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Tag: portrait
Vera is our Shorkie (half Shih Tzu and half Yorkshire Terrier). Now, when friends first meet Vera, their first thought is, that she is an ugly dog. After they get to know Vera and witness her personality, they say, she is adorable and they don't feel that she is an ugly dog. It's hard to get a good picture of Vera, because she is do dark, the details are lost, unless you have the proper lighting. I used my studio Elinchrom BX250RI strobes for this shot. The lights really show the detail that are normally lost in our snap shot pictures we take. Now if I could just get her to smile and relax more.
Bummer, I missed a post yesterday! It totally slipped my mind. We attended a meeting and on the way home I made a mental note to get the camera out. When we arrived at the house, I turned on the MLB All-Star Game, then sat down to watch. After watching a while, I grew tired, then retired to my comfortable bed and was out like a light. I awoke from a deep sleep, panicking, "oh no, I didn't post a photo!" Oh well… For today's photo, I wanted to compare an effect from On One's Perfect Frame 4.6. I shot the photo, processed in Lightroom 4 as a black and white photo and then exported to Photoshop CS5. The first picture I added a black outline by extending the canvas. For the second photo, I added an effect called, "Aged Wood" from Perfect Frame 4.6. The dark frame adds a grungy, dramatic, and mysterious feel, by adding to the shadows, it leaves more to the imagination. I feel bad that I missed yesterday's post, I will try to keep on track the rest of the way!
Day 85 Justin Senior Year
Justin and I went out and shot some more location photos. It is hard to believe Justin is wrapping up his senior year. It seems like only yesterday we were taking pictures of his first day of kindergarten. The years have flown by and our little guy has grown into a man. We are very proud of you Justin. Keep up the good work!
I finally persuaded my lovely wife Vicki to pose for a portrait. In addition to being my wife, she is my best friend, and I enjoy our time together. So it was great that Vicki was willing to pose. I wanted practice lighting with 1 light. I'm glad Vicki posed for me, I really like this photo. I like the way the shadows create a mysterious feel. My light source was a Nikon SB900 in a soft box. I have been shooting mostly in manual mode lately. I have found it helps me become more aware of the light and depth of field in my photos; which in turn helps me to become more involved with each shot. The result is that there is more of me in the photos, I am more intimately involved with each picture. I used to just set it on aperture or shutter priority and let the camera decide what the best setting was. I have come to enjoy manual mode.
Today we scheduled Amelia's Winter photo shoot. We figured we better make it happen since we had snow on the ground. After all in Michigan, you never know how long the present conditions will remain. We could have 50 degrees and sunny tomorrow, which would put an end to any winter snow shots. So off we trekked to the Gugel Bridge, a Frankenmuth favorite. We were blessed with another beautiful sunny day, although it was very cold and windy. We had to act fast so we wouldn't freeze. Amelia was a real trooper braving the cold. So we shot some photos on the bridge. Amealia's friends decided to join in the fun. So we had everyone pose for a group shot.
A few more photos and then I suggested that we go to the Hart One Room School house and take a few more at that location. I really like that setting with the snow, and the rustic feel, with the split rail fence and the weathered wood. So we jumped in our vehicles, cranked the heat up, and headed over to the One Room Schoolhouse for a few more shots, and then we called it a day. We were just too cold to continue. It was worth braving the cold though, we ended up with some very nice photos. Thanks Amelia.


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subset: commoncrawl


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HOLDEN � Anticipating what may happen if member towns fail to pass a budget anytime soon, Superintendent Darryll McCall outlined for the Wachusett Regional School District Committee what will happen if the budget request is reduced or fails altogether.
Rutland and Paxton are the two holdouts on the $83 million budget request that needs approval by four of the five district towns in order to be adopted. Both towns are asking for reductions to the budget of approximately $2 million.
Paxton is scheduled to reconvene its annual town meeting on June 22 to consider a new school budget if one is available, and also to take up its general operating budget.
McCall told the committee that at this point, the district will begin Fiscal Year 2016 that starts on July 1, with no budget.
He outlined various scenarios where the proposed $83 million district budget is slashed by $750,000, $1.25 million and $2 million.
Reductions can be made through cutting personnel, saving on salary and attendant benefits, McCall said. Also on the line are instructional support, equipment and technology as well as operations and maintenance.
Potential personnel reductions ranged from four Applied Behavioral Analyst (ABA) positions, up to nine existing teachers and the proposed restoration of the deputy superintendent's position. In-between steps included the elimination of several new teachers and guidance counselors.
The ABA positions are first on the line, McCall said, because there will be fewer district students with Individualized Education Plans that generally require aides in the upcoming school year.
Instructional support and technology cuts would range from $70,000 in a best case scenario to $311,000 in the worst case. This includes textbooks, hardware and annual contracts for things like Powerschool, the web-based student information system currently used by the district.
"We should be able to handle it for the time being," McCall said.
He cautioned that in order for the district to pay for two full-time unemployment claims, it needs to lay off three staff members.
There are parts of the school district budget that cannot be reduced, he said, including pupil services, special education tuitions, transportation and debt service.
Operations and management cuts would affect building and grounds maintenance, custodial supplies and services as well as utility services. Those reductions could range from $117,000 to $285,000, McCall said.
If there is no budget in place by June 30, the state requires that the district move to a budget where each month they can expend an amount that is approximately one-twelfth of last year's $81 million budget. The district has followed this procedure for the past two years while waiting for a budget to be approved.
While it would be possible for the district to hobble through the summer on the one-twelfth budget, Business and Finance Director Joseph Scanlon said, "in October, we hit a wall."
That could mean mid-year staff layoffs with the possibility that they could take place before the school year.
The district could pursue the option of seeking a Revenue Anticipation Note (RAN) to obtain the cash flow needed to operate, he said.
Committee members voiced dismay that the district found itself once again in this situation.
"I'm distraught and disappointed that we're talking about this again," Christine Smith of Holden said. "People begged that we not cut teachers. What happened to that support?"
Smith said she would not vote to reduce the original budget request, even though she understood the repercussions of moving to a one-twelfth budget.
"We need to stop this pattern," she said.
Other members brought up the possibility of declining property values as the result of an under-funded school system.
The district has to pass some type of budget by July 2, according to state mandate, WRSDC chair Lance Harris said.
At that point, member towns have 45 days to pass or reject the budget.


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20.06.2019, Tony212
CNN In April , scientists used a global network of telescopes to see and capture the first-ever picture of a black hole, according to an announcement by researchers at the National Science Foundation Wednesday morning. They captured an image of the supermassive black hole and its shadow at the center of a galaxy known as M Chat with us in Facebook Messenger. Find out what's happening in the world as it unfolds. More Videos


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Since the days of ancient Greece, people have used plants and their essential oils to obtain important benefits for healing mind and spirit. Essential oils are so named because they refer to the "essence" or spirit of a plant and are thought to be a concentrate of its healing powers stored for convenient use.
Advent of Aromatherapy
In the last century, this healing practice has been dubbed "aromatherapy", but the use of essential oils traces back to Dioscorides in the first century A.D., who documented the healing powers of oils extracted from flowers, bark, stems and all other parts of a wide plant assortment.
Though the steam distillation of essential oils was first perfected in the 11th century, it was early 20th century France that witnessed a new intense interest in herbal treatment and aromatherapy. In fact, the French were the first to use essential oils to treat wounded soldiers in World War II.
How Essential Oils Work
People use essential oils to treat or prevent diseases, induce relaxed or alert moods, increase energy and reduce pain. The pleasant, potent fragrances of volatile oils reach the brain through the olfactory and limbic systems. The oils are never ingested — they are for external use only. Mounting evidence suggests that the molecules of essential oils and compounds stimulate the brain, help the body to strengthen its immune response and promote natural healing. Some studies suggest that many essential oils act as antiseptics.
The essential oils used in aromatherapy are applied in two basic modalities — through inhalation and physical contact.
Inhalation of Essential Oils
Aromatherapy can be as simple as unscrewing a bottle cap. However, a diffuser or vaporizer is a much more effective and economical way to inhale the pleasant-smelling molecules wafting up from essential oils. A diffuser creates a very fine vapor without heating the delicate oils. You can also use a vaporizer by putting a few drops of oil onto a tissue and placing in front of the escaping steam. Never put essential oils into the vaporizer — it will not mix properly and may damage the device. Another simple approach is to sprinkle some oil into a bowl of hot water and then inhale as you use a towel over your head to create a tent.
Essential Oils on the Skin
Because of their potency, you usually dilute essential oils with a vegetable oil complex before skin contact. You place a few drops of the diluted oil in your hands and apply to the desired body location. Massage the oil in with a circular motion. For the ultimate delightful experience, have a masseur apply essential oils as part of a full-body massage. Your choice of oils will dictate your response — anything from deep relaxation to enhanced mental alertness.
You can also purchase soaps, shampoos, conditioners, and creams that are formulated with essential oils. These provide the benefits of these natural products in a convenient form.
Forms of Essential Oils
Essential oils are isolated and concentrated through a variety of methods:
- Distillation — plants and plant materials are suspended over boiling water, allowing the steam to separate out the volatile oils, which are then condensed into concentrated extracts.
- Water solutions — items such as rosewater, chamomile and lemon balm are examples of herbal distillates or infusions in an aqueous solution. They are called hydrosols or herbal distillates and are the condensed water-fractions resulting from distillation.
- Expression — items such as grapefruit oil are cold-pressed from fruit peels.
- Supercritical extraction — delicate flowers and other plant parts are extracted using solvents under high temperature and/or pressure.
- Dried and crushed — some essential oils are retrieved by drying, crushing and warming plants with high oil content. These oils are best suited for use in a vaporizer.
Popular Essential Oils
Eucalyptus oil is perhaps the best known essential oils. There are many different species of eucalyptus, some of which are well suited for aromatherapy. Rose oil and its water-based version, rose absolute, are well loved for their remarkable scents. Lavender essential oil is mild and produces a relaxed mood. Tea tree oil is popular for aromatherapy but is also used in many cosmetic products and shampoos.
There are dozens of essential oils for consumers to choose. It's fun to research which oils are most appealing for your own personal use. Whether wafting through your rooms, enlivening a bubble bath or delighting your skin with soaps, shampoos and conditioners, high-quality essential oils can make a positive contribution to your life and your lifestyle.
Don't Be Fooled
Many soaps, hair products and face creams have artificial fragrances not derived from essential oils. These are often chemical concoctions that can have undesirable side effects, such as allergic reactions and drying. When purchasing shampoos, cosmetics and soaps, always make sure you understand what ingredients have been added to provide the fragrance. Once you experience essential oil products, you'll never settle for less.


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- Opinion
- Chanticleer
'We're living in dog years': Transurban's road toll conundrum
Scott Charlton has a delicate balancing act to perform to keep investors happy, workers employed and customers on the tarmac.
- James Thomson
Transurban eyes US expansion as Australian traffic weakens
Investors have been putting money in high-yielding infrastructure stocks due to low interest rates.
- Updated
- Jenny Wiggins
- Opinion
- Chanticleer
Transurban sees growth at end of tunnel drama
Scott Charlton is confident of resolving the dramas surrounding the Westgate Tunnel project, and is eyeing the biggest pipeline of opportunities he's seen.
- James Thomson
Transurban on congestion charge alert
Transurban has back up plans for some of its tollroads if Australian cities eventually charge motorists to drive into CBDs.
- Jenny Wiggins
April 2019
'ZEVs will dominate,' says Transurban CEO Scott Charlton
The time has come for zero-emission cars, and they won't need subsidies, says the Transurban boss.
- Jenny Wiggins


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It's the new school year and Sarah's class are going on a field trip to a museum! That sounds like more fun than just studying books and Sarah can't wait to check out the dinosaur displays and exhibits. But Sarah's strict teacher is tagging along and and she just loves to catch naughty students messing around and slacking... Will Sarah be able to outwit her teacher and have a great time at the museum? Or will her slacking days be put back into the prehistoric era?
Game Controls:
Game Controls:
This game is played with mouse.


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Swarovski - Austria
Sale price
Regular price
Genuine Swarovski, Aquamarine Glacier Blue, 29ss - 6mm rhinestone in a sew on setting - choose 2 or 4 piece lots. These Swarovski crystal stones are set in a high quality 4 pronged, open back 4 hole - one hole on each side - sew on setting. Settings available in silver or gold plated - your choice. Finished jewelry shown are design suggestions only, and not included.These sparklies are Swarovski's Peridot green with an AB finish - truly sparkly and dazzling. They are a fabulous size for stud earrings or for use in my Fancy Clasp Ring Tutorial.If you would like to make the earrings shown, you can find that tutorial here Easy Peasy Post Stud Earrings and the tutorial for the Clasp Ring is here Simply Fancy Ring with a Clasp.Looking for more equally yummy gems and crystals in settings? You can find them here Crystals in Settings And don't forget to check what new goodies I've added here New Items
A small note about foil-backed stones: Swarovski's sew-ons are excellent quality; I have at least 3 I rarely take off. However, there are chemicals and some oils and lotions that can eat at the foil on the back of the crystal. If in doubt, whether something will hurt the foil, take the ring (or other piece of jewelry off).thanks for stopping by - hope to see ya again soon.....~Bobbi


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text: Javier Ballester20 de oct de 2013sanity is a phantom"The distinction between sanity and insanity is narrower than a razor's edge, sharper than a hound's tooth, more agile than a mule deer. It is more elusive than the merest phantom. Perhaps it does not even exist; perhaps it is a phantom. " Philip K. Dick.#philipkdick


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Home / Music / Which artist had a hit with the simple title "Yeah!"?
Which artist had a hit with the simple title "Yeah!"?
The song was released on January 27, 2004. "Yeah!" topped the US Billboard Hot 100 chart for 12 consecutive weeks, before being dethroned by Usher's follow-up single "Burn".
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The Peoples Democratic Party (PDP) has rejected what it called the "fictitious" votes; being collated by the Independent National Electoral Commission (INEC) in favour of the All Progressives Congress (APC) in the Kogi governorship elections.
The PDP further called for the nullification of the votes garnered by the APC and Governor Yahaya Bello in Okene.
This was contained in a statement signed by its National Publicity Secretary, Kola Ologbondiyan; on Sunday, November 17.
Ologbondiyan said, "nobody can accept the provocative alteration of results from polling units; as well as the allocation of fictitious votes, particularly the awarding of 112, 000 votes to APC in Okene."
Also Read: Fashola fears 'Brexit experience' over restructuring
According to the party, the people of Kogi State have glaring evidence of alterations of figures; to reduce the vote of the PDP and allocate "imported figures" to the APC.
The party, therefore, advised INEC not to ever declare such "mutilated results"; as they will not be accepted by the people.
The statement reads: "Our party hopes that this apparent moves to allocate fictitious results to the APC; has no bearing with the allegations in the public space that; Governor Yahaya Bello and the APC had compromised highly placed INEC officials with huge sums of money to alter the result in favour of the APC.
"The PDP, therefore, cautions INEC to note that any attempt to declare the figures; being bandied for the APC will be a validation of the bribery allegation."
Meanwhile, the election on Saturday left no fewer than three persons dead; with several others injured. The incident occurred when suspected political parties thugs invaded Adankolo and Lokongoma primary schools in Lokoja.
Reports revealed that the thugs disguised as police officers shot at two electorates in Adankolo; the other in Lokongoma when they wanted to snatch the ballot box.
It was also gathered that the thugs wounded several other persons, leaving them with degrees of injuries including a woman.
This forced many voters to scamper for safety following the ugly development in those polling units.


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Gate Opening Times
Please find 2019 Gate Times below. Please email Customer Service with any questions.
Our parking attendants are on duty 3 hours prior to the performance gate times.
Tanglewood Box Office in Lenox, MA
Monday – Friday 10am-6pm or Intermission
Saturday 9am- 6pm or Intermission
Sunday 10am-6pm or Intermission
For most Ozawa Hall concerts the Box Office opens 3 hours in advance.
The Box Office Will Call opens at the Linde Center in the Marignetti Lobby 60 minutes prior to Tanglewood Learning Institute Events.
Please note: The Tanglewood Box Office closes at the end of intermission, or 30 minutes after the start of a performance if there is no intermission. After the Box Office closes, any remaining will-call tickets will be moved to the Main Gate for late pick-up. The Main Gate is located at 297 West Street. You may not enter the grounds without a ticket. Tickets cannot be purchased after the box office closes.
Download Tanglewood Campus Map (PDF)
September 1 3:30PM - GATES OPEN AT 1PM


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Posted by Sun Devil P.A.T. on November 15, 19 at 16:44:25:
In Reply to: Re: Myles Garrett video posted by PhiDevil on November 15, 19 at 16:35:59:
You mean the same player's union that screams about "player safety," sticking up for a guy who uses a helmet as a weapon, on a quarterback who just had a severe concussion about one month ago. The player's union would look like big time hypocrites, if they take that stance, in my opinion. Go Sun Devils!
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Apocalypse by the Srinagar Hydro Electric Project
Translated from Sitaram Bahuguna's report from Srