in olfactory synaesthesia and a member of the IAO Board of Advisors, will guide our way to smelling shapes and colors in his experiential, interactive workshop.
This event will be strictly limited to 20 people, and is functioning as a fundraiser for the IAO.
A select amount of early bird tickets are available, until April 1, 2013. 6pm (PST).
When & Where
The Institute for Art and Olfaction
1933 Broadway
Los Angeles,
Tuesday, April 23, 2013 from 7:00 PM to 9:00 PM (PDT)
Add to my calendar
The Institute for Art and Olfaction
The Insitute for Art and Olfaction is devoted to creative experimentation with a focus on scent. Our goal is to create new points of entry for people interested in the field of olfaction, while accommodating creative ideas in all mediums.


text: "No risk, no reward." Hey everyone! I'm Aiden Russo, and you probably know my best friend Michelle. I'm not really a writer but I might as well give it a shot. I am almost 15, but not quite. I play the guitar, the piano, and a funny instrument Michelle taught me how to play called the dijeridoo. I've known Michelle all my life pretty much. She's totally awesome and I love her to death, in a friend way. I am usually labeled as 'emo' but whatever. I'm just glad Michelle sees me differently. Hmmm. There really isn't much else to say!


text:
ED Magedson – Founder
Ripoff Report
The Law Office of William Trantham1710 W. University Drive Denton, Texas United States of America
Does your business have a bad reputation?
Fix it the right way.
Corporate Advocacy Program™
This report was posted on Ripoff Report on 04/22/2012 12:58 PM and is a permanent record located here: http://www.ripoffreport.com/reports/the-law-office-of-william-trantham/denton-texas-76201/the-law-office-of-william-trantham-bill-trantham-law-firm-warning-rip-off-law-firm-be-w-872128. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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text:
Bourree And Hornpipe
AATB saxophones - Grade 4
From Water Music Suite In F Major. Composed by George Frideric Handel (1685-1759). Arranged by Frank Halferty. Saxophone Quartet. Duration ca. 3:50. Published by Kendor Music Inc (KN.15936).
Item Number: KN.15936
George Frederic Handel wrote the Water Music Suite at the outset of his English career, and it is one of his best known works. This edition at the grade 4 level is filled with all the spirit and stylistic nuance of Handel's original. It will delight audiences and present great individual and ensemble challenges for the performers. Total duration ca. 3:50.


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- 802.11b/g/n 150Mbps
- 10/100 Mbps Ethernet Port x4
- Compatible with old 802.11b/g
Some manufacturers place restrictions on how details of their products may be communicated.
Amazingly... So far.. So good 07/12/2010
This review is from: Linksys WRT120N Wireless Router 802.11b/g/n up to 150Mbps/ 10/100 Mbps Ethernet Port x4
CHEAP. After reading numerous scathing reviews, the original owner actually GAVE this device to me. Firmware upgrade a snap. Setting up WEP security through the Linksys utility was easy.
The router does run hot to the touch. I have the router raised up a bit as the extra height is provided by four added rubber feet so that more air can pass easily underneath the unit for ventilation. Perhaps heat is the cause of all the earlier negative reviews?
Other Thoughts:
The prior owner of my WRT120N reported NOTHING but trouble with this router. I reset the device to factory defaults/original firmware Version 1.0.01 using the reset button. I had no problem downloading the bin file for the newest firmware from October of last year... 2009. Firmware Version 1.0.04. Upgrading using the Linksys firmware upgrade utility went easily.
There are reports that this latest firmware doesn't support typical "N" router speeds. "G" speeds at best. I've never measured the wireless speed after the firmware upgrade. I running for a week now with the new firmware. All four wired devices are connecting. My iPhone 4G would not connect using the default wireless Auto Channel 6 setting. So I changed the broadcast channel to Channel 1 and the iPhone connects.
I was really prepared to slam this device with a one star review. However, so far I have nothing to complain about. Still, after all the bad reviews I'll not likely be buying another one of these.
Did you find this review helpful? Yes / No Thanks for the valuable feedback you provided!


text:
Current versions:
300 Wallpaper for free download: Easy & fast original pictures from the movie on the desktop.
On the PC to install the software "300 wallpaper", is very easy. You simply click on the download link provided or goes on with a right click "save target under". The download for the 300 wallpaper is available as an .exe file, which then can be found in the "Downloads" folder. After a quickly guided through installation of the software a compressed zip folder with name "300 wallpaper" now located in the specified directory. Can you unpack this easily by other applications or the operating system software. Already, a new folder will appear and you can look at the pictures.
In which 300 wallpaper folder will find it so every fan's heart: six different images in JPEG format with different motivations - including the actor - available as 300 wallpaper. The images can be seen again in three different resolutions. The 300 wallpaper are located in the sizes of 1024 x 768, 1280 x 1024 and 1600 x 1200 in the folder, you can choose from so just the right size for his desktop. Unnecessary image edits are no longer necessary.
If you have found the appropriate 300 wallpaper, simply highlight and right click "use as desktop background". A window of the software installed on the PC, to once again the desktop to be able to fit the image. If the correct size is selected, here also, no other changes to the images of the software 300 wallpaper are needed, and the desired image immediately lands on the desktop. Using the 300 wallpaper package can get so easily, the impressions on its screen, without long search in search engines for matching images to. Six different 300-wallpaper files can be changed arbitrarily often also.
Steps to install of the software of 300 combined wallpaper
• Download link follow, save the installation file on your PC
Software installation • start "300 wallpaper"
• Zip folder in the directory open and unzip it
• Opening images, choose appropriate subject and corresponding resolution to the desktop
• Right click on desired image as desktop background save
• Sustainable use and change the images possible
Background to the film
The successful movie of the year 2007 is a film adaptation of the fight of the Spartans against the Persians. Under the leadership of King Leandros, the battles are impressively documented, although it is rather an action as to a fact-based historical film. After the cinema breakthrough original cuts as 300 wallpapers were put online and in form of zip files for free download made public.
