text:
Footys wars!! builders Battle.
We have in no particular order.
Heron Composites, Laja
Scale sailing ,Kittiwake.
Victor Models, V12
GoSpectre, 507
Sparkman Micro Marine, BearFoot
Dennis Deprois,American Footy
Others I belive are Tooling up to build these amazing little boats,The Footy class is growing fast and builders everywhere see a Future.
Who will win the battle of the kit builders? Is there room for them all?
when will we see the first mass marketed Chinese/Asian model?
What will shakeout when all these models meet on the racecourse? or will the One offs appearing from the drawing boards of the class faithful prevail?
More questions than answers right now.
The market has so many different aspects that I believe there will be enough room for anyone who wants to stick at it. Also footys are so cheap that an enthusiast is likely to own a number of different manufacturers products, a very different situation than the $1000 1 Metre. Woodies, glassies, vaccies... we have a nice mix developing for all tastes.
I for one will be here for the long haul, I have been actively designing/manufacturing model kits for 10 years now. That means selling a good number each week, I guess I should have all of my ducks in a row by now. In the aircraft business I saw many dramatic start-ups fizzle within the year, that's just the way it is and is just as likely to happen here.
I do wish everyone on that list the very best because if we can all make it to three years (the key year they say) the footy world will be the better for it.
Cheers everyone....
In a Worldwide marketplace*, there is surely enough of a customer base to keep afloat ( please pardon the pun ) a half-a-dozen Footy model yacht production companies...?
* = essentially what we have, with the internet and PayPal...
I am sure there is room for other modellers to get on board... ( Oh dear - another one... ) and still eek out a meagre income.
no war here!! think we all just want to get people sailing and share the joy of sailing!!!
i have only been delivering model yachts since 1996 (we dont call it selling , but more helping owners take on a new baby boat!!), but the footys seem a way to get more kids into sailing, keep more adults that are sailing and give enjoyment to heaps of people!!!
as a low volume builder there are always issues, shall see how the footys go, as long as it doesnt cut into my sailing time too much!!!
I assume therefore, that there is no initial charge for the kits ?
Yours is a service - a charge is levied for that, later ?
I'll take one on those terms, please !
( PS - Of course Akela exists - I made her - as an experiment !!!
http://www.rcgroups.com/forums/showthread.php?t=658136 )
Last edited by keven64; Jun 01, 2007 at 01:21 AM.
Since you bring this up on a public forum I will answer you here as this an obvious attempt to discredit me.
Steve I sent your boats to the address you supplied ( Your Sisters Church I belive)on no less than 3 occasions as you are well aware. The NZ post office tells me the packages made it to the USA on all 3 occasions,what has happened when the parcels are given to US Post I have no control of nor does my carrier ,NZ Post.
I belive the problem could be the lead(bulbs) in the shipments to New York,since 9.11 this seems to have been a problem when shipping from Obscure places like New Zealand which many of your Countrymen don't even know exist!
I am out of pocket 2 lots of the product and 2 lots of Shipping and handling.I cannot recover these expenses and have no proof that you never received any of these shipments.
Do I just keep sending product until you say you have them.
|Thread Tools


text:
Hello, Hello. Sign In
Please enter a valid city, state or zip code
124 reviews | 4.3 out of 5
Purchased this to supplement a smaller Brother machine that I thought couldn't be beat. This one does it! Love the variety of stitches, smooth operation, easy set-up. It's what I've come to expect from Brother. One hitch: There was a $30 manufacturer's rebate advertised under the price on the site. This was in error. The rebate for this machine, if purchased between the dates advertised, is $10, enough to cover the extended warranty. Still a good bargain, anyway you look at it.
I am 32 years old and decided after 10 years, to pick sewing back up. I ran down to the local Wal-mart and purchased this machine. It is so much easier to use than the old Singer machine I used as a teenager and into my early adulthood. Changing out the foot is simple as is changing out the needle. Threading is also very easy. I love the machine and the quality it affords me. I would highly recommend this product to anyone, even if just beginning.
I had never used a sewing machine in my entire life before purchasing this product, but wanted to teach myself to sew for alterations, dress-making, etc. The instruction booklet is precise, easy to follow, and had me starting the first stitch within 45 minutes! The bobbin winder is so easy, they really made this machine beginner-friendly. I think this will be the perfect machine to handle my larger tasks down the road once my skill level progresses.
The only reason I did not rate it a "5" is because I have not yet tried all of the features; although I am very satisfied thus far. Sidenote: I ordered this item from walmart.com on a Saturday evening, and it was already on my doorstep Tuesday afternoon. Very quick delivery!
got this as a gift from my grandmother. I used to borrow my mom's older kenmore sewing machine and was worried that it being digital would make it more difficult to use. It is so easy to use! I love that there are pictures of the stitches on the machine so I don't have to keep checking in the manual. My one complaint is that it says size 15 plastic bobbins on the box, but they are just a tiny bit smaller. So in order to get extra bobbins (comes with 4), I had to find a special brother sewing machine dealer (or order online and pay shipping). As far as sewing, it does a great job, and was easy to operate and troubleshoot. (I always end up getting something stuck and getting it unstuck was simple). The bobbin thing is why it doesn't get a 5 star from me.
I have been eyeballing this machine and one other one for a while now and today I broke down and bought this machine. I was a little intimidated at first because I have been using a Brother XL-5340 for the past 8 yrs and this one has a lot more features. I brought it home, unpacked it, read the manual, and got started. OMG, I am in heaven and have no idea why I was intimidated by this machine. It is super easy to use and I simply love, love, love it. The functionality is so great. I basically have the serge capability that I needed as well as a great way to do appliques now on shirts to go with pj pants that I make and sell. This is sure to increase my business potential greatly. Thank you Wal-mart for having it in stock and at a great price. I would highly recommend this machine to anyone who is looking to upgrade from a standard sewing machine. Awesomeness, with a cherry on top!
I made this purchase for my fiance who wanted to create embroidered shirts and scarves for her daughter.
The pattern selections are great and programing is relatively easy once you READ THE MANUAL and learn how to.
The most difficult item she had learning is how to correctly use the automatic needle threader. The directions are a little vague, the diagram is of some help and the lighting could be a little brighter in this area.
Overall she is thrilled with how easy it is to operate. When she got the needle threader figured out there was no stopping her! The speed control is very easy to work, the patterns are looking wonderful and the items she has created thus far look great!
If you're looking for a fun machine to get creative with, this is one she and I readily and fully recommend for both novice and experienced machine operator!
I sew for a living and recently started to teach my future daughter-in-law how to sew, too. My husband and I decided to buy her a new sewing machine for Christmas. We bought one at another store and tried it out when we got home. It was horrible and we brought it back. I found this one and read the reviews and we bought it. I tried it and loved it so much that I bought one for myself, too and recommended it to friends at an online quilting group. It sews like a dream, is relatively quiet and easy to use. The variety of stitches gives you a lot of bang for the buck. If I were able to change anything it would be that the stitch panel could light up and that there was a beep when the bobbin or needle thread ran out but not having these features is easy to live with. This is a fabulous machine. Get one while they're around!
GREAT machine, especially for the price. My mom is an advanced quilter and uses a Janome 7700, which she spent $1999 on, and even she has given this machine her seal of approval!
Coming from my first machine being a top of the line Elna - that stayed in the shop quite a bit, to a Pfaff machine that I purchased off of Ebay that was a disaster to this Brother machine that I love. I am not a seamstress, but I enjoy sewing when I want to sew! I love this machine, quiet, easy to use, and affordable. I would have never thought that I would be happy on a machine that was so inexpensive - boy was I wrong! This machine is a jewel!!!
I received this machine as a gift, after having looked at another brother machine for several months. I haven't really ever done any sewing in, well, ever. Totally a sewing newbie. I have to say that the sewing machine was a little intimidating initially because it has far more bells and whistles than the one I was looking at. But the machine itself is very simple to use. The manual is very straight forward, and easy to understand. Though I often supplement the manual's instructions with google, just to be sure I have my bases covered, I usually find the information in the manual would have been sufficient. While I've only had the machine for a week, I have used it quite a bit--including mending several things, making a tote bag, making a hat, a pair of spats, and a plushie. The machine comes with an assortment of accessories (including a seam ripper, which is great since I sort of, accidentally, snapped my previous one. Oops! It was nice to have an instant replacement on hand.). Most of these are things like extra bobbin spools, as well as all the extra presser feet. So far, I have used the "default" foot, as well as the button hole foot and blind hem stitch foot. Changing the presser foot is incredibly simple, and the button hole foot and stitches were amazing. Maybe I'm just easily impressed, but it was great to be able to do button holes so easily on my very first 'from scratch' project. I've also played around with some of the decorative stitches--some of which look pretty awesome, and some of which look...somewhat less awesome. But they seem like they would be really fun to use for different home decorating type projects. And the 'less awesome' ones are possibly due to my lack of skill in working the machine, rather than the stitch itself. With all the projects I've done, I've only had the bobbin tangle once, and I'm fairly certain it was an 'operator error' so to speak. The machine runs really well. It's easy to control the speed, it's easy to work all the functions, and choose stitches, as well as alter their length and width. The manual is really great at explaining the different functions, as well as how to do certain things (like blind hem stitching). I haven't had any issues with it, and in this first week, I've done a variety of different projects, and worked with a variety of different fabrics from thick, fluffy fleece, to polyester, to cotton, to craft felt. The machine has handled them all, without any issues, sometimes sewing through up to four layers of thick fabric. Over all, I think it's a great machine so far. I'm really looking forward to all the projects I have in mind, and I have no doubt the machine is going to be able to handle them all quite beautifully.
I purchased this on wednesday and it ran fine until I tried to use swimsuit knits to make a bikini bottom(1 day), the machine got a knot under the feed dogs and the engine stalled. I took apart the bobbin casing and presser foot (just like the manual said to do), and put it all back together. The diagrams in the manual are nice. I got the error message on the screen to stop and it "works" but has no speed now and makes too much noise in the upper part of the machine now so I am returning it. I will be buying a heavy duty sewing machine with metal gears made by a better brand. Plus it sounds like a copy machine when you flip it on. Bottom line, if you plan on quilting or using broadcloth then its a good machine, but if you work with a lot of knits but still want to make household items then not the right machine.
I was amazed at how easy this machine is to use. The stitch choices are all at your fingertips. Super easy to thread and load a bobbin. Doesn't get easier than this. Sews smoothly. Price is good too. I really like it A LOT!!!!
I decided I wanted to teach myself how to sew and started looking for sewing machines. I had went into several sewing shops and did a lot of online research. Then I came across this sewing machine. The reviews were great. As a beginner it has been wonderful to use. The value was the best part though. The machine started out at over $650 and is still selling at some conventions and expos for $500. Mine was brand new for only $144.
I was so excited when I received this sewing machine for Christmas. When I finally got around to trying it out, it was awesome, lots of stitch functions and I was excited. Once I started sewing, however, it was only a couple of weeks before it didn't cooperate. I wanted it to zigzag and it only did straight stitching. I would turn it off and then turn it back on, trying to reset it, and this worked f0r a couple of times. Now, it just doesn't understand at all. I don't get an error. I only get straight stitching. It might sew another stitch, but I need a zigzag for my elastic waist bands, I make tutus. So, I am taking it back tomorrow and hoping to exchange it for something else. Not sure if I want to go digital or not. May just go right back to plain ol plain ol.
Although I own several Brother machines, this is not one I would recommend. The tension was off from the very beginning and the only answer was to put it in the repair shop before I had even used it. Not a great start when you are anxiously waiting to begin projects designated for that machine. I sent it back immediately. I have fund once a machine has a tension problem it always will.
This was the best sewing machine ever!!! I bought this for my Mom and how did she thank me? She made me a dress!!!
LOVE this machine! I upgraded from a starter Brother machine. It is more machine with more functions than I ever could imagine using. I am primarily sewing costumes and theater garb. Not quilting or challenging embroidery. It runs smoothly and the Drop-In bobbin feature is great. It comes with a wonderful assortment of extra accessories like a cleaning brush and 4 bobbins (one pre-threaded with white thread) and they all self-store in the machine. My old machine had tons of tension issues. Not this one. It set up in moments. You don't even have to pull lower tension thread up, this machine automatically just loads and sews. As my needs and uses grow, this machine will grow with me and may even be a machine I will pass on to my kids. Better performance and comparable features to machines I have seen for more than $300. Great bargain for this high level of performance. Glad my hubby got it for my birthday.
The tension was off right out of the box. I tried rethreading and adjusting the tension, but could never get it right. I am returning it tomorrow.
I am a professional seamstress and have been working at my craft for over 40 years. I chose a Brother sewing machine because of Brother's reputation for quality. Since I've enjoyed watching "Project Runway," it tickled me to find a machine with the name.
This machine is terrific! It meets all of my expectations for quality stitches and ease of use. It is necessary to carefully read the manual several times, because of the different features.
I have used expensive Viking machines through the years and found myself at a loss after a move to another state. That's when I thought, "Let's see what a Brother machine does."
Well, I am very happy with this Brother "Project Runway Limited Edition" machine; and, it's cost for the quality amazes me every time I use it.
This is my 2nd Brother sewing machine. My first was about 10 years old when I bought it and it worked well enough. However, I wanted a NEW machine and I wanted a computerized machine. With this machine I got the best of both worlds. I find myself wanting to do nothing but sew and make things on this machine. It is SEW EASY to work with!! :) I am now looking for a quilting table attachment that might work with it because I am going to start my very first quilt. I LOVE quilts and bedspreads so I'll be starting with the quilt. Not a fan of comforters though. Quilts, bedspreads, that's the ticket.

