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Ten Reasons
Hire An Artist
Peter Funt, 6/9/2014 [
Hillary's Puzzling Demo Gap
By Peter Funt
The headline on a new poll about Hillary Clinton's presumed presidential bid is that fallout over Benghazi hasn't hurt her chances. The data show 67 percent of all voters believe she is a "strong leader" and 60 percent find her to be "trustworthy."
With the release of her book, "Hard Choices," this week, in which she addresses Benghazi and other challenges during her time as secretary of state, Clinton is oiling the machinery for a run. The ABC News and Washington Post poll results seem to fit perfectly: despite Benghazi and relentless attacks by Republicans, her numbers remain just about where they were when she entered the 2008 race as the overwhelming favorite among Democrats.
But deep inside the poll -- so far down that the Post didn't even mention them in its story -- are some details that the Clinton team might find troubling. It seems that as people become older, wealthier and more educated they are somewhat less likely to support Mrs. Clinton.
Put another way: Clinton is weakest among people who are most like her.
Take, for example, the question of whether Clinton "has new ideas for the country's future." Among adults 18-39, an impressive 61 percent believe she does. But when those over 40 respond, the positive responses plunge to 49 percent. Within her own age group of people over 65, only 48 percent believe she has new ideas.
Among those who, like her, have a college degree, only 47 percent believe she has new ideas. However, among people with no degree, it's 56 percent.
An even more puzzling result involves the statement, "She understands the problems of people like you." Among people under 40, there was a 59 percent positive response. But affirmative answers among those over 40 yielded only 51 percent.
A remarkable 75 percent of people under 40 believe Clinton is "a strong leader." Yet, among Clinton's own age group the number slips by 14 percentage points. If she runs and wins, Hillary Clinton would be 70 when she takes the oath of the office.
The question regarding leadership provides similarly surprising data among those who, like Clinton, are more affluent. Those earning less than $50,000 a year, affirm her leadership by 73 percent; those with higher incomes by only 63 percent.
The Post-ABC poll does not cross-reference demographic factors, such as age, education and wealth, with party affiliation. It could be that older, wealthier Republicans account in part for the gap in Mrs. Clinton's numbers. However, in 2012, President Obama beat John McCain handily among the most educated voters as well as the most affluent. McCain's greatest strength was among middle-age males.
In her book Clinton insists she remains undecided about running. If she does seek the presidency, she'll have to work harder than might have been expected to gain support among her peers. Why are those who've known her longest and share some of her demographic traits, beyond gender, the most skeptical?
Answers may be provided in the coming days as Mrs. Clinton makes as big a media splash as modern book-promotion can provide. She'll be taking questions from all corners and on virtually all networks about "Hard Choices."
She writes, "Ultimately, what happens in 2016 should be about what kind of future Americans want for themselves and their children -- and grandchildren. I hope we choose inclusive politics and a common purpose to unleash the creativity, potential, and opportunity that makes America exceptional."
So far, those with the longest, toughest road ahead seem most willing to buy in.
Peter Funt is a writer and speaker. His book, "Cautiously Optimistic," is available at Amazon.com and CandidCamera.com. © 2014 Peter Funt. Columns distributed exclusively by Cagle Cartoons, Inc., newspaper syndicate.
This column has been edited by the author. Representations of fact and opinions are solely those of the author
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Noticing These 3 Symptoms in Your Dog? They May Have Food Allergies.
As you probably know by now, your dog's diet has a lot to do with their activity levels and overall well being. Food is your dog's first source of energy and nutrition. It's also their first source of irritants and pollutants. Often, your dog will let you know that they've eaten something they don't like by vomiting or displaying other gastrointestinal distress. Sometimes, however, symptoms may show up differently than you'd expect.
Beef, dairy, and wheat are among the three most common food allergies dogs experience. Here's how you can tell your dog's system doesn't agree with some of the ingredients in their food:
Constantly itchy skin. Itchiness is one of the most commonly cited symptoms of food allergies, usually accompanied by pink skin underneath. If your pup seems to itch without relief or is losing patches of hair due to too much scratching, there's a good chance the cause is a food allergy - especially if over-the-counter shampoos and solutions don't seem to work.
Reddish-brown stains around the mouth and paws. Generally, these red patches can be attributed to yeast colonies, which are better able to grow at infection sites. These patches are often itchy and you may notice your dog licking its paws more than usual.
Frequent ear infections. While dogs that are frequent swimmers are prone to swimmer's ear, food allergens can also cause recurring infections in your furry friends. Check the ears for dark brown or black, stinky buildup. This, too, is usually caused by a yeast infection triggered by the appearance of an allergy.
What's the next step if you think your dog has a food allergy? The most reliable test is through an elimination diet, which can take about 4-6 months to complete. In a program like this, you'll take your dog's diet back down to the very basics and slowly reintroduce each ingredient to determine how it affects their overall wellness. As always, consult your veterinarian before going on a program such as this.
Human-Grade Dog Food for Healthy Coat
Many pet parents find that their fur babies have healthier skin and coat after switching to a human-grade diet. Keeping your dogs on a simple, natural diet makes it easy to tell what ingredients work in harmony with their systems. Get human-grade dog food delivered to your door from Pawsitively Pure. Click here to browse our products.

subset: commoncrawl


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North Star Roller Derby held a closed Mini Tournament last night, (2/17/18) At their practice space in Northeast Minneapolis. Thank you to Wijadi Jodi for inviting me to help cover the event.
I had only been to the NSRD practice space once before and since I had been invited to shoot on short notice I didn't really know what the plan was for the evening. Soon as I walked in the door I got excited though. Everyone was doing their part getting the space ready for the night's bout.
The first thing I noticed was the light. Normally NSRD hosts their bouts at the Lee & Rose Warner Coliseum and I always find myself cringing a little. The light there is not pretty, it's harsh top-down lighting that always gives off a weird greenish yellow cast that makes everything garish. The practice space offered a very welcome change of scenery, space is smaller and all the light was being provided by work lights so everything was much softer, more shadowy and in my opinion much more interesting.
I didn't realize how dark it was in the space until I started talking with Wijadi, and taking test photos. But I found it to be an excellent opportunity to really push the abilities of my camera which was quite exciting for me. I really loved shooting in the practice space, it is a much more intimate setting then the Coliseum and the way I ended up covering the action I feel shares that intimacy. I found myself shooting tighter, trying to get the struggle of players moving through the crush of bodies in the pack. The breakaways feel more earned, and the low light and noise gave everything more of an edge and mood then I have found myself getting from my previous NSRD bout coverage. I was also FINALLY able to catch a few good Starpass shots that I have been aiming for this season!
I hope you enjoy these photos as much as I do. Thank you again to Wijadi Jodi for inviting me to help shoot.
See you at the next bout!


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Launch of Apollo 16, Kennedy Space Centre, April 1972
Launch of Apollo 16, Kennedy Space Centre, April 1972
Vintage chromogenic print, 20.2 x 25.4 cm
[NASA description] The huge, 363-feet tall Apollo 16 (Spacecraft 113/Lunar Module 11/Saturn 510) space vehicle is launched from Pad A, launch Complex 39, Kennedy Space Centre (KSC), Florida, at 12:54:00.569 p.m.(EST), April 16, 1972, on a lunar landing mission. Aboard the Apollo 16 spacecraft were astronauts John W. Young, commander; Thomas K. Mattingly II, command module pilot; and Charles M. Duke Jr., lunar module pilot.
About the Apollo 16 mission here.
All of our photographs are sold unframed.
--> Add framing to your order here.


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Liberty of London: Tana Lawn Wiltshire A in Multi
Liberty of London: Tana Lawn Wiltshire A in Multi
Wiltshire Tana Lawn from the Liberty Art Fabrics collection.
Wiltshire is a leaf and berry pattern, designed for Liberty in 1933. Like Betsy, it was created by the designer with the initials DS. It was redesigned by Tana in 1968, and has been on classic Tana since 1979.
Composition: 100% cotton
Width: 53" (137 cm)
Weight: 76 g/m²
Sold in ¼ yard increments. Additional increments will be cut continuously. For example, for 1 yard of fabric, select a quantity of 4. You will receive a piece of fabric that measures 53" wide by 1 yard long.
In 1875, Arthur Lasenby Liberty began selling imported fabrics from a small shop on London's Regent Street. The popularity of these beautiful textiles meant the Liberty name soon became synonymous with a very distinct and highly fashionable style of fabric. Today, Liberty is London's best-loved department store and its cutting edge design studio remains at the forefront of fashion across the world.


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She shows up and in my judgmental but silent voice I thought "Gee Girl, why'd you even bother?"
I was even more surprised to see an administrator and her assistant hustling around to get her chair put back in place. Where was their scowl?
Later, learned that this young graduate was late because first she went to our school (we have the ceremony at a local university arena, a good 30 minutes from where the school is). It was locked up and she didn't know what to do.
You see, this young graduate has been living in her car since her mom took off for who knows where. This young graduate has attended 5 different high schools in 4 years following her mom and apparently this time she decided she was staying put. Yes, even if it meant she lived out of her car until she finished up her classes and earned her credits.
Why did she go to the school in the first place? Seems she went there in hope of being able to take a shower in the locker room, something our administrators have been aware of and assisting with.
It breaks my heart to think of the determination it took for her to make it to graduation. What thoughts she must have had as she raced downtown, hoping to make it in time. And how heart warming it was to see a usual gruff-by-the-book administrative assistant smile at this young lady and hurry off to make sure a chair got placed back in her row.
What a timely reminder for me...I may know their name (as was the case today), but that doesn't mean I know their stories. I may have heard what she had done, but clearly I had no idea what she had been through.
Congratulation to this young graduate, and the 550 classmates that received their diplomas today!
Here's a few of my favorite photos from today's graduation...
Way to go ILC graduates!
This young man, was one of the first to arrive...he just couldn't stop smiling the whole time...
The beauty of working in the neighborhood school, I've known this fella since he was in 1st grade!
Twins...this young man has a heart of gold and the love he shows in helping care for his sister...
well, you'd just have to see it to believe it!
We shouldn't have favorites...but this one is one my favorite clinic visitors!