You get 6 high-quality images that are available in the resolutions 1024 x 768, 1280 x 1024 and 1600 x 1280 with the official wallpapers for the hit movie "300". All images are graphically very complex and reflect the dense and dark atmosphere of the film very well, so that one feels transported directly in the film. All images to establish people at the Center, which can be seen in typical situations from the film.
Features in the current release of 300 wallpaper
• 6 high-quality wallpapers from the movie "300"
• Each available in 3 different resolutions
• Realistically reflect the atmosphere of the film


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Shin Megami Tensei IV Apocalypse 3DS Game
£32.99 Out of stock Ask us a question about this product
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Shin Megami Tensei IV Apocalypse 3DS Game Product Description
Debuting as the top seller for its launch in Japan, Shin Megami Tensei IV Apocalypse is headed to the Americas. The story of Shin Megami Tensei IV Apocalypse depicts the remnants of Tokyo, ravaged by a catastrophe. Demons and humans co-exist, but in this state, the remnants of humankind are just a prize being fought over by the higher deities. One lone Hunter, the game's protagonist, meets an early demise and through a pact with a lesser god, is brought back to life in exchange for his pledge of servitude. From there, the protagonist allies with other Hunters in the ruins of post-apocalyptic Tokyo beginning a plot that crosses over with the events of the original Shin Megami Tensei IV.
- The SMT series, known for exploring the tenuous symbiosis between man and deity, reaches deeper than ever, as the band of main characters must decide which side of peace or anarchy they are on.
- Demons Aplenty: Shin Megami Tensei IV Apocalypse has nearly 450 demons to fight, fuse, or recruit to player parties, with demons from a wide selection of cultures and religions.
- Fuse for Affinities: Demon fusion, combining two or more demons to form a more powerful one, is one of the cornerstones of Shin Megami Tensei games
- Language: The game features English voice-overs and English text.
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text:
The £1.6 billion Cassini craft performed a balletic series of twists to slow from more 70,000mph and slip between two of the planet's famous rings.
A miscalculation or misfire of its engine, which has remained unused through its seven-year journey, would have seen the probe shoot past or smash into its mysterious target.
But the 2.2 billion-mile journey ended successfully with the craft safely in orbit and ready to embark on a four-year feast of scientific discovery.
The joint European and US probe, named after the 17th-century Italian astronomer Jean-Dominique Cassini who discovered four of Saturn's moons, is now 12,000 miles above the planet's surface and preparing to send back its first data.
British scientists have provided eight of the 18 critical instruments which will be used in the mission.
Professor Ian Halliday, chief executive of the Particle Physics and Astronomy Research Council, said: "Right now we stand on the threshold of a wealth of scientific discovery."
Planned for more than 15 years, Cassini represents the most ambitious unmanned space expedition in history and is the first time any craft has seen Saturn since Voyager 1 and Voyager 2 flew past in the early Eighties.
The mission will also send Huygens, a European Space Agency lander, down to the surface of Saturn's cloud-covered moon Titan early next year. It will be the most distant landing on an interplanetary body ever attempted.
Titan has a surface temperature of minus 180C, and is thought to be too cold to sustain life - but scientists believe the conditions there mirror those that existed on Earth billions of years ago.
Cassini, the largest interplanetary spacecraft ever built, will orbit Saturn 76 times and have 52 close encounters with seven of Saturn's 31 known moons.
Scientists also hope to discover the origins of Saturn's famous rings. It is thought they may have been formed from larger moons that were shattered by impacts of comets and meteorites, or they may be composed of mini-icebergs.
- More about:
- Astronomer
- Astronomy
- Italy
- Physics
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text: Plot: Freshly hatched bee Maya is a little whirlwind and won't follow the rules of the hive. One of these rules is not to trust the hornets that live beyond the meadow. When the Royal Jelly is stolen, the hornets are suspected and Maya is thought to be their accomplice. No one believes that she is the innocent victim and no one will stand by her except for her good-natured and best friend Willy. After a long and eventful journey to the hornets hive Maya and Willy soon discover the true culprit and the two friends finally bond with the other residents of the opulent meadow. Genre: Animation, Adventure, Comedy IMDB Rating: 6,0/10 from 570 users Metascore: 51/100 Directed by: Alexs Stadermann Starring: Kodi Smit-McPhee, Noah Taylor, Richard Roxburgh Release Name: Maya.the.Bee.Movie.2014.LIMITED.1080p.BluRay.x264-AN0NYM0US Size: 6.66 GB Video: MKV | 1920 x 1040 | 9154 kbps Audio: English | DTS | 1509 Kbps Runtime: 87 mins Information: Code: http://www.imdb.com/title/tt3336368/ Code: Download from NITROFLARE http://www.nitroflare.com/view/CBB311A6DAD3941/Maya.the.Bee.Movie.2014.LIMITED.1080p.BluRay.x264-AN0NYM0US.mkv Code: Download from REDBUNKER http://www.redbunker.net/ehuqo2yen6j8/Maya.the.Bee.Movie.2014.LIMITED.1080p.BluRay.x264-AN0NYM0US.mkv.html Screens: NFO:


text:
Ler livro online, o resumo Livro Virus Separation And Purification Methods – Polson, Alfred – ISBN: 0824791495. Confira preço em loja online confiável.