subset: commoncrawl


text:
A Love Song Revisited
Like us on Facebook
Watch homeboy take Patsy Cline's "Crazy" and rip it a new one. How? He high-kicks into action and then screams himself into a near heart attack while delivering a visually and audibly painful performance. Love may be blind, but for his sake, let's hope it's tone deaf, too.


text:
Tags: Asian, cute, couple, wild, hiddencam, fuck, amateur, homemade, brunettes
Categories: Amateur, Asian, Hardcore, Teens, Webcam
Submited: 10 months old
Views: 89933
Copyright © 2006 - 2015 DeviantClip.com - All Rights Reserved.
Support / Contact
Webmasters Area
Site Map
All actors and models appearing on this website are 18 years or older.
Section 2257


text: I recently bought a steel frame. I found after the purchase that the rear dropouts are not stainless. Will those pose future rust problems? I do not ride in rain; but, just how much moisture can penetrate the dropouts between the frame and the wheel? Can anyone give me some advice or suggestions?


text:
Posted on 27/11/2011 by Jenny | 252 comments
252 comments
I followed you on Twitter as katyakate and tweeted about your giveaway. Thanks
I liked Freebie Spot on Facebook as Mommy Growing up (on the comment), also liked in the side bar
I followed your cooking blog on Facebook as Mommy Growing up (in the wall comment). Thanks
I liked Anti Monkey Butt Powder on Facebook as Mommy Growing up
The Christmas stocking to receive this freeby is my mom's. She always do all the decorations. Thanks
Hello Jenny, I dropped by from vB blog hop. I followed you through FB and I'm thinking of joining this give away too. Thanks!


text:
San Diego San Diego school board candidates are shoring up their campaigns with last-minute endorsements from top officials at City Hall.
On Friday, Mayor Jerry Sanders endorsed Bill Ponder in the race to replace Shelia Jackson and represent a southeastern swath of the city in San Diego Unified's Sub-District E. He also endorsed Mark Powell in the contest to unseat board President John Lee Evans in north-inland Sub-District A.
Also Friday, City Council President Tony Young announced his endorsement of Evans, two days after he endorsed Ponder.
"Under Dr. Evans' inclusive leadership, the teachers and the school district came to a historic agreement that brought back 1,500 teachers for the first day of school this year," said Young, who has worked with Evans on education issues in the past year.
Sanders made his endorsements without comment to the public. Registered voters are scheduled to hear from Sanders this weekend in a blitz of recorded phone messages.
"When you have the mayor of the city endorse your campaign, that's major," Ponder said.
A retired university administrator, Ponder is competing against community college educator Marne Foster to represent an area that includes Paradise Hills, Encanto and City Heights. Foster has support from the teachers union and wants to strengthen community partnerships and expand opportunities for student internships.
Powell, a former San Diego Unified vice principal and National University instructor, is running against Evans, a psychologist, to represent University City, Clairemont, Mira Mesa and other communities.
Evans is trying to continue efforts to promote reforms that would eliminate the need for parents to bus their children across the city in search of a decent education.


text:
Image Unavailable
- Sorry, this item is not available in
- Image not available
- To view this video download Flash Player
Product Details
Gives me gas and stomach pain. It tastes really good but I guess it's not for me :(Published 16 months ago by JC SD
Hard to believe but this stuff works as advertised, I have been using it for several years now. I completely agree with the website on all points with this product, great meal... Read morePublished 20 months ago by Amazon Customer
I usa Vega protein powders all the time. I use them to make protein shakes, use it in recipes and even in my oatmeal. I love it.Published on November 13, 2013 by Cheryl Harbinson
LOVED this formula. I still use the new formula but it doesn't sit as well with me and does cause an upset to my GI tract. Read morePublished on September 18, 2013 by Lilly Provocateur
VEGA is incredible. I drink it each morning. This way I get all the nutrition I need without animal products and wasting time!Published on September 14, 2013 by Marti J. Barrett
I have IBS so this product actually gave me a lot of GI problems, but I do have a very sensitive stomach. I am fairly certain it's the inulin. Read morePublished on March 27, 2013 by andrea
This is good stuff, straight up! Easy on the digestion and tastey to boot! A little different texture than regular PP, but good tasting. Read morePublished on January 13, 2013 by Gluten Intolerant
all the nutrients, minerals, herbs, protein, micronutients, etc. you'd want for peak health and performance without the nastiness of dairy, whey, or soy. i love the taste, esp. Read morePublished on January 4, 2013 by A. Baum


text:
Bank Tax and TARP Funds
Steve Bartlett talked about President Obama's proposed "Financial Crisis Responsibility Fee" for banks that received… read more
Steve Bartlett talked about President Obama's proposed "Financial Crisis Responsibility Fee" for banks that received Troubled Asset Relief Program (TARP) money, and he responded to telephone calls and electronic communications. Mr. Bartlett believes the new fee would "stifle economic recovery."
C-SPAN's Nancy Calo read news headlines at the end of the program. close
People in this video
- Harry "Steve" S. Bartlett President and CEO Financial Services Roundtable
- Nancy Calo Producer C-SPAN->Radio
- Bill Scanlan Host C-SPAN
Hosting Organization
More Videos From
Washington JournalMore Videos
Related Video
Impact of Foreclosure Freeze
Steve Bartlett talked about financial sector opposition to the push for a national foreclosure moratorium and the…
Tax-Free Investment Accounts
Ms. Novack talked about President Bush's proposal for two new tax-free investment accounts. She…
Washington Journal: Tuesday
Current news events and political issues were examined through reviews of the morning newspapers; interviews with…
Washington and Wall Street
Greg Ip talked about economic policy-making in Washington and its impact on Wall Street, including President…


text:
It takes a moment. As �Calvary� opens, a small-town Irish priest sits to hear a confession. A few beats have likely come and gone before a viewer realizes that the image isn�t cutting away, that the audience is being asked to watch a man listen. It�s unusual but also unexpectedly riveting.
Written and directed by John Michael McDonagh, �Calvary� reveals itself over and over to be a movie of such surprises, a serious-minded, lightly comedic rumination on life, death, faith and community. In its steady assemblage of details over an incidental, episodic structure it accrues a building sense of moral gravity.
That the priest is played by an actor with the natural, compelling presence of Brendan Gleeson certainly helps. That what the priest hears from an unseen victim of priestly sexual abuse is a threat to kill him in one week�s time ups the ante as well, alongside the would-be villain�s rationale that killing a good, innocent priest such as Gleeson�s Father James will pack more of a wallop than killing an abuser. The film deftly avoids becoming some kind of whodunit in reverse, as McDonagh shows little particular interest in who might actually be the would-be killer. Rather, the inevitably of what�s coming gives a sense of clarity and purpose to everything the good father does in the time he has left.
McDonagh�s previous film, �The Guard,� which starred Gleeson alongside Don Cheadle, was something of a dark comedy, fish-out-of-water police procedural. There, variances in tone often came across as uncertainty, while in �Calvary� the story darts and dives with more assuredness, moving from serious to silly from scene to scene in what might be described as purposeful meandering.
Often dressed in an anachronistic cassock and with a rather astonishing sweep of leonine hair, Gleeson cuts an imposing and authoritative figure, like something from a rough-edged spaghetti Western dropped on the dramatic west coast of Ireland. The role provides a fantastic showcase for the actor, as he captures the inner conflict and outward placidity of the character. Written with him in mind, it is hard to imagine anyone else in the part.
McDonagh often pulls tension from the postcard imagery of the settings, such as when Father James and his daughter Fiona (Kelly Reilly), born before he joined the priesthood, have an intense, emotional conversation while walking along grand, picturesque cliffs. The film was shot by Larry Smith, cameraman on Stanley Kubrick�s dreamlike �Eyes Wide Shut� and more recently the garish, hypnotic �Only God Forgives,� and so the images have a dynamic quality to match the forceful storytelling.
The town has an assorted cast of characters � small-town eccentrics of various stripes � many of whom become possible suspects, played by actors including Aidan Gillen, Dylan Moran, Isaach De Bankole, Orla O�Rourke and the seemingly immortal American character actor M. Emmet Walsh. Perhaps to keep viewers off-balance, McDonagh continues throwing in new characters fairly late in the story, such as Gleeson�s real-life son Domhnall Gleeson in a single scene as an imprisoned serial killer. Chris O�Dowd, as the local butcher and cuckold, syncs well with McDonagh�s sensibility, moving from comically funny to funny odd to outright unnerving.
McDonagh is the older brother of playwright and filmmaker Martin McDonagh, writer-director of the dark thriller �In Bruges� and the inside-out killer comedy �Seven Psychopaths.� The two seem to share a creative DNA in the intense self-awareness of their storytelling � Gleeson at one point expressly refers to a �third-act revelation� � but �Calvary� has a humane compassion that the films of Martin McDonagh keep more buried.
As the end credits roll on �Calvary� there is a series of images of the spaces from the film devoid of people � empty tables and open tableaux. It gives the life-or-death decisions of the movie a whole new framing, the world existing on after the travails of the story.
The film is then not so much a meditation but a reverie, a swirl of emotions and ideas, managing to be both calmly reflective and skittishly anxious at the same time. �Calvary� is a serious comedy, a funny drama, a ruminative film about life and a lively film about death. From the jolting simplicity of the opening scene right through those final shots, �Calvary� is never quite the film you expect it to be. It sneaks up on you.
Starring: Brendan Gleeson, Chris O'Dowd, Kelly Reilly, Aidan Gillen
Rating: R for sexual references, language, brief strong violence and some drug use
Running time: 1:44
Location, ST | website.com