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Real estate developer La Vista has launched an OOH campaign to announce a new residential project in the upcoming Administrative Capital of Egypt.
Using dark backgrounds and white typo, the ads present the logo and name of La Vista City, informing viewers that the project will be located in the Golden Square Extension of the New Capital.
The campaign uses a wide variety of large-format OOH types, such as double-decker poles, mega signs and uni-poles in gate and serial combinations of enhanced visibility across the Greater Cairo.
Industry: Real Estate
Brand: La Vista City
Advertiser: La Vista Developments
OOH Size: Double-decker Pole | Gate | Large uni-pole | Mega-Sign | Serial
Country: Egypt
Region: North Africa
Tags: New Administrative Capital | Product Launch | Golden Square Extension
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Canada Markets
Canada's February Trade Data
Statistics Canada's Canadian international merchandise trade report shows the country's total exports fell 5.4% to $43.7 billion in February, a disappointing follow-up to the record $46.2 billion reported in January. A quote from CIBC suggests "after a string of stronger-than-expected Canadian data releases, February trade data spoiled the party." After reaching a trade deficit with the world of $628.4 million in January, a six-month low, Canada's trade deficit rose to $1.9 billion in February, a four-month high.
Also disappointing is Canada's overall trade with the U.S. While the U.S. also reported disappointing trade data on Tuesday which showed that country's trade deficit widening from $45.9 to $47.1 billion in February, Canada's trade with the U.S. waned during the month, with our country's trade surplus shrinking from a six-month high in January to $2.678 billion in February, a three-month low.
Statistics Canada's Farm, fishing and intermediate food products group shows total exports at $2.775 billion was up .9% from February 2015 while reaching a two-month high (seasonally adjusted). A narrower classification, the Farm and fishing products group, showed February exports at $2.170 billion, down 4.5% from year-ago data and at a three-month low. The dollar value of wheat exports reached a two-month high while the dollar value of canola exports reached a five-month high. Statistics Canada commentary points to exports of "other food products" falling 10.4% from last month which is largely tied to a drop in lentil exports.
Here's a look at miscellaneous pulse and special crop export data from today's report:
Lentil exports in February totaled 109,092 metric tons, the lowest monthly volume shipped since August. Year-to-date exports total 1.82 million metric tons, 35% higher than the same date last year and 75.6% higher than the three-year average. With cumulative exports now at 75.9% of the current AAFC export target of 2.4 mmt, exports are well-ahead of the pace needed to reach this target given data representing 58% of the crop year.
As seen on the attached chart, monthly activity has fallen sharply from the 526,630 mt shipped in October, with February volume falling below the volume shipped in February 2015 as well as the three-year average. As expected for some time, this year's activity is expected to be a test of overall supplies with current estimates suggesting ending stocks will slide to just 100,000 mt or just 4% of annual demand. The Stocks of principal field crop report, due to be released May 6, will be watched closely, given prices remaining at record highs leaving little or no incentive to store product. Today's Statpub.com price report shows large greens averaging $75.94/cwt delivered to Saskatchewan plants ($41.69) while small red lentils are averaging $44.53/cwt ($33.32), with approximate year-ago prices in brackets.
Dry pea exports in February totaled 150,467 mt, a three-month low and the second-lowest monthly volume moved this crop year. Cumulative crop year exports as of August 1 total 1.823 mmt, down 13.6% from year-ago volumes and 5.7% above the three-year average. Cumulative exports are just above the pace needed to achieve the 3 mmt export target set by AAFC, with 60.8% of the target reached as of February, which represents the first 58% of the crop year.
Chickpea exports for February totaled 14,269 mt, the second highest monthly volume shipped this crop year. Year-to-date, 83,773 mt has been shipped, 172% higher than the volume shipped as of February 2015 while 181% higher than the three-year average. Total exports are estimated at 110,000 mt for the year by AAFC, with year-to-date exports reported at 76.2% of the annual target with data covering 58% of the crop year. The current pace of exports suggests that export volumes will exceed this target and ending stocks will be much tighter than the 50,000 mt that current AAFC estimates would suggest.
Monthly data suggest that 11,384 mt of mustard was shipped in February, the highest monthly volume shipped this crop year. Year-to-date, 68,278 mt has been shipped, 4.4% behind year-ago volumes and 6.3% behind the three-year average. Year-to-date volumes suggest current exports at 56.9% complete, just slightly behind the necessary pace needed to reach the annual export target given that current data represents 58% of the crop year.
A total of 11,935 mt of canary seed was shipped in February, the third-highest monthly volume shipped this crop year. Year-to-date, 87,167 mt has been shipped, 16.2% behind year-ago volumes and just slightly behind the three-year average. Current AAFC estimates suggest that 145,000 mt will be shipped this crop year, with current volumes representing 60.1% of the current target, while just slightly ahead of the pace needed to reach this volume.
DTN 360 Poll
This week's poll asks what you think about the recent release of the final report released by the Canadian Transportation Agency review panel. You can weigh in with your thoughts on this week's poll, found at the lower-right of your DTN Home Page. We thank you for your input!
Cliff Jamieson can be reached at email@example.com
Follow Cliff Jamieson on Twitter @CliffJamieson
© Copyright 2016 DTN/The Progressive Farmer. All rights reserved.


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Reward for Not Investing
Not Investing Can Increase Your Purchasing Power
What this means is that, during these turbulent financial and monetary times, deciding to defer investing part or all of your money and opting instead to save it for the time being by simply exchanging it for gold or silver bullion (which are monetary metals or another form of money yet to be fully recognised again as such), can be quite rewarding. This is the case, in our opinion, because our current global monetary system is failing and our fiat currencies are losing their purchasing power. In other words, it is no longer necessary to invest to increase one's purchasing power.
The two monetary metals are priced in US dollars, but investors in any country can purchase bullion by converting their currency. The price in USD of gold has now increased on average by 9.0% per annum or a total of 369% over eighteen years. The silver price has done less well during the same period, but still rose 252% or 7.2% per annum. How do these gross (before any tax and fees) rates of return compare with your investments? If your base currency is not the USD, then the following document will be of particular interest to you. It shows what those gross rates of return were across ten major currencies and for each of the past eighteen calendar years, as well as cumulatively.
The document presents two tables, one for gold and one for silver, showing the reward for hoarding the metals in ten major currencies, based on their price change in those currencies. For example, you'll see that for 12 consecutive calendar years, the price of gold in USD (and broadly across all ten currencies) increased; but it dropped significantly (by 28.1%) in 2013. In 2014, the price in USD stayed much the same (rising only 0.1%). In 2015, it dropped again; this time by 12.1%. Then, in 2016 and in 2017, the gold price resumed its course of rising prices. In 2018, the gold price fell by less than 1% in USD, but rose in 7 of the 10 currencies. After 18 years, the gold price is up on average across all currencies by 8.3% per annum or by 333% cumulatively. This price appreciation represents a significant increase in purchasing power which can be acheived by simply hoarding the physical gold metal.
For an investment to have outperformed gold hoarding, its performance measured in currency units must be greater, before tax and fees. For US based investors for example, an investment must have earned more than 9.0% p.a. during the past 18 years to have been more rewarding than simply hoarding gold. On the other hand, some currencies have been much stronger over this period than the US dollar: for example, the NZ dollar. At the beginning of 2001, it took more than two NZ dollars, i.e. NZ$2.25, to buy one USD; at the end of 2018, it took one-third less (i.e. NZ$1.49) to buy the same USD!
As a result, hoarding gold (or silver) has not yet been as rewarding for New Zealanders as it has been for Americans or indeed most anyone else based in the countries of the eight other currencies considered. But what if the NZD/USD exchange rate was to fall back to US$0.50 (after all, it did fall from US$0.80 to US$0.50 in 2008 and another crisis could be just around the corner...)? Then the gold price in NZ dollars would rise by 33%, even without any increase in the price of gold in USD, and the reward or cumulative increase in purchasing power for the NZ hoarder of gold would rise from 6.4% per annum or 206% (as indicated in the table) to 8.2% per annum or 311% cumulatively.
In other words, although gold is indeed priced in USD and its price is normally expressed or quoted in USD only, the true value of hoarding gold is the increase in the purchasing power you get from it in your own currency. The question for investors to ask themselves is: how confident are you that your country's central bank will preserve the purchasing power of your currency? In fact, since no central bank anywhere has ever succeeded at preserving the purchasing power of its fiat currency over time, the question then becomes: do you prefer to place your full faith in central bankers or do you also want to protect yourself from their prolonged and systemic debasement of fiat money? The choice is yours.