This reference presents many contributions made by Dr Alfred Polson during his 41 years of research into the physicochemical properties of plant and animal viruses – detailing his timesaving approaches to the characterization, extraction, separation, concentration and purification of viruses, proteins, antibodies and biopolymers.;Describing successful laboratory techniques featuring the Beckman preparative centrifuge, this reference; examines the electro-extraction process for isolating the maximum amount of viruses from infected plant material; analyzes zone electrophoresis in sugar concentration gradients as a powerful tool for purifying entero-, insect born and insect viruses; contains material on the use of a reorienting gradient centrifuge rotor to separate components of the haemolymph of the mollusc Turbo sarmaticus as a model for virus separation; explains the construction of a modified thin-layer ultracentrifuge rotor for concentrating viruses into suspension rather than pellets; elucidates the use of inserts in ultracentrifuge tubes in order to decrease the time of centrifuging; and introduces a method for determining pure viruse suspension densities.;This resource is intended for microbiologists, virologists, biochemists, molecular and cell biologists, immunologists, biochemical and bioprocess engineers, chemical engineers and graduate students in these disciplines.


text:
Home >> 6mm to 6mm Hose Push in Touch Right Angle Connector Quick Pneumatic Fittings 20pcs
|Product Name
|Pneumatic Quick Fitting
|6mm to 6mm ( Right Angle)
|Main Material
|Dimension (Approx)
|26 x 26 x 13mm/1" x 1" x 0.5"(L*W*T)
|Main Color
|Black, Blue
|Package Content
|20 x Pneumatic Quick Fittings
- It is designed for using to connect with polyethylene, nylon and PV tubing.
- Simple push in design for connecting to the tube and provide secure connection, requires no extra sealing.
- Elliptical release ring on two ends facilitates pneumatic installations in confined spaces
|Battery Included
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text:
Lady Cassidy's RetortMature
Lady Cassidy
Mairim was always the virgin. Her 'no sex' policy was cataclysmically Catholic, and the only thing remotely Catholic about her views, its genesis being a mystery to me. There was nothing wrong was her life of seclusion, aside possibly from the fact that her estate deserved more attention than that of a Lady and her servants, the few of them there were, but there was a part of me that was concerned for Mairim's lack of desire. She had always been a temperate squib, but to put herself out of display when everything around her was the best of a display (surely if one has the money, one should proclaim it so?)...it was close to madness.
Our lives had been that way since birth. I, the elder, willed myself to flirt into the company of others, be they necessary to my advancement of not. I liked to talk; Mairim liked to play dumb. When we were teenagers, we had the most of differences. Mairim became slowly introverted to an extreme where she would spend hours staring into her thoughts, fingers interlocking with their air. On the other hand, I began to shine. First, there was the captaincy of the Lacrosse team, next the chance at being Head Girl. Naturally, I declined. It wouldn't be appropriate to be seen to have achieved the post through relations; though he cared not for the title he would soon inherit, I doubted that the school would be proud to display the fact that the Head Students were shagging.
I remember once upon a time, when Mairim had spouted long sentences of what she thought was elegant English. As soon as she entered that school, however... It was as if the one trigger of change in her life had sent a silence to consume her. From that moment on, she could be found in the library, her head deep in some Philosophy book.
Of course, it went on to prove the right move for her. Her studies of the sky and of the frantic desires of the human minds were possibly what she was known for, far more than the title she had inherited when she had turned eighteen. Despite my follies, I had already joined union with someone who I knew would do better for me, money-wise, than I would for him. Not a former Head Boy, nor a politician, he simply lived as proud Lord, rather like I supposed I would soon be, even without marrying him. I had considered a life of leisure already definite. And so the conditional clause of my mother's will was my downfall: the remaining estate of Lady DeStanlia to go to her daughters equally after their eighteenth birthday; should one marry into lineage before the time of death, the estate shall go to the bachelorette; should both marry: the estate shall be split in a way befitting, that those less well-off should gain the most.
My mother was always picky with her rules; it was always my opinion (duly rejected by Geoffrey, my beloved) that my mother had altered the will on learning of my engagement; her joy being placed with that of silent Mairim, and her success in education.
Mairim also liked to travel. It was patronage and 'missionarianism' that mother's brother, newly Marquised, said began the trail of Mairim's illnesses. It was overwork and being out in those dastardly poor countries. It was the change in weather, the excuses for meals. Never did he remark the obvious: it was fate, her penitence for taking the birth-right that was mine. As a matter of course, I never wanted my sister to fall so ill that she was on her deathbed by the time she turned thirty-eight...but there was some pleasure inside me that rejoiced in the fact of her demise. It would have meant, I had always assumed, that I would have gained the final piece of my inheritance.
Well, imagine my shock, when I was presented with merely words and those material flashes of a title! 'Short-changed' might not cut into the deep fury that resided over me those weeks after Mairim's funeral. Yet, there was something more, another piece that lingered over my mind, followed up with headache after headache.
I didn't think that my sister told me everything. I would have, however, thought that she would consider me a confidante in any important motion of her life. Mairim was always meant to be the virgin. So how could she have a child, mysteriously concealed from us all when she had been a part of society's eyes for so very long? And what gave either of them the right to take away the estate which so naturally belonged to me?