text:
If you find there are broken links, misssing pages, wrong chapters or any other problems in a manga/manhwa, please comment here. We will try to solve them the first time.
Please enter the email.
Kamikaze 7 released in mangahere fastest, recommend your friends to read Kamikaze 7 now!
Tags: read Kamikaze 7 english, Kamikaze 7 raw manga, Kamikaze 7 online, Kamikaze 7 chap, Kamikaze 7 chapter, Kamikaze 7 high quality, Kamikaze 7 manga scan
Next Chapter: Kamikaze 8
Previous Chapter: Kamikaze 6


text:
Salado at Walnut Creek
2104 E Anderson Ln, Austin, TX
Are you the manager of this property?Click here to claim this listing Check Your Credit Now
About Salado at Walnut Creek
Enjoy this beautiful apartment community located in North Austin. Salado at Walnut Creek we offer you one and two bedroom apartment floor plans. If you are looking for a townhome, we have the perfect two bedroom floor plan to suit your needs. Our community is rich in amenities including two beautiful pools, lighted sports court, sand volleyball and a laundry center. Many of our apartment homes overlook Walnut Creek. Our floor plans include Berber carpet, frost free refrigerators, large kitchens with plenty of cabinets and full size washer/dryer connections (most homes). Salado at Walnut Creek is located just off of Hwy. 183 with easy access to Hwy. 290 and IH 35. Call us today and make your appointment to tour our grand community.
As you search for your perfect apartment, please keep in mind that rental rates (including specials and discounts) and availability are very fluid in the apartment industry. Based on many factors, the rental rates (including specials and discounts) and availability can actually change not just daily, but hourly, if not even more quickly. While our advertisers do take steps to keep their information current, the rental rates or availability you may find here now at ForRent.com may not be the same as when you contact the communities.
Here are some similar properties you might be interested in
Towne Oaks I
Austin, TX
From $729
1 - 3 Beds
The Park at Crestview
Austin, TX
From $765
1 - 3 Beds
Chateau Avalon of Austin
Austin, TX
From $900
2 Beds
Avesta Solano
Austin, TX
From $665
Studio - 2 Beds


text:
FCFBlazer - I was frozen and had all my posts deleted at one time due to outing their bullshit! So did a lot of others including the old school members of the AL UG, plus stranglehold26's newb ass!
Joe Rogan - Wow. That's some sad shit.
Stranglehold26 -FCFBlazer - I was frozen and had all my posts deleted at one time due to outing their bullshit! So did a lot of others including the old school members of the AL UG, plus stranglehold26's newb ass! <img src="/images/phone/post_tag.png" alt="Phone Post" border="0" style="vertical-align:middle;"/>
yes, every time I called her loud mouth ass out for being a fraud, and her husband for being a fraud, Sally would have Kirik ban or freeze me. Fuck Skip Hall.
Daniel Carver -That's obviously just my opinion and I still do listen to his show but he is always talking about how hard-hitting he is and quite frankly I see no hint of it at all, I'm talking about his shows over the past year or so.RavenFighter -I used to listen to Carson's Corner when my brother-in-law hosted files and provided bandwidth, but I haven't for a while now. If Carson's giving softball interviews, that's sad to hear. He ws the only MMA Media member that would ask tough questions the last time I listened.Ethan Boyle -Bring it Skip. Carson throws softballs so if you can't get past him then you're probably something of a fibber.Carson's Corner radio show - Skip, if you want a fair venue to get your side out let us at Carson's Corner know.
Voted up.
williepep - I am getting the feeling, that you don't think this could all be one big (well not one, but multiple) misunderstanding ?
Daniel Carver -Bigwormy - Oh yeah and a big FUCK YOU to all the mods here that took that piece of shit's word over ours.
Sofa King Cool - LM was a massive cunt back in the day.
Mulletron5000 - makes sense that they defended Mark Bailey.
akiyonomotu - I'm glad this is airing out.Why does Zipplokk come into this out of nowhere? The guy has been nothing but good to me. Nothing but love for Zipplokk from me.
LawdyMama was a huge see you next tuesday. Skip is a joke. This site is quick to ban members that question frauds like this. How long til zipplokk freezes this thread?
jkennedy -William Colosimo - Expo$ed
but to be fair, this line from the OP's referenced article needs to be exposed also:
"And he was the oldest fighter until 71-year-old Dr. John J. "Gray Wolf" Williams came on the scene a few years ago."
IIRC, this guy was a fucking scammer also, everyone realized it when we saw footage of that "fight"
Yeah the Williams work was nothing even close to a fight. More like two old fucks fighting over a parking spot at a 7/11. "Grey Wolf" is another guy with a laundry list of claims that reak of BS.
Jason Reinhardt - Skip hall has my uptmost respect for stepping his 2 feet inside the cage at his age against much younger fighters. I didn't read all the stuff about the allegations of the military, but to me, Skip Hall deserves a little respect for having the courage, fighting in the cage time and time again. just my 2 cents. Jason Reinhardt
This thread has been frozen.


text:
Magic by Dr. Wilderness Show, mad and zany hats, spinning 3-D projects with The Stardust Arts. There is something for everyone that brings excitement and joy to all ages at the art booth.
Take part in one of Southern California's Premier Outdoor Arts Events! 14th Annual Art in the Village in Carlsbad, CA will be held on Sunday, August 12, 2012. It is a grand stopping point during the event, which annually has become the anticipation of children and parents alike.
Last year, Stardust had 941 children and visitors stop by the art activities booth manned by three volunteers at the stations of art. The street scene is full of art, vendors and music, hosting over 150 artists and craftspeople and around 20,000 attendees. Sponsored by the Carlsbad Village Association, we hope you can join us! It is "free" to the public .
Art activities provided by Stardust Arts are sponsored by local participants, HomeLife Village Realtors and the Carlsbad Village Association, to make this a success.
The Stardust Arts is recognized by the California State Assembly and the San Diego County Board of Supervisor's and participates in local school after school programs providing the development of creativity and imagination in art. Proudly co-partnered with the non-profit, Dr Wilderness Show, jointly we have touched over two million youth with the Magic of Art at http://youtu.be/
For more information on the event please contact, Cheryl Ehlers at 760-519-1551 or firstname.lastname@example.com. . The Stardust Arts a www.thestardustarts.com.


text:
Princess Diaries 2
*Original movie grossed $108 million in the U.S. *The sequel releases August 2004 *The all-star cast and director are back *Cine-Manga will release with the DVD
Mia Thermopolis is now a college graduate and on her way to Genovia to take up her duties as princess. The fun begins as her best friend Lilly also joins her for the summer. However, it's not all fun and games as Mia must continue her 'princess lessons'-- riding horses side-saddle, archery, and the like. But will she be able to take on her biggest duty yet... getting married!
Please visit Help Page for Questions
regarding ISBN / ISBN-10 / ISBN10, ISBN-13 / ISBN13, EAN / EAN-13, and Amazon
Home | ISBN Directory | Affiliate
Program | Contact
Copyright © 2003-2014 AllDiscountBooks.net All rights reserved.


text:
Belk & Co. Sterling Silver with 14k Gold Blue Topaz Earrings
$130.00 Original Price $325.00
Savings: $195.00 (60%)
Sterling silver and 14k yellow gold Blue Topaz square earrings.
- 1/3 ct. t.w.
- Sterling silver;14k gold
- Clean with a soft bristle brush or cloth and warm soapy water
Shipping & Returns
Shipping your purchase is easy on belk.com. Depending on the destination of your package, you can expect it to arrive within 4-7 business days. Shipping Information
Returns are easy on belk.com. Easy returns