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Summer Holiday in the French Alps? Mais Oui, Merci!
Ah, France. It's been a busy few months. Arthur's gig is complete, and we are conveniently close to the Bourgogne, where cycle paths wind through ripening summer vineyards dotted with scenic villages along the Route de Vin. We have no commitments for the next few weeks, hurrah! This doesn't happen very often, time for a break in the warm summer sunshine.
I tap my way through the maps on Tom, our GPS, looking for a nearby village campsite, when a conspicuous blob of green appears on the left of the screen. Nature!! Uh-oh, our proximity to the Bourgogne also means we are within a days drive of the Alps, and I'm not the only one who has noticed. There on Tom's screen is a thick red tell-tale, pre-planned route, snaking across the GPS leading from down here in the valley, to up there, on the mountain.
Arthur grew up in the Netherlands, or, en français, Pays-Bas, literally translated, Lands-Low. If you've ever met a Dutchman anywhere in the world outside of the Netherlands, I'm guessing it was on Lands-High. Lowlanders are so attracted to mountain-tops, I didn't even try to plead my case. The apparition I had conjured up of me basking in the sunshine, feet up, book in one hand, chilled glass of Chardonnay in the other quickly evaporated. I pulled out a few layers of merino wool, cranked up the seat-heater, and we began to wind our way toward higher ground.
I have a 4-season wardrobe of merino wool that travels with me to all destinations, I have my seat-heater set to high many months of the year, and I yearn for 25 degrees of warmth. Arthur, on the other hand, is the guy in a t-shirt on a frosty morning, and he positively brightens up at the thought of a walk in the rain. So, as in much of what we do together, we have to find compromise when it comes to vacation. The Alps in summer was not exactly what I had in mind for my holiday in France, but I was willing to give it a go for a few days.
If at all possible, Lowlanders will avoid taking tunnels through mountains, always assuming that there must be a pre-tunnel, single-track hair-pinned hair-raising cobbled "pass" rising to the summit, and more often than not, there is. After a few hours of ascent on a winding goat track, we burst out above the tree-line somewhere around the village of Beaufort. Rather than looking out the window, I had been sitting in a slump, reading through my guide book trying to find some good reason within its pages to be spending my summer holiday in frigid temps (yes, I was sulking). When I looked up, I saw a bright turquoise lake set in a brilliant green meadow littered with wildflowers and stone cottages. I dropped my book. The scene was spectacular.
I jumped out of the van into the chilled air and gazed around looking for something that would indicate a nearby campsite. Ha! Not a chance – the hoards of holiday-makers were in the Bourgogne. No market for campsites up here. Electric wire fencing was a continuous barrier to the meadow, and finding a small, unblocked road leading into the hills, proved futile, so we pulled the van into a nook on the side of the road. Our view overlooked an alpine bowl, about a kilometre wide and a kilometre long, with sides reaching up toward the sky and disappearing into the clouds. All around us were cows. So many cows!
There was some daylight left, so I pulled on more wool and my best wind-breaking jacket, laced up my hiking boots and we headed off to explore. We followed a rushing creek through the meadow, past a vacant stone cottage, and up a rocky hillside trail. I stopped many times to catch my breath and look down into the valley. Electric Blue was far away, a tiny toy on the edge of the bowl. We descended a small hill and came across a local farmer who had come to milk his many cows at a moveable milking station. Arthur spoke to him in French and then filled me in; The farmer came up from the valley every day at dawn and dusk to milk. He was the chief of the Beaufort Dairy Cooperative, a group of 184 farmers. He explained that these were the Beaufort Cows grazing in their summer meadow, their milk used exclusively for Beaufort Cheese. He told us to come back with a jug for fresh milk.
We spent three days here among the cows, hiking higher and higher into the alpine, never tiring of the challenge or the view. One day I slid down a green hill on my bum to find a crevice out of the chilly wind for our picnic, and lay there in the flowers, marvelling that there could be such a gorgeous spot on earth. Each day we drank warm, sweet milk straight from the cows, and awoke to the sound of hundreds of heavy bells donging and clanging as a herd came to check out the curious, bright blue object with the tent on the roof. It was magical.
While Arthur threw himself into the freezing cold creek each morning for a wash. I did not. I put my odor-resistant merino to the test, and after three days, it was time for a plunge into a warm lake. We left the cows behind and wound our way down the mountain to Lac d'Aiguebelette near Annecy. A summery place with sunshine and warm water! I put up my feet, read my book, and sipped Chardonnay while I munched on Beaufort Cheese.
The Alps in summer? I wouldn't have missed it for the world.
Hover over the pictures (or choose landscape on mobile) for some images of my summer holiday in the French Alps


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New Summer Hours/
Starting today, Fogama will be on a new summer schedule: on Sundays (ONLY) we will be Open from 5pm-10pm
Starting today, Fogama will be on a new summer schedule: on Sundays (ONLY) we will be Open from 5pm-10pm
This Mother's Day, stop by Fogama where every mother will receive a free glass of sparkling wine. Make your reservations today!
HAPPY MOTHER'S DAY from your friends at Fogama!
Roses are Red, Violets are Blue, Come to Fogama for a Special Dinner for Two!
Tonight's Valentine's Day Special includes:
From the Kitchen: Lobster Bisque Soup & Tilapia with Black Bean Sauce
From the Sushi Bar: Lover's Roll (spicy crunch crab meat with avocado on the inside wrapped with tuna)
All couples will receive a complimentary sparkling wine from the house. Happy Valentine's Day!
There are two sample dishes to pick from:
Small Tray for 4-5 people $110
House Dumplings
Fried Calamari
Rock Shrimp Tempura
6 Regular Rolls
2 Special Rolls
Large Tray for 6-8 people $130
Same Appetizer
7 Regular Rolls
3 Special Roll
Please call for information
While chefs require razor-sharp carbon steel knives in their kitchens, restaurant owners need nerves of steel. Just ask Steve Ma, who just opened his Japanese restaurant Fogama at 92 Purchase Street, the former site of Café Livorno and Umberto's before that.
After arriving in this country from his native Hong Kong and successfully operating Asian fusion restaurants in midtown Manhattan, he signed a long-term lease for a new one in Penn Plaza – in the summer of 2001. But, even that serving of bad timing did not deter Ma from his chosen profession.
Eventually, he arrived at another Rye restaurant, Koo, where he watched over sushi-lovers' needs for several years.
Fogama's customers can sample the specialty — sushi and sashimi — at the first-floor bar. There is ample seating and a bar serving liquor upstairs in an understated, attractive setting.
For those not in a mood for sushi, Fogama fuses numerous alternative appetizers and entrees into its in-house and take-out menus, such as black pepper grilled salmon, poussin with Japanese curry sauce, or Seafood Pad Thai.
© 2011 Rye Record. All rights reservedSource
From our family to yours,
Fogama would like to wish you a very happy and healthy new year!
On New Year's Eve, we will be open from
Reservations are highly recommended
On Christmas Day, we will be open from:
5pm - 10pm
Please make reservations!
Happy Holidays
Hooray! Fogama has finally acquired a liquor license!
Please stop in to enjoy a drink (or two) while also enjoying our delicious cuisine
While our LIQUOR LICENSE
is still PENDING,
Please Enjoy our Cuisine,
And Feel Free to BYOB


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Hey! We would love to have you join the Parent Teacher Student Association of CFI 70!
Please fill out the form below and pay your dues via Paypal. We will accept cash or check in person at school events.
Please make sure you have joined by 3/10/18 to be eligible to run for an elected position.


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SurveyMonkey, Qualtrics, Zoomerang, SurveyGizmo, QuestionPro, or any other form of data collection or organization that can get your data to a spreadsheet and you are ready to import your data for analysis in Dedoose. We've been listening to the many happy users who've taken advantage of this Dedoose feature or any of the other migration tools we've created for Atlas.ti, NVivo, HyperResearch, MaxQDA or other tools and our importers are more robust than ever.
As, many Dedoose users have already learned, making use of the Dedoose survey importer is a huge time saver when you have data or can export/convert data to an Excel spreadsheet. So if you are using tools for your qualitative research data or a study that involves mixed method research where you can get to a spreadsheet, you can move from this…
..to this…
…with just a few clicks of your mouse.
Awesome! Yes! The Dedoose survey importer:
- Scans your file to identify which columns contain closed-ended descriptor data and which contain open-ended narrative responses
- Creates a descriptor set
- Creates descriptor fields (columns) by grabbing the column headers for each closed-ended response field
- Creates descriptors (rows) for each case
- Creates a code system by grabbing the column header for each open-ended response field
- Grabs all the open-ended content for each case, compiles into one Dedoose document
- Links each document to the appropriate case (descriptor) and
- Excerpts and codes all the open ended-responses with the appropriate code.
All automatically and, thus, in just seconds, you are ready to start taking advantage of all Dedoose analytic features.
But now there is more!
In its first life, the Dedoose Survey Importer was great for single imports. Yet, users don't always collect all their data at once and importing additional sets of data into existing project required some clean up.
Not any more!
The Dedoose Survey Importer graduated to a higher class of smartness. As long as your first column of data is a unique values—like in our sample 'ID' field we just numbered the cases from '1' to '196,' Dedoose can be much more clever. The importer is now smart enough to:
- Recognize if a particular row of qualitative data or quantitative data has been imported before—so if you simply add new cases to a big file you are building from your data collection you won't get any repeat cases when imported to Dedoose
- Add the new descriptors to the existing set—before a new set was created with duplicate fields and
- Map codes to the existing code tree so no duplicates are created
So much more efficient and this re-programming has also opened a door for the development of another awesome new Dedoose feature. Hush hush, but stay tuned for that one coming soon to Dedoose near you!;


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Hosted by Edmonton's Nextgen, Pecha Kucha Night is a networking event where presenters share their ideas in a unique format of 20 images, each shown for only 20 seconds. Pecha Kucha Night has quickly become a popular event in Edmonton, and I was happy to have the opportunity to create the graphics for its 32nd iteration.Read More
KEEN in the Kitchen was a self promotion project. It was a collaborative project that included working with other designers, photographers, videographers and letterers. I had the opportunity to design and lay out the book, produce a shot list, and assist in the food typography…Read More
Starling at Big Lake is a unique neighbourhood in Northwest Edmonton developed by Rohit Land Development. I had the opportunity to develop marketing materials for their spring event. These materials feature modern and playful illustrations highlighting the unique benefits of living close to nature in Starling at Big Lake. The illustrations were applied to online advertisements, social media accounts, newspaper advertisements and other marketing materials.Read More
This is a quarterly newsletter for the Edmonton Valley Zoo. This newsletter was intended to be an extension of the Zoo branding, allowing people to get closer to the animals at the Zoo…Read More


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Brian Crower Connecting Rods - 93-98 Toyota Supra 2JZGTE/GE - 5.590 - BC625+ w/ARP Custom Age 625+ Fasteners
Brian Crower Connecting Rods - 93-98 Toyota Supra 2JZGTE/GE - 5.590 - BC625+ w/ARP Custom Age 625+ Fasteners
Part Number: BC6309
Description: Connecting Rods; BC625+ Series; w/ARP Custom Age 625+ Fasteners; Set of Six
Rod Length: 5.590"
Notes: Fits 93-98 Toyota Supra both N/A and Turbo
Made from premium, 4340 steel billet material, Brian Crower connecting rods are available in two distinct styles depending on horsepower, budget and engine application, but both include the exclusive Brian Crower oiling groove in the pin bushing for added oiling.
The bROD Series is intended for moderate boost applications and all motor race applications where peak horsepower is not going to exceed 700 at the flywheel. Utilizes venerable ARP2000 fasteners.
The BC625+ Series rods are designed for high horsepower, high boost and extreme rpm applications where nothing but the best rod will do. Features include ARP's premium Custom Age 625+ bolt rated to 280,000 p.s.i. for ultimate fastening capabilities as well as aluminum-bronze bushings to insure long, reliable service, no matter what the boost, nitrous or RPM.