text: Turkish Journal of Biology Turk J Biol (2014) 38: 633-639 © TÜBİTAK doi:10.3906/biy-1401-92 http://journals.tubitak.gov.tr/biology/ Research Article Cloning, purification, and characterization of a thermophilic ribulokinase from Anoxybacillus kestanbolensis AC26Sari 1 2 1 3 1, Müslüm TOKGÖZ , Kadriye İNAN , Ali Osman BELDÜZ , Öznur GEDİKLİ , Sabriye ÇANAKÇI * 1 Department of Biology, Faculty of Sciences, Karadeniz Technical University, Trabzon, Turkey 2 Department of Molecular Biology and Genetic, Faculty of Sciences, Karadeniz Technical University, Trabzon, Turkey 3 Department of Physiology, Faculty of Medicine, Karadeniz Technical University, Trabzon, Turkey Received: 31.01.2014 Accepted: 29.05.2014 Published Online: 05.09.2014 Printed: 30.09.2014 Abstract: The gene encoding ribulokinase araB from Anoxybacillus kestanbolensis AC26Sari was cloned and sequenced. The recombinant protein was expressed in Escherichia coli BL21 under the control of isopropyl-β-D-thiogalactopyranoside-inducible T7 promoter. The enzyme, designated as AC26RK, was purified with the MagneHis Protein Purification System. The molecular mass of the native protein, as determined by SDS-PAGE, was about 61 kDa. AC26RK was active throughout a broad pH (pH 5.0–10.0) and temperature (50– 75 °C) range, and it had an optimum pH of 9.0 and optimum temperature of 60 °C. The enzyme displayed about 90%–100% of its original activities after a 30-min incubation at a pH interval of 5.0–10.0. The enzyme exhibited a high level of D-ribulose activity with apparent Km, Vmax, and Kcat values of 0.94 mM, 3.197 U/mg, and 3.31 s–1, respectively. AC26RK activity was strongly inhibited by Zn2+ but increased by Mg2+. The effects of some chemicals on the ribulokinase activity revealed that Anoxybacillus kestanbolensis AC26Sari does not need metallic cations for its activity. In this paper, we describe for the first time the cloning and characterization of a thermophilic ribulokinase from thermophilic bacteria. Key words: Anoxybacillus, ribulokinase, thermophilic, expression 1. Introduction Anoxybacillus is a relatively new genus compared to the well-studied genera Geobacillus or Bacillus. The genus Anoxybacillus represents aerobic or facultatively anaerobic, neutrophilic, obligately thermophilic, endospore-forming bacteria (İnan et al., 2011). Most of the reported data have revealed that the members of this genus produce interesting enzymes that are thermostable and tolerant to alkaline pH. Some of the well-studied enzymes were discovered through partnerships with industry; for example, the raw starch-degrading amylase was discovered by a Novozyme team (Viksø-Nielsen et al., 2006), and the BfιI RE was discovered by New England Biolabs (D'Souza et al., 2004; Goh et al., 2013). L-ribulokinase (RK; EC 220.127.116.11) is 1 of 3 major enzymes of the arabinose catabolic pathway. L-arabinose is 1 of the major polysaccharide components in plant cell walls and among the most abundant monosaccharides in nature. Furthermore, its utilization pathway in bacteria has been investigated extensively (Zhang et al., 2012). The arabinose regulon is 1 of many gene systems in Escherichia coli and the regulon consists of 4 operons, araBAD, araC, *Correspondence: [email protected] araE, and araFGH, which are responsible for L-arabinose catabolism, gene regulation, low-affinity transport, and high-affinity transport, respectively (Englesberg and Wilcox, 1974; Lichenstein et al., 1987). In the lowaffinity transport system, the transporter, the araE gene product, is bound to the inner membrane and utilizes the electrochemical potential to transport arabinose. The araFGH genes encode arabinose-specific components of a high-affinity transport system, ABC transporters. These are 3 proteins of the ATP-binding cassette transporter family. AraF is the periplasmic arabinose-binding protein, AraG is the ATP-binding component, and AraH is the membrane-bound component (Schleif, 2010). AraC acts directly as an inducer or an activator of gene expression. The araBAD operon encodes 3 different enzymes required for catabolism of L-arabinose, which are responsible for the conversion of L-arabinose into D-xylulose-5-phosphate. AraA, as an isomerase (L-arabinose isomerase), converts arabinose to L-ribulose; AraB, as a kinase (L-ribulokinase), phosphorylates L-ribulose; and AraD, as an epimerase (L-ribulose-5phosphate 4-epimerase), converts L-ribulose-phosphate 633 TOKGÖZ et al. / Turk J Biol to D-xylulose-phosphate. This final product is then transferred to the pentose phosphate pathway (Schleif, 2010; Agarwal et al., 2012). L-ribulokinase is unusual among kinases since it phosphorylates all four 2- ketopentoses (L- or D-ribulose and L- or D-xylulose) with almost the same Kcat values (Lee et al., 2001). Despite the central role of L-ribulokinase in arabinose catabolism, its 3-dimensional structure and mechanism of action as well as the bases of this broad substrate selectivity have not been elucidated (Agarwal et al., 2012). This is the first report on thermophilic L-ribulokinase. In this paper, we describe for the first time the cloning and characterization of thermophilic L-ribulokinase from Anoxybacillus kestanbolensis AC26Sari. 2. Materials and methods 2.1. Bacterial strains, plasmids, and growth conditions Dulger et al. (2004) isolated and identified Anoxybacillus kestanbolensis AC26Sari from the Çamköy mud hot spring in the province of Çanakkale, Turkey. A. kestanbolensis AC26Sari was used as the source of chromosomal DNA and thermophilic L-ribulokinase. E. coli strains used in this study were BL21 (DE3):pLysS (Novagen) and JM101 (NEB). pGEM-T Easy (Promega) and pET 28a(+) vectors (Novagen) were used for cloning and overexpression. A. kestanbolensis AC26Sari was grown aerobically at 55 °C and pH 7.0 in either Luria-Bertani medium (LB) or minimal medium with the addition of 2 g L–1 casamino acid (M9CA) (Sambrook et al., 1989). All E. coli strains containing recombinant plasmids were cultured in LB medium supplemented with 50 µg/mL ampicillin or kanamycin for selection, as appropriate, at 37 °C and pH 7.4, unless otherwise stated. 