text:
Erwinia pyrifoliae is a newly described necrotrophic pathogen, which causes fire blight on Asian (Nashi) pear and is geographically restricted to Eastern Asia. Relatively little is known about its genetics compared to the closely related main fire blight pathogen E. amylovora.
The genome of the type strain of E. pyrifoliae strain DSM 12163T, was sequenced using both 454 and Solexa pyrosequencing and annotated. The genome contains a circular chromosome of 4.026 Mb and four small plasmids. Based on their respective role in virulence in E. amylovora or related organisms, we identified several putative virulence factors, including type III and type VI secretion systems and their effectors, flagellar genes, sorbitol metabolism, iron uptake determinants, and quorum-sensing components. A deletion in the rpoS gene covering the most conserved region of the protein was identified which may contribute to the difference in virulence/host-range compared to E. amylovora. Comparative genomics with the pome fruit epiphyte Erwinia tasmaniensis Et1/99 showed that both species are overall highly similar, although specific differences were identified, for example the presence of some phage gene-containing regions and a high number of putative genomic islands containing transposases in the E. pyrifoliae DSM 12163T genome.
The E. pyrifoliae genome is an important addition to the published genome of E. tasmaniensis and the unfinished genome of E. amylovora providing a foundation for re-sequencing additional strains that may shed light on the evolution of the host-range and virulence/pathogenicity of this important group of plant-associated bacteria.
Fire blight caused by the enterobacterium Erwinia amylovora, a quarantine pathogen in Europe, is the most important global threat to pome fruit production (i.e., apple, pear) and to a wide-variety of Rosaceae (Maloideae), including amenity and forest species [1]. E. amylovora is native to the North Eastern USA, and it was the first phytopathogenic bacterium described (see [2]). This invasive pathogen was first introduced into the UK in the late 1950s and has spread across Europe over the past three decades, with continuing advance eastward that threatens the native origin of apple in Central Asia [3]. Fire blight symptoms include recurvature of shoots (shepherd's crook), necrosis, ooze, and cankers. The pathogen can enter through nectaries, hydathodes and wounds resulting in blossom, shoot or rootstock blight syndromes [4]. E. amylovora growth is highly dependent on weather conditions, which has been used to develop disease forecasting models, and it is passively vectored by flower-foraging insects [5]. Epidemics can develop rapidly and result in death of individual plants or entire orchards within a single season, leading to severe economic losses [2]. E. amylovora has poor ecological fitness away from host or surrogate plants [4].
Erwinia pyrifoliae is a newly described pathogen, closely related to the main fire blight pathogen E. amylovora. E. pyrifoliae is primarily a pathogen of Asian or Nashi pear (Pyrus pyrifolia) and is considered to have a restricted geographic distribution in East Asia (Korea and Japan) [6-8]. Monitoring to detect E. pyrifoliae is rarely conducted, although a quantitative PCR method based on differential 16S rRNA and 16S-23S ITS sequences [9,10] has recently been developed; thus the precise distribution of this pathogen is somewhat uncertain. E. pyrifoliae causes fire blight disease symptoms essentially indistinguishable from those of E. amylovora infection. Limited host germplasm screening with E. pyrifoliae strains indicates a slightly wider host-range than originally thought, with virulence observed on selected commercial pear (Pyrus communis) and apple (Malus domestica, 'Idared') varieties [9]. The level of virulence on non-Asian pear hosts is markedly lower than that typically observed in E. amylovora, suggesting that important genetic differences remain to be elucidated.
In contrast to the considerable genetic information available for E. amylovora [11,12], relatively little is known about the genetic basis of virulence and environmental fitness in E. pyrifoliae [13]. As for E. amylovora, E. pyrifoliae intra-species genotypic diversity appears low, with strain differences primarily observed in plasmid content [14]. Thus far, only minor genetic differences have been found between E. pyrifoliae and E. amylovora [13,15].
The objectives of our work were to sequence the complete genome of the type strain of E. pyrifoliae, strain DSM 12163T, and then compare it to the recently sequenced genome of the non-pathogenic pome fruit epiphyte, E. tasmaniensis Et1/99 [16]. Identifying differential sequences among these related bacteria may provide useful insights into host-pathogen interactions that could eventually be exploited for fire blight control.
Results and discussion
General genome description
The genome of the type strain of E. pyrifoliae, strain DSM 12163T (also archived as Ep16/96T, CFBP 4172T, and CIP 106111T [7]), was sequenced using 454 pyrosequencing and Solexa sequencing, and assembled. The genome contains a circular chromosome of 4,026,286 bp (Fig. 1), and four plasmids of 35,901 bp (pEP36), 4,960 bp (pEP5), 3,070 bp (pEP3), and 2,610 bp (pEP2.6) (Table 1).
Figure 1. Circular representation of the chromosome of E. pyrifoliae DSM 12163T. Circles (from outside to inside) First: scale bar in kb; second and third: predicted coding sequences of E. pyrifoliae DSM 12163T chromosome on the leading and lagging strand, respectively (colors according to COGs); fourth: coding sequences of the hrp/hrc T3SS (red), the inv/spa T3SS (green), the T6SS clusters (blue), the flagellar genomic island (yellow), dispersed flagellar gene clusters (purple) and the EPS biosynthetic cluster (orange); fifth, G+C content; sixth, G+C skew.
Table 1. General features of the genome of E. pyrifoliae DSM 12163T.
Annotation was completed using the genome annotation system GenDB [17], and manually optimized. In total, 4,038 open reading frames (ORFs) were predicted. The chromosome contains seven rRNA operons, and 74 tRNAs, which falls within the range observed for most free-living enterobacterial genomes. Several low G+C regions indicated the presence of horizontally acquired sequences. Inspection of some of these indicated the presence of several prophages in the chromosome.
Earlier reports identified four plasmids in a large group of E. pyrifoliae strains, including strain DSM 12163T [18,19]. Recently, the complete sequences of plasmids pEP36 and pEP2.6 from E. pyrifoliae Ep1/96 were published [8]. These two plasmids were found to be present in the genome of E. pyrifoliae DSM 12163T with slightly differing sizes (35901 bp and 2610 bp, respectively). We sequenced and annotated two additional small plasmids of sizes 4,960 bp (pEP5) and 3,070 bp (pEP3).
From the coverage of the individual plasmids, it is possible to estimate the copy number of each plasmid in E. pyrifoliae DSM 12163T (Table 1). According to this calculation, plasmid pEP36 and pEP5 are present in 5-6 copies per cell, while the small plasmids pEP3 and pEP2.6 have a medium copy number ranging from 18-20 copies per cell. This explains the high yield of plasmids reported in earlier studies, whereas it is more difficult to obtain plasmid pEA29 from E. amylovora present at approximately two copies per cell (T.H.M. Smits and B. Duffy, unpublished data).
Plasmid pEP3 shares a region (Fig. 2) with that of ColE1-type plasmid pEP2.6 [8], having 99.2% sequence identity over approximately 800 bp including the RNA I modulator gene but not covering the oriV region. The rest of the plasmid contains a large region that is similar (78% identity) to a part of plasmid pEP5 (see Fig. 2). The latter plasmid contains mobCABD genes similar to those on the ColE1-type plasmid pSW100 found in the phytopathogenic bacterium Pantoea stewartii subsp. stewartii SW2 [20], indicating that it may be a transmissible element. The region that is shared between pEP3 and pEP5 encodes three ORFs (see Fig. 2), two of which are hypothetical proteins and one which encodes Hcp, a putative type VI secretion system (T6SS) effector protein (see below).
Figure 2. Comparison of the annotated sequences of E. pyrifoliae DSM 12163T plasmids pEP5, pEP3 and pEP2.6. Genes with high homology are indicated in the same color. Grey shading indicate regions with DNA sequence identity.
rpoS RNA polymerase sigma factor deletion
During genome assembly, it was observed that E. pyrifoliae DSM 12163T has a 140 bp deletion in the rpoS gene (EPYR_03050), when compared to the E. amylovora OT1 or E. tasmaniensis Et1/99 rpoS gene sequence. This 140 bp deletion results in a frameshift and a coding interruption after the RpoS subregion 1.2 [21]. After this deletion, the remaining sequence of the rpoS gene resumes in a different frame, starting from subregion 2.4 in RpoS [21]. This deletion thus encompasses a highly conserved region that putatively interacts with the core RNA polymerase.
Using PCR primers designed in the rpoS region, we confirmed that this deletion is present in E. pyrifoliae DSM 12163T (Fig. 3). However, the rpoS gene of E. pyrifoliae CFBP 4174 has the same size as the amplicon for E. amylovora strains CFBP 1232T and OR29 and Erwinia billingae LMG 2613T, indicating that the deletion is specific for E. pyrifoliae DSM 12163T. This mutation may have occurred prior to submission to the culture collection, or represents an environmental adaptation in this strain, possibly associated with host-pathogen interactions or ecological fitness [22-24].
Figure 3. Electropherogram of amplified rpoS genes from different Erwinia species. Lane 1: E. amylovora CFBP 1232T; lane 2: E. amylovora OR29; lane 3: E. pyrifoliae DSM 12163T; lane 4: E. pyrifoliae CFBP 4174 and lane 5: E. billingae LMG 2613T. A minus sign denotes the negative control (-); M denotes the marker (Fermentas GeneRuler DNA Ladder Mix). Relevant marker sizes (in bp) are indicated at the left side of the figure. The rpoS gene was amplified using primer set Ep-rpoS-F (5'-AGTACTGGCACGAGTTCTGTTAGA-3') and Ep-rpoS-R (5'-TGCAGTATTTCACGCAGACGACGC-3'). The expected amplicon size of an intact rpoS gene is 1109 bp; for the 140 bp deletion the amplicon is 969 bp.
Type III secretion system operons
Type III secretion systems (T3SSs) are widely distributed among proteobacterial pathogens of plants, animals, and humans, and constitute a fundamental virulence determinant [25]. In other bacteria, T3SSs were found to be critical for the establishment of non-pathogenic host-relationships with plants [26,27] and insects [28]. Mutations or absence of T3SS genes interfere or abolish bacterial-host interaction [29]. Generally, genes encoding elements of the T3SS machinery are conserved among bacterial species, but each system includes specific effectors targeting respective hosts. Proteins secreted by the Hrp complex of E. amylovora and other plant pathogenic bacteria are required for virulence on susceptible host plants and for the elicitation of a hypersensitive response in resistant or non-host plants [25,30].
The genome of E. pyrifoliae DSM 12163T contains two distinct T3SSs. One T3SS is closely related to the hrp/dsp cluster of E. amylovora, while the second T3SS is more similar to the inv/spa cluster of Salmonella typhimurium [31]. The
- h
- r
- p
The genomic arrangement of the hrp region in E. pyrifoliae DSM 12163T is identical to the T3SS arrangement found in E. pyrifoliae strain WT3 [33]. In E. pyrifoliae DSM 12163T the hrp region is bordered by the
- e
- e
- a
- e
Genes of an inv/spa system are organized in a single gene cluster (EPYR_02139-EPYR_02160) in the genome of E. pyrifoliae DSM 12163T (Fig. 4). This cluster displays similarity with those present in E. amylovora Ea273 [36], E. tasmaniensis Et1/99 [16], and the insect endosymbiont Sodalis glossidinius str. morsitans [28]. The highest sequence and structural similarity is to the PAI-3 system of E. amylovora Ea273 [36], where all the genes are also condensed into a single cluster. In E. tasmaniensis Et1/99, the inv/spa system is composed of two separate clusters complementing themselves to form a complete injection apparatus similar to that encoded by the Salmonella typhimurium pathogenicity island 1 [16]. Compared to E. amylovora Ea273 [36], only the hypothetical orf43 was absent in E. pyrifoliae DSM 12163T. The inv/spa system of E. pyrifoliae DSM 12163T includes genes with no currently known function in plant pathogenicity or plant associations, and which are more related to T3SSs from endosymbionts and animal pathogens. It is proposed that E. pyrifoliae DSM 12163T may utilize this second T3SS to facilitate vectoring associations with insects.
Figure 4. Comparison of inv/spa-type Type III Secretion Systems in E. pyrifoliae DSM 12163T and E. tasmaniensis Et1/99. For the comparison, the annotation of the E. tasmaniensis Et1/99 clusters was manually checked. Previously denoted pseudogenes were shown to have close orthologs in E. pyrifoliae DSM 12163T, while "missing" genes in E. tasmaniensis Et1/99 [16] could be found. Blocks of related genes are shaded grey. Putative T3SS core genes are colored green, with low homology genes in light green. Effectors are colored red, regulatory genes black and chaperones blue. Genes with no homology are in white.
Type VI secretion system
T6SS gene clusters have recently been found to be widespread among pathogenic and non-pathogenic Gram-negative bacteria [37]. Bacteria living in close association with eukaryotic cells are purported to use the T6SS as a mechanism for maintaining pathogenic or symbiotic interactions with their hosts. T6SS gene clusters consist of a set of 14 currently recognized core genes and conserved genes that vary in composition between species [38]. Two putative effector proteins in the human pathogen Vibrio cholerae, Hcp (haemolysin co-regulated protein) and Vgr (Val-Gly repeats), are secreted by a T6SS [39]. In the plant pathogenic bacterium Pectobacterium atrosepticum SCRI1043 (syn. Erwinia carotovora subsp. atroseptica), vgr and hcp genes are up-regulated when this pathogen is grown in the presence of potato extract, suggesting a potential role in virulence [40]. Two mutants, each defective in a gene of the T6SS gene cluster, showed reduced virulence compared to wild-type P. atrosepticum in potato tuber and stem virulence bioassays [41]. In the pea symbiont Rhizobium leguminosarum, a locus linked to the T6SS was discovered that negatively affects pea nodulation [42].
