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Quality Service of Window Replacement Company in Brooklyn Park
Do you have a broken or cracked window that needs replaced immediately? Are your current windows old and in need of a quick upgrade? Do you think that energy efficient windows are the way to go? These are just some of the many reasons why you should consider window replacement services. A window replacement is needed for a variety of reasons, but the result should always be the same. A window replacement should be high quality and affordable, but also add value and visual appeal to your home. We are the window replacement company in Brooklyn Park that is ready and willing to help with all of your varying window needs.
Why Choose Us?
We are a window replacement company in Brooklyn Park that is dedicated to providing customers with the windows that they need. We offer high quality at reasonable prices that the competition just can't beat. Not only do you need a high quality replacement window, but you also need installation that is expert-level. We offer both and strive to provide the entire Brooklyn Park region with replacement window services that you can count on. Choosing us I simply choosing the very best in the industry.
Old Windows Let In Cold Air and Let Money Out
The most common reason for needing replacement windows id due to windows that are old. Do you know that if you have old windows in your home you are letting money go right out the window? Old windows will create a draft in your home that only raises the cost of your energy bills. However, by choosing our window installation services, you can save money on your energy bills right away.
Cost Effective Choice
This means that new windows are a cost effective solution that will pay off over time. Don't let money just fly out your old window. Now is the time to call on a window company that you trust to give you a more energy efficient option. A draft or leak is the sign of a problem and you should call o us when you first notice this type of issue.
Call The Window Replacement Company in Brooklyn Park
You should not wait any longer to get your very own replacement windows installed. Call us today at (763)-271-8700 for a no pressure, no hassle price quote that will blow your mind. High quality at affordable prices is what we are known for. You can also visit our website Perfectexteriorsmn.com to know more about window replacement company in Brooklyn Park.


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At GSI, we know that implementing new systems and managing your company's workflow can be a challenge, so we're committed to providing you expert service through every stage of the process. We aim to make our software solutions easy to use and seamlessly integratable with your current systems. Our support staff has two decades of expertise in assisting companies like yours troubleshoot their onboarding and integration process, so don't hesitate to ask, we're always happy to offer a helping hand!
If you have an existing support ticket, and wish to view the status of your ticket, please login by clicking the link below:
Please Fill Out the Following Support Form:
GSI Distribution Design Studio & LD-Pro
Technical assistance on GSI Distribution Design Studio & LD-Pro will be available by telephone during normal business hours, Monday to Friday, 8 a.m. to 5 p.m. PST or PDT.
GSI Forester, Scout, and Collector
Technical assistance on GSI Forester, Designer, Collector, & Scout will be available by telephone during normal business hours, Monday to Friday, 8 a.m. to 5 p.m. CST or CDT.

subset: commoncrawl


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Sydney - Melbourne
1800 251 440
Email us...
Sydney :
[email protected]
[email protected]
Brisbane :
[email protected]
Adelaide :
[email protected]
Perth :
[email protected]
Canberra :
[email protected]
Mr.Fingerfood On Google+
2. Do you have vegetarian/vegan selections?
Yes, we are able to accommodate most dietary preferences & restrictions.
3. Do you have staff to serve our guests?
Of course we have chefs in uniform to cook and prepare waitresses /waiters to mix and mingle with the guests serving the food on platters and even bar people to serve your drinks.
4. Do you deliver cold food only?
We can gladly provide you with menu items to include delivery & full instructions
on food handling. and heating instructions A delivery charge of $40.00 within
the metropolitan area is applicable.
5. Can I as a client pick up my order?
Yes you can pick up your order from our party showrooms and save $40 and a
further 10% of the total price Pick ups prepaid by credit card can be picked
up Mon-Saturday 10am-6pm.
6. Can you provide drinks table's chairs, table linens, marquees?
We can provide or help you coordinate all of these different services.
We have suppliers all over the metropolitan area.
7. Do you take credit cards?
Yes, we accept VISA , American Express & Mastercard You
may also pay by company cheque EFT or cash on the day.
8. Do you require a deposit?
Yes a deposit of $100 is requested to confirm the date of
your event. We then will ask you for a guest guarantee 7 days
before your event. However, we can accommodate any additional
guests up 48 hours prior to the event.
if you have a question we haven't covered
please email your nearest office
Sydney Melbourne


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FORT WORTH — The next steps he takes are in Iowa, in the Big Ten, the big time.
But on Tuesday night Nic Shimonek took his final steps as a high school quarterback — and they led him right into the end zone.
"He's a heck of a player, a heck of a player,'' said Stephenville football coach Joseph Gillespie moments after Shimonek scored to nail down a 39-29 win for the North in the 79th annual Texas High School Football Coaches Association All-Star Game.
Gillespie was the head coach of the North, and moments after the game he was Shimonek's biggest fan.
Hard not to be...
The kid from Mildred had just staged one of those hold-your-breath, game-on-the-line drives. You know the kind — Elway, Manning, Staubach drives. Those guys made last minute drives famous.
Shimonek was just saying farewell, just waving good-bye to a high school career at Mildred no one in this part of Texas will soon forget.
It was the perfect wave.
By the time it was over, Shimonek had completed 8-of-9 passes for 172 yards and had thrown for a TD (a beauty on a 69-yard pass to Whitehouse's Dylan Cantrell that lifted the North to an 18-14 lead in the third) and he had run for a TD.
He was brilliant on the final drive — going 3-for-3 in the air for 76 yards before slipping into the end zone with a 9-yard TD run that would have made Peyton Manning proud.
It was an audible.
"It was just a read. The play was going to the running back, but I saw the end pinch in and knew that play wouldn't work,'' Shimonek said. "He (the running back) went left and I kept the ball and went to the right. I don' think I scored because of my speed. It was more because of deception. I don't think they knew I had the ball. The running back on the play asked me, 'how did you find that hole?' ''
With just 2:12 left in the game, Shimonek's missing-ball-trick sealed the win.
"You're going to laugh when I tell you this,'' Shimonek said afterward. "All week I was telling myself that if I scored a touchdown I was going to spike the ball, cause you can never spike the ball, but this is an all-star game. I was going to spike it.''
But he went spikeless.
"Then when I scored, I forgot to do it,'' Shimonek said with one of those can-you-believe-that smiles. "I guess I just got caught up in the moment.''
It was his moment.
He took over at his own 25 with a 32-29 lead and 4:48 left in a see-saw game that had the crowd of 12,185 at Amon Carter Stadium riding right along up-and-down all night.
The South fans were up — way up after the first play from scrimmage on that last drive. Shimonek was sacked, and just-like-that, he faced a second and 20 from his own 15. It was the third time the South sacked him, and the sixth of the night for a defense that was coming fast and hard in the final minutes.
"That was a great drive,'' Gillespie said. "And he had to start out in a hole after that sack.''
It seemed like Shimonek was starting in the hole all night — because he was. He completed his first pass on his first play from scrimmage in the first quarter only to see it get wiped out with a block in the back penalty that happened after the catch. It left Shimonek facing a first-and-20.
On the third quarter drive that ended with the bomb to Cantrell, the first play was a pitchout to the tailback, who fumbled and took a four-yard loss. Shimonek was swarmed on the second-and-14 play, but managed to avoid the sack and pick up three yards. Then on third-and-11, he stepped out of the pressure and sent a strike to Cantrell, who and waltzed in untouched.
So when Shimonek's last drive started with a sack, he just picked himself up and started finding a way to win. He connected with Cantrell over the middle to the 26 and then on third-and-11 he found Lufkin's Jabryce Taylor, who made a remarkable leaping catch between two defenders. Taylor was incredible all night, setting a record for receiving yards with 226 on 10 receptions. But his high-flying circus 29-yard catch at the 45 was his only reception from Shimonek.
"He ran the wrong route on the play,'' Shimonek said. "I told him, 'you better learn that playbook when you get to SMU and run the right route.' And I told him it was a great catch and told him he's got great God-given ability to make a catch like that. He is really gifted.''
Shimonek got hammered after he released the ball, and even the player who hit him — El Campo defensive end Cole Hunt — gained respect for the Mildred QB.
"I hit him hard, and he got back up,'' Hunt said. "Man that guy can throw. He puts the ball right on the money. He's a great quarterback. All their quarterbacks can throw.''
Shimonek felt the it and was on the ground when he looked up and saw Taylor make the catch.
"He hit me hard. I thought my back popped out,'' Shimonek said.
Two plays later, he got hammered again, but managed to throw a perfect strike to Cantrell over the middle for a 36-yard gain to the 9-yard-line. Then Slick-Nic gave the South the slip — into the end zone.
"Did you see those hits he took? He took two big hits on that drive,'' Gillespie said. "But he got back up. He got roughed up and kept going. That drive was huge.''
Shimonek got hit even harder after the game.
Mix in a little history, a little ground-breaking stuff. It packs a wallop.
Shimonek is the first player from Mildred to play in this game, this 79-year-old classic that brings some of the best in Texas together every July.
"I didn't know that,'' said Shimonek.
Then he went silent. He wasn't staggered, just hit hard with the news. He picked himself and threw a completion to Mildred.
"It goes a long way...It's an honor, such a honor to be here,'' Shimonek said. "But what it really says, is it says volumes about Mildred, volumes about our program and how far we've come. We won our first playoff game in 2A as sophomores, and as juniors we went to the state semifinals, and then this year we got to the state title game. It just speaks volumes about the program.''
It hit him hard.
Consider Shimonek's feat: Corsicana has sent 20 to the THSCA All-Star Game, including Otha Langston and Paul Harshaw, who played in the first one back in 1935, and Jon Bauer, who was the Tigers' last player in this game in 2005.
But Shimonek is by himself, the lone Eagle.
And that made every snap he took, every handoff and every pass he threw mean so much more. It's always like that when history mingles with sports.
He smiled and said he wouldn't be the last Eagle to play in this game. He meant it.
That's part of his legacy.
With one foot pointed toward Iowa and his future, Shimonek wanted to talk more about Mildred than what he had done Tuesday night.
Sure, there's all the awards, district MVPs, back-to-back all-state honors, and the Daily Sun's Del Thrash Player of the Year Award as the Golden Circle's best football player. There are numbers stacked as high as Big Tex — 8,215 career passing yards, 99 TDs and a senior season (35 TDs and 2,718 yards and a trip to Cowboys Stadium) no one will forget. But Shimonek's love for the program and what it meant to him — and will always mean to him — to be an Eagle is the best part of that final drive, that last wave good-bye.
That's his real legacy.