2.2. Cloning and sequencing of the ribulokinase gene A ribulokinase gene was amplified directly from the genomic DNA of A. kestanbolensis AC26Sari using a pair of primers (RibkF: 5'-CGC TAG CAT GGG GAA AAA GTA TGT CAT TGG-3'; RbikR: 5'-CAA GCT TAA TCA CGA TAA ACT TAT AGA TTT TTT C-3') designed according to the sequence of ribulokinase from Anoxybacillus flavithermus. The restriction sites NheI and HindIII were incorporated into the forward and reverse primer sequences, respectively. Taq DNA polymerase was used to perform PCR with A. kestanbolensis AC26Sari genomic DNA as the template. The PCR conditions were as follows: 1 initial denaturation step at 95 °C for 2 min, 36 cycles at 94 °C for 1 min, annealing at 55 °C for 1.5 min, and extension at 72 °C for 2 min, except in the final cycle, where the extension proceeded for 5 min. The PCR fragment was cloned into the pGEM-T Easy vector and sequenced by Macrogen (Amsterdam, the Netherlands). Similarity analyses of the sequence were carried out with 634 the advanced BLAST program of GenBank (NCBI, NIH, Washington, DC, USA). 2.3. Overexpression and purification of the ribulokinase A 1695-nt fragment was released from pGEM-T digested with NheI and HindIII restriction enzymes and then was ligated into pET28a(+) having overhanging ends digested with NheI and HindIII. The ligation products were transformed into E. coli BL21 (DE3):pLysS. The recombinant plasmid was designated pAC26RK and expressed in the same cell in 1 L of LB containing 50 µg/ mL kanamycin. The expression products were designated as in AC26RK and contained a His tag at the N-terminal. The transformed cells of E. coli BL21 were incubated up to an optical density of about 0.6 at 600 nm. The expression of recombinant proteins was induced with 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG), and the strain was cultured for 4 h at 37 °C. The cells were harvested by centrifugation at 11,000 rpm for 10 min, and were then resuspended in 50 mM phosphate buffer (pH 7.0) followed by sonication with a Sartorius Labsonic M (70% amplitude, 0.6 cycles for 5 min) to release intracellular proteins. The cell-free extract was centrifuged at 14,800 rpm for 15 min to remove cell debris. The purification procedure was performed at room temperature. Crude extract was heated at 65 °C for 15 min, and the precipitated proteins were removed by centrifugation at 14,800 rpm for 15 min. The ribulokinase protein containing a His tag was purified with the MagneHis Protein Purification System (Promega, Madison, WI, USA). 2.4. Ribulokinase activity assay Ribulokinase activity was assayed by modified cysteinecarbazole reaction. The activity system (200 µL) contained 1.5 mM ribulose, 50 mM Tris-HCl buffer (pH 9.0), 12 mM ATP, 20 mM magnesium acetate, 1 mM NaF, 20 mM β-mercaptoethanol, and the enzyme preparation. After incubation at 60 °C, 0.4 mL of 5% ZnSO4.7H2O and 0.4 mL of 0.3 N Ba(OH)2 were added to the reaction mixture. The precipitate containing phosphorylated sugar was removed by centrifugation, and the supernatant was assayed for free ribulose by the cysteine-carbazole method. Next, 0.9 mL of 70% H2SO4 was added to 0.2-mL aliquots of the incubation mixture, and the solution was subjected to the cysteinecarbazole reaction at room temperature for 20 min (Lim and Cohen, 1966). After this ribulokinase activity assay, all experiments were carried out in triplicate and the average values were taken. 2.5. Optimum temperature and thermostability of ribulokinase The optimum temperature was measured by performing the ribulokinase activity assay for 20 min at temperatures ranging from 50 to 75 °C at pH 8.0 with an increment of 5 °C. The effect of temperature on ribulokinase stability was determined by measuring the residual activity (%) at 60 °C TOKGÖZ et al. / Turk J Biol after 120 min of preincubation ranging from 50 to 85 °C with an increment of 5 °C. 2.6. Optimum pH and pH-dependent stability of ribulokinase The optimum pH of the enzyme was measured at 60 °C using buffer solutions with different pH values, and relative activity (%) was also measured. The following buffers (50 mM) were used: 50 mM sodium acetate (pH 5.0–6.0), 50 mM potassium phosphate (pH 6.0–7.0), 50 mM Tris-HCl (pH 7.0–9.0), and 50 mM glycine buffer (pH 9.0–10.0). In order to identify the stability of the enzyme at pH intervals of 5.0 to 10.0, a variety of buffer systems such as 50 mM sodium acetate (pH 5.0–6.0), 50 mM potassium phosphate (pH 6.0–7.0), 50 mM Tris-HCl (pH 7.0–9.0), and 50 mM glycine buffer (pH 9.0–10.0) were tested. Preincubation was performed for each pH value at the optimum temperature for 120 min. The residual activities (%) were measured in Tris-HCl (pH 9.0) at 60 °C for 20 min. 2.7. Effect of metal ions and other reagents To determine of the effect of metal ions and other reagents on ribulokinase activity, metal ions were removed from the purified ribulokinase by treatment with 5 mM EDTA at 4 °C for 1 h followed by dialysis against 50 mM TrisHCl (pH 8.5) overnight with several changes of buffer. The dialyzed enzyme was centrifuged at 14,800 rpm for 15 min. The effect of various ions and other reagents on the transferase activity were assayed at 60 °C (pH 9.0) using D-ribulose as substrate for ribulokinase, and then 1 mM metal ions were added to the reaction. The ribulokinase activity of the enzyme without metal ions was defined as the 100% level. The residual activity (%) was assayed spectrophotometrically. 2.8. Kinetic parameters The values of Michaelis–Menten kinetic parameters, Vmax and Km, of the enzyme were calculated using D-ribulose as the substrate. Vmax, Km, and Kcat parameters were calculated using OriginPro 8.1 (OriginLab Data Analysis and Graphing Software, Northampton, MA, USA). 