The T6SS gene cluster of E. pyrifoliae DSM 12163T is composed of 30 genes (EPYR_00645-EPYR_00674; Fig. 5A), of which 15 were identified as core genes, four identified as conserved genes between species, two identified as putative signal transducers, and nine remaining as hypothetical genes. Conserved blocks of genes were observed among the different bacterial species, and were interspaced by hypothetical genes. Both E. pyrifoliae DSM 12163T and E. tasmaniensis Et1/99 have closely related gene organizations with two exceptions: the genes between hcp and COG3456 are different (EPYR_00656-EPYR_00659, ETA_06210-06250 respectively) and the VgrG2 of E. pyrifoliae DSM 12163T is more closely related to VgrG1 of E. tasmaniensis Et1/99. These genes (EPYR_00656-EPYR_00659) have a lower G+C content than the backbone of the chromosome, indicating an acquired foreign origin of these genes. The genes downstream of vgrG2 in E. pyrifoliae DSM 12163T are homologous to genes downstream of vgrG1 in E. tasmaniensis Et1/99 (EPYR_00676, EPYR_00677, EPYR_00679, ETA_06380-06400).
Figure 5. Gene organization of Type VI Secretion System (T6SS) gene clusters. (A) Large T6SS gene clusters. (B) Small T6SS gene clusters. Blocks of related genes are shaded grey. Putative core genes are colored green, putative effectors red, putative signal transducers black, conserved genes between clusters grey and genes without related or homologues in all other clusters white. The frameshift in EPYR_02110/EPYR_02111 is indicated by a red flash.
The Hcp encoded in the T6SS cluster of E. pyrifoliae DSM 12163T is 91.9% identical to its ortholog in E. tasmaniensis Et1/99. Apart from this hcp gene, E. pyrifoliae DSM 12163T possesses two additional hcp genes on the plasmids pEP3 and pEP5 (Fig. 2). The Hcp proteins encoded on the E. pyrifoliae DSM 12163T plasmids are more related to each other (82.5% identity) than to the Hcp proteins of the large chromosomal T6SS clusters. The related enterobacterium Serratia proteamaculans 568 has a similar gene organization for its T6SS (Fig. 5A), but has no additional genes downstream of hcp1, an additional hcp gene (hcp2), and only one vgrG gene. In comparison, Hcp1 is more closely related to the Hcp genes of E. pyrifoliae DSM 12163T and E. tasmaniensis Et1/99 than to Hcp2. The P. atrosepticum SCRI1043 T6SS gene cluster is completely different from that in E. pyrifoliae DSM 12163T regarding its organization [37,43].
A second set of designated T6SS-related genes was found in the genomes of E. pyrifoliae DSM 12163T (EPYR_02109-EPYR_02113) and E. tasmaniensis Et1/99 (ETA_18657-ETA_18659) (Fig. 5B) which are absent in S. proteamaculans 568. In E. pyrifoliae DSM 12163T, three putative core genes (EPYR_02109, EPYR_02110/02111, EPYR_02113) and one conserved gene were found (EPYR_02112), while in E. tasmaniensis Et1/99 one of these core genes was absent. Genes in this second set show low homology to each other and to the large T6SS clusters. A frameshift in COG3523 (EPYR_02110/02111) within this T6SS cluster of E. pyrifoliae DSM 12163T was identified which leads to an early stop-codon in the gene. It can be deduced that this gene cluster is inactivated, and prone to mutations and deletions.
Flagellar genes
Two sets of genes encoding for flagellar assembly and chemotaxis related proteins were found in the genome of E. pyrifoliae DSM 12163T. One set is tightly clustered (EPYR_00976-EPYR_01028) and the encoded proteins show higher identity with the corresponding proteins of Salmonella and Escherichia spp. than with those of E. tasmaniensis Et1/99 [16]. This suggests that the entire region was acquired as a genomic island via horizontal genetic transfer. Only the right boundary of the genomic island is found in E. pyrifoliae DSM 12163T, as discernible by a reduced G+C-content bordered by an IS2-type integrase (EPYR_01040) flanking smpB (EPYR_01041), a gene also found the E. tasmaniensis Et1/99 genome (ETA_09720). The left border of the genomic island does not display distinctive insertion features. The first upstream-gene with high homology to E. tasmaniensis Et1/99 is tsx (EPYR_00974), which matches a gene that encodes a nucleoside-specific channel-forming protein in E. tasmaniensis Et1/99 (ETA_09440). Notably, in E. tasmaniensis Et1/99 the region included between ETA_09440 and ETA_09720 contains quorum-sensing genes expRI which are absent in E. pyrifoliae DSM 12163T.
A second complete set of flagellar genes is present but split among different clusters in the genome of E. pyrifoliae DSM 12163T (EPYR_01609-EPYR_01614, EPYR_01651-EPYR_01668, EPYR_02267-EPYR_02282, EPYR_02322-EPYR_02336). This second set closely resembles the flagellar genes found in E. tasmaniensis Et1/99 and appears to be ancestral in E. pyrifoliae DSM 12163T. Compared to the acquired flagellar assembly apparatus, the native gene cluster contains two extra genes (EPYR_01615 and EPYR_02267) encoding FliZ, a putative alternative sigma factor regulatory protein and the putative CheV chemotaxis signal transduction protein, respectively.
Sorbitol metabolism
Metabolism of sorbitol has been described as a contributing virulence factor in E. amylovora [44] on pome fruits which utilize sorbitol as a primary carbon storage compound [45]. The genome of E. pyrifoliae DSM 12163T contains a complete sorbitol gene cluster (EPYR_00602-EPYR_00606), and the organism is able to utilize sorbitol as a sole carbon source [19]. The non-pathogenic E. tasmaniensis Et1/99 lacks a sorbitol gene cluster in its genome [16], and may thus be disadvantaged in competitive interactions in environments rich in sorbitol. The exact role of sorbitol utilization in virulence is unresolved, since sorbitol content in apple trees has no specific effect on fire blight disease severity [46]. Associations between sorbitol and fire blight [47] suggest rather an osmotic-potential effect which could be due to various carbon compounds. Transgenic apple trees with down-regulated sorbitol synthesis compensate by increasing production of other sugars (i.e., sucrose, glucose) [45], and both of these are also substrates for E. pyrifoliae and E. amylovora which carry the respective metabolic genes.
Exopolysaccharide biosynthesis
E. pyrifoliae produces an exopolysaccharide (EPS) that is similar to amylovoran produced by E. amylovora [48]. In E. amylovora, mutation of one of the exopolysaccharide genes, wceB, impaired pathogen EPS biosynthesis and reduced virulence on apple [48]. The EPS biosynthetic gene cluster of E. pyrifoliae DSM 12163T (EPYR_01479-EPYR_01490) is more related to the gene cluster of E. amylovora Ea7/74 [49] than that of E. tasmaniensis Et1/99 [16]. The latter has different glycosyltransferases in the central part of the cluster (WceNM; Fig. 6) and is more similar to the stewartan biosynthetic cluster found in P. stewartii subsp. stewarii DC283 [50]. Overall, proteins encoded by the EPS cluster found in E. pyrifoliae DSM 12163 T are 87.7 - 96.3% identical to the corresponding proteins in E. amylovora Ea7/74, while the identities to E. tasmaniensis Et1/99 range between 68.2 and 92.8%, with no match for WceN and WceM (Fig. 6). Thus the E. pyrifoliae amylovoran genes are anticipated to have a role in virulence similar to that previously demonstrated in E. amylovora.
Figure 6. Comparison of the exopolysaccharide (EPS) biosynthetic clusters of E. pyrifoliae DSM 12163T (middle), E. amylovora Ea7/74 (top) and E. tasmaniensis Et1/99 (bottom). Identical colours indicate identical predicted functions. White arrows indicate flanking genes probably not involved in EPS biosynthesis. The numbers between the gene clusters indicate the sequence identity of the translated gene products indicated.
A second EPS produced by E. amylovora strains, levan, is also reported to contribute to virulence [51]. E. pyrifoliae DSM 12163T does not have levan biosynthetic genes and does not produce levan, which has also been reported to be lacking in the related fire blight Erwinia sp. from Japan [9]. It is proposed that the lack of levan may contribute to the restricted host-range in these species compared to E. amylovora, although levan biosynthesis in vitro has been reported in non-pathogenic E. tasmaniensis Et1/99.
Quorum-sensing apparatus
Quorum-sensing (QS) refers to the ability of bacteria to regulate gene expression in accordance with the presence of extracellular signalling molecules, or autoinducers (AI), that are produced in a cell density-dependent manner [52]. Two principal QS systems are known in Gram-negative bacteria and are defined by the chemical nature of the AI involved [53].
The AI-1 system utilizes N-acyl homoserine lactones (AHLs), produced by the LuxI family of proteins, as signal molecules. This system was first described in the marine bacterium Vibrio fisheri [54], where it controls bioluminescence production. Above a certain concentration, the secreted AHL binds the LuxR receptor and the resulting complex activates target gene transcription by binding a specific promoter site (lux box). Several AHLs have since been described in a wide-variety of Gram-negative pathogenic and non-pathogenic bacteria, differing principally in the N-acyl side chain moiety of the AHLs, with each unique AHL signal controlled by its own pair of luxRI gene orthologs [55].
In E. pyrifoliae, proteins encoded by one gene set (EPYR_02385-EPYR_02386) have low sequence identity with the autoinducer biosynthetic and receptor proteins PhzI/PhzR of Pseudomonas chlororaphis (22 and 26% respectively) and ExpI/ExpR of P. atrosepticum SCRI 1043 (17% and 22%). However, EPYR_02386 also shows 45% identity with the DNA-binding transcriptional activator SdiA, which regulates cell division in Escherichia coli and S. typhimurium [56], and which is known to respond to AHLs generated by other microbial species. For SdiA, no cognate gene encoding a corresponding signal-generating enzyme is present in either E. coli or in S. typhimurium [57]. Thus, SdiA may have a similar activity/non-activity in E. pyrifoliae and other Erwinia species.
Given the very low identity of EPYR_02385 to known AHL synthesis protein and the failure of E. pyrifoliae DSM 12163T to induce a positive reaction in AHL-biosensors Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NTL/pZLR4 (data not shown), it is hypothesized that the gene set composed of EPYR_02385 and EPYR_02386 contributes to production and detection of a yet unknown extracellular signal not obviously related to known QS signals.
A second QS system, based on production of an AI-2 signal molecule and controlled by the LuxS protein, is also widespread among Gram-negative and Gram-positive bacteria [58], and putatively involved in cross-species bacterial communication [59]. LuxS, a S-ribosylhomocysteine lyase, is however also known to be a central component of the activated methyl cycle (AMC), a metabolic cycle responsible for recycling methionine and generation of the major methyl donor S-adenosyl-L-methionine (SAM) in bacterial cells [60,61]. It is thus critical that the decision of whether or not the AI-2 system is present in certain species is not only based on the presence of luxS, but also on presence and functionality of the genes coding for the AI-2 receptors [61]. E. pyrifoliae DSM 12163T has a functional luxS gene (EPYR_03027) but it lacks orthologs for both of the two known AI-2 receptors: the LuxPQ-receptor of Vibrio harveyi [62] and the Lsr ABC-transporter of Salmonella typhimurium [63]. Thus, as in E. amylovora Ea273 [64], which also carries luxS but lacks AI-2 receptors, this gene probably has a metabolic rather than QS role in E. pyrifoliae.
Iron uptake determinants
Iron is an essential nutritional factor for plant pathogenic bacteria, which have high-affinity iron acquisition systems in order to supply this need [65]. Iron regulation of a wide range of genes is coordinated by Fur (encoded by EPYR_02663), which specifically regulates biosynthesis and uptake of iron-affinity siderophores [66]. The genome of E. pyrifoliae DSM 12163T contains four TonB-dependent receptor genes, one of which is also a putative copper receptor (oprC; EPYR_01920).
Another TonB-dependent receptor gene (EPYR_03492) encodes an ortholog of the E. amylovora ferrioxamine-receptor FoxR [67]. The gene product is directly involved in uptake of ferrioxamines into the periplasm. In E. amylovora, foxR mutants retain ability to synthesize desferrioxamine E, but lose their ability to utilize this as an iron substrate, are impaired in growth under iron-limited conditions such as found on flower surfaces, have reduced virulence, and are less resistant to plant defence responses [67,68]. Adjacent to foxR, E. pyrifoliae also has a biosynthetic gene cluster dfoJAC (EPYR_03489-EPYR_03491) containing an ortholog of dfoA, one of the biosynthetic genes for desferrioxamine in E. amylovora CFBP1430 [67], indicating that E. pyrifoliae DSM 12163T synthesizes desferrioxamine E and has an iron-acquisition system similar to E. amylovora [69]. In E. pyrifoliae DSM 12163T, a ferrihydroxamate TonB-dependent receptor and ABC transport system fhuACDB; EPYR_00903-EPYR_00906) is also present, which may contribute to uptake of ferrihydroxamate siderophores (Stockwell et al. 2008).