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Currently, your company is operating on a reactive hiring strategy, but that probably isn't working too well. Waiting to start searching for talent until there's an opening on your roster promises you'll either be left shorthanded until you're able to find the perfect fit or forces you to hire a subpar candidate just to fill the gap. Clearly, it's time to institute some major changes.
It's time to make the shift from a reactive hiring strategy to a proactive approach once and for all. Use these tips to begin the transition:
- Plan for the Future
A reactive hiring strategy focuses on short-term fixes, while a proactive plan centers on long-term goals. Figure out where you want your company to be one, two and even five years from now, and use this to plan your hiring needs. For example, if you want to double your client list in the next two years, you might need to hire more sales staff.
- Define Your Ideal Candidate
Whether you're filling an existing position or creating a new one, things are often crazy busy when it's time to hire a new employee. Simplify the task by going through every position on your lineup and determining what you want in the perfect candidate. This gives you an excellent guideline to reference when it's time to add a new person to the team.
- Never Stop Recruiting
Proactive managers are always searching for potential hires, and you need to adopt this strategy. Whether you simply accept resumes on an ongoing basis or hire outstanding passive candidates when the opportunity arises largely depends on your budget; just make sure you're not limiting the search process to when you need someone immediately.
- Work With a Staffing Firm
Working with a staffing partner is an easy way to develop a passive hiring strategy because they'll take the lead. Your recruiter will work with you to learn more about your company and where you're headed, then start searching for candidates that fit the profile of your perfect hire. They'll vet the candidates and create a talent pool you can access when you're ready to fill an open position — it's that simple.
Partner with Sparks Group to change your hiring strategy for the better. We're here to help make your company more productive, efficient and ready to take on anything the future has in store. Contact us today to learn more about our recruiting services in the Washington D.C. area.


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A few years ago the idea of buying an $80,000+ electric car that had limited range, few places to refuel, and no dealer network was a laughable proposition. To top it all off, the company that wanted to sell the car had very little automotive design or manufacturing experience.
Around the same time, one of the world's top automakers had plans for a new electric car that promised to be affordable, good looking, and easy to take on a road trip anywhere roads exist without worrying about running out of range.
The winner?
Like we even have to say it.
There are myriad reasons why the Tesla Model S has kicked the electric pants off GM's Chevy Volt. It's better looking, lighter, more fun to drive, and has that "cool" factor that leads buyers to fall in love. The Model S isn't just squeaking out a victory over the Volt this year, it's winning by a count of about 6,400 sales to 2,800. Not bad for an underdog carmaker with a big dream!
One of the biggest reasons for the success of the Model S, I think, is the fun-to-drive factor. Its crazy-fast acceleration is unmatched by the comparably lethargic Volt.
That reasoning goes against the opinion of Audi chairman Rupert Stadler, who recently said that he believes the way a car drives will take a back seat to other qualities, such as in-car technology. That might become the case as self-driving cars hit the market and vehicles become more a transportation device than a source of fun.
One of the most insane cars on the market is the $127,000 Model S P85D, an electric car with acceleration so fast Tesla labeled it "Insane Mode." Check out this video to get a sense of what it's like:
Even the well-loved Tesla isn't without problems, though. Consumer Reports purchased a P85D only to have the door handle fail and completely lock all drivers out until Tesla could send a technician to repair the problem.
Considering how new Tesla is in the world of auto manufacturing, the fact that a failed door handle makes the news says a lot about its cars. Until Mr. Stadler's prediction comes true, we can look forward to Tesla remaining the unlikely leader in fun-to-drive electric cars.
What's more your style, the Tesla Model S or the Chevrolet Volt?
Find Used Cars in Your Area at CarGurus


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Reclaimed Men's Silver Hook Pendant With Optional Silver Necklace
This beautiful reclaimed silver hook pendant is an absolutely perfect piece of jewelry for the man looking for an accessory that represents the fisherman or outdoorsman lifestyle! It's made of reclaimed sterling silver and comes with an optional 20in or 22in silver anchor chain necklace. This is the perfect Father's Day, birthday, graduation, or Christmas gift for that special man in your life.
Pendant is aprox. 1.25 inches in length.


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Color Correction Recommendation: X-Rite's Color Munki
X-Rite recently updated its product line for the color munki display and i1 display pro, for gaining accurate color representation in your digital images. This is really important because you cannot accurately adjust color, if you are not sure you are even looking at your image's true initial color projection. For example, if the image looks cold and blue to you, how do you know the image is actually blue, or if your monitor is adding a blue color cast? You don't know unless you calibrate. I love love love the color munki display, and at Rayko Photo Center, we just got the i1 display and have been getting amazing results. The Munki is $189.00 and the i1 is $269.00. Which to get? If you are making a living off of photography you should have the i1, but if you are a prosumer to occasional photo money maker the Munki still works fantastic. I would say it is about a 10% improvement for perfection between the two. Click on the image below to see the Munki sales pitch video. You should take a shot of tequila or espresso every time she says variations on the word "perfection." If that is too easy, try a shot every time she blinks. Yes this is how I have fun. - Carlos


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|Two armed police officers are at one of check points on expressways and roads that connect Beijing with the neighboring cities and provinces on May 20, 2014. [Photo/CFP]
The city has deployed more than 100 armed police officers to 17 checkpoints on expressways and roads that connect Beijing with neighboring cities and provinces since Tuesday. Each point will be staffed by five groups of officers who work a 24-hour shift until mid-June.
Ma Desheng, head of the branch of Beijing armed police corps where the military officers come from, said they were picked two weeks ago and received training to handle emergencies.
On Tuesday afternoon, several vehicles were waiting for inspection at a checkpoint in Shunyi district by a police officer and two armed officers.


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I live in Portland, Oregon where coffee is a food group. A new coffee shop advertised free coffee for their Grand Opening and cars lined up around the block. No surprise. What caught my eye was Starbucks across the street. They too had a line of cars around the block. What the what?? Huh? Then after some thought I said, "wow." That's the power of the brand. It rivals free. It transcends price. I wonder if they could sell a $10 Venti Whatsahoosit… and then I stopped thinking. Of course they could!
I'm intensely interested in this dynamic, the choices consumers make, and the motivations for those choices. This hit home while exploring the agenda for TMRE 2012. Looking at the tracks it's ridiculously full of action. How do I choose? Which path will inspire me and ignite my next idea? Do I look for big name speakers? Do I focus on a track area where I am less adept so that I learn more? Or do I let my curiosity guide me – making decisions on a whim? Probably the latter.
I find that two principles hold true at these events: 1) whatever sparks my curiosity leads to my next big idea; and 2) attending a session because I think I should unleashes my inner rebel. She listens poorly and filters badly. So, instead of doing that, I'll seek out presentations that made me go "What the what??" So hopefully after I can say "wow."
** ** **
Today's guest post is from Kelley Styring. Styring is principal of InsightFarm Inc. a market research and consumer strategy consulting firm. She has led insights for Procter & Gamble, Pepsico, Black & Decker and NASA prior to founding her own firm in 2003. Kelley is a published author and has been featured in USA Today, ABC News, Good Morning America, Brandweek, Fortune, Quirk's Marketing Research and The Market Research Daily Report from RFL Online. She will be live blogging from The Market Research Event 2012 this November 12-14 in Boca Raton, Florida. If you'd like to join her, register today and mention code TMRE12BLOG to save 15% off the standard rate!


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Consistent with the
formula--warm blood, no gills, two pairs of hands and a few
is a mammal; there he sits in his own habitat,
serge-clad, strong-shod.
--from "The Pangolin"
Warm blood? Check. No gills? Check. Two pairs of hands and a few hairs? Check and check. Strong-shod? Check. Serge-clad? Beyond your pitiful understanding, little ants of philosophy, and beyond your wildest dreams.


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This section applies to you if you have ever been registered/licensed to practice in another Canadian jurisdiction (province/territory).
The information in this section pertains to:
Other Resources:
Overview of Registration Process
- Review the Requirements for Registration for Canadian Applicants
- Complete the Application for Registration/Provisional Registration including application fee ($189.75 includes HST). Application fee will not be refunded once the fee has been processed by ARNNL.
- Request a Verification of Registration(s) from all jurisdictions where you are currently registered/licensed be sent directly to ARNNL.
- Request a Statement Current/Most Recent Employer(s) for all RN employers within last five years. Statement must be sent directly to ARNNL.
- Submit a Certificate of Conduct.
- Allow two to four weeks to process your application. All required documents and forms must be received before an application is reviewed.
- ARNNL will notify you by email that your application for registration has been approved.
- To obtain a license to practice nursing in NL you must apply for initial licensure or membership through MyARNNL before commencing employment, which includes orientation.
Canadian Applicants who practice without a valid practicing license will be subject to an Unauthorized Practice Fee in accordance with ARNNL Bylaw Article II, section 2(C) ($100/shift worked with no maximum) and discipline in accordance with section 21 of the RN Act where the period of practice exceeds 3 shifts from and including the 1st shift.
Nursing in Newfoundland and Labrador is governed by the Registered Nurses Act (2008), Registered Nurses Regulations (2013) and By-Laws.
Registered Nurses in Newfoundland and Labrador must adhere to the Standards of Practice for Registered Nurses and Canadian Nurses Association Code of Ethics. In addition, all RNs wishing to renew a practicing license with ARNNL must complete all components of the ARNNL Continuing Competency Program CCP.


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Organizational wellbeing made tangible: space, furniture and tools designed to nurture individual wellbeing. read more
374 Congress Street 6th Floor Boston Tel: 6177232100View company website
Digital Lumens is driving the industrial IoT revolution through its cloud-based IIoT platform, SiteWorx, which ...
5 Holland Building #219 Irvine, CA 92618 Tel: 248-890-8991View company website
The main benefit of buying from our company is that we can offer low prices ...
3612 Ocean Ranch Blvd Oceanside, CA 92056 Tel: 877-724-8100View company website
Pestgon, Inc. has been responding to the needs of commercial property managers for over 30 ...
743 S. Dudley ST. MEMPHIS, TN 38104 Tel: 901-774-5771View company website
Lumalier offers a complete selection of UVGI fixtures for upper room air and total building ...
2 Interchange Pl Ste D York, PA 17406
Kelly Overlander, Account Executive 717-767-1850
10005 Muirlands Blvd Ste I Irvine, CA 92618
Mike Post, Account Executive 949-214-8963
12 Gibson Drive Hilton Head Island, SC 29926
Warren Lynn, President 843-785-8699
6249 Harlem Rd Ste 101 New Albany, OH 43054
David Pierce, Principal 847-516-7008
212 South Mt. Zion Road Lebanon, IN 46052
Meredith Stines, President 765-483-9514
20413 Crimson Place Leesburg, VA 20175
John Younts, President | http://www.criticalpowergroup.com 703-443-1717


text: Man, Linux is cooking nowadays!Spent the last couple of days tweeking and tuning Ubuntu 10.10 <binary geek chuckle> and it really is a very very serious OS. Okay I might be a couple of months late realising this, but hey – I'm finally almost converted! I've been toying with Linux since Windows was still wetting the bed, and I've always liked what I see. Other than the obvious rebellion to Windo$e, I've always preferred it as an OS. It's stable and basic, an "un-fucked with" OS that does exactly what it says on the tin, without treating its users like fucktards. You have to know what you are doing to use it, so it is educational from the minute you switch on. Now, I'm almost at the stage that I can go primary with it. I just need a binary newsreader, par2 and rarchive – all of which are out there. finally I will kiss goodbye to M$ – It'll be a proud day! Linux – Live Long and Prosper 🙂 ❤ penguins


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CLEAR LAKE, IN LUCERNE, 3830 Highland Dr in Lucerne, CALIFORNIA
General Information
- Price:
- $2,200
- Acreage:
- 0.14 acres
- House Size:
- n/a
- 18
- Address:
- n/a
- City / State / Zip:
- Lucerne, CA
- Bed / Bath:
- n/a
- MLS Link:
- n/a
IN LUCERNE, 3830 Highland Dr.,LAKE COUNTY, CA . Owner will finance the property.
Lake County in Northern California is next to Napa and Sonoma Valleys, and picturesque wine country. Lake County has the cleanest air in the state and is the only county in California to have met state and federal air quality standards for over 10 years in a row. Lake County is a destination for the outdoor enthusiast with a wide range of trails for hiking, biking, and off-road adventures and many lakes for fishing, kayaking, and wakeboarding; for the nature lover with parks, preserves, and open spaces for viewing many types of birds, animals, and native plants; for the history buff with numerous museums, landmarks, pioneer towns, and Pomo culture; and it is a destination for the wine lover with some of Californias newest appellations, award-winning wines, and friendly wineries. Major national bass tournament tours are held at Clear Lake, as well as local and amateur tournaments, where the average tournament-winning weight can be more than 5 pounds, a weight unmatched by any other lake in the nation. Clear Lake has earned a reputation for being home to monster bass, and those in the know refer to Clear Lake as the Bass Capital of the West.
Owner financing available. $200 down. Monthly Payments of only $97./ mo.
*note: This lot is not buildable at this time, there are no sewer, water, or electrical infrastructure in place at this time, this is good to hold for future appreciation.
Private financing is available.
Contact Information
- Company:
- Pat Liske Realtor
- Contact:
- Patrick Liske
- Phone:
- (415) 990-0070
- State:
Property Details
- Listing ID:
- 973694
- 18
- Property Type:
- Land - Undeveloped
- Date Posted:
- July 28, 2018
- Views:
- 422
Contact Seller
Send to Friend
More Properties From This Seller
- Quintette, CA
5.76 acres, $59,700
- 3 Lots in CLEAR LAKE AREA LOT, IN NICE
0.21 acres, $1,900
- Clearlake 10th Ave.
0.2 acres, $5,900
- Nice, Lake County, CA Land For Sale - .13 A
0.13 acres, $5,900
- 10.88 Acres Garden Valley, CA
10.88 acres, $89,700
- View all properties from seller
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Disclaimer: All information listed is provided by the seller and is deemed reliable, but should be independently verified. Price and specifications are subject to change without notice. Our site provides this service without representations or warranties of any kind either expressed or implied.


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Posted by Unknown at 1:22 PM Read our previous post
I've talked about the wedding table numbers a little while back (click) and i was thrilled when i got the first order. The lovely client who wanted our flat table numbers also threw a challenge. And we love challenges! Here's the whole story!
I've listed the table numbers on our Etsy shop quite recently and i had a pretty good feeling about them. Here's why:
- there are 4 types of calligraphy you can choose from
- they are printed on beautiful, glossy, white cardstock of 300 gr
- they are simple and elegant
- they are perfect for picture frames, but you can just as easily use them with table number holders
- the beautiful calligraphy is emphasized by the printed border
- they are 100% customizable - you can choose the colors of the writing and border, choose the size, choose if you want them flat or tent, choose if you want them in digital form or the actual physical items!
Here's where you can find them in our Etsy shop - click!
Considering all that, the lovely customer who wanted them told me that her wedding's color theme was going to be navy blue and gold. Therefore, her perfect flat wedding table numbers must have included those colors. As for the size, she wanted them 4 x 6".
We really had fun working on these table numbers and the result was really beautiful! Since i haven't had the chance to take some proper photos of them (me and my "fear" of photography again hehe!), here's part of the digital proof!
©Funky Box Studio - table numbers with a touch of gold
We're really proud of how these table numbers turned out and even more so that it was the very first order for them. We hope there will be many more to go!
Speaking of weddings and paper goodies for weddings, we're also considering putting up for sale the invitations + place cards design set that we had for our own wedding. I created them to fit my own ... funky style, but hopefully they'll be well liked by other happy couples as well!
In the mean time, don't forget about July's kraft promotion! Time's running out and you know you need those kraft supplies! Check out the details here - click!
Funky Box Studio wishes you a colorful day!
Visit our Etsy shop - Funky Box Studio
Twitter I Pinterest I Picsity I StumbleUpon


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"I know Stu and Eric Weber, and I know that they love God and those in pastoral ministry. Their dream of a retreat center in beautiful central Oregon, a get-away for pastors and their wives, could be used of God to bring refreshment, encouragement and new zeal for ministry to the battle-weary. Stu and Eric's vision for Kerith Springs Lodge resonates with me in light of Christ's invitation to his disciples: "Come with me by yourselves to a quiet place and get some rest" (Mark 6:31). This sort of quiet place, one that offers not only rest but spiritual enrichment, isn't easy to find in this hyper-busy culture. Kerith Springs has the potential to bring help, healing, perspective and renewed energy. I join Stu and Eric in praying that if God so desires, He would raise up this refuge of refreshment. By the grace of Jesus, may He use it to make an eternal investment in pastors' lives, and thereby revitalize His church.
I've thought and written about eternal perspective, and years ago named our organization Eternal Perspective Ministries. I believe that it's not only right, but smart to invest our resources--which really belong not to us but to God--in what will last for eternity. What we try to hang onto will ultimately perish, but whatever we put into God's hands will have eternal impact.
I don't presume to know God's direction for your life, but I believe Kerith Springs Lodge could be an eternal investment God wants you to consider. Supporting Kerith Springs could encourage the hearts and strengthen the hands of those who lead, teach, and guide God's people. And that's an investment opportunity worth prayerfully considering.
Jesus said, "It is more blessed to give than to receive" (Acts 20:35). May God bless you as you seek his guidance as to where to invest his assets, to honor Him and further His kingdom."
Randy Alcorn
Eternal Perspectives Ministries
"If Stu and Eric are going into battle, I'd like to tag along to see what will happen. In this case they are battling for pastors who are on the front lines of spiritual warfare 24/7. Every pastor and his wife need time away to be refreshed and encouraged. I believe that Stu and Eric are being led by the Lord in this venture. That's why I'm motivated to line up behind them and support them in whatever way that I can. I hope that you will as well."
Steve Farrar
Point Man Ministries
"Pastors and wives spend their lives caring for the souls of others...that's what they are called to do! But the sad reality is that often the needs of their own hearts go unnoticed and very few seem to care for the barrenness of their souls. Bravo for Stu and Eric who have this passion for "Kerith Springs Lodge"...The time has come to refresh the weary hearts of these special people!"
Joe Stowell
Former Moody Bible President
"The local church pastor faces trials and challenges that few other Christians face. They are on the frontlines fighting for the Kingdom and can become weary and wounded from their daily battles. Kerith Spring Lodge will offer pastors a vital time of renewal, refreshment, and reconnecting with God and with their spouse. Please prayerfully consider how you might be a part of this vision to build into the lives of pastors through your financial giving."
"Come to me, all who labor and are heavy laden, and I will give you rest." Matthew 11:28
Dennis Rainy
Family Life Ministries
"Anytime an attempt is made to reach out and provide a place of shelter for spiritual leaders to be encourages and refreshed is making the right decision. The vision set forth for Keith Springs is a God idea and is greatly needed for the Northwest. May God touch the dream on the paper and make it a reality."
Pastor Frank Damazio
Portland Bible Church


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Dr Andrew Lim
MBBS(Hons) | B.Pharm
NTRI Research Fellow
Dr Andrew Lim has completed the graduate MBBS(Hons) program at Monash University, having previously completed a Bachelor of Pharmacy and Monash University's pharmacy intern program.
His previous research project aimed to find improvements in intercostal catheter viability by using innovative technologies, and identify new procedural modifications to prevent complications of intercostal-catheter insertions.
His current research project's key objective is to improve outcomes from trauma presentations by looking into identification of surgically inaccessible bleeding. His research interests include improvement of procedural/surgical techniques and improvement in patient outcome, particularly in Emergency, Trauma and Surgery.
Furthermore, Andrew has assisted in the Trauma Verification process of The Alfred with the Royal Australian College of Surgeons.