2.9. Nucleotide sequence accession number The nucleotide sequence of the A. kestanbolensis AC26Sari ribulokinase gene was deposited in GenBank under accession number KC905743. 3. Results and discussion 3.1. Cloning of the ribulokinase gene PCR fragments amplified by using the previously described primers were cloned into the pGEM-T Easy vector and then sequenced. A BLAST search revealed that the amplified fragment is a ribulokinase-encoding gene. The analysis of the whole gene revealed the presence of a 1695-bp open reading frame encoding a hypothetical 564-amino acid protein with a molecular mass of 61 kDa (calculated by ProtParam, www.expasy.org), and the protein was identified by a database enquiry (BLAST program) as a ribulokinase (Figure 1). A. kestanbolensis AC26Sari ribulokinase was found to have 99% amino acid and 99% DNA identity with Anoxybacillus flavithermus WK1 ribulokinase, 96% amino acid and 90% DNA identity with Geobacillus thermodenitrificans NG80-2 ribulokinase, and 95% amino acid and 86% DNA identity with Geobacillus kaustophilus HTA426 DNA ribulokinase. Based on the sequence similarities, 14 key residues responsible for the catalysis and substrate-binding interactions of the enzyme were identified as conserved: Asp10, Gly12, Thr13, Thr93, Ala94, Met97, Trp124, Glu158, Lys206, Asp272, Ala273, Thr294, Cys297, and Glu327. These data strongly supported the conclusion that AC26RK should be included in the FGGY family of carbohydrate kinases (FGGY carbohydrate kinase domain-containing). As previously stressed, AC26RK reveals similarities to the reported ribulokinases based on amino acid and nucleotide sequences, yet there are some differences among them. The putative ATP-binding motif of Bacillus halodurans ribulokinase was recently specified as 447GGLPQK452 (Agarwal et al., 2012). This motif is also conserved in AC26RK and positioned in a slightly different location (between 445 and 450 amino acids instead of 447 and 452 amino acids). On the other hand, O1, O2, O3, and O4 of L-ribulose are responsible for interaction with the Glu329, Lys208, Ala96, and Asp274 residues of Bacillus halodurans ribulokinase, respectively (Agarwal et al., 2012). These residues were located in the AC26RK aa sequence as Glu327, Lys206, Ala94, and Asp272, respectively. 3.2. Expression of ribulokinase gene and purification of the recombinant ribulokinase pAC26RK, which places the ribulokinase gene under the control of a T7 promoter, was transformed into E. coli BL21 (DE3). The overexpression of the ribulokinase protein induced by the addition of IPTG resulted in a high expression pattern of soluble ribulokinase, with activity of 2.17 U/mg; no activity was identified from the control cells harboring only the empty vector pET28a(+). The ribulokinase expressed in the E. coli BL21 (DE3) strain was purified with the MagneHis Protein Purification System (Promega) (Figure 2). The purified enzyme yield was 90%. The molecular weight (MW) of the protein was 61 kDa through SDS-PAGE. Parallel to A. flavithermus WK1 ribulokinase (accession number: YP_002314899) and Deinococcus maricopensis DSM 21211 ribulokinase (accession number: YP_004169580), which both consist of 564 amino acids and have 61 kDa MW, AC26RK protein also has 61 kDa MW. 635 TOKGÖZ et al. / Turk J Biol Figure 1. Gene and protein sequences of ribulokinase of Anoxybacillus kestanbolensis AC26Sari. 1 2 3 4 5 Figure 2. SDS-PAGE analysis of AC26RK. Lane 1: Crude extract from E. coli BL21 DE3 including only pET28a(+) vector, lane 2: protein marker, lane 3: AC26RK after heat shock and Ni column, lane 4: AC26RK after heat shock, lane 5: crude extract from E. coli BL21 DE3 including pAC26RK. 636 TOKGÖZ et al. / Turk J Biol 3.3. Characterization of the cloned ribulokinase 3.3.1. Effects of temperature on activity and stability Based on the effect of temperature on ribulokinase activity, AC26RK exhibited optimal activity at 60 °C (Figure 3). The enzymatic activity decreased significantly at temperatures below 50 °C and above 75 °C (Figure 4). The enzyme was active in a broad temperature range (50–75 °C). The thermostability assays indicated that the enzyme revealed about 45%–50% of its original activity after 30 min at 50, 55, and 60 °C, whereas incubations at 65–85 °C for 30 min led to a loss of activity around 90%. The enzyme displayed about 40% residual activity at 50, 55, and 60 °C after 1 h of incubation, yet the enzyme activities were about 1% of their original activity at other temperatures for the same incubation period (Figure 5). In the literature, it was reported that E. coli (Lee and Bendet, 1967) and Lactobacillus plantarum (Burma and Horecker, 1957) ribulokinases revealed optimum activity at 37 °C. The optimum temperature for AC26RK activity, however, was determined at 60 °C. To the best of our knowledge, there is no previously reported ribulokinase with such a high optimum temperature. 3.3.2. Effects of pH on activity and stability When assayed at various pH values at 60 °C, it was found that the recombinant ribulokinase reveals higher activity in alkaline conditions. The optimum pH for enzyme activity was 9.0 (Figure 4). However, the enzyme maintained its high activity across a broad pH range, with >80% of maximum activity observed at pH values of 8.0 to 9.5. The recombinant ribulokinase stabilities were studied between pH 5.0 and 10.0 at 60 °C, and the enzyme exhibited about 90%–100% of its original activity after 30 min. The enzyme had 70% residual activity at pH 10.0, and the enzyme activities were about 80%–95% of their original activity at other pH levels tested after 1 h of incubation. Moreover, in 24 h the enzyme maintained its activity and exhibited about 90% stability at pH 6.0–7.0 and about 40%–80% stability at pH 7.5–10 (Figure 6). It was reported that the pH optima of E. coli and Lactobacillus plantarum ribulokinases were 7.0 and 7.5, respectively (Burma and Horecker, 1957; Sedlak and Ho, 2001). The optimum pH of AC26RK was 9.0. Thus, we suggest that AC26RK operates at higher pH conditions than other ribulokinases. 