A fourth TonB-dependent receptor (EPYR_01969) found in E. pyrifoliae DSM 12163T is predicted to belong to the Transport Classification DataBase (TCDB; http://www.tcdb.org/ webcite) group TC 1.B.14.2.-, which consists primarily of heme and porphyrin uptake systems. An inactivated homolog of the TonB-dependent receptor yncD (EPYR_02731-EPYR_02732) is also present in the E. pyrifoliae DSM 12163T genome, but is inactivated by frameshifts. The ent-fep-fec gene cluster encoding enterobactin biosynthesis and transport in many Enterobacteriaceae, is absent in the genome of E. pyrifoliae DSM 12163T, as has been observed for E. tasmaniensis Et1/99 [16].
Comparative genomics to non-pathogenic Erwinia tasmaniensis Et1/99
The chromosome of E. tasmaniensis Et1/99 is 3,883,467 bp [16], compared to the 4,026,286 bp chromosome of E. pyrifoliae DSM 12163T. Genomic comparison of the complete genomes of the two related bacteria was performed using a Mauve progressive alignment [70], and large gaps were examined against the annotation (Fig. 7). With the exception of the plasmids, the two genomes are essentially collinear, with only a few genomic blocks exchanged.
Figure 7. Mauve progressive alignment of the genomes of E. pyrifoliae DSM 12163T (top) and E. tasmaniensis Et1/99 (bottom). Some relevant features within regions that have large differences in the alignment are indicated with arrows, plasmids are indicated as horizontal bars. Abbreviations: GI: genomic islands; T1RS: type 1 restriction system; T2RS: type 2 restriction sytem; Tn: transposase, T3SS: type 3 secretion system, T4SS: type 4 secretion system.
The difference in genome size between E. pyrifoliae DSM 12163T and E. tasmaniensis Et1/99 is largely due to the insertion of mobile genetic elements in E. pyrifoliae DSM 12163T. At least three regions contain phage genes, indicating insertion of prophages or remnants thereof. Additionally, a large number of genomic islands have inserted into the genome of E. pyrifoliae DSM 12163T, as indicated by the large number of possibly active or inactivated transposases identified. Of the remaining areas, two regions encode restriction systems and one cluster carries a complete additional set of flagellar genes (EPYR_00976-EPYR_01028; see above).
In contrast, E. tasmaniensis Et1/99 contains a large number of fimbrial gene clusters spread across the chromosome that are lacking in E. pyrifoliae DSM 12163T. Additionally, the nas/nir cluster is present in E. tasmaniensis but absent in E. pyrifoliae DSM 12163T, which explains the inability of E. pyrifoliae to generate nitrite from nitrate [7,19]. We observed that the chromosome of E. tasmaniensis Et1/99 encodes part of the central region of E. pyrifoliae plasmid pEP36 carrying the thiOSGF and the betB genes, but not the entire plasmid. E. tasmaniensis Et1/99 also carries genes for the ferric citrate uptake system fecRABCDE and for achromobactin uptake, which are all lacking in E. pyrifoliae DSM 12163T. From the alignment (Fig. 7), it was observed that E. pyrifoliae DSM 12163T contains the same set of T6SS genes as found in E. tasmaniensis Et1/99 but not previously reported. The incomplete inv/spa T3SS cluster, located close to the mutS gene in E. tasmaniensis Et1/99 (Fig. 4), is absent in E. pyrifoliae DSM 12163T.
Overall, the large-scale analysis of genomic differences between E. pyrifoliae DSM 12163T and E. tasmaniensis Et1/99 does not clearly reveal why E. pyrifoliae and not E. tasmaniensis is pathogenic. Mutational analysis of different genomic features is needed to determine their role in pathogenicity compared with non-pathogenic E. tasmaniensis. Our results also suggest that comparison of the genome sequences of E. pyrifoliae and E. amylovora, and different strains within each species, could reveal host-range determinants.
Compared to E. amylovora, the genome of E. pyrifoliae DSM 12163T encodes many of the same virulence factors, including two T3SSs, sorbitol metabolism, exopolysaccharides, and desferrioxamine biosynthesis. However, in E. pyrifoliae levan production and a third T3SS cluster are absent. Whether these factors contribute to the reduced host-range of this pathogen remains to be elucidated. Comparison to the genome of the non-pathogenic E. tasmaniensis Et1/99 indicated an additional flagellar gene cluster, absence of AHL biosynthesis genes, and a modified range of iron acquisition systems which may play a role in pathogenicity. As more of the species that have been identified with fire blight-like symptoms or as epiphytes on Rosaceae [9,13,18,36,71-77] are sequenced and can be compared on a whole-genome scale, further clues to the evolution and origin of necrotrophic Erwinia, and insights into host-pathogen interactions, can be anticipated.
Whole-genome sequencing
Genomic DNA was isolated using the Wizard Genomic DNA Purification Kit (Promega, Madison WI, USA). Whole-genome sequencing was performed by GATC (Konstanz, Germany) using both a 454 GS-FLX and a Solexa sequencer.
From a single run with the 454 GS-FLX, a total of 533,966 high-quality filtered sequence reads were generated with an average read length of 232 bp (Table 2). Coverage was equivalent to 31 times. Quality filtered sequences from whole-genome shotgun-sequencing were assembled using the 454 Newbler assembler. In total, 727 contigs were generated, of which 171 contigs larger than 500 bp. The average size of the large contigs was 5,480 bp. Reassembly from the individual data sets of each of the regions on the PicoTitrePlate (Roche) generated less contigs that were longer at average, and assembly of all contig sets allowed the exclusion of bad or contaminant sequences, that mainly constituted the majority of small contigs, under the assumption that the newly generated contigs should be covered by assemblies from all data sets.
Table 2. Overview of sequence data for the genome of E. pyrifoliae DSM 12163T.
A total of 4,835,468 high-quality filtered sequence reads with an average read length of 36 bp were generated by Solexa sequencing (Table 2). Average coverage was equivalent to 43.5×. Reads were assembled using the E. pyrifoliae Ep1/96 chromosome, plasmid pEP36 and pEP2.6. All reads were additionally de novo assembled using the program EDENA [78] with manually optimized settings. This yielded 3,162 contigs (N50 = 22,267; mean length = 1,206; maximum length = 12,612; minimum length = 100). The assembled contigs were aligned to the sequence using the SeqMan program of the LASERGENE package (DNASTAR, Madison, WI, USA).
Assembly of the E. pyrifoliae DSM12163T genome sequence was done in two steps. For the chromosome, GenBank accession FP236842 was used as reference. The sequences for plasmids pEP36 and pEP2.6 (AY123045, AY123048) were published [8], and used as a template for assembling these sequences. In a first step, the longer pyrosequencing reads were mapped to the available references using Roche's gsMapper application. When both sequence coverage and quality indicated a difference between the two strains with high confidence, the reference sequence was modified to reflect this change. Afterwards, both 454 and the shorter Solexa reads were mapped to the modified sequences with several custom scripts. In most cases, the Solexa reads supported the information from the 454 sequences and in some cases the Solexa data could be used to close gaps in the references where no 454 data was available. Based on the mapping information from both 454 and Solexa reads, contigs were created where coverage was high enough and at least 75% of all mapped reads agreed about a base for a given position. When no consensus base could be determined, an 'N' was inserted.
The reference sequence was also used as a template for the previously assembled contigs using SeqMan. Most contigs aligned to the template, and the non-aligned contigs were mainly due to false assembly in an earlier phase and could be resolved by realignment. The leftover contigs were checked by BlastN versus the template, and two contigs that were not covered by any part of the template were extended by several rounds of assembly versus the 454 reads until they were closed. This assembly was then confirmed by assembly using the Solexa reads, and by shifting the zero-point in the newly assembled plasmid sequences (pEP5 and pPE3).
Genome annotation
Genes were predicted using a combined strategy [79] based on the CDS prediction programs Glimmer [80] and Critica [81]. Subsequently, the potential function of each predicted gene was automatically assigned using the GenDB annotation pipeline [17]. The resulting genome annotation was curated manually, and metabolic pathways were identified using the KEGG pathways [82] tool in GenDB.
Routine sequence manipulations were done using several subroutines of the LASERGENE package. Whole-genome comparisons were done using the progressive alignment option of the Mauve comparison software (Version 2.0 [70]).
For the detailed analysis of the E. pyrifoliae DSM 12163T genome sequence, the Transport Classification DataBase (TCDB) http://www.tcdb.org/ webcite was used for improving the automated annotation of transporters. For comparative classification of the T3SS effectors, the Pseudomonas syringae Hop database http://pseudomonas-syringae.org/Hop_database.xls webcite was blasted on protein level versus the annotation of the E. pyrifoliae DSM 12163 T genome.
Accession numbers
The genome sequence of E. pyrifoliae DSM 12163T has been deposited at EBI and received the accession numbers EMBL:FN392235 (chromosome), EMBL:FN392238 (pEP36), EMBL:FN392239 (pEP5), EMBL:FN392237 (pEP3) and EMBL:FN392236 (pEP2.6).
Authors' contributions
THMS and SJ conducted the genome assembly. THMS performed the annotation, comparative genomics and wrote the manuscript. SJ, AG, FR and TK participated in the genome annotation, pathway identification and in writing the manuscript. JEF contributed to the project conception. BD conceived of and supervised the project and participated in writing the manuscript. All authors read and approved the final manuscript.
This work was supported by the Swiss Secretariat for Education and Research (SBF C07.0038) and the Swiss Federal Office for Agriculture (BLW Fire Blight Research - Pathogen). It was conducted within the European Science Foundation research network COST Action 864.
Duffy B, Schärer H-J, Bünter M, Klay A, Holliger E: Regulatory measures against Erwinia amylovora in Switzerland.
EPPO Bull 2005, 35:239-244. Publisher Full Text
Bonn WG, Zwet T: Distribution and economic importance of fire blight. In Fire blight the disease and its causative agent, Erwinia amylovora. Edited by Vanneste JL. Wallingford, UK CAB International; 2000:37-53. Publisher Full Text
Jock S, Donat V, López MM, Bazzi C, Geider K: Following spread of fire blight in Western, Central and Southern Europe by molecular differentiation of Erwinia amylovora strains with PFGE analysis.
Environ Microbiol 2002, 4(2):106-114. PubMed Abstract | Publisher Full Text
Thomson SV: Epidemiology of fire blight. In Fire blight the disease and its causative agent, Erwinia amylovora. Edited by Vanneste JL. Wallingford, UK CAB International; 2000:9-36. Publisher Full Text
Johnson KB, Stockwell VO: Management of fire blight a case study in microbial ecology.
Annu Rev Phytopathol 1998, 36:227-248. PubMed Abstract | Publisher Full Text
Geider K, Auling G, Jakovljevic V, Völksch B: A polyphasic approach assigns the pathogenic Erwinia strains from diseased pear trees in Japan to Erwinia pyrifoliae.
Lett Appl Microbiol 2009, 48:324-330. PubMed Abstract | Publisher Full Text
Kim W-S, Gardan L, Rhim S-L, Geider K: Erwinia pyrifoliae sp. nov., a novel pathogen that affects Asian pear trees (Pyrus pyrifolia Nakai).
Int J Syst Bacteriol 1999, 49:899-906. PubMed Abstract | Publisher Full Text
McGhee GC, Schnabel EL, Maxson-Stein K, Jones B, Stromberg VK, Lacy GH, Jones AL: Relatedness of chromosomal and plasmid DNAs of Erwinia pyrifoliae and Erwinia amylovora.
Appl Environ Microbiol 2002, 68(12):6182-6192. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Kim W-S, Hildebrand M, Jock S, Geider K: Molecular comparison of pathogenic bacteria from pear trees in Japan and the fire blight pathogen Erwinia amylovora.
Microbiology 2001, 147:2951-2959. PubMed Abstract
Lehman SM, Kim W-S, Castle AJ, Svircev AM: Duplex real-time polymerase chain reaction reveals competition between Erwinia amylovora and E. pyrifoliae on pear blossoms.
Phytopathology 2008, 98(6):673-679. PubMed Abstract | Publisher Full Text
Eastgate JA: Erwinia amylovora the molecular basis of fireblight disease.
Mol Plant Pathol 2000, 1(6):325-329. Publisher Full Text
Oh C-S, Beer SV: Molecular genetics of Erwinia amylovora involved in the development of fire blight.
FEMS Microbiol Lett 2005, 253:185-192. PubMed Abstract | Publisher Full Text
Triplett LR, Zhao Y, Sundin GW: Genetic differences between blight-causing Erwinia species with differing host specificities, identified by suppression subtractive hybridization.
Appl Environ Microbiol 2006, 72(11):7359-7364. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Shrestha R, Lee SH, Kim JE, Wilson C, Choi S-G, Park DH, Wang MH, Hur JH, Lim CK: Diversity and detection of Korean Erwinia pyrifoliae strains as determined by plasmid profiling, phylogenetic analysis and PCR.
Plant Pathol 2007, 56:1023-1031. Publisher Full Text
Shrestha R, Tsuchiya K, Baek SJ, Bae HN, Hwang I, Hur JH, Lim CK: Identification of dspEF, hrpW and hrpN loci and characterization of the hrpNEp gene in Erwinia pyrifoliae.
J Gen Plant Pathol 2005, 71:211-220. Publisher Full Text
Kube M, Migdoll AM, Müller I, Kuhl H, Beck A, Reinhardt R, Geider K: The genome of Erwinia tasmaniensis strain Et1/99, a non-pathogenic bacterium in the genus Erwinia.
Environ Microbiol 2008, 10(9):2211-2222. PubMed Abstract | Publisher Full Text
Meyer F, Goesmann A, McHardy AC, Bartels D, Bekel T, Clausen J, Kalinowski J, Linke B, Rupp O, Giegerich R, et al.: GenDB - an open source genome annotation system for prokaryote genomes.
Nucleic Acids Res 2003, 31:2187-2195. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Maxson-Stein K, McGhee GC, Smith JJ, Jones AL, Sundin GW: Genetic analysis of a pathogenic Erwinia sp. isolated from pear in Japan.