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DNA Polymerase I
More general concepts than this:
Filter by:
Content Type:
- Chapter [36] http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://pub2web.metastore.ingenta.com/ns/Chapter
- Article [9] http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://pub2web.metastore.ingenta.com/ns/Article
- Backmatter [2] http://www.w3.org/1999/02/22-rdf-syntax-ns#type http://pub2web.metastore.ingenta.com/ns/Backmatter
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- 2013 [1] http://pub2web.metastore.ingenta.com/ns/yearOfPublication 2013
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- 2011 [2] http://pub2web.metastore.ingenta.com/ns/yearOfPublication 2011
- 2010 [1] http://pub2web.metastore.ingenta.com/ns/yearOfPublication 2010
- 2009 [4] http://pub2web.metastore.ingenta.com/ns/yearOfPublication 2009
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- 1995 [2] http://pub2web.metastore.ingenta.com/ns/yearOfPublication 1995
Direct DNA Lesion Reversal and Excision Repair in Escherichia coli
- Authors: Sophie Couvé, Alexander A. Ishchenko, Olga S. Fedorova, Erlan M. Ramanculov, Jacques Laval, and Murat Saparbaev
Citation: Couvé S, Ishchenko A, Fedorova O, Ramanculov E, Laval J, Saparbaev M. 2013. Direct DNA Lesion Reversal and Excision Repair in Escherichia coli, EcoSal Plus 2013; doi:10.1128/ecosalplus.7.2.4
- DOI 10.1128/ecosalplus.7.2.4
- More Less
Cellular DNA is constantly challenged by various endogenous and exogenous genotoxic factors that inevitably lead to DNA damage: structural and chemical modifications of primary DNA sequence. These DNA lesions are either cytotoxic, because they block DNA replication and transcription, or mutagenic due to the miscoding nature of the DNA modifications, or both, and are believed to contribute to cell lethality and mutagenesis. Studies on DNA repair in Escherichia coli spearheaded formulation of principal strategies to counteract DNA damage and mutagenesis, such as: direct lesion reversal, DNA excision repair, mismatch and recombinational repair and genotoxic stress signalling pathways. These DNA repair pathways are universal among cellular organisms. Mechanistic principles used for each repair strategies are fundamentally different. Direct lesion reversal removes DNA damage without need for excision and de novo DNA synthesis, whereas DNA excision repair that includes pathways such as base excision, nucleotide excision, alternative excision and mismatch repair, proceeds through phosphodiester bond breakage, de novo DNA synthesis and ligation. Cell signalling systems, such as adaptive and oxidative stress responses, although not DNA repair pathways per se, are nevertheless essential to counteract DNA damage and mutagenesis. The present review focuses on the nature of DNA damage, direct lesion reversal, DNA excision repair pathways and adaptive and oxidative stress responses in E. coli.
Stress-Induced Mutagenesis
- Authors: Ashley B. Williams, and Patricia L. Foster
Citation: Williams A, Foster P. 2012. Stress-Induced Mutagenesis, EcoSal Plus 2012; doi:10.1128/ecosalplus.7.2.3
- DOI 10.1128/ecosalplus.7.2.3
- More Less
Early research on the origins and mechanisms of mutation led to the establishment of the dogma that, in the absence of external forces, spontaneous mutation rates are constant. However, recent results from a variety of experimental systems suggest that mutation rates can increase in response to selective pressures. This chapter summarizes data demonstrating that,under stressful conditions, Escherichia coli and Salmonella can increase the likelihood of beneficial mutations by modulating their potential for genetic change.Several experimental systems used to study stress-induced mutagenesis are discussed, with special emphasison the Foster-Cairns system for "adaptive mutation" in E. coli and Salmonella. Examples from other model systems are given to illustrate that stress-induced mutagenesis is a natural and general phenomenon that is not confined to enteric bacteria. Finally, some of the controversy in the field of stress-induced mutagenesis is summarized and discussed, and a perspective on the current state of the field is provided.
The DNA Exonucleases of Escherichia coli
- Author: Susan T. Lovett
Citation: Lovett S. 2011. The DNA Exonucleases of Escherichia coli, EcoSal Plus 2011; doi:10.1128/ecosalplus.4.4.7
- DOI 10.1128/ecosalplus.4.4.7
- More Less
DNA exonucleases, enzymes that hydrolyze phosphodiester bonds in DNA from a free end, play important cellular roles in DNA repair, genetic recombination and mutation avoidance in all organisms. This article reviews the structure, biochemistry, and biological functions of the 17 exonucleases currently identified in the bacterium Escherichia coli. These include the exonucleases associated with DNA polymerases I (polA), II (polB), and III (dnaQ/mutD); Exonucleases I (xonA/sbcB), III (xthA), IV, VII (xseAB), IX (xni/xgdG), and X (exoX); the RecBCD, RecJ, and RecE exonucleases; SbcCD endo/exonucleases; the DNA exonuclease activities of RNase T (rnt) and Endonuclease IV (nfo); and TatD. These enzymes are diverse in terms of substrate specificity and biochemical properties and have specialized biological roles. Most of these enzymes fall into structural families with characteristic sequence motifs, and members of many of these families can be found in all domains of life.
Homologous Recombination—Experimental Systems, Analysis, and Significance
- Author: Andrei Kuzminov
Citation: Kuzminov A. 2011. Homologous Recombination—Experimental Systems, Analysis, and Significance, EcoSal Plus 2011; doi:10.1128/ecosalplus.7.2.6
- DOI 10.1128/ecosalplus.7.2.6
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Homologous recombination is the most complex of all recombination events that shape genomes and produce material for evolution. Homologous recombination events are exchanges between DNA molecules in the lengthy regions of shared identity, catalyzed by a group of dedicated enzymes. There is a variety of experimental systems in Escherichia coli and Salmonella to detect homologous recombination events of several different kinds. Genetic analysis of homologous recombination reveals three separate phases of this process: pre-synapsis (the early phase), synapsis (homologous strand exchange), and post-synapsis (the late phase). In E. coli, there are at least two independent pathway of the early phase and at least two independent pathways of the late phase. All this complexity is incongruent with the originally ascribed role of homologous recombination as accelerator of genome evolution: there is simply not enough duplication and repetition in enterobacterial genomes for homologous recombination to have a detectable evolutionary role and therefore not enough selection to maintain such a complexity. At the same time, the mechanisms of homologous recombination are uniquely suited for repair of complex DNA lesions called chromosomal lesions. In fact, the two major classes of chromosomal lesions are recognized and processed by the two individual pathways at the early phase of homologous recombination. It follows, therefore, that homologous recombination events are occasional reflections of the continual recombinational repair, made possible in cases of natural or artificial genome redundancy.
DNA Probes for Culture Confirmation and Direct Detection of Bacterial and Fungal Infections: a Review of Current Technologies and Assays
- Authors: Julie Kingery, Karen C. Carroll
Source: Molecular Microbiology , pp 21-30
Publication Date :
January 2011
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This chapter focuses on nonamplified nucleic acid probes and their current uses in the clinical laboratory. DNA probes are pieces of nucleic acid that are labeled in some way and are designed to seek out and bind to stretches of DNA or RNA that have sequences that are complementary to the probe. In hybridization reactions, a double-stranded DNA molecule is denatured to single strands. Several formats for the hybridization reactions exist: solid phase; in solution (liquid phase); in situ; or by use of a Southern hybridization procedure after gel electrophoresis. In sandwich hybridization assays, one probe is attached to a solid support such as a nitrocellulose filter in single-stranded form and ''captures'' homologous nucleic acids in liquid samples; a second probe, which recognizes a contiguous area of the nucleic acid, carries the reporter molecule such as a radioisotope or biotin. The target and probe nucleic acids are free to move in solution, maximizing chances that complementary sequences will bind. Southern hybridization involves using purified DNA that is cleaved with restriction endonucleases. There are numerous methods for detecting the binding of probe to target nucleic acid. Commercially available DNA probes used for culture confirmation of bacteria, mycobacteria, and fungi are discussed in the chapter. Of the assays discussed in the chapter, probes for the detection of mycobacteria have had the greatest clinical impact. The chapter describes the utility of nonamplified probes for the diagnosis of sexually transmitted diseases, vaginal infections, and streptococcal pharyngitis.
Initiation of DNA Replication
- Authors: Alan C. Leonard, and Julia E. Grimwade
Citation: Leonard A, Grimwade J. 2010. Initiation of DNA Replication, EcoSal Plus 2010; doi:10.1128/ecosalplus.4.4.1
- DOI 10.1128/ecosalplus.4.4.1
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In recent years it has become clear that complex regulatory circuits control the initiation step of DNA replication by directing the assembly of a multicomponent molecular machine (the orisome) that separates DNA strands and loads replicative helicase at oriC, the unique chromosomal origin of replication. This chapter discusses recent efforts to understand the regulated protein-DNA interactions that are responsible for properly timed initiation of chromosome replication. It reviews information about newly identified nucleotide sequence features within Escherichia coli oriC and the new structural and biochemical attributes of the bacterial initiator protein DnaA. It also discusses the coordinated mechanisms that prevent improperly timed DNA replication. Identification of the genes that encoded the initiators came from studies on temperature-sensitive, conditional-lethal mutants of E. coli, in which two DNA replication-defective phenotypes, "immediate stop" mutants and "delayed stop" mutants, were identified. The kinetics of the delayed stop mutants suggested that the defective gene products were required specifically for the initiation step of DNA synthesis, and subsequently, two genes, dnaA and dnaC, were identified. The DnaA protein is the bacterial initiator, and in E. coli, the DnaC protein is required to load replicative helicase. Regulation of DnaA accessibility to oriC, the ordered assembly and disassembly of a multi-DnaA complex at oriC, and the means by which DnaA unwinds oriC remain important questions to be answered and the chapter discusses the current state of knowledge on these topics.
Making, Breaking, and Sensing of Cyclic Di-GMP: Structural, Thermodynamic, and Evolutionary Principles
- Authors: Dorothy Kim, John F. Hunt, Tilman Schirmer
Source: The Second Messenger Cyclic Di-GMP , pp 76-95
Publication Date :
January 2010