120 80 Relative activity (%) 120 100 Relative activity (%) 100 60 40 20 0 50 °C 55 °C 60 °C 65 °C Temperature (°C) 70 °C 80 60 40 20 0 75 °C Figure 3. Optimum temperature for activity of AC26RK. 5 5.5 6 6.5 7 7.5 pH 8 8.5 9 9.5 10 Figure 4. Determination of optimum pH for activity of AC26RK. 100 80 60 70 °C 75 °C 80 °C 85 °C 100 Residual activity (%) Residual activity (%) 120 50 °C 55 °C 60 °C 65 °C 40 20 0 80 60 pH 5 pH 5.5 pH 6 pH 6.5 40 20 0 20 40 60 Time (min) 80 100 120 Figure 5. Effect of temperature on thermostability of AC26RK. 0 0 0.5 pH 7 pH 7.5 pH 8 pH 8.5 pH 9 pH 9.5 pH 10 1 1.5 Time (h) 2 2.5 Figure 6. Effect of pH on thermostability of AC26RK. 637 TOKGÖZ et al. / Turk J Biol 120 Relative activity (%) 100 80 60 40 20 0 D-ribulose L-ribulose D-xylulose Substrate Relative activity (%) Figure 7. Substrate specificity of AC26RK. L-xylulose Unit (µmol/min) Hill Fit of Sheet1 Unit 0.03 Unit (µmol/min) 3.3.3. Substrate specificity The substrate specificity of the enzyme was determined by performing the assay with different substrates. AC26RK enables phosphorylation of D-ribulose, L-ribulose, D-xylulose, and L-xylulose. The phosphorylation assay results indicated that AC26RK exhibits maximal activity toward D-ribulose and decreasing affinities to L-ribulose, D-xylulose, and L-xylulose (Figure 7). Substrate saturation curves for D-ribulose showed that A. kestanbolensis AC26Sari ribulokinase follows a simple Michaelis–Menten kinetic pattern. The substrate saturation curve was obtained by decreasing substrate concentrations gradually. 3.3.4. Kinetic parameters Kinetic parameters were measured by employing a cysteine-carbazole assay, which requires the use of D-ribulose as a substrate. The initial rates were calculated by measuring the absorption at 540 nm for several concentrations of D-ribulose over a range of 0–1 mM. The ribulokinase exhibited simple Michaelis–Menten kinetics for D-ribulose (Figure 8). The values of Km and Vmax were 0.94 mM and 3.197 U/mg, respectively. However, the Kcat value for E. coli was 3.31 s–1 and the Km value was 0.39 mM (Lee et al., 2001). The analysis results suggest that AC26RK exhibits better or moderate Km values compared to E. coli ribulokinase. 0.02 0.01 0 1 Substrate (mM) 3 Figure 8. Michaelis–Menten plot. The measurements were performed under optimum pH and temperature conditions. 3.3.5. Effect of metal ions and other reagents The effects of various metal ions and other reagents on phosphorylation activity of ribulokinase were assayed at 60 °C and pH 9.0 using D-ribulose as a substrate. The activity measured without additional metal ions and reagents was considered as 100%. Independent Mg2+ addition increased ribulokinase activity up to 633%. Mg2+ ion plays a role in the stability of all polyphosphate compounds in the cells (Bachelard, 1971). An independent Mg2+ addition stabilizes ATP, which is a substrate of ribulokinase, so that it induces the activity of ribulokinase. Treatment of AC26RK with β-mercaptoethanol induced activity by approximately 175%. Cysteine, which favors an S-S to -SH interchange, suggests that ribulokinase is an enzyme that does not contain disulfide linkages (Ay et al., 2011). On the other hand, Zn2+ strongly inhibited AC26RK activity. K+, Li+, Ca2+, and EDTA additions did not induce any variation in AC26RK activity (Figure 9). The chelating agent EDTA 800 700 600 500 400 300 200 100 0 Metal ions and reagents Figure 9. Effect of some metal ions and reagents on AC26RK activity. In the presence of metal ions or reagents, the enzyme activities were compared with a control including no metal ion or reagent whose activity was considered 100%. 638 2 TOKGÖZ et al. / Turk J Biol did not affect the activity of AC26RK, suggesting that the enzyme has good activity without metallic cations and that metallic cations are not necessary for its activity. This study provides the first report on the cloning, purification, and characterization of a ribulokinase from thermophilic bacteria. In conclusion, AC26RK reported from thermophilic bacteria is the first thermophilic and alkaline ribulokinase belonging to the FGGY family of the carbohydrate kinase classification system. References Agarwal R, Burley SK, Swaminathan S (2012). Structural insight into mechanism and diverse substrate selection strategy of L-ribulokinase. Proteins 80: 261–268. Lee LV, Gerratana B, Cleland WW (2001). Substrate specificity and kinetic mechanism of Escherichia coli ribulokinase. Arch Biochem Biophys 396: 219–224. Ay F, Karaoglu H, Inan K, Canakci S, Belduz AO (2011). Cloning, purification and characterization of a thermostable carboxylesterase from Anoxybacillus sp. PDF1. Protein Expres Purif 80: 74–79. Lee N, Bendet I (1967). Crystalline L-ribulokinase from Escherichia coli. J Biol Chem 242: 2043–2050. Bachelard HS (1971). Allosteric activation of brain hexokinase by magnesium ions and by magnesium ion-adenosine triphosphate complex. Biochem J 125: 249–254. Burma DP, Horecker BL (1957). Pentose fermentation by Lactobacillus plantarum. III. Ribulokinase. J Biol Chem 231: 1039–1051. D'Souza DR, Morgan RD, Parashar V, Capalash N, Sharma P (2004). Characterization of BfιI- a thermostable, Co ++-requiring isoschizomer of BfιI from Anoxybacillus flavithermus. World J Microb Biot 20: 593–598. Dulger S, Demirbag Z, Belduz AO (2004). Anoxybacillus ayderensis sp. nov. and Anoxybacillus kestanbolensis sp. nov. Int J Syst Evol Micr 54: 1499–1503. Englesberg E, Wilcox G (1974). Regulation: positive control. Annu Rev Genet 8: 219–242. Goh KM, Kahar UM, Chai YY, Chong CS, Chai KP, Ranjani V, Illias R Md, Chan KG (2013). Recent discoveries and applications of Anoxybacillus. Appl Microbiol Biot 97: 1475–1488. İnan K, Çanakçı S, Beldüz AO (2011). Isolation and characterization of xylanolytic new strains of Anoxybacillus from some hot springs in Turkey. Turk J Biol 35: 529–542. Lichenstein HS, Hamilton EP, Lee N (1987). Repression and catabolite gene activation in the araBAD operon. J Bacteriol 169: 811–822. Lim R, Cohen SS (1966). D-Phosphoarabinoisomerase and D-ribulokinase in Escherichia coli. J Biol Chem 241: 4304–4315. Sambrook J, Fritsch EF, Maniatis T (1989). Molecular Cloning: A Laboratory Manual. 2nd ed. New York, NY, USA: Cold Spring Harbor Laboratory Press. Schleif R (2010). AraC protein, regulation of the L-arabinose operon in Escherichia coli, and the light switch mechanism of AraC action. FEMS Microbiol Rev 34: 779–796. Sedlak M, Ho NWY (2001). Expression of E. coli araBAD operon encoding enzymes for metabolizing L-arabinose in Saccharomyces cerevisiae. Enzyme Microb Tech 28: 16–24. Viksø-Nielsen A, Andersen C, Hoff T, Pedersen S (2006). Development of a new α-amylases for raw starch hydrolysis. Biocatal Biotransfor 24: 121–127. Zhang L, Leyn SA, Gu Y, Jiang W, Rodionov DA, Yang C (2012). Ribulokinase and transcriptional regulation of arabinose metabolism in Clostridium acetobutylicum. J Bacteriol 194: 1055–1064. 639


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Joe Cole returns to competitive action at Chelsea tomorrow evening in his favoured central berth behind the strikers, the playmaking position he has christened "the sausage role". He will be taking his first step towards regaining his England place in time for the summer's World Cup.
The midfielder has been absent since rupturing his cruciate knee ligaments in January's FA Cup victory at Southend, his return delayed further when he damaged the meniscus in the joint before the club's pre-season tour of the United States in July. He will start tomorrow night's Carling Cup third-round tie against Queens Park Rangers, when a number of first-team regulars will be rested, although the Russia international Yuri Zhirkov, an £18m signing from CSKA Moscow, will make his first appearance for the club.
Cole has not played for his country since he left the 4-1 victory in Croatia a little over a year ago on a stretcher, though he had previously been a regular in the national side. Chelsea should benefit from his eagerness to return to the fold. "Joe can make a difference because he does offer that bit of 'fantasy'," said the club's assistant manager, Ray Wilkins. "He can go both sides, left and right, plays a pass and his work ethic is quite special. The World Cup is a lovely carrot for him, to work his way back into Fabio Capello's reckoning.
"He's had a few problems with his knees but, with the reconstruction that has happened, the knee is far stronger and better prepared for football now. He'll be a bit ring rusty but, given the amount of work he's put in to get back, it's lovely to see him back. It's been tough for him but the workload he's taken on has been phenomenal. The cruciate injury is difficult to recover from, not physically – medicine has really come on in that respect – but mentally. You get down in the dumps sometimes and you always worry it might go again, wondering about that first kick you get. But he's received a number of kicks in training and he's coped.
"He can provide something for us and for England ... A fully fit Joe Cole will be in and around the England squad. Hecan play in any of the three midfield roles in the system we have but he likes that central one behind the forwards. He calls it the 'sausage role in the hole'. That's Joe. It's nice to see he's bubbling. We're playing with the midfielders more 'inside' these days, so he'll enjoy that, and he's been looking forward to this for a long time."
Zhirkov, too, could be forgiven a certain frustration, having seen his Chelsea career limited to date to a match-winning display in July's friendly victory over Milan in Baltimore. He scored the winner that night only to be hampered by a knee injury since. "He's been desperate to show what he can do," said Wilkins. "He's an extremely intelligent young lad and a great footballer. You want to show people what you can do when you join a new club, so naturally there's been a bit of frustration on his behalf, but it's onwards and upwards now."
Chelsea will give a number of squad players a start against the Championship side, resting, for example, Didier Drogba – a scan on the forward's calf indicated he had been suffering from severe cramp against Tottenham Hotspur on Sunday – and offering the academy graduates Sam Hutchinson, Jeffrey Bruma and Fabio Borini the chance to feature at some point.
The Londoners have still to receive Fifa's full written reasoning behind its decision to impose a 12-month transfer ban on the club, although, as Chelsea prepare to appeal against that sanction to the Court of Arbitration for Sport, there is an acceptance within the club that an opportunity has presented itself for the younger players.
"Carlo [Ancelotti] has been to watch the majority of the reserve games since he's been here and those guys have impressed him," said Wilkins. "This gives us an opportunity to bring these players in and use them later in the season. If we progress, hopefully we'll be able to continue using them in the Carling Cup. The ban, although we'll appeal it, offers the younger players at the club a great opportunity to impress. They'll be in the squad. Whether they'll start is another matter."


text:
Forgotten Dreams
Sometimes, in our lives, we forget the things we once wanted to do. We forget the people we once wanted to be.
But I don't think we should, you know. And so I've created this as a place you can remind yourself of your childhood fancies. We can laugh together at the strange jobs we wanted when we were five, and we could sigh together over the whimsical hopes that we lost when we matured and grew older.
This is a place for sharing, but it's also a place for caring. Though we laugh together, we never laugh at each other. We laugh with each other.