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Status: I get by with a little help from my friends. Mood: grateful :)
I don't know how many of you use MySpace regularly but I can tell you that imho, the new version is quite attrocious. Cluttered, chaotic and teeming with even more useless advertisements and information than ever before, to put it bluntly, I hate it. However, one thing that I'm glad they saved from the devastation was the "status and mood" bar. I always felt that the addition of "mood" as opposed to Facebook's sole "status" set it apart from other social networks.
It's quite rare that I go to MySpace nowadays. But when I do, I usually put something like the title of this blog as my status and mood. Because every time I listen to music or have a concert or hold a workshop or have a lesson or sing one single note, I think about all the people who held my hand along the way. And am very, very grateful to them for helping me come to this point. I spoke to my dear teacher Baldo a couple of weeks ago and thanked him for giving me the wings to fly towards my dreams.
I dedicate this blog to all of you, to all my friends both real and virtual, who have given me positive energy to keep going the distance, for giving me a perch to land on when I've needed a rest, for helping me heal my wings when they've been injured. I thank you for listening to our music, visiting this website, coming to our concerts, dropping me the occasional line. It means the world to me.
There are still many heights for me to reach and I know that I'll get by with a little help from you, my friends.


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Some reviewers are smitten nonetheless. Charles Spencer of The Daily Telegraph gives Radcliffe the most ecstatic praise:
Daniel Radcliffe brilliantly succeeds in throwing off the mantle of Harry Potter, announcing himself as a thrilling stage actor of unexpected range and depth.
Michael Billington of The Guardian also raves about Radcliffe, giving the play itself four stars mostly on the strength of Radcliffe's performance:
The revelation of this revival is that Daniel Radcliffe really can act, proving that his screen appearances as JK Rowling's boy-hero are no flash in the magic pan.
The Independent's David Lister seems to think Radcliffe shows promise but stops short of calling him "revelatory":
In the event, Radcliffe acquits himself well. He is not the most expressive of actors, and his stage presence will take time to evolve; but from the moment he enters the psychiatrist's office, shoulders hunched, eyes narrowed, and singing advertising jingles to avoid questioning, he cuts a compelling figure. As the evening goes on, there are moments when he touches, even if not tugs at, the heart strings. One feels for this boy because one senses from his performance a repression hiding a reservoir of feelings desperate to burst.
The Times' Benedict Nightingale is lukewarm and even a bit critical of Radcliffe's interpretation of the difficult role:
Radcliffe proves an assured actor and makes a perfectly able equimaniac. He can do aggression and pain, and, oddly, is lacking only in the sense of magic and wonder the part demands…
…Radcliffe has only one obvious weakness. His Alan is pale, vulnerable, defensive, surprisingly tough; but he's supposed also to find an exhilaration bordering on religious ecstasy in the company and, especially, the secret riding of horses. This, Radcliffe misses."
The notoriously discerning Evening Standard theater critic, Nicholas de Jongh, gives Radcliffe a near-pan:
[P]layed by Radcliffe with an air of pathetic, small-scale belligerence…[his] touching, little-boy lost Alan never convinces you he is wild with desire for horses or girls.
Hey, don't let de Jongh get you down, Danny. You can always rely on those world-renowned intellectual giants, Peaches and Pixie Geldof, to offer fresh insights:
Peaches Geldof said: "I think Daniel acted brilliantly. I wasn't shocked by the nudity."
Younger sister Pixie gasped: "I was so shocked! I love him, I think he is beautiful."
In other news:
- The Guardian highlights some of Britain's most breathtaking and seemingly out-of-place landscapes.
- Victoria Beckham goes blonde. Not the full Monroe, but a skunky two-toned platinum blonde front and brunette rear. Yuck. (The Sun)
- Victoria Beckham has obviously adapted to a certain way of L.A. life – she's drinking and playing dominoes – but it hasn't been easy. The Mirror reports she's "struggling with LA's demand for 24-hour-a-day glamour." She could start by fixing that jacked-up dye job. I'm just sayin.'
- Haha, this announcement doesn't even warrant a lead spot on The Mirror page, but Jamiroquai's Jay Kay says he's quitting the music business. He says, "All I'm going to do now is fly my helicopter and look for the right lady to have children with." I wish him as much success in those endeavors as he received as a musician.
- Dame Helen Mirren has already been photographed erotically sucking in hamburgers at Oscar parties. Now the 61-year-old actress has revealed her Oscar speech was a "commando" performance. Mirren told Oprah that she wore no knickers under her glittering gown Sunday evening. I'm waiting with bated breath for Perez Hilton to post the tell-tale coochie-cam limo shot.
- Jade Goody's passage to India: if you believe The Sun, it's not going over so well. Supposedly, Goody staged a photo shoot inside a small hut in Delhi, but people waiting outside to be photographed were crushed as they tried to get in. "Women were knocked flying and children trampled underfoot," claims The Sun. I'm sure a couple of people tripped or scuffed their shoes, but you know the drama queens at The Sun. They get snaps, though, for this quote from a witness: "Mum Anu, 26, was knocked to the ground with her three-month-old tot. She said: 'My baby is hurt – who is this awful person? She is not welcome here if she is going to behave like this.'"
- Beth Ditto and Noel Gallagher are a mutual admiration society.(NME)
- "If you find yourself tied up and doused in petrol, don't worry if all your assailant has is a lighted cigarette: scientists have proved you won't end up as a human fireball."(Guardian)
- Will Justin Hawkins pen the UK's Eurovision tune now that Morrissey (and apparently every other good songwriter in Britain) has passed on it? (Mirror)
- Deep Purple slag off their own live album and discourage fans from buying it. Ian Gillan says this about the live concert behind the album: "It was awful. It was one of the lowest points of my life – all of our lives, actually." I admire the honesty and integrity of this statement. Some old rockers would have taken the paycheck and said, "F*ck the fans."(Mirror)
- Will Mick Jagger collaborate with new Oscar winner, Martin Scorsese, on a film about the music business?(The Sun)
- Naomi Campbell confesses a long-time struggle with cocaine to GQ magazine.(Daily Mail)
- "War"…what is it good for? A top 10 hit for the band Ugly Rumours, it seems. (Guardian)
- Is there a Go Fug Yourself for the roadways? Cuz Kristin Scott Thomas' hideous tiger-striped electric car is just itching for a fug.(Daily Mail)


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Welcome to my Riviera construction project
[ Home ]
[ Layout Area ]
[ Laminations ]
[ Mechanical Installation ]
[ Deck Framing ]
[ Seating and Interior ]
[ Site Map ]
[ Tools and Shop ]
[ Schedule ]
Line Drawing of the Glen-L Riviera, by Glen-L Marine Designs
Mark Bronkalla
Here I am tracing the blueprints for the
frame pieces onto the work table.
Why is this site here?
This site covers the construction process of my boat, a Glen-L Riviera.
The Riviera is a 20 foot double cockpit traditionally styled wooden runabout.
It was built with cold molded construction technique consisting of and
wood laminations and epoxy. More on
This site is organized in rough chronological order as a tour of the construction
process. It has been updated over time due to questions from other builders
or as experience with the materials and techniques dictates. Start the tour
The Site Map provides topic oriented navigation
through the site. It also has the site search at the bottom.
This is a picture intensive site. Please be patient while the photos download.
I have compressed them as much as possible while retaining reasonable
image quality. If you want to see a high resolution version of an image,
let me know and I can probably e-mail it to you.
The boat is complete and has been used extensively for seven summers. It
is a great ski and wakeboard boat. We have also used it for fishing as
well. Top speed as measured by a GPS is 53 mph!
I have a number of pictures in my office showing the boat construction.
Several co-workers have asked: "How I can take something as pretty
as this fishing or skiing, as it looks like a piece of furniture".
My reply was:"Not many pieces of furniture can go over 50 MPH".
The boat is now referred to as a piece of "50 mph furniture" TM.
Look for a number of my columns in the Glen-L
Webletters. Sign up for a free subscription at www.Glen-L.com.
Newer sections:
Many of my newer projects and adventures are documented in photos
on Facebook. Here is a link to one of the albums
Many of my newer projects and adventures are documented in photos
on Facebook. Here is a link to one of the albums
If you would like to contact me, send e-mail to:
builders at bronkalla.com. Replace the at with @.
Just trying to dodge a few spammers.


text: Accent On Achievement [Trumpet] Book 1 John O'Reilly £7.95 Calculate shipping price Add to Basket Quantity Buy Now Currently sold out. Order now and we will ship when availableLive stock levels are a guide only. Full details Reviews Item Details Item Details Category: Band Methods Publisher: Alfred Publisher's reference: 17090 Our Stock Code: 913867 Media Type: Paperback; includes accompaniment CD


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How Using Adwords Can Boost your Business
The internet contains millions of websites – how can you lead potential customers to yours? When individuals need to find a product or service, they search for it on Google, the most popular search engine in the world. If your business is not included on the first page of Google search results, it's likely no one will ever click on your link. In fact, the higher up the chain you are, the more traffic you will get. One statistic stated that website listed as one of the first results got 36.4% more traffic than those listed below. Utilizing inbound marketing, you can help online users find your site or social media page, and AdWords is the solution you need to help you boost online traffic and increase your sales!
What is AdWords?
AdWords is Google's own advertising service, which allows business owners to create ads that will be shown as "Sponsored Links" above and to the right of the organic (non-sponsored) Google search results. When a person searches for a specific keyword, your ad will show on their screen, encouraging them to visit your site.
What are Keywords?
We've mentioned keywords a few times in this article, but what exactly are they? They are the words or phrases that online users utilize to search for something online. Keywords can be as simple as a business name or as complicated as "affordable Italian restaurant near me." Each and every business likely has hundreds, if not thousands, of applicable keywords.
AdWords allows business owners to pay for search engine optimization (SEO), which tells Google to list your site in its top results. AdWords has a pay-per-click (PPC) system that charges you every time an online user clicks on your link when it's in the sponsored section. Keywords are not created equal, and some cost more than others.
Many business owners don't know which keywords can help them improve their online presence; however, Google has a free tool that allows them to conduct keyword research. Google's Keyword Planner lets online users search for a keyword, get suggestions for other related terms and provides information about how competitive that keyword is, how many people search for it, and how much it would cost in a PPC campaign.
What are the Biggest Advantages of Google AdWords?
Compete with the Big Guys
While it's not necessary to pay for campaigns in order to utilize SEO as part of your inbound marketing, growing an online reputation organically can take a long time. Plus, small businesses have to compete with others who may have expert marketing professionals on their team, promoting their website through blogs, articles, social media outreach, etc. AdWords levels the playing field by allowing anyone to bid on a keyword to advertise their brand.
Target Your Audience
If your business only caters to local customers, there's no point for you to pay for clicks from people outside of your targeted area. AdWords solves this issue by allowing businesses to geo-target which locations their ads get seen in. You can specify which city you want your ad featured in, or even a specific proximity around your local business.
Retarget Your Visitors
Google AdWords has a feature that follows your online visitors and features ads for something they searched for on your site on other websites they visit, a strategy called retargeting. If you've ever searched for a pair of shoes, only to have them pop up for weeks after on various sites you visit, you've seen retargeting work its magic.
Online marketing with Google AdWords can be planned according to your budget; however, if you want to make a real impact, you may need to invest in this endeavour. If you can benefit from financial help to start this process, IOU Financial is committed to fueling the growth of small businesses. Contact us today!