Phytopathology 2003, 93(11):1393-1399. PubMed Abstract | Publisher Full Text
Rhim S-L, Völksch B, Gardan L, Paulin J-P, Langlotz C, Kim W-S, Geider K: Erwinia pyrifoliae an Erwinia species different from Erwinia amylovora causes a necrotic disease of Asian pear trees.
Plant Pathol 1999, 48:514-520. Publisher Full Text
Fu J-F, Chang H-C, Chen Y-M, Chang Y-S, Liu S-T: Sequence analysis of an Erwinia stewartii plasmid, pSW100.
Plasmid 1995, 34(2):75-84. PubMed Abstract | Publisher Full Text
Lonetto M, Gribskov M, Gross CA: The s70 family sequence conservation and evolutionary relationships.
J Bacteriol 1992, 174(12):3843-3849. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Anderson M, Pollitt CE, Roberts IS, Eastgate JA: Identification and characterization of the Erwinia amylovora rpoS gene RpoS is not involved in induction of fireblight disease symptoms.
J Bacteriol 1998, 180:6789-6792. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Andersson RA, Kõiv V, Norman-Setterblad C, Pirhonen M: Role of RpoS in virulence and stress tolerance of the plant pathogen Erwinia carotovora subsp. carotovora.
Microbiology 1999, 145:3547-3556. PubMed Abstract
Mukherjee A, Cui Y, Ma W, Liu Y, Ishihama A, Eisenstark A, Chatterjee AK: RpoS (Sigma-S) controls expression of rsmAa global regulator of secondary metabolites, harpin and extracellular proteins in Erwinia carotovora.
J Bacteriol 1998, 180:3629-3634. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Hueck CJ: Type III protein secretion systems in bacterial pathogens of animals and plants.
Microbiol Mol Biol Rev 1998, 62(2):379-433. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Freiberg C, Fellay R, Bairoch A, Broughton WJ, Rosenthal A, Perret X: Molecular basis of symbiosis between Rhizobium and legumes.
Nature 1997, 387:394-401. PubMed Abstract | Publisher Full Text
Göttfert M, Röthlisberger S, Kündig C, Beck C, Marty R, Hennecke H: Potential symbiosis-specific genes uncovered by sequencing a 410-kilobase DNA region of the Bradyrhizobium japonicum chromosome.
J Bacteriol 2001, 183(4):1405-1412. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Dale C, Young SA, Haydon DT, Welburn SC: The insect endosymbiont Sodalis glossinidius utilizes a type III secretion system for cell invasion.
Proc Natl Acad Sci USA 2001, 98(4):1883-1888. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Alfano JR, Collmer A: Type III secretion system effector proteins double agents in bacterial disease and plant defence.
Annu Rev Phytopathol 2004, 42:385-414. PubMed Abstract | Publisher Full Text
Jock S, Kim W-S, Barny M-A, Geider K: Molecular characterization of natural Erwinia pyrifoliae strains deficient in hypersensitive response.
Appl Environ Microbiol 2003, 69(1):679-682. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
McClelland M, Sanderson KE, Spieth J, Clifton SW, Latreille P, Courtney L, Porwollik S, Ali J, Dante M, Du F, et al.: Complete genome sequence of Salmonella enterica serovar Typhimurium LT2.
Nature 2001, 413:852-856. PubMed Abstract | Publisher Full Text
Oh C-S, Kim JF, Beer SV: The Hrp pathogenicity island of Erwinia amylovora and identification of three novel genes required for systemic infection.
Mol Plant Pathol 2005, 6(2):125-138. Publisher Full Text
Shrestha R, Lee SH, Cho S, Lim CK, Hong S, Cho JM, Hwang I, Hur JH, Kim W-S: The hrp genes cluster in Erwinia pyrifoliae and determination of HR active domain in HrpNEp protein.
Boureau T, ElMaarouf-Bouteau H, Garnier A, Brisset M-N, Perino C, Pucheu I, Barny M-A: DspA/E, a type III effector essential for Erwinia amylovora pathogenicity and growth in planta, induces cell death in host apple and nonhost tobacco plants.
Mol Plant-Microbe Interact 2006, 19(1):16-24. PubMed Abstract | Publisher Full Text
Petnicki-Ocwieja T, van Dijk K, Alfano JR: The hrpK operon of Pseudomonas syringae pv. tomato DC3000 encodes two proteins secreted by the type III (Hrp) protein secretion system HopB1 and HrpK, a putative type III translocator.
J Bacteriol 2005, 187(2):649-663. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Bocsanczy AM, Beer SV, Perna NT, Biehl B, Glasner JD, Cartinhour SW, Schneider DJ, DeClerck GA, Sebaihia M, Parkhill J, et al.: Contributions of the genome sequence of Erwinia amylovora to the fire blight community.
Boyer F, Fichant G, Berthod J, Vandenbrouck Y, Attree I: Dissecting the bacterial type VI secretion system by a genome wide in silico analysis what can be learnt from available microbial genomic resources?
BMC Genomics 2009, 10:104. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text
Bingle LEH, Bailey CM, Pallen MJ: Type VI secretion a beginner's guide.
Curr Opin Microbiol 2008, 11(1):3-8. PubMed Abstract | Publisher Full Text
Pukatzki S, Ma AT, Sturtevant D, Krastins B, Sarracino D, Nelson WC, Heidelberg JF, Mekalanos JJ: Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system.
Proc Natl Acad Sci USA 2006, 103(5):1528-1533. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Mattinen L, Nissinen R, Riipi T, Kalkkinen N, Pirhonen M: Host-extract induced changes in the secretome of the plant pathogenic bacterium Pectobacterium atrosepticum.
Proteomics 2007, 7:3527-3537. PubMed Abstract | Publisher Full Text
Liu H, Coulthurst SJ, Pritchard L, Hedley PE, Ravensdale M, Humphris S, Burr T, Takle G, Brurberg M-B, Birch PRJ, et al.: Quorum sensing coordinates brute force and stealth modes of infection in the plant pathogen Pectobacterium atrosepticum.
PLoS Pathog 2008, 4(6):e1000093. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Roest HP, Mulders IHM, Spaink HP, Wijffelman CA, Lugtenberg BJJ: A Rhizobium leguminosarum biovar trifolii locus not localized on the Sym plasmid hinders effective nodulation on plants of the pea cross-inoculation group.
Mol Plant-Microbe Interact 1997, 10(7):938-941. PubMed Abstract | Publisher Full Text
Bell KS, Sebaihia M, Pritchard L, Holden MTG, Hyman LJ, Holeva MC, Thomson NR, Bentley SD, Churcher LJC, Mungall K: Genome sequence of the enterobacterial phytopathogen Erwinia carotovora subsp atroseptica and characterization of virulence factors.
Proc Natl Acad Sci USA 2004, 101(30):11105-11110. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Aldridge P, Metzger M, Geider K: Genetics of sorbitol metabolism in Erwinia amylovora and its influence on bacterial virulence.
Mol Gen Genet 1997, 256(6):611-619. PubMed Abstract | Publisher Full Text
Teo G, Suzuki Y, Uratsu SL, Lampinen B, Ormonde N, Hu WK, DeJong TM, Dandekar AM: Silencing leaf sorbitol synthesis alters long-distance partitioning and apple fruit quality.
Proc Natl Acad Sci USA 2006, 103(49):18842-18847. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Duffy B, Dandekar AM: Sorbitol has no role in fire blight as demonstrated using transgenic apple with constitutively altered content.
Suleman P, Steiner PW: Relationship between sorbitol and solute potential in apple shoots relative to fire blight symptom development after infection by Erwinia amylovora.
Phytopathology 1994, 84:1244-1250. Publisher Full Text
Kim W-S, Schollmeyer M, Nimtz M, Wray V, Geider K: Genetics of biosynthesis and structure of the capsular exopolysaccharide from the Asian pear pathogen Erwinia pyrifoliae.
Microbiology 2002, 148:4015-4024. PubMed Abstract
Bernhard F, Schullerus D, Bellemann P, Geider K, Nimtz M, Majerczak DR, Coplin DL: Genetics and complementation of DNA regions involved in amylovoran synthesis of Erwinia amylovora and stewartan synthesis of Erwinia stewartii.
Coplin DL, Majerczak DR, Bugert P, Geider K: Nucleotide sequence analysis of the Erwinia stewartii cps gene cluster for synthesis of stewartan and comparison to the Erwinia amylovora ams cluster for synthesis of amylovoran.
Gross M, Geier G, Rudolph K, Geider K: Levan and levansucrase synthesized by the fireblight pathogen Erwinia amylovora.
Physiol Mol Plant Pathol 1992, 40(6):371-381. Publisher Full Text
Pierson LS III, Wood DW, Pierson EA: Homoserine lactone-mediated gene regulation in plant-associated bacteria.
Annu Rev Phytopathol 1998, 36:207-225. PubMed Abstract | Publisher Full Text
Miller MB, Bassler BL: Quorum sensing in bacteria.
Annu Rev Microbiol 2001, 55:165-199. PubMed Abstract | Publisher Full Text
Nealson KH, Hastings JW: Bacterial bioluminescence its control and ecological significance.
Microbiol Rev 1979, 43(4):496-518. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Lerat E, Moran NA: The evolutionary history of quorum-sensing systems in bacteria.
Mol Biol Evol 2004, 21(5):903-913. PubMed Abstract | Publisher Full Text
Wang X, de Boer PAJ, Rothfield LI: A factor that positively regulates cell division by activating transcription of the major cluster of essential cell division genes of Escherichia coli.
EMBO J 1991, 10(11):3363-3372. PubMed Abstract | PubMed Central Full Text
Michael B, Smith JN, Swift S, Heffron F, Ahmer BMM: SdiA of Salmonella enterica is a LuxR homolog that detects mixed microbial communities.
J Bacteriol 2001, 183(19):5733-5742. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Xavier KB, Bassler BL: Interference with AI-2-mediated bacterial cell-cell communication.
Nature 2005, 437:750-753. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Winans SC: Bacterial esperanto.
Nat Struct Biol 2002, 9(2):83-84. PubMed Abstract | Publisher Full Text
Winzer K, Hardie KR, Williams P: LuxS and autoinducer-2 their contribution to quorum sensing and metabolism in bacteria.
Adv Appl Microbiol 2003, 53:291-396. PubMed Abstract | Publisher Full Text
Rezzonico F, Duffy B: Lack of genomic evidence of AI-2 receptors suggests a non-quorum sensing role for luxS in most bacteria.
BMC Microbiol 2008, 8:154. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text
Reading NC, Sperandio V: Quorum sensing the many languages of bacteria.
FEMS Microbiol Lett 2006, 254(1):1-11. PubMed Abstract | Publisher Full Text
Taga ME, Miller ST, Bassler BL: Lsr-mediated transport and processing of AI-2 in Salmonella typhimurium.
Mol Microbiol 2003, 50(4):1411-1427. PubMed Abstract | Publisher Full Text
Rezzonico F, Duffy B: The role of luxS in the fire blight pathogen Erwinia amylovora is limited to metabolism and does not involve quorum sensing.
Mol Plant-Microbe Interact 2007, 20(10):1284-1297. PubMed Abstract | Publisher Full Text
Expert D: Withholding and exchanging iron interactions between Erwinia spp. and their host plants.
Annu Rev Phytopathol 1999, 37:307-334. PubMed Abstract | Publisher Full Text
Dellagi A, Brisset M-N, Paulin J-P, Expert D: Dual role of desferrioxamine in Erwinia amylovora pathogenicity.
Mol Plant-Microbe Interact 1998, 11(8):734-742. PubMed Abstract | Publisher Full Text
Kachadourian R, Dellagi A, Laurent J, Bricard L, Kunesch G, Expert D: Desferrioxamine-dependent iron transport in Erwinia amylovora CFBP 1430 cloning of the gene encoding the ferrioxamine receptor FoxR.
BioMetals 1996, 9(2):143-150. PubMed Abstract | Publisher Full Text
Expert D, Dellagi A, Kachadourian R: Iron and fire blight role in pathogenicity of desferrioxamine E, the main siderophore of Erwinia amylovora. In Fire blight the disease and its causative agent, Erwinia amylovora. Edited by Vanneste JL. Wallingford, UK CAB International; 2000:179-195. Publisher Full Text
Feistner GJ: Proferrioxamine synthesis in Erwinia amylovora in response to precursor or hydroxylysine feeding metabolic profiling with liquid chromatography-electrospray mass spectrometry.
BioMetals 1995, 8(4):318-327. PubMed Abstract | Publisher Full Text
Darling ACE, Mau B, Blattner FR, Perna NT: Mauve multiple alignment of conserved genomic sequence with rearrangements.
Genome Res 2004, 14:1394-1403. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Roselló M, Ferrer S, Llop P, López MM, Christen R, Gardan L: Description of Erwinia piriflorinigrans sp. nov., causal agent of pear blossom necrosis.
Chung YR, Brenner DJ, Steigerwalt AG, Kim BS, Kim HT, Cho KY: Enterobacter pyrinus sp. nov., an organism associated with brown leaf spot disease of pear trees.
Hao MV, Brenner DJ, Steigerwalt AG, Kosako Y, Komagata K: Erwinia persicinus a new species isolated from plants.
Int J Syst Bacteriol 1990, 40:379-383. PubMed Abstract | Publisher Full Text
Stockwell VO, Hockett K, Marie C, Duffy B: Pink Erwinia amylovora colony discoloration in diagnostic isolations by co-cultured bacteria.
Jock S, Geider K: Molecular differentiation of Erwinia amylovora strains from North America and of two Asian pear pathogens by analyses of PFGE patterns and hrpN genes.
Environ Microbiol 2004, 6(5):480-490. PubMed Abstract | Publisher Full Text
Geider K, Auling G, Du Z, Jakovljevic V, Jock S, Völksch B: Erwinia tasmaniensis sp. nov., a non-phytopathogenic bacterium from apple and pear trees.
Int J Syst Evol Microbiol 2006, 56(12):2937-2943. PubMed Abstract | Publisher Full Text
Mergaert J, Hauben L, Cnockaert MC, Swings J: Reclassification of non-pigmented Erwinia herbicola strains from trees as Erwinia billingiae sp. nov.
Int J Syst Bacteriol 1999, 49:377-383. PubMed Abstract | Publisher Full Text
Hernandez d, François P, Farinelli L, Østerås M, Schrenzel J: De novo bacterial genome sequencing millions of very short reads assembled on a desktop computer.
Genome Res 2008, 18:802-809. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
McHardy AC, Goesmann A, Pühler A, Meyer F: Development of joint application strategies for two microbial gene finders.
Bioinformatics 2004, 20(10):1622-1631. PubMed Abstract | Publisher Full Text
Salzberg SL, Delcher AL, Kasif S, White O: Microbial gene identification using interpolated Markov models.
Nucleic Acids Res 1998, 26(2):544-548. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Badger JH, Olsen GJ: CRITICA coding region identification tool invoking comparative analysis.
Mol Biol Evol 1999, 16(4):512-524. PubMed Abstract | Publisher Full Text
Kanehisa M, Goto S, Kawashima S, Nakaya A: The KEGG databases at GenomeNet.
Nucleic Acids Res 2002, 30(1):42-46. PubMed Abstract | Publisher Full Text | PubMed Central Full Text