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This chapter begins with a discussion on a cyclic-di-GMP (c-di-GMP), a self-associating messenger molecule. The proteins involved in c-di-GMP formation, recognition, and degradation are drawn primarily from large gene families which have multiple representatives encoded by most bacterial genomes. Degradation of c-di-GMP is mediated by two families of specific PDEs called EAL and HDGYP proteins based on a stereotyped amino acid sequence and these proteins once again have a broad and frequently redundant phylogenetic distribution. motif present in their active sites. The chapter focuses on the EAL proteins that are most abundant and structurally best characterized. The exact separation between the c-di-GMP switch and the N-terminal β-strand of the PilZ domain varies in different family members, as does the conformation of this β-strand in some family members, which enables the switch to effect distinct changes in interdomain interactions in different PilZ domain-containing receptors. Ironically, neither the PilZ protein nor the XC1028 protein binds c-di-GMP in biochemical assays, and the structure of XC1028 does not contain either c-di-GMP binding motif described. Nuclear magnetic resonance (NMR) spectroscopy was used to determine the structure of PA4608 from Pseudomonas aeruginosa, which provided the first view of PilZ domain architecture. c-di-GMP is an exquisite multivalent molecule that can immobilize disordered protein segments or change the relative orientation of protein domains.
The Nucleoid: an Overview
- Author: Akira Ishihama
Citation: Ishihama A. 2009. The Nucleoid: an Overview, EcoSal Plus 2009; doi:10.1128/ecosalplus.2.6
- DOI 10.1128/ecosalplus.2.6
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This review provides a brief review of the current understanding of the structure-function relationship of the Escherichia coli nucleoid developed after the overview by Pettijohn focusing on the physical properties of nucleoids. Isolation of nucleoids requires suppression of DNA expansion by various procedures. The ability to control the expansion of nucleoids in vitro has led to purification of nucleoids for chemical and physical analyses and for high-resolution imaging. Isolated E. coli genomes display a number of individually intertwined supercoiled loops emanating from a central core. Metabolic processes of the DNA double helix lead to three types of topological constraints that all cells must resolve to survive: linking number, catenates, and knots. The major species of nucleoid core protein share functional properties with eukaryotic histones forming chromatin; even the structures are different from histones. Eukaryotic histones play dynamic roles in the remodeling of eukaryotic chromatin, thereby controlling the access of RNA polymerase and transcription factors to promoters. The E. coli genome is tightly packed into the nucleoid, but, at each cell division, the genome must be faithfully replicated, divided, and segregated. Nucleoid activities such as transcription, replication, recombination, and repair are all affected by the structural properties and the special conformations of nucleoid. While it is apparent that much has been learned about the nucleoid, it is also evident that the fundamental interactions organizing the structure of DNA in the nucleoid still need to be clearly defined.
Transition Metal Homeostasis
- Authors: Dietrich H. Nies, and Gregor Grass
Citation: Nies D, Grass G. 2009. Transition Metal Homeostasis, EcoSal Plus 2009; doi:10.1128/ecosalplus.220.127.116.11
- DOI 10.1128/ecosalplus.18.104.22.168
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This chapter focuses on transition metals. All transition metal cations are toxic—those that are essential for Escherichia coli and belong to the first transition period of the periodic system of the element and also the "toxic-only" metals with higher atomic numbers. Common themes are visible in the metabolism of these ions. First, there is transport. High-rate but low-affinity uptake systems provide a variety of cations and anions to the cells. Control of the respective systems seems to be mainly through regulation of transport activity (flux control), with control of gene expression playing only a minor role. If these systems do not provide sufficient amounts of a needed ion to the cell, genes for ATP-hydrolyzing high-affinity but low-rate uptake systems are induced, e.g., ABC transport systems or P-type ATPases. On the other hand, if the amount of an ion is in surplus, genes for efflux systems are induced. By combining different kinds of uptake and efflux systems with regulation at the levels of gene expression and transport activity, the concentration of a single ion in the cytoplasm and the composition of the cellular ion "bouquet" can be rapidly adjusted and carefully controlled. The toxicity threshold of an ion is defined by its ability to produce radicals (copper, iron, chromate), to bind to sulfide and thiol groups (copper, zinc, all cations of the second and third transition period), or to interfere with the metabolism of other ions. Iron poses an exceptional metabolic problem due its metabolic importance and the low solubility of Fe(III) compounds, combined with the ability to cause dangerous Fenton reactions. This dilemma for the cells led to the evolution of sophisticated multi-channel iron uptake and storage pathways to prevent the occurrence of unbound iron in the cytoplasm. Toxic metals like Cd2+ bind to thiols and sulfide, preventing assembly of iron complexes and releasing the metal from iron-sulfur clusters. In the unique case of mercury, the cation can be reduced to the volatile metallic form. Interference of nickel and cobalt with iron is prevented by the low abundance of these metals in the cytoplasm and their sequestration by metal chaperones, in the case of nickel, or by B12 and its derivatives, in the case of cobalt. The most dangerous metal, copper, catalyzes Fenton-like reactions, binds to thiol groups, and interferes with iron metabolism. E. coli solves this problem probably by preventing copper uptake, combined with rapid efflux if the metal happens to enter the cytoplasm.
Oxidative Stress
- Author: James A. Imlay
Citation: Imlay J. 2009. Oxidative Stress, EcoSal Plus 2009; doi:10.1128/ecosalplus.5.4.4
- DOI 10.1128/ecosalplus.5.4.4
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The ancestors of Escherichia coli and Salmonella ultimately evolved to thrive in air-saturated liquids, in which oxygen levels reach 210 μM at 37°C. However, in 1976 Brown and colleagues reported that some sensitivity persists: growth defects still become apparent when hyperoxia is imposed on cultures of E. coli. This residual vulnerability was important in that it raised the prospect that normal levels of oxygen might also injure bacteria, albeit at reduced rates that are not overtly toxic. The intent of this article is both to describe the threat that molecular oxygen poses for bacteria and to detail what we currently understand about the strategies by which E. coli and Salmonella defend themselves against it. E. coli mutants that lack either superoxide dismutases or catalases and peroxidases exhibit a variety of growth defects. These phenotypes constitute the best evidence that aerobic cells continually generate intracellular superoxide and hydrogen peroxide at potentially lethal doses. Superoxide has reduction potentials that allow it to serve in vitro as either a weak univalent reductant or a stronger univalent oxidant. The addition of micromolar hydrogen peroxide to lab media will immediately block the growth of most cells, and protracted exposure will result in the loss of viability. The need for inducible antioxidant systems seems especially obvious for enteric bacteria, which move quickly from the anaerobic gut to fully aerobic surface waters or even to ROS-perfused phagolysosomes. E. coli and Salmonella have provided two paradigmatic models of oxidative-stress responses: the SoxRS and OxyR systems.
DNA Methylation
- Authors: M. G. Marinus, and A. Løbner-Olesen
Citation: Marinus M, Løbner-Olesen A. 2009. DNA Methylation, EcoSal Plus 2009; doi:10.1128/ecosalplus.4.4.5
- DOI 10.1128/ecosalplus.4.4.5
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The DNA of Escherichia coli contains 19,120 6-methyladenines and 12,045 5-methylcytosines in addition to the four regular bases, and these are formed by the postreplicative action of three DNA methyltransferases. The majority of the methylated bases are formed by the Dam and Dcmmethyltransferases encoded by the dam (DNA adenine methyltransferase) and dcm (DNA cytosine methyltransferase) genes. Although not essential, Dam methylation is important for strand discrimination during repair of replication errors, controlling the frequency of initiation of chromosome replication at oriC, and regulation of transcription initiation at promoters containing GATC sequences. In contrast, there is no known function for Dcm methylation, although Dcm recognition sites constitute sequence motifs for Very Short Patch repair of T/G base mismatches. In certain bacteria (e.g., Vibrio cholera and Caulobactercrescentus) adenine methylation is essential, and in C.crescentus it is important for temporal gene expression which, in turn, is required for coordination of chromosome initiation, replication, and division. In practical terms, Dam and Dcm methylation can inhibit restriction enzyme cleavage,decrease transformation frequency in certain bacteria,and decrease the stability of short direct repeats andare necessary for site-directed mutagenesis and to probe eukaryotic structure and function.
The Biology and Pathogenicity of Vibrio anguillarum and Vibrio ordalii
- Authors: Jorge H. Crosa, Luis A. Actis, Marcelo E. Tolmasky
Source: The Biology of Vibrios , pp 251-265
Publication Date :
January 2006
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Vibriosis is one of the most prevalent fish diseases caused by bacteria belonging to the genus Vibrio. Vibriosis caused by Vibrio anguillarum has been particularly devastating in the marine culture of salmonid fish. The correlation between serotype and virulence may reflect the ability of the bacterial surface antigens to interact with the host tissues. The ferric-anguibactin receptor FatA is an 86-kDa protein that is essential for anguibactin transport. The FatA amino acid sequence is similar to other receptors involved in iron transport, e.g., FhuA and FepA of Escherichia coli, and a TonB box can be identified at its amino-terminal end. V. ordalii causes vibriosis in wild and cultured marine salmonids in the Pacific Northwest of the United States and Japan. The determination and analysis of the complete nucleotide sequence could provide information about the role of pMJ101 in the pathobiology of this fish pathogen. Juvenile salmon exposed to V. ordalii by parenteral challenge developed a systemic infection, and the bacterium was recovered from liver, kidneys, spleen, and blood immediately after the infection. However, the number of bacteria in the liver declined after 1 h and then increased 22 h after infection; bacterial numbers were high in all the organs, and 100% mortality occurred 6 days after infection. The latter observations demonstrate that artificial infection of juvenile salmon is a valid experimental model to study the mechanisms and the bacterial virulence factors involved in the pathogenesis of the infections caused by V. ordalii.
Alternative Excision Repair of DNA
Source: DNA Repair and Mutagenesis, Second Edition , pp 379-388
Publication Date :
January 2006
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This chapter discusses repair pathways of DNA such as alternative excision repair (AER). An AER pathway that has been most extensively defined by studies of E. coli involves a specific endonuclease called endonuclease V. Endonuclease V-mediated incision of DNA is presumably followed by other biochemical events that complete the excision repair of deaminated bases and other substrates. Early studies demonstrated enzymatic activity primarily on DNA containing deoxyinosine, a product of the deamination of deoxyadenosine. This activity was thus designated deoxyinosine 3' endonuclease of E. coli. A series of genetic studies indicate that endonuclease V of E. coli also incises DNA containing the purine analog N-6-hydroxylaminopurine (HAP). Then, the chapter talks about AER mediated by other endonucleases. It is evident from the examples provided in the chapter that nature has been versatile in evolving mechanisms that afford the repair of various types of spontaneous and environmentally generated base damage in DNA. In summary, the chapter provides a cogent reminder that DNA damage is a pervasive phenomenon in living cells and it is obvious that there is no dearth of strategies adopted by nature to mitigate the lethal and mutagenic effects of such damage.
Diseases Associated with Defective Responses to DNA Strand Breaks
Source: DNA Repair and Mutagenesis, Second Edition , pp 919-946
Publication Date :
January 2006
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This chapter focuses on the clinical disorders associated with defective DNA strand break repair and the relationship between the defective genes and the phenotypes observed in patients and their cells, where the disorders are Ataxia Telangiectasia (AT) (Louis-Bar Syndrome), Nijmegen breakage syndrome (NBS), AT-like disorder (ATLD), DNA ligase IV mutations, seckel syndrome, severe combined immunodeficiency, and spinocerebellar ataxia with axonal neuropathy (SCAN1). Identification of the defective genes responsible for these disorders has proven to be extremely valuable in elucidating pathways for DNA strand break repair and, importantly, for cell cycle checkpoint mechanisms.
The Mitochondrion
- Author: Akhil B. Vaidya
Source: Molecular Approaches to Malaria , pp 234-252
Publication Date :
January 2005
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This chapter provides an overview of the mitochondrion in malaria parasites in the light of what has been learned from the genome sequence. Historically, functions of the mitochondrion in malaria parasites were unclear because of its sac-like appearance and paucity of cristae. Prior to successful culture of Plasmodium falciparum, work on mitochondrial biochemistry also suffered from contamination with host components. The need to be aware of host mitochondrial contamination is still valid when using rodent malaria parasites as a source. Although mitochondria are called the powerhouse of cells for providing ATP, it is the generation of proton motive force across their inner membrane that justifies this moniker. Gene expression profiles of blood-stage parasites revealed an apparently coordinated expression of genes for the tricarboxylic acid (TCA) cycle enzymes. Mitochondrial ATP synthesis is carried out by a multiprotein rotary enzyme, F0F1ATP synthase (complex V), located within the inner membrane by utilizing the proton motive force. The assembly of iron-sulfur [Fe-S] clusters is a complex process requiring participation of several enzymes, chaperones, and transporters. In metazoa, mitochondria are central to the process of programmed cell death or apoptosis, releasing several of proapoptotic molecules such as cytochrome c in response to a number of different apoptotic signals, including the collapse of mitochondrial membrane potential. DNA replication, recombination, and repair would require participation of a large number of proteins in these complex processes. Mitochondrial translation machinery is necessary for the synthesis of three proteins encoded by mitochondrial DNA (mtDNA).
DNA Elongation
- Authors: Manju M. Hingorani, Mike O'Donnell
Source: The Bacterial Chromosome , pp 193-216
Publication Date :
January 2005
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This chapter describes the structure and mechanisms of action of bacterial DNA replication proteins, and how their activities are coordinated for efficient duplication of chromosomal DNA. The proteins holoenzyme/replicase function together as catalytic or structural components of the chromosomal DNA replication machinery known as a replisome. At the heart of the replisome is the replicase, or DNA polymerase holoenzyme-itself a complex protein machine comprising DNA polymerase and the accessory clamp and clamp loader proteins. Prokaryotic single-stranded DNA-binding protein (SSB) and eukaryotic replication protein A (RPA) are known to bind several proteins involved in DNA metabolism. The interactions between Escherichia coli SSB and primase and between SSB and χ protein of γ/τ complex are critical for assembly of DNA polymerase onto primed DNA. Each replisomal protein provides a specific function or interaction at a specific point in the DNA replication pathway, enabling highly efficient DNA synthesis. Cell survival depends on the ability of the replisomal protein machinery to overcome such blocks in the path of DNA elongation. Bypass polymerase activity appears to be distributive, suggesting that once the polymerase traverses the lesion it "falls off," allowing reassembly of the replicative DNA polymerase and continued DNA elongation. The processes described clearly indicate that the pathways of DNA replication, recombination, and repair are intimately intertwined.
Overview of Homologous Recombination and Repair Machines
- Authors: Andrei Kuzminov, Franklin W. Stahl
Source: The Bacterial Chromosome , pp 349-368
Publication Date :
January 2005
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The study of homologous recombination between plasmids, or between a plasmid and the chromosome, revealed that the RecFOR pathway is less of a poor cousin than first thought. When the exquisite sensitivity to DNA damage of the first recombination-deficient mutants was found, it became clear that homologous recombination might be the only way to repair certain DNA lesions. Generally, the stronger the defect in homologous recombination, the higher the sensitivity to DNA damage. In Escherichia coli, chromosomal lesions are repaired by homology-guided strand exchange between sister chromatids. The evidence in support of this notion comes in three forms. First, physical connections between parental and daughter strands, associated with lesion repair, can be detected. Second, repair of chromosomal lesions is not observed in recA mutants. Third, DNA damage stimulates homologous recombination although the structure of chromosomal lesions in this case is unspecified. Single-stranded DNA-binding protein (SSB) complexes single-stranded DNA (ssDNA), facilitating its subsequent use in replication and in degradation and repair pathways of DNA metabolism. Chromosomal dimerization in E. coli creates a chromosomal lesion, because it prevents segregation of the replicated chromosomes into daughter cells. The understanding of the formation of replication-dependent chromosomal lesions is still primitive. There is one in vivo study on the structure of stalled replication forks, a report documenting replication fork reversal in vivo, as well as a few reports of replication fork reversal in vitro, likely to be an artifact of DNA isolation.
Recombination Machinery: Holliday Junction-Resolving Enzymes
- Author: Malcolm F. White
Source: The Bacterial Chromosome , pp 405-412
Publication Date :
January 2005
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Holliday junctions are resolved into recombinant duplex DNA species by a class of structure-specific endonucleases known as the Holliday junction-resolving enzymes. The primary cellular resolving enzyme in bacteria is RuvC, which is the main focus of this chapter. The author also talks about the RusA protein, which may act as an alternative to RuvC in some bacterial species, and attempts to place RuvC in a wider context based on our knowledge of other junction-resolving enzymes. The first cellular Holliday junction-resolving enzyme identified was RuvC from Escherichia coli. Homologous recombination is ubiquitous among cellular life forms and many prokaryotic and eukaryotic viruses, and wherever Holliday junctions are formed, junction-resolving enzymes can be confidently expected. Resolving enzymes recognize the branched structure of the Holliday junction and introduce paired phosphodiester bond cleavages in opposing strands to collapse the junction, releasing nicked duplex DNA products. The study of homologous recombination and the Holliday junction was for many years the realm of geneticists. Holliday junction migration work has largely been driven by studies of the E. coli RuvABC resolvasome, emphasizing the continuing utility of bacteria as a model system to study some of the most interesting problems in biology.
Dr. Jekyll and Mr. Hyde: How the MutSLH Repair System Kills the Cell
- Author: M. G. Marinus
Source: The Bacterial Chromosome , pp 413-430
Publication Date :
January 2005
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This chapter concentrates on the repair mechanism in Escherichia coli, but the lessons learned in this organism should also apply to analogous systems in other organisms. Although there are several distinct DNA mismatch repair systems, in this chapter the term is used to denote the MutSLH system. DNA polymerase III, the replicative enzyme, catalyzes resynthesis of nucleotides and ligation followed by Dam methylation to complete the process. An alternative to the futile cycling model based on double-strand DNA breaks (DSBs) recombinational repair is described in the chapter to explain how mismatch repair sensitizes E. coli dam mutants (and human cells) to methylating agents and cisplatin. In dam mutants there is constant repair of DSBs, and the recombinational capacity of the cell is probably near its maximum. This conclusion is based on the higher basal level of transcription of certain SOS genes in dam cells, suggesting that one or more of the RecA or RuvA or RuvB proteins is limiting. The hypothesis that dam bacteria are sensitive to these agents because of inability to repair all DSBs is quite plausible. An important common theme is the requirement for replication forks to stall or collapse at lesions. The hyperrecombination phenotype is explained by the increased number of DSBs leading to increased initiation of recombination. Together with the roles for Dam methylation in controlling transcription initiation and its role in regulating initiation of chromosome replication and its synchronization, almost all the phenotypic properties can now be explained.
Misalignment-Mediated Mutations and Genetic Rearrangements at Repetitive DNA Sequences
- Author: Susan T. Lovett
Source: The Bacterial Chromosome , pp 449-464
Publication Date :
January 2005
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Strand mispairing interactions between repeated DNA sequences provoke a host of mutations and genetic rearrangements in bacteria. Repeated DNA sequences in either direct or inverted orientation can promote misalignment-mediated mutations and rearrangements by very similar mechanisms. This chapter catalogs several types of misalignment-mediated genetic changes in bacteria, with an emphasis on the mechanisms by which they occur. Systematic study of misalignment-mediated mutation and genetic rearrangements has revealed many elements of the mechanisms of these processes, including the integral role for DNA replication. The impact of these misalignment processes on bacterial physiology and genomic evolution is also discussed in this chapter. Sister chromosome exchange (SCE)-associated misalignment has been studied only on plasmid replicons. When recA-independent deletions or expansions between tandem repeats are selected on plasmids, concomitant replicon dimerization is often seen. Single-strand annealing (SSA) contributes as a major pathway to rearrangements in eukaryotes and in bacteriophage-infected Escherichia coli but may occur efficiently only under restricted circumstances in normal bacterial growth. Short spurious repeats can be sites for deletion, duplication, or inversion; these processes create larger repeats that can lead to even higher rates of rearrangements. These rearrangements can occur independently of the bacterial homologous recombination pathways and are dependent on the length and perfection of the repeats, as well as their proximity.