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Class D β-lactamase-mediated resistance to β-lactams has been increasingly reported during the last decade. Those enzymes also known as oxacillinases or OXAs are widely distributed among Gram negatives. Genes encoding class D β-lactamases are known to be intrinsic in many Gram-negative rods, including Acinetobacter baumannii and Pseudomonas aeruginosa, but play a minor role in natural resistance phenotypes. The OXAs (ca. 150 variants reported so far) are characterized by an important genetic diversity and a great heterogeneity in terms of β-lactam hydrolysis spectrum. The acquired OXAs possess either a narrow spectrum or an expanded spectrum of hydrolysis, including carbapenems in several instances. Acquired class D β-lactamase genes are mostly associated to class 1 integron or to insertion sequences.
Class D β-lactamases, also known as oxacillinases or OXA-type β-lactamases (OXAs), are active-serine-site enzymes like Ambler class A and class C β-lactamases, differing from class A and C enzymes in amino acid structure, whereas class B β-lactamases are metalloenzymes with a Zn2+ ion(s) in the active site (4, 71, 78). Even though class D includes mostly enzymes with higher hydrolysis rates for cloxacillin and oxacillin than for benzylpenicillin (hence the name oxacillinases), not all class D β-lactamases have this characteristic. Most of the class D enzymes belong to group 2d of the Bush functional classification scheme for β-lactamases (23). Among the four β-lactamase molecular classes, class D β-lactamases are the most diverse enzymes (107). This diversity is observed at both the genetic and biochemical levels, with enzymes possessing either a narrow or expanded spectrum of hydrolysis. In addition, several class D β-lactamases have an expanded spectrum of activity resulting from point mutations.
Although many class D β-lactamase genes are embedded into class 1 integrons, recent reports indicated that other specific genetic structures, including insertion sequences and transposons, may be associated with class D β-lactamase genes. Numerous class D β-lactamase genes have been identified as a source of acquired resistance in gram-negative bacteria, but recent studies have shown that class D β-lactamases are also naturally produced in clinically significant pathogens and environmental species (107).
This review focuses on the diversity and substrate profiles of class D β-lactamases, their sources, and the genetics of acquisition of the corresponding genes. All the class D β-lactamases for which a sequence is available in the GenBank databases are listed in Table 1.
OXAs are β-lactamases belonging to class D according to the molecular nomenclature based on amino acid identity comparison. Similar to class A and C β-lactamases, they possess an active-site serine which is located at position 70 in the class D β-lactamase numbering system (the DBL numbering system) (72, 97) (Fig. 1). The DBL numbering system has been established to analyze the molecular structures of the class D β-lactamases, in analogy to the numbering system adopted for class A enzymes (5, 32). The two groups of enzymes share the serine and lysine residues which are part of the highly conserved motif S-T-F-K found at positions 70 to 73 (79). The Y-G-N motif (positions 144 to 146) and the K-T-G motif (positions 216 to 218) are highly conserved in class D β-lactamases, but the F-G-N motif may replace Y-G-N in several OXAs (Fig. 1).
Class D β-lactamases are usually not inhibited by clavulanic acid, tazobactam, and sulbactam, whereas their activities may be inhibited in vitro by sodium chloride (NaCl). This property is not shared by β-lactamases of other classes, thus defining it as a useful characteristic for in vitro identification. NaCl at a concentration of 100 mM inhibits totally the activities of most class D β-lactamases (9, 54, 65, 127, 130). This property is not clearly explained, but it has been related at least to the presence of a Tyr residue at position 144. In vitro mutagenesis showed that the replacement of a Tyr residue by a Phe at that position makes a mutant resistant to NaCl inhibition (65).
Class D β-lactamases were first defined as β-lactamases hydrolyzing cloxacillin and oxacillin faster than benzylpenicillin (23). This definition does not seem valuable anymore since recently described enzymes actually inactivate cloxacillin and oxacillin poorly, even sometimes sparing those substrates. However, all class D β-lactamases significantly hydrolyze amino- and carboxypenicillins.
Although all the OXAs first characterized corresponded to acquired genes, it has been demonstrated lately that many gram-negative species naturally possess genes in their genomes coding for OXAs, which are therefore considered to be resident enzymes.
ACQUIRED NARROW-SPECTRUM CLASS D β-LACTAMASES
The OXA-1 subgroup.As mentionned by Boyd and Mulvey (20), β-lactamases OXA-1 and OXA-30 are the same enzyme, since an original sequencing error in the OXA-1 sequence had introduced a mistake (113). The blaOXA-1 gene has been found in plasmid and integron locations in a large variety of gram-negative organisms (see below). OXA-1 β-lactamase, like most OXAs, hydrolyzes amino- and ureidopenicillins significantly and hydrolyzes narrow-spectrum cephalosporins weakly (9). In addition, OXA-1 slightly hydrolyzes broad-spectrum cephalosporins, conferring reduced susceptibility to cefepime and cefpirome (9). It may therefore be classified as a β-lactamase possessing either a narrow or an expanded spectrum of hydrolysis (see below).
The blaOXA-1 gene has often been identified in ampicillin-resistant enterobacterial strains such as isolates of Escherichia coli, Shigella flexneri, and Salmonella spp., with spread occuring among community-acquired enterobacterial species (44, 62, 148). It has been identified in human and animal isolates. It was, for instance, identified in Shigella dysenteriae strains from diverse geographical origins (45) and in Salmonella enterica serovar Typhimurium isolates from humans and food products of animal origin in Portugal (6). Noteworthily, it seems to be a frequent source of resistance to ampicillin in Shigella spp. and Salmonella spp. isolated from humans in the United States (63).
The blaOXA-1 gene has frequently been found to be associated with genes encoding extended-spectrum β-lactamases (ESBLs). Plasmids coharboring the blaOXA-1 and blaCTX-M-1 genes have been identified in animal E. coli isolates from Spain (31), and recent studies have reported very frequent association of blaOXA-1 with the worldwide-spread CTX-M-15 ESBL determinant in human E. coli isolates from diverse geographical origins (29). This association of blaOXA-1 with blaCTX-M genes makes isolates resistant to β-lactam-β-lactamase inhibitor combinations. Some OXA-1 variants such as OXA-47, carrying up to seven substitutions compared to OXA-1 (none located inside the conserved motifs of class D β-lactamases), possess a narrow spectrum of hydrolysis (127). The blaOXA-47 gene has been identified as plasmid borne and is carried on a gene cassette inserted into an integron in a Klebsiella pneumoniae isolate from Turkey coproducing OXA-48 (123) (see below).
Among the OXA-1 derivatives that have been identified, no known variant possesses the ability to hydrolyze ceftazidime. OXA-1 possesses the ability to hydrolyze cefepime and cefpirome slightly, and OXA-31, which differs from OXA-1 by two amino acid substitutions and has been identified in a Pseudomonas aeruginosa isolate from France (9), also possesses that property. We have demonstrated previously that a transfer of the natural plasmid harboring the blaOXA-31 gene into a P. aeruginosa recipient strain confers resistance to cefepime and cefpirome but that susceptibility to ceftazidime is not modified. By comparing the MICs of β-lactams for E. coli recombinant clones producing OXA-1 or OXA-31, as well the kinetic parameters, we showed that OXA-1 and OXA-31 possess similar hydrolysis profiles, including the ability to hydrolyze cefepime and cefpirome (9). Therefore, OXA-1 and OXA-31 may be considered to be class D β-lactamases characterized by expanded-spectrum profiles with significant impacts on MICs for bacterial species that possess high-level intrinsic impermeability (such as P. aeruginosa) and not on those for bacterial species with low-level intrinsic impermeability (such as E. coli).
All the genes for the OXA-1-like β-lactamases described above have been identified in the form of gene cassettes inserted into class 1 integrons (9, 44, 107, 127).
The OXA-2 subgroup.OXA-2 β-lactamase is another narrow-spectrum β-lactamase that shares ca. 30% amino acid identity with OXA-1 (77). It constitutes, together with its derivatives OXA-3, OXA-15, OXA-21, OXA-32, OXA-34, OXA-36, and OXA-53, a distinct genetic cluster (Fig. 2). The blaOXA-2 gene has been identified often in P. aeruginosa and S. Typhimurium isolates producing the ESBL PER-1 (46), as has the variant OXA-3 (142). Recently, blaOXA-2 has been identified in P. aeruginosa isolates from Serbia and Hungary that also produced the ESBL PER-1 (81). In addition, it has been identified in other species such as Morganella morganii in Argentina (135), K. pneumoniae in Uruguay (161), and E. coli in France (GenBank accession no. CAJ13583). Surprisingly, it has also been identified in distantly related species such as Bordetella bronchiseptica (GenBank accession no. ABD63309) and Aeromonas hydrophila (GenBank accession no. ABF69297) and even the gram-positive species Corynebacterium amycolatum (GenBank accession no. CAI40608) (unpublished data). The OXA-21 variant, a point mutation derivative of the OXA-3 enzyme, has been identified once in Acinetobacter baumannii (163). The genes for these β-lactamases have been also identified in the form of gene cassettes inserted into class 1 integrons.
The OXA-10 subgroup.OXA-10 β-lactamase (formerly known as PSE-2) also possesses the ability to hydrolyze cephalosporins, hydrolyzing cefotaxime, ceftriaxone, and aztreonam at low levels but sparing ceftazidime, cephamycins, and carbapenems (70). The blaOXA-10 gene is encountered in a large variety of gram-negative species, being the blaOXA gene identified most frequently in P. aeruginosa (8, 25, 37-39, 46, 60, 99, 125).
There are some point mutation derivatives of OXA-10, such as OXA-11, OXA-13, OXA-16, OXA-28, OXA-35, and OXA-74, possessing increased activities toward expanded-spectrum cephalosporins (see below) (Fig. 2). Very few OXA-10-like enzymes possessing a narrow spectrum of hydrolysis have been described, one of which is OXA-7, encoded by a gene reported to be plasmid borne in E. coli. OXA-7 has 95% amino acid identity to OXA-10 (91, 145).
Other subgroups.Among the narrow-spectrum OXAs, some variants are not, or are weakly, related to the OXA-1, OXA-2, and OXA-10 subgroups (Fig. 2). This is the case for OXA-9, which shares 45 and 54% amino acid identity with the most closely related enzymes, OXA-12 and OXA-18 (155). OXA-9 has the property, uncommon for an OXA enzyme, of being inhibited by clavulanic acid and cloxacillin but not by NaCl (16). Its hydrolytic profile remains to be precisely determined, even if OXA-9 would likely correspond to a narrow-spectrum β-lactamase (our unpublished data). The blaOXA-9 gene was first identified on a plasmid in a K. pneumoniae isolate (155). The blaOXA-9 gene has been identified in a Pseudomonas putida isolate from France coexpressing the metallo-β-lactamase VIM-2 (122), in an Enterobacter cloacae isolate from Canada (119), and in an Enterobacter aerogenes isolate from France (120). Recently, it has also been identified in a carbapenem-resistant E. cloacae strain originating in the Untied States and coproducing the class A carbapenemase KPC-3 (43) and in a carbapenem-resistant K. pneumoniae isolate in Turkey coproducing the class D carbapenemase OXA-48 (127).
Another narrow-spectrum class D β-lactamase is LCR-1, which was one of the very first class D β-lactamases to be identified on the basis of biochemical properties only (146), was recovered from P. aeruginosa, and hydrolyzes only penicillins, nitrocefin, and oxacillin (168). Its recently determined amino acid sequence (32) shows that it belongs to a separate cluster among class D enzymes (Fig. 2), sharing 40% or less amino acid identity with other OXAs (40% with OXA-55 and 37% with OXA-48). NPS-1 is another narrow-spectrum enzyme which is related to LCR-1 (carrying eight amino acid differences) (114). The blaNPS-1 gene was identified on a conjugative plasmid from P. aeruginosa isolates in the United Kingdom (83).
The product of the blaOXA-5 gene represents another subgroup of narrow-spectrum class D β-lactamases (Fig. 2). After the gene was phenotypically described as plasmid borne in a P. aeruginosa isolate recovered in the United Kingdom (91), its sequence was determined (32). The OXA-5 determinant shares less than 81% identity with OXA-10 derivatives. The blaOXA-5 gene was found as a gene cassette inserted into a class 1 integron and was later found in association with the blaGES-2 gene, encoding a peculiar ESBL with carbapenemase properties, in carbapenem-resistant P. aeruginosa isolates from South Africa during an outbreak period (134). The hydrolysis spectrum of OXA-5 is limited to narrow-spectrum penicillins, oxacillin, and cephalothin (cefalotin). Its activity is inhibited by NaCl but not by clavulanate (91). Recently, OXA-129 was identified in S. enterica subsp. enterica serovar Bredeney porcine isolates from Brazil (95). It differs from OXA-5 by 12 amino acids.
OXA-20 is another narrow-spectrum class D β-lactamase that has weak amino acid identity to other class D β-lactamases (108). It shares less than 75% identity with the most closely related enzyme, OXA-2. OXA-20 hydrolyzes penicillins, cephalothin, and cephaloridine but spares expanded-spectrum cephalosporins. Interestingly, OXA-20 is inhibited by clavulanic acid (108). The blaOXA-20 gene was identified in a P. aeruginosa isolate from Sicily, Italy, that coexpressed OXA-18, the latter OXA possessing an expanded-spectrum profile (118) (see below). Noteworthily, the blaOXA-20 gene was also identified in A. baumannii isolates from France and Italy and was also located inside integron structures (53, 58, 121, 169). A point mutation derivative of OXA-20, OXA-37, has been identified in an A. baumannii isolate from Spain (109).
OXA-46 was identified in a P. aeruginosa isolate from Italy. In fact, this determinant was identified in a multidrug-resistant isolate that coexpressed the metallo-β-lactamase VIM-1 (57). OXA-46 shares 78% amino acid identity with OXA-2 and OXA-53, the most closely related enzymes. It hydrolyzes mostly penicillins, oxacillin, and narrow-spectrum cephalosporins. Interestingly, it is inactivated by tazobactam and carbapenems but exhibits low susceptibility to NaCl (57).
ACQUIRED ES-OXAs
Whereas most of the previously mentioned class D β-lactamases exhibit a substrate profile limited mostly to penicillins, nitrocefin, oxacillin, and cloxacillin, class D β-lactamases able to hydrolyze expanded-spectrum cephalosporins have also been identified (though their distribution remains quite limited). There are two main types of expanded-spectrum class D β-lactamases (ES-OXAs) (Table 1). Some ES-OXAs are just point mutation derivatives of narrow-spectrum class D β-lactamases. Others may be completely different enzymes with weak amino acid sequence identity. Interestingly, most of the ES-OXAs identified so far are from P. aeruginosa.
ES-OXAs as point mutation derivatives of narrow-spectrum OXAs.The first variant described to possess an extended-spectrum profile was OXA-15, a point mutation derivative of OXA-2 (with a change from Asp to Gly at position 150 in the DBL numbering system). The blaOXA-15 gene was identified inside a class 1 integron structure in a P. aeruginosa isolate from Turkey (36). OXA-15 has increased ability to hydrolyze ceftazidime compared to that of OXA-2, but it also hydrolyzes cefepime and aztreonam at a lower level than OXA-2. OXA-32 is another OXA-2 derivative possessing an expanded-spectrum hydrolysis profile, identified in a ceftazidime-resistant P. aeruginosa isolate recovered from a patient originating from Guadeloupe in the French West Indies (124). The blaOXA-32 gene was integron borne and located on a 250-kb conjugative plasmid. OXA-32 differs from OXA-2 by a Leu169Ile substitution (DBL numbering) that extends its hydrolysis spectrum toward ceftazidime but not cefotaxime (124). Interestingly, this substitution also modifies the behavior of the class D β-lactamase OXA-32 toward inhibitors, since imipenem and cefoxitin were found to significantly inhibit the activity of OXA-32 compared to that of OXA-2.
A large number of class D β-lactamases with expanded-spectrum activity have been identified among OXA-10 variants. OXA-11, identified in a P. aeruginosa isolate from a Turkish patient, was found to be encoded by a plasmid (60). Compared to OXA-10, it possesses two substitutions (Asn146Ser and Gly167Asp [DBL numbering]) that extend its ability to hydrolyze ceftazidime. Other OXA-10-like ES-OXAs have been identified in P. aeruginosa strains from Turkey, including OXA-14 (carrying a Gly167Asp change) (38), OXA-16 (with Ala114Thr and Gly167Asp changes) (39), and OXA-17, the latter having an Asn76Ser substitution conferring an increased hydrolysis rate for cefotaxime and a decreased rate for ceftazidime (37). OXA-35, which has been identified in a P. aeruginosa isolate from France (8), is a point mutation derivative of both OXA-19 (95) and OXA-28 (125). OXA-35 possesses hydrolytic activity toward penicillins and cefsulodin but spares narrow- and expanded-spectrum cephalosporins. In contrast, OXA-19 and OXA-28, differing from OXA-10 by 9 and 10 amino acid substitutions, respectively, confer resistance to ceftazidime (98, 125).
Other ES-OXAs.Several ES-OXAs are not structurally related to narrow-spectrum OXAs. OXA-18 was the first enzyme of this type to be identified, in a P. aeruginosa isolate obtained in 1995 in Paris from a patient previously hospitalized in Sicily (118). OXA-18 shares less than 50% amino acid identity with the other OXAs. It is peculiar since it confers high-level resistance to expanded-spectrum cephalosporins and is surprisingly well inhibited (for a class D β-lactamase) by clavulanic acid (118). Its substrate profile does not include cephamycins and carbapenems. OXA-18 can therefore be considered a typical ESBL. The blaOXA-18 gene is very likely chromosomally located. Other OXA-18-producing P. aeruginosa isolates have been recovered recently in Belgium, as sources of outbreaks (105). The structures surrounding the blaOXA-18 gene in those strains were identical to those discovered in the OXA-18-producing Italian isolate (105).
OXA-45 is another ES-OXA, identified in a multidrug-resistant Texan P. aeruginosa isolate coexpressing the class B β-lactamase VIM-7 (153). OXA-45 shows the highest degrees of identity to OXA-18 (66%), OXA-9 (43%), and OXA-22 (40%). The blaOXA-45 gene, located on a 24-kb plasmid, was not in the form of a gene cassette but was associated with two copies of an ISCR5-like element. OXA-45 includes in its substrate profile expanded-spectrum cephalosporins, such as ceftazidime, cefotaxime, and aztreonam, but spares carbapenems and cephamycins. The activity of OXA-45, like that of OXA-18, is inhibited by clavulanic acid (153).
Finally, OXA-53 is another ES-OXA that has been identified recently in an S. enterica serovar Agona isolate from Brazil (103). It shares 90% amino acid identity with the narrow-spectrum OXA-2 but exhibits unusual properties since it is well inhibited by clavulanic acid and confers reduced susceptibility to ceftazidime. The blaOXA-53 gene was plasmid and integron borne (103).
ACQUIRED CHDLs
Some acquired class D β-lactamases may also hydrolyze carbapenems. None of these carbapenem-hydrolyzing class D β-lactamases (CHDLs) (131, 133) significantly hydrolyze expanded-spectrum cephalosporins, therefore indicating that currently known class D β-lactamases are unable to combine extended-spectrum and carbapenem-hydrolyzing properties. Most of these enzymes are from Acinetobacter spp.
As noticed by Queenan and Bush (137), the level of hydrolysis of carbapenems by CHDLs remains low, due to poor turnover of these β-lactams. In general, the hydrolysis of imipenem, even if slow, is faster than that of meropenem. Usually, the Km values for imipenem are low, indicating that CHDLs have very high apparent affinities for that substrate (65, 130). Due to these peculiar properties, the exact contribution of these enzymes to phenotypic resistance has been debated. However, it has been demonstrated recently using knockout or complementation experiments that acquired CHDLs such as OXA-23, OXA-40, and OXA-58 identified in A. baumannii significantly contribute to carbapenem resistance (67).
The first reported acquired class D β-lactamase with carbapenemase activity was OXA-23 (also known as ARI-1 [43, 115]), detected in an A. baumannii isolate from Scotland and found to be plasmid mediated after its transfer to Acinetobacter junii (143). The ability of OXA-23 to hydrolyze carbapenems was weak, since hydrolytic measurements showed only low-level activity against imipenem. However, compared to the activities of other class D β-lactamases, this hydrolysis was significant and was allowed to define the novel subgroup of OXAs currently named CHDLs (133). OXA-23, weakly related to other class D β-lactamases with only 36% amino acid identity to OXA-5 and OXA-10, belongs to the first group of CHDLs. Although kinetic parameters of hydrolysis have been determined with crude extracts of OXA-23 (115), precise parameters are still not available. The other member of the first group is OXA-27, which possesses two substitutions (Thr86Ala and Asn250Lys [DBL numbering]) compared to OXA-23 (1). The corresponding blaOXA-27 gene, identified in a carbapenem-resistant A. baumannii isolate from Singapore, was likely chromosomally located (1). Whereas blaOXA-27 has been identified in only a single isolate, blaOXA-23 is widespread and identified only in Acinetobacter spp., with a single exception, since this gene was also identified as chromosomally carried in a carbapenem-resistant Proteus mirabilis isolate from France (17). Recent data indicate that OXA-23-positive A. baumannii strains have spread throughout the world, to locations including France (30), Bulgaria (149), Iran (48), the United Arab Emirates (101), Tunisia (86), Brazil (35), and Australia (160). In addition, OXA-23 producers have been at the origin of hospital outbreaks in French Polynesia (104), Colombia (163), the United Kingdom (28), Turkey (92), China (166, 170), and Korea (74). OXA-23-positive A. baumannii isolates and one A. junii strain have been also identified in Romania (87).
A second group of CHDLs is made up of OXA-25, OXA-26, OXA-40, and OXA-72 (an original sequencing error in OXA-24 makes it now OXA-40 [see the Lahey website at http://www.lahey.org/Studies/ ]). These enzymes differ by a few amino acid substitutions only. OXA-24/OXA-40 was originally identified as chromosomally encoded in a carbapenem-resistant A. baumannii isolate recovered from Spain (18, 19). The blaOXA-40 gene has since been identified in different areas, especially in Portugal and Spain (40, 116, 138, 141), but also in the Untied States (84, 136). Interestingly, the blaOXA-40 gene has been identified as either chromosomally carried or plasmid borne (138). Surprisingly, it has been very recenty identified as plasmid located in two carbapenem-resistant P. aeruginosa isolates in Spain (146). OXA-40 is not inhibited by NaCl (65). OXA-25 and OXA-26 are point mutation derivatives of OXA-40 that have been identified in carbapenem-resistant A. baumannii isolates recovered from Spain and Belgium, respectively (1). OXA-72 has been identified in A. baumannii isolates from China, South Korea, Taiwan, and Bahrain (80, 85, 102, 166).
A third identified group of CHDLs is represented by OXA-58, which has also been detected only in Acinetobacter spp. so far. OXA-58 was first identified in a multidrug-resistant A. baumannii isolate from France (130), that strain being at the origin of an outbreak in the local hospital (64). OXA-58 shares 35, 33, and 18% amino acid identity with OXA-5, OXA-10, and OXA-1, respectively. OXA-58 hydrolyzes penicillins and carbapenems at low levels. Whereas weak hydrolysis of cefpirome was detected, hydrolysis of cefepime, ceftazidime, and cefotaxime was not. The rate of hydrolysis of imipenem was 10-fold lower, and that of meropenem was 100-fold lower, than that of benzylpenicillin (130). The blaOXA-58 gene was found to be plasmid borne in A. baumannii MAD (130), as well as in most of the OXA-58-producing A. baumannii isolates reported worldwide, including those in Europe, Argentina, Australia, and the United States (26-28, 69, 87, 117); OXA-58 is often associated with hospital outbreaks and has been involved in such outbreaks in France, Belgium, Italy, Turkey, Greece, and the United States (12, 15, 53, 64, 69, 128, 159). Noteworthily, OXA-58 has also been identified in other Acinetobacter species, such as A. junii in Romania (87) and Australia (117), Acinetobacter genomospecies 3 in Spain (88), and Acinetobacter phenon 6/ct13TU in Spain (89). OXA-97 is a point mutation variant of OXA-58 that shares the same hydrolytic properties and has been recently identified in A. baumannii isolates from Tunisia (129). The blaOXA-97 gene was plasmid carried and identified in a clonal strain at the origin of an outbreak at the Salhoul Hospital in Sousse, Tunisia. Another point mutation derivative is OXA-96, identified in A. baumannii in Singapore (75).
Very recently, the novel CHDL OXA-143 was identified in a clinical A. baumannii isolate that had been recovered in Brazil (68a). It shares 88% amino acid identity with OXA-40, 63% with OXA-23, and 52% with OXA-58. It may therefore correspond to a new subgroup of CHDLs encountered in A. baumannii and involved in carbapenem resistance in that species. Its substrate profile was similar to those of other CHDLs, and the corresponding gene was not integron or transposon carried but was likely acquired by a homologous recombination process.
Another unrelated CHDL is OXA-48, which has been identified in a carbapenem-resistant K. pneumoniae isolate from Istanbul, Turkey (127), thus, in a member of the Enterobacteriaceae. That isolate coexpressed several β-lactamases, including the class A ESBL SHV-2a and the narrow-spectrum β-lactamases TEM-1 and OXA-47. OXA-48 β-lactamase hydrolyzes penicillins and, at a lower level, imipenem but is not active against expanded-spectrum cephalosporins (127). This enzyme is the class D β-lactamase with the highest known catalytic efficiency for imipenem (having a kcat value of 2 s−1) (127). It shares less than 46% amino acid identity with other class D β-lactamases and has weak identity to OXA-23 and OXA-40 (36 and 32%, respectively). The blaOXA-48 gene was found to be plasmid located in K. pneumoniae 11978 (127). Very recently, we reported an important outbreak of infection with OXA-48-producing K. pneumoniae that occurred in Istanbul from May 2006 to January 2007 (24). Two distinct clones differing from the first OXA-48 producer and producing different ESBL determinants (SHV-12 and CTX-M-15) were identified in the same hospital. In addition, the blaOXA-48 gene has been identified in E. coli and Citrobacter freundii strains in Turkey (59, 110). The spread of the blaOXA-48 gene may be much more important than expected. Indeed, the gene has been found in K. pneumoniae isolates from Lebanon and from Belgium (34, 90). Its detection may be difficult since the level of acquired resistance to carbapenems may remain quite low.
Overall, the hydrolysis spectra of these class D β-lactamases are quite consistent, all enzymes hydrolyzing imipenem at low levels but not broad-spectrum cephalosporins and aztreonam.
NATURALLY OCCURRING CLASS D β-LACTAMASES
Recent studies have shown that many bacterial species possess chromosomally located and naturally occurring class D β-lactamase genes. The first such OXA gene to be identified was that from Aeromonas jandaei (formerly A. sobria). This species naturally produces the class D β-lactamase OXA-12, which has strong activity against cloxacillin and oxacillin and whose expression is inducible (3, 139). Another class D β-lactamase, AmpS, lacking significant activity against cloxacillin and oxacillin but hydrolyzing penicillins, was also detected in A. jandaei (A. sobria) (165). AmpS shares 96% amino acid identity with OXA-12. However, analysis using current GenBank databases indicates that AmpS likely corresponds to the intrinsic class D β-lactamase of another Aeromonas species whose genome sequences have been released, namely, A. hydrophila (GenBank accession no. YP_858675). β-Lactamases OXA-12 and AmpS both possess the property, peculiar for a class D enzyme, of being inhibited by clavulanic acid (139, 165).
Ralstonia pickettii is naturally resistant to penicillins, narrow-spectrum cephalosporins, ceftazidime, and aztreonam or shows intermediate susceptibilities to these agents. Two types of class D β-lactamases have been identified in that species. OXA-22 exhibits narrow-spectrum β-lactam hydrolysis activity that does not explain resistance to monobactams in its natural host (112). The gene encoding OXA-22 is chromosomally located, and induction experiments demonstrated that its expression is inducible (linked to a peculiar open reading frame [ORF] located upstream of blaOXA-22) but also revealed the overexpression of another β-lactamase gene. Further cloning experiments identified a second class D β-lactamase gene, blaOXA-60, which is also located on the chromosome of R. pickettii (55). PCR screening of multiple R. pickettii isolates identified variants of both blaOXA-22 and blaOXA-60 genes. OXA-60 possesses only 19% amino acid identity to OXA-22 and is weakly related to the other OXAs. It exhibits a narrow-spectrum hydrolysis profile, including activity against penicillins and nitrocefin and weak activity against carbapenems (55). The molecular basis of the induction process requires the presence of ORF RP3, located 190 bp upstream of blaOXA-60 and divergently transcribed. Disruption of ORF RP3 abolishes induction of both β-lactamases, suggesting that ORF RP3 may be a global regulator, although the 532-amino-acid-long product displays no obvious sequence homology to known regulatory proteins (56).
The blaOXA-61 gene was identified on the chromosome of Campylobacter jejuni and encodes a narrow-spectrum β-lactamase not inhibited by clavulanic acid (2). Considering the MICs of β-lactams for the original C. jejuni isolates, it seems likely that OXA-61-like enzymes are poorly expressed in their natural host and consequently play a minor role in susceptibility to β-lactams (2).
The blaOXA-62 gene was identified on the chromosome of a carbapenem-resistant Pandoarea pnomenusa isolate recovered from a cystic fibrosis patient (145). Pandoraea spp. are gram-negative, non-glucose-fermenting rods. The blaOXA-62 gene, which possesses an unusually high GC content of 65.3%, encodes a CHDL that is well inhibited by NaCl and hydrolyzes penicillins and oxacillin, as well as imipenem and meropenem at low levels, but does not hydrolyze expanded-spectrum cephalosporins. OXA-62 shares weak amino acid identity with other class D β-lactamases, the highest degree of similarity being that to OXA-50 (43% amino acid identity). Related blaOXA-62 genes have been identified in other P. pnomenusa isolates but not in other Pandoraea species (144).
The causative agent of melioidosis, Burkholderia pseudomallei, naturally produces OXA-42 (111) and derivatives, such as OXA-43 (111) and OXA-57 and OXA-59 (73). OXA-57, which has also been identified in Bulkholderia thailandensis, is the only enzyme among these variants that has been studied biochemically, and it hydrolyzes penicillins, oxacillin, cephalothin, and nitrocefin but spares ceftazidime. It is uninhibited by clavulanate and uninhibited by NaCl, the latter property being quite uncommon (73).
The blaOXA-63 gene was identified in a clinical isolate of the anaerobic spirochete species Brachyspira pilosicoli (94). It encodes a narrow-spectrum β-lactamase that possesses the unusual property of being well inhibited by clavulanic acid and resistant to NaCl inhibition. OXA-63 shows the closest identity (53%) to FUS-1 (OXA-85) (see below). Very recently, Mortimer-Jones and colleagues (96) analyzed B. pilosicoli srains recovered from humans and pigs in Australia and Papua New Guinea. They identified genes encoding OXA-136 and OXA-137, sharing 94 to 95% amino acid identity with OXA-63 and conferring similar patterns of resistance once the genes were cloned and expressed in E. coli. Interestingly, in that study blaOXA-63-like genes were identified in penicillin-resistant strains only and not in penicillin-susceptible isolates (96).
FUS-1 (also named OXA-85) corresponds to the naturally occurring class D β-lactamase of Fusobacterium nucleatum subsp. polymorphum, a gram-negative anaerobic rod (164). OXA-85 shares only 25 to 44% identity with other class D β-lactamases. It also possesses a narrow spectrum of β-lactam hydrolysis and is not inhibited by NaCl or by clavulanate.
The blaOXA-29 gene was identified in a Legionella (Fluoribacter) gormanii strain recovered from soil (52). OXA-29 shares less than 50% amino acid identity with other OXA enzymes (44% with OXA-18) and hydrolyzes oxacillin, penicillins, cefazolin, and at lower levels, cefuroxime, cefotaxime, ceftazidime, and aztreonam, sparing carbapenems. OXA-29 activity is inhibited by tazobactam but not by clavulanic acid and is also surprisingly resistant to NaCl (52).
The blaOXA-114 gene was cloned from the genome of Achromobacter xylosoxidans (formerly Alcaligenes denitrificans subsp. xylosoxydans) (41). It encodes OXA-114, which shares 56% amino acid identity with the naturally occurring class D β-lactamase of Burkholderia cepacia (as determined by GenBank in silico analysis) and 42% with β-lactamases OXA-9 and OXA-18. OXA-114 has a narrow-spectrum hydrolysis profile, although it shows a very low level of activity against imipenem. Several blaOXA-114-like genes have been identified in all five isolates of A. xylosoxidans tested, making such genes a feature of that species (41). In contrast to the expression of some other natural blaOXA genes, the expression of blaOXA-114 is not inducible (41).
Interestingly, we recently identified Acinetobacter radioresistens, a nonpathogenic and environmental Acinetobacter species, as the progenitor of the acquired CHDL-encoding gene blaOXA-23 (122). Similar blaOXA genes identified in other A. radioresistens isolates encode OXA-23 variants (namely, OXA-102, OXA-103, and OXA-105) exhibiting no more than six amino acid substitutions compared to OXA-23 and conferring similar levels of resistance to carbapenems once expressed in E. coli (123). Since both A. baumannii and A. radioresistens naturally possess class D β-lactamase genes, we believe that other Acinetobacter species may act as reservoirs for other clinically relevant class D β-lactamase genes. Preliminary investigations identified the blaOXA-134 gene in the chromosome of Acinetobacter lwoffii (51). blaOXA-134 encodes a CHDL that shares 63, 58, 57, and 53% amino acid identity with OXA-23, OXA-51, OXA-24, and OXA-58, respectively. The substrate profile of OXA-134, including penicillins and carbapenems, is similar to those of other CHDLs.
Expansion of the knowledge of entire genome sequences leads to the identification of putative ORFs that may encode class D β-lactamases. Some of these ORFs have subsequently been shown to encode functional β-lactamases, while the products of others are not β-lactamases even if such designations are falsely used in the databases, constituting a source of confusion. Some other products are good candidates for β-lactamases on the basis of in silico analysis and require further investigations, in particular accurate enzymatic analyses.
Sequencing analysis of the entire genome of A. baumannii isolate AYE revealed a chromosomal gene encoding OXA-69 that was further characterized (66). OXA-69 shares 62 and 56% amino acid identity with OXA-40 and OXA-23, respectively, and 97% identity with OXA-51, identified in several unrelated A. baumannii isolates from Argentina (22). OXA-69 hydrolyzes penicillins and, at low levels, carbapenems. OXA-51/OXA-69 enzymes do not contribute significantly to the natural resistance pattern observed in A. baumannii. The blaOXA genes for these enzymes are neither inducible nor regulated. Genes encoding OXA-51/OXA-69 derivatives were identified in different collections of A. baumannii isolates recovered from widespread geographical areas (66, 93). Some of these variants had also been identified in carbapenem-resistant A. baumannii isolates disseminated worldwide (21, 47, 158). Up to 45 OXA-51/OXA-69 variants are now known (Table 1), and most if not all probably have identical biochemical properties. Due to the very large and increasing number of OXA-51/OXA-69 variants identified, we propose here to define an alternative nomenclature for these class D β-lactamases specific for A. baumannii, which may include OXA-AB (for OXA-A. baumannii) followed by a number and which would start with OXA-51's being consequently renamed OXA-Ab1, with the next enzyme being called OXA-Ab2, etc. (this nomenclature is presented in Table 1). Interestingly, even though the role of OXA-51-like β-lactamases in conferring the natural resistance pattern is small, they may be at least partially the source of acquired resistance. Indeed, it has been reported previously that insertion sequences ISAba1 and ISAba9 may sometimes be located upstream of the blaOXA-51-like gene in carbapenem-resistant A. baumannii isolates, providing promoter sequences for high-level expression of OXA-51-like enzymes (49, 50, 157).
In silico analysis has identified a putative class D β-lactamase gene in the chromosome of P. aeruginosa. The gene, blaOXA-50, was cloned and expressed in E. coli (54). OXA-50 (also referred to as PoxB [76]) hydrolyzes β-lactams at a very low level (54) and plays only a minor role in the phenotype of natural resistance to β-lactams in P. aeruginosa. OXA-50 variants have been identified in diverse P. aeruginosa clinical isolates recovered from different geographical areas (54). However, the expression of blaOXA-50 in P. aeruginosa is not inducible, as opposed to that of the blaOXA-22 gene in R. pickettii (56).
In silico analysis revealed that the chromosome of Shewanella oneidensis strain MR-1, an environmental gram-negative isolate from lake sediment, harbored a class D β-lactamase gene (126). This blaOXA-54 gene was cloned and expressed in E. coli. OXA-54 shares 92% amino acid identity with OXA-48, the gene for which was identified in K. pneumoniae (see above). OXA-54 possesses enzymatic properties similar to those of OXA-48, significantly hydrolyzing imipenem (with even a higher kcat/Km ratio due to better affinity for this substrate) and not being inhibited by clavulanic acid. Thus, S. oneidensis may be considered a reservoir of CHDLs that are able to spread among Enterobacteriaceae (126).
More generally, the Shewanella genus may be considered to be a progenitor of other related enzymes, since OXA-55 (with 55% amino acid identity to OXA-54) was further identified in Shewanella algae and possesses biochemical properties similar to those of OXA-54, in particular the ability to hydrolyze carbapenems (68). In addition, with the recent release of many genome sequences, it appears after in silico analysis that Shewanella-related species are often (if not always) potential sources of class D β-lactamase genes, as observed for Shewanella pealeana, Shewanella loihica, Shewanella baltica, and Shewanella putrefaciens (http://blast.ncbi.nlm.nih.gov/Blast.cgi ).
In addition, in silico analyses of several other genomes of very unrelated species, such as Limnobacter sp. (Burkholderiaceae family), Idiomarina loihiensis (Idiomarinaceae family), Bradyrhizobium japonicum and Rhodopseudomonas palustris (Bradyrhizobiaceae family), Delftia acidovorans (Comamonadaceae family), Geobacter uraniireducens (Geobacteraceae family), Agrobacterium tumefaciens (Rhizobiaceae family), and Cytophaga hutchinsonii ("Flexibacteraceae" family), reveal some ORFs that may correspond to class D β-lactamases. However, due to the lack of adequate cloning experiments, the ability of all these putative proteins to act as β-lactamases has not yet been demonstrated. These observations, however, suggest that such ORFs may play a physiological but still unidentified role in many gram-negative organisms.
GENETIC PLATFORMS FOR ACQUIRED CLASS D β-LACTAMASE GENES
Most of the class D β-lactamase genes identified in Enterobacteriaceae and Pseudomonas species are located on large, transferable plasmids. Some class D β-lactamase genes found in P. aeruginosa, such as those encoding OXA-13, OXA-17, OXA-18, and OXA-20, have been found to be chromosomally located (37, 99, 108, 118). However, negative results from mating-out assays and the lack of plasmid detection in P. aeruginosa do not rule out plasmid origins for a class D β-lactamase gene.
Most plasmid-located class D β-lactamase genes have been found in association with genetic vehicles responsible for their mobilization. These vehicles are mobile elements, often transposons belonging to the Tn3 family (10, 82). These transposons enable integrons (which are not self-mobile elements) and, consequently, their associated resistance gene cassettes to diffuse and spread. Integrons are genetic structures capable of integrating individual genes in the form of gene cassettes (61).
Among class D β-lactamase genes which have been identified in the form of gene cassettes (140), the most common are the blaOXA-1-like, blaOXA-2-like, and blaOXA-10-like genes (107). There are peculiar features associated with the blaOXA-9 gene identified in a plasmid from a K. pneumoniae isolate (155), since the blaOXA-9 gene was in the form of a gene cassette but, instead of being associated with an integron, was part of the Tn3-type transposon Tn1331 also carrying the blaTEM-1 gene and lacking an integrase gene.
Besides all these blaOXA gene cassettes, there are other blaOXA genes which have been identified in other genetic environments. The blaOXA-23 gene originating from the chromosome of A. radioresistens may be mobilized onto plasmids diffusing into A. baumannii through different genetic structures, such as the composite transposon Tn2006, formed by two ISAba1 elements (30, 100). Alternatively, ISAba4 was found upstream of the blaOXA-23 gene in a single copy (14, 30).
The blaOXA-48 gene has been identified in association with insertion sequence IS1999 in K. pneumoniae (127). The blaOXA-48 gene was part of composite transposon Tn1999, made of two copies of IS1999, with the copy located upstream of blaOXA-48 providing promoter sequences for blaOXA-48 expression (7).
The blaOXA-58 gene, which has been identified only in Acinetobacter sp. isolates, has been associated with a variety of different genetic structures. By analyzing a series of OXA-58-positive A. baumannii isolates recovered in different countries (France, Spain, Romania, Greece, and Turkey), 11 distinct structures were identified (128, 130, 132). Nevertheless, there are always common features, since blaOXA-58 is always bracketed by two ISAba3 elements. The ISAba3 copy upstream of blaOXA-58, as well as other insertion sequences, such as IS18, ISAba1, and ISAba2, may provide promoter sequences playing a role in blaOXA-58 expression (132). For the A. baumannii isolate MAD, which has been at the origin of a clonal outbreak in a French hospital (64, 130), we demonstrated that the entire structure encompassing the ISAba2-blaOXA-58-ISAba3 fragment had very likely integrated into the plasmid backbone by a recombination process (132). In addition to being dependent on IS elements, the level of expression of the blaOXA-58 gene may be related to the gene copy number, as exemplified in one Italian clone exhibiting various levels of resistance to imipenem as a consequence of the blaOXA-58 copy number (13).
Finally, another mechanism of acquisition of class D β-lactamase genes has been identified recently. It is mobilization by peculiar IS elements named ISCRs, which mobilize adjacent sequences by rolling-circle (RC) transposition (151, 152). blaOXA-18 gene acquisition is likely the result of RC transposition mediated by the ISCR19 element (105), and the aquisition of the blaOXA-45 gene is likely mediated by ISCR5 (154).
DETECTION OF CLASS D β-LACTAMASE GENES
One of the major concerns for controlling the spread of class D β-lactamase producers is the absence of phenotypic tests that could contribute to their easy recognition. Although tools are available for the detection of class A ESBLs (clavulanic acid-based synergy tests), metallo-β-lactamases (EDTA-based approaches), and AmpC enzymes (cloxacillin-based tests), there is no phenotypic tool allowing rapid and easy detection of class D β-lactamase producers. However, some class D β-lactamases possess peculiar properties that may facilitate their detection. For example, OXA-13 and its extended-spectrum variant OXA-19 both possess the ability to be inhibited by imipenem (98). Once the enzyme is produced in P. aeruginosa, it is possible to demonstrate this characteristic by placing a disk of imipenem next to one of cefsulodin, a substrate well hydrolyzed by OXA-13 (99). We have observed that this specific property is shared by other class D β-lactamases, such as those belonging to the OXA-10 subgroup (unpublished data). Also, the production of some OXAs whose activity is inhibited by clavulanate or tazobactam, such as OXA-12 from A. jandaei (139) and OXA-18, OXA-45, and OXA-46 from P. aeruginosa (57, 118, 153), may be identified by synergy tests using clavulanic acid-containing disks. However, the result of these synergy tests may be misinterpretated and the corresponding isolates may be considered to be class A ESBL producers.
One alternative approach that may be performed in specialized laboratories is spectrophometric analysis. By using crude extracts and UV spectrophotometry, it is possible to evaluate the abilities of β-lactamase extracts to hydrolyze oxacillin. It is also possible to use the NaCl inhibition property with a referenced substrate such as benzylpenicillin. However, as indicated above, there are some limits in this approach since (i) some class D β-lactamases do not hydrolyze oxacillin, (ii) the in vitro inhibition of their activities by NaCl is not always observed, and (iii) OXA-positive clinical isolates often express additional non-class D β-lactamases. In addition, many CHDLs hydrolyze carbapenems at a low level, interfering negatively with the sensitivity of the technique. PCR-based methods remain, therefore, the "gold standard" for the identification of class D β-lactamases (133, 167).
OXA β-lactamases are distributed in a large variety of gram-negative species. Whereas their acquisition is detected mostly in Enterobacteriaceae, Pseudomonas spp., and Acinetobacter spp., naturally occurring chromosomal genes are increasingly discovered in gram-negative organisms, such as Aeromonas, Legionella, Shewanella, and Campylobacter species. The identification of the progenitor of a clinically significant OXA is an exciting perspective, since it may help to better control the sources of emerging class D β-lactamases, to better evaluate their dissemination, and eventually to better control their spread.
According to Barlow and Hall (11), this diversity of class D β-lactamases, at least those whose genes have been incorporated onto plasmids, is the result of ancient events that occurred millions of years ago. That hypothesis is debatable, since for example, the widely distributed OXA-23 determinant has been found with almost perfect identity in the chromosome of an A. radioresistens strain which corresponds to its natural reservoir. It is very likely that the diversity of plasmid-borne blaOXA genes simply reflects the diversity of their natural progenitors.
It is likely that antibiotic selective pressure is a driving force for the spread of several blaOXA genes. The blaOXA genes identified as having been aquired are often expressed at high levels, and in contrast, the genes are very often silent in their natural progenitors. Mobilization elements (e.g., integrons and IS elements) contribute to the spread and expression of these genes by providing strong promoters. Future work may be directed toward the role of antibiotic concentrations in given environments as a source of enhancement of the spread of blaOXA genes from their reservoirs to secondary hosts.
From a clinical point of view, these OXA β-lactamases should be considered a threat similar to β-lactamases of the other Ambler classes, since many of them possess an expanded spectrum of activity. Currently, CHDLs in A. baumannii, ES-OXAs in P. aeruginosa, and the OXA-48 CHDL in Enterobacteriaceae represent potential sources of clinical failure for many β-lactams (70a). Lack of detection may enhance their hidden and rapid spread among clinical isolates.
The work performed in our unit and acknowledged in the review has been funded partly by a grant from the Ministère de l'Education Nationale et de la Recherche (UPRES-EA3539), Université Paris XI, Paris, France, and mostly by grants from the European Community (DRESP2 grant no. LSHM-CT-2005-018705 and TROCAR grant no. HEALTH-F3-2008-223031) and the INSERM (Paris, France).
- Copyright © 2010 American Society for Microbiology
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text: A big congratulations to long-time Elite Performance Academy client Meredith Paci and Coach Donnie Ruiz on Meredith's outstanding debut at the GNC AllMax Cobourg Naturals OPA Show. Meredith took 2nd Place in Masters Women's Bikini, and 3rd Place in the Open Women's Bikini! She has qualified for Provincials and is laser focused on big things ahead!…Read more


text: Yesterday we showed you Lady Gaga's cover for the June 2011 issue of Rolling Stone. Now, here's a peek inside the magazine as fashion and music's favorite lady takes the issue by storm. Inside the magazine. Gaga looks, well, gentle and graceful, by Gaga standards! Soft lighting, pretty clothes, and a very pretty Gaga. What do you think?


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I've got two bits of news regarding Ligne Bretagne pipes - One good and one not so good. Hitting the good first, buyers purchasing LBs starting this week will notice a little improvement - We've got in our first shipment of stamped pipe bags. Over the past couple of years we've gradually upgraded the entire LB line in various small ways, from improved finishing to improved staining to the move from what started as simple cloth bags and progressed to a nice green velvet bag and now to the same velvet bags embossed with the Ligne Bretagne logo, as seen here:
This move helps further my goal of really establishing Ligne Bretagnes as a quality midrange pipe brand, and less as the red-headed stepchild of Talbert Briars. BUT... and there is always a "but", it brings one major problem and that is the bad news. Part of doing our year-end closeout was running over the price and profit calculations for the various different pipes we sell. Talbert Briars, Ligne Bretagne Collectors, and Goblins all came out good, but standard Ligne Bretagnes, not so good. Unfortunately, they are by far the least profitable pipe we make - We're talking about a whopping hourly wage drop for any time spent working on, selling and shipping Ligne Bretagnes. Given that I like to eat, and am carrying a lot of debts still from our French relocation, I can't afford to allot much of my time to the lowest paid work. Something has to give somewhere. Since pipe folks are often keenly interested in production/price/quality equations, I thought I would put the question to you all, and see what the buyers favored. The choices are three:
* I can make a lot fewer Ligne Bretagnes. This means essentially making them in my spare time, or when I just need a change of pace from making Talberts. Availability would drop off a LOT.
* I can retain current quality and quantity and raise the prices across the standard line to make the pipes profitable to produce. This would entail a pretty sizable price hike, bringing them back into the price range they were when priced in euros. Currently they run $119-$165 on average. Increasing to an equal profit point would put them at $145-195 average.
* I could keep them at the current prices and current output, but scale back the working time invested. Things that would go away would be multi-layer staining, two tone contrasted sandblasts, etc. The bulk of our LB labor is in finishing, so it would basically mean doing much simpler stains. No more golden contrast-stained super bird's-eye fests and that sort of thing. I'm not willing to compromise the engineering, internals, or fit & assembly, so finishing work is basically the only point of fluctuation.
If you're a pipe buyer, think about the three options above and then vote in the poll below. Let me know your preference among the three possible options. And it really does have to be one of the three, too - The only other possibility is to hire someone to help keep production up, and I have no room nor sufficient income to afford to hire an employee.
Which direction would you prefer we take with Ligne Bretagnes?
I look forward to seeing how people vote.
In other news, you've probably noticed that the blog has changed its look! I've been meaning to spiff it up for a long time now, to make it better match the website. Since we're talking about polls, cast your vote in the poll up in the top right to let me know what you think of the new look. The two biggest changes are the re-instatement of the comments system, which had been broken for some time, and the new "Popular Posts" category over in the right-hand column, where Blogger will automatically post links to entries I've written that were most visited.


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Federal District Court Order Granting Defense Motion to Dismiss FCRA Action Against Consumer Reporting Agency Reversed
A consumer (Stephen Levine) opened a store credit card account with a clothing retailer (Structure) through its financial affiliate (World Financial National Network Bank) that the consumer closed in 1998. Credit reports at a consumer reporting agency (Experian Information Solutions) showed that the account had been paid in full and closed. Nonetheless, in 2002 Structure requested credit reports on Levine from Experian, stating that it needed them for purposes of "account review." Structure had not reported Experian with any new information on Levine's account during the preceding four years, and Levine had not made any inquiries to Experian or Structure concerning his closed account. Experian provided copies of Levine's credit report to Structure in May 2002 and in August 2002. Levine sued Experian and Structure for violating the federal Fair Credit Reporting Act (FCRA). 15 U.S.C. §§ 1681 et seq. Experian's defense attorneys moved to dismiss the complaint. The district court granted the motion on two grounds: (1) "FCRA does not suggest that a credit report may only be permissibly obtained for account review during particular points in the parties' relationship" and "Experian had not duty to investigate a facially valid request for a consumer report," and (2) Levine failed to adequately allege damage. Levine v. World Financial Network Nat'l Bank, 437 F.3d 1118, 1119-20 (11th Cir. 2006). The Eleventh Circuit reversed.
The Circuit Court began by summarizing the operative allegations of the complaint: "(1) Levine's account with Structure had been voluntarily closed and paid in full for years; (2) Levine could no longer use the account; (3) Levine had not disputed any information on the account; (4) Structure did not ask solely for information regarding its own trade line but instead requested the entire credit report; and (5) Structure twice requested his credit report from Experian within the span of about three months." Levine, at 1120. Levine argued that Structure did not have any permissible purpose to review his credit report because his account had been closed long before, and that Experian negligently or willfully produced reports in violation of the FCRA because it receives a fee for each requested report. Id., at 1120-21.
The Court of Appeals held that the FCRA prohibited Experian from producing Levine's credit report to Structure "if it ha[d] reasonable grounds for believing that the consumer report will not be used for a purpose listed in section 1681b," Levine, at 1121 (quoting § 1681e(a)). Experian argued that so long has it receives a facially valid request for a consumer report it may produce it "notwithstanding any reasonable grounds to believe that the request is instead made for an impermissible purpose." Id. The Circuit Court soundly rejected this argument, explaining at pages 1122:
Levine alleges that his report was twice requested within the span of a few months despite his account being closed for years with an undisputed zero balance. A simple recitation of "account review" by a former creditor does not automatically absolve Experian of its duty to protect confidential information when there are reasonable indications that the request was for other purposes. In light of the legislative findings in FCRA and considering the standard of review, we conclude that the question of whether Experian had "reasonable grounds" to believe that Structure intended to use Levine's consumer report for an impermissible purpose, or whether Experian made "reasonable efforts" to verify the validity of Structure's request, is a fact intensive one that is not resolved by the pleadings.
The Court also held that Levine had adequately pleaded damages. Id., at 1123-25.
NOTE: The Eleventh Circuit expressly reserved deciding "whether there is an absolute prohibition against such requests [for consumer reports] by former creditors for accounts that are closed and paid in full." Levine, at 1122.
Download PDF file of Levine v. World Financial Network National Bank
Comments are closed.


text:
WTO Public Forum: Gender Responsive Trade Policy Drives Growth
How does Gender responsive trade policy drive growth?
Join Women@theTable at the WTO Public Forum 17
26 September, 11:30 - 13:00, Room S-1
with an extraordinary panel from the private sector and member states who are leading as we explore
How Trade Can Help Achieve Sustainable Development Goal #5: Gender Equality
Moderated by:
Caitlin Kraft-Buchman – CEO/Founder, Women@theTable
Moderated by:
Caitlin Kraft-Buchman – CEO/Founder, Women@theTable
The recent addition of a chapter on trade and gender to modernize the 20-year-old Canada-Chile Free Trade Agreement, and the chapter on trade and gender between Chile-Uruguay have set a new standard. Will the EU now follow suit and reinforce the coordination and supervision of women's rights in trade? Share in our dynamic discussion with private sector and member states articulating how gender is a genuine core trade issue and why it is of increasing importance to negotiators, governments, and the business community.
How do we apply gender perspective to economic and trade issues to ensure that economic growth benefits everyone? What are the modern frameworks on issues related to trade and gender, including women's entrepreneurship and development of gender-focused indicators? How do we encourage women's participation in national and international economies including but going beyond entrepreneurship ? How can free open progressive trade be harnessed to help us achieve Sustainable Development Goal #5?
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text:
If more golf carts looked like this custom Batman Tumbler golf cart spotted on the Warner Bros. studio lot, I'd sure play a lot more golf!
HA, HA! Just kidding. It'd take a hell of a lot more than fancy carts to get me on the golf course. We're talking about attractive topless ladies at every tee offering free massages* and magical iPhone 4s that work on Verizon.
Then we can talk about your stupid put-the-tiny-ball-in-the-tiny-hole game.
Via This L.A. Life, who invented a game about putting a medium-sized ball in a tiny hole. So far it hasn't taken off.
* Wiiiiink.
Find LEGO Batman Tumbler kits on Ebay


text:
Click the link below to submit a question:
Submit a Question
The purpose of the $1,500 assignment fee is to cover administrative and legal costs incurred by the EDC each time the counterparty chooses to assign the PSA. This fee is applied on a per assignment basis and is applicable each time the PSA is assigned to a new party, including assignments related to, or in connection with, a sale of the property and/or the project. Please check the PSAs for further information.
On the Webinar, you mentioned "Developer Caps". Where can I learn more about what a Developer Cap is?
The Developer Caps are described in Section 5.1.9 – 5.1.12 of the RFP and only apply to projects in the ACE and JCP&L service territories only. The Developer Caps set maximum percentages for amounts awarded to developer/majority owners by segment and by EDC.
Does the Interconnection Application need to be accepted by the utility prior to submitting the EOI and Bid Package?
No. The RFP does not require the Interconnection Application be accepted by the utility prior to submitting an EOI or Bid Package.
Due to the time required for issuance of an SRP #, would an EOI without an SRP # be acceptable? (if it were to be accompanied by proof that the SRP # process has been initiated)
No, an SRP # must accompany the EOI. Please see the announcement posted to the RFP Web Site on 3/31/2015 which provides information furnished to us by the SRP Program Administrator:
1 - Can the system be built after the EOI and still qualify, even if it is allowed to be connected PTO prior to the awards announced?
The system can be constructed after the EOI and still qualify.
Per Section 5.1.9, the following Developer Caps (kW) for the first solicitation round apply unless the RFP is undersubscribed:
Yes, the following summarizes the total fees:
With respect to an original submission of a Proposal, the successful proposers must provide a cash deposit within 14 days of the Board's written order approving a bid selection. If the cash deposit is not made, then the project will be rejected. If the cash deposit is made and the PSA is not executed within the prescribed time, then the cash deposit will become property of the EDC. Beyond the cash deposit and application fees, there are no other costs or damages in the event the PSA is not executed within the prescribed time. The prescribed time is within 5 business days of being given a Final Notice of Award by the EDC per Section 4.1.7 of the RFP rules. In the case of a subsequent re-submittal of a Proposal, the successful re-submitting proposer, again, would be required to comply with the same requirements including application fees and providing a cash deposit within 14 days of Board's subsequent approval as evidenced by a subsequent written Board Order.
I note that there is a requirement to submit SRP registration by 4/13 at 4pm. One of the requirements of SRP registration is an executed contract for the system. For the project I am applying for, I do not yet have an executed contract for the purchase of the system; however I have an executed PPA dated 3/13/15. Will this project meet the requirement for SREC registration? Please advise.
Please direct all questions with respect to the SRP registration process to the SRP Program Administrator:
I cannot find the summary of average winning bids in prior solicitations. I thought I saw it somewhere but I cannot find it anymore.
Average winning bids from prior solicitations are available in the "Announcements" section of the www.njedcsolar.com web site.
When will Round 2 of be announced and due for bid? Thank you.
There will be approximately 100 days between solicitations. No date has been announced yet.
Can any project be included as long as it has not received a "permission to operate" yet from the utility company, even if it received an initial SREC registration application number last year?
Provided that the initial SRP registration continues to be valid at the time, the response to question #5 above (as repeated here) would still be applicable:
Does every project that gets accepted pay a $1500 project assignment fee?
The fee is payable if the awarded PSA is assigned to another party. Otherwise, the fee is not applicable to parties who have not assigned the PSA.
In the event that the Host and Owner are different legal entities with a common parent company, Appendix C (set forth in Appendix 5 to the RFP) would still need to be completed by the Host that owns the premises or facility where the project will reside. Sharing the same parent company does not change the distinction between the Host and the project Owner and the need for Appendix C.
The Supplement Form has been posted to the Documents section of the RFP web site.
I submitted the SREC RFP Registration by the 4pm 4/13 deadline specified on the SREC II Program website. For reference, the account number for the Site Host on my Registration Form is XXXXXXX. It is not specified in the RFP materials as to when I should expect to be notified if my project cleared the process and if I will receive a NJCEP Application Number to submit in time for the EOI due on Monday, 4/20. Please advise how this process is expected to work. Thank you.
Please submit the EOI with the NJCEP Application number once you have received it from the SRP Program Administrator. The SRP process is separate from the RFP and managed by the SRP Program Administrator. EOIs are due on Monday, April 20 and require an NJCEP Application Number.
We are very interested in responding to the RFP under the SREC- Based Financing II. However we have not submitted an application for the SRP #. Does this mean that we are disqualified from the project?
Per the instructions posted in the announcements section of the RFP web site, the SRP Administrator has indicated that it needed submittals by April 13th at 4 PM in order to process the SRP # in time for the April 20th deadline established for the EOI. To the extent you are able to receive an SRP # in the meantime, you may bid into this first round. If unable, you will still have the opportunity to bid into subsequent Rounds.
I'm interested in submitting a bid for the SREC II RFP. However, one question I have is that is this RFP just for Round 1? If so, if I don't respond to this RFP then will I not be allowed to submit a proposal for Round 2?
This RFP is Round 1. Bidders who do not respond to this RFP will be able to bid into subsequent rounds. There will be a total of 9 rounds over a span of 3 years.
Yesterday at the NJ Renewable Energy meeting I asked a question concerning construction of a system timing. You were not able to hear my question and Scott Hunter said to look on your web site and if it is not clear, to email you. I hope you receive this, since the web site has no "contact us" page. Having been on the site, it is even more ambiguous. I asked can we begin construction on a Segment 1 system, now that the bid has been submitted. It appears in 1.2.10 of the RFP that it is permitted to be built, (after bid is submitted) but if it is interconnected before the SREC PSA is executed, than any SREC earned during that time will not be purchased by the EDC. Am I correct in assuming that these exempt SRECs are earned and able to be sold on the open market? You stated some time lines that you hope to meet. These help us to determine when we can build and apply for the interconnection that is needed. On the web site under "Calendar" the dates are different than what you stated yesterday. On the site it states the Board meeting will be May 27, and you said June 17. Your stated schedule is 3 weeks later than what is posted. Looking at the "Announcement" page, there is no mention of these later dates. I am also subscribed through the "Subscribe for updates" page, and nothing has come to me through that.
In response to your question, a Segment 1 project cannot (i) request final interconnection approval from the EDC or (ii) be interconnected prior to the Board Order approving the results of the solicitation. Projects that are either interconnected or request final interconnection approval from an EDC prior to the Board's approval of awards will be ineligible to enter into a PSA with the EDC. See section 1.2.10 of the RFP for further detail.
How can I learn of future RFPs?
The next RFP is tentatively scheduled for late June. Please register for updates on our mailing list here and periodically check the RFP web site located at www.njsolarprogram.com.
I have two questions:
The next RFP is tentatively scheduled for late June. Please register for updates on our mailing list here and periodically check the RFP web site located at www.njsolarprogram.com
It has been submitted in accordance with the schedule posted.
I want to confirm if there is a required first priority lien placed on the solar equipment for those systems that participate in the EDC SREC II solar financing program?
I know that there is such a lien to those facilities that participate in the PSEG Solar Loan III program but just clarifying if that is the same case for this program as well.
the SREC II Program, there is no lien placed on the solar equipment. See
the applicable SREC Purchase and Sale Agreement (the "SREC-PSA") for details
regarding the EDC's purchase of Transferred SRECs from projects that enter into
the SREC-PSA.
Is there any way for me to see what prices were bid for the First Solicitation. It says in the Board Order that average price per SREC will not be disclosed due to too few distinct price offers – but we are interested in participating in the next solicitation, and are looking for some benchmark or guidance regarding where bids should be.
Any help in this regard would be very much appreciated. Even if a comparison to cleared bid prices from the first round of EDC SREC solicitations would be helpful.
As noted in the July 23, 2015 Board Order, the results from the first round
will not be disclosed.
Is there an actual application package? I cannot find it. Just submit first page of SREC registration form and send it in? No average price? How about a range for residential? Can someone who is about to start construction and already have an SRP # apply?
Timing of Build and Interconnection – Is it correct that a project that winds a bid in Round 2 would not be able to receive 'Permission To Operate' until January 2016 once its PSA has been executed? Is it also correct that a project cannot request "Permission To Operate' until December 2015 when winners have been announced? If these dates are not correct, when is the earliest that a project could request PTO (ie. request final interconnection approval from the EDC), and receive PTO (ie. approval to operate).?
Please see Section 2.3.1 (second and third bullets) of the RFP which refers to the timing of construction completion, interconnection (i.e., receipt of permission from the EDC to operate) and requesting final interconnection approval from the EDC. Also see, Table 2-2 (RFP Round 2 Target Schedule).
Post-PSA Process – After a PSA has been executed, what additional paperwork/processes are required?
With respect to the RFP, no additional paperwork/processes are required after the PSA has been executed. Thereafter, all paperwork/process requirements will be as set forth in the SREC PSA.
Automatic Transfers – How do we set-up automatic transfers of created SRECs from GATS to the EDC's accounts?