text:
This site is targeted at medical and radiology professionals, contains user contributed content and material that may be confusing to a lay audience. Use of this site implies acceptance of our Terms of Use.
The Article "chromic lymphocytic leukaemia" doesn't exist
If this article should exist but doesn't, you can start it: create new article
If the article exists but is called something different you can link to it
Search results for "chromic lymphocytic leukaemia":
- Richter tranformation (Article)
- Chronic lymphocytic leukaemia (Article)
- Small lymphocytic lymphoma (Article)
- Rai staging system for chronic lymphoid leukaemia (Article)
- Binet staging system for chronic lymphocytic leukaemia (Article)
- WHO classification of tumours of haematopoietic and lymphoid tissues (Article)
- Deauville five-point scale (Article)
- Intradural spinal involvement in leukaemia (Article)
- Tree-in-bud sign (Article)
- Thymolipoma (Article)
- Malignant pleural disease (Article)
- Autosomal recessive osteopetrosis (Article)
- Neurosarcoidosis (Article)
- Metaphyseal radiolucent lines in leukemia (Case)
- Chronic lymphocytic leukaemia (Case)
- Pericardial effusion (Case)
- Hamburger sign (Case)
- Tree-in-bud sign - normal CT virtual bronchoscopy (Case)


text:
- Product Highlights
- Helps Strengthen Hair, Skin & Nails With 2,000 mcg of Biotin Per Serving
- Vegetarian Formulated Without Most Common Allergens
- Provides Nutritive Support for Healthy Hair, Skin & Nails
- Delivered in Time Release Tablets to Ensure Optimal Efficacy
SKU 015794050292 Quantity 90 Tablets Serving Size 2 Tablets Servings Per Container 45 Dietary Needs Dairy Free, Free of Artificial Colors, Free of Artificial Flavors, Free of Artificial Sweeteners, Gluten Free, Milk Free, Soy Free, Vegetarian, Wheat Free Directions Adults take two (2) tablets of Country Life Maxi-Hair daily. For best utilization, take with food. Warnings As a reminder, discuss the supplements and medications that you take with your health care providers. Do not accept if seal is broken. Store between 59-86F. Keep out of the reach of children. If you are pregnant or nursing, taking medication or planning a surgery, consult your doctor before using this product. Not recommended for people who regularly consume liver. If any adverse reactions occur, stop taking the product and consult your doctor. Ingredients Vitamin A (as retinyl palmitate), Vitamin C (as ascorbic acid, rose hips), Vitamin E (as d-alpha tocopheryl acid succinate), Thiamin (vitamin B-1)(as thiamine hydrochloride), Riboflavin (vitamin B-2), Niacin (as niacinamide), Vitamin B-6 (as pyridoxine hydrochloride)…for product label information including a complete list of ingredients, click all product images above. Other Ingredients Cellulose, stearic acid, oligosaccharides, magnesium stearate, cellulose & glycerin coating, silica, calcium silicate, natural vanilla, vegetable glaze.
- Product Details
Country Life Maxi-Hair is a unique vitamin, mineral and amino acid complex for hair, skin and nails.
Biotin helps support healthy hair, skin and nails.
MSM provides essential sulfur, important to hair, skin and nails.
Certified Vegetarian
Time Release
Nourishment for hair, nails and skin
Dietary Supplement
Certified Gluten-Free
Maxi-Hair Brand: Country Life4.4 stars, based on 3 review(s).Rating:Price: $17.99 - In stock
- Why SuperSupplements?
Unbeatable Selection ~ We carry an unbeatable selection of the very best nutritional supplements, sports nutrition and natural personal care products. We continually refresh our selection so you always have access to the newest, most effective products right here in one convenient spot.
Satisfaction Guarantee ~ Your complete satisfaction with any Super Supplements purchase is guaranteed. You may return any item that does not meet your expectations within 30 days of purchase for a full refund of the product price. Guaranteed!
Free Shipping ~ Why pay for shipping when you can use those dollars for more great products? Spend $25 and enjoy free, fast shipping to most locations.
Passionate People ~ We love what we do. We work hard to bring you the very best products at the very best overall value every single day. If there is anything we can do to make your experience with Super Supplements a better one, we want to hear from you - with a passion!
Country Life Maxi-Hair, 90 Tablets Reviews Showing 3 out of 3 reviews for Country Life Maxi-Hair, 90 Tablets
great stuff
Maxi-Hair review posted by M. Toner on 9/5/13
My nails wouldn't grow for the last 2 years and now they are thanks to this product. I am very happy.
Maxi-Hair review posted by La'Quan on 1/26/13
So far so good. I've been taking two tablets daily, my bowl movements have been pretty regular, my nails look fabulous and my hair is fuller-thicker.
I haven't noticed any improvements in my skin yet. Maybe it's because I don't have bad skin but I like the product and will continue to buy it.
Noticeable Difference
Maxi-Hair review posted by GJ Carlson on 10/15/12
My hair is thin to the point if it's layered there is only one layer. Maxi-hair give me a noticeable improvement. A person I admire asked me if I was cutting it differently because it looked fuller and thicker.
Hope they alway have this one on the shelves!
Hair Nutrients
120 Capsules
Country Life
Maxi Hair
60 Tablets
Super Supplements | 750 S Michigan St, Seattle, WA 98108 | (877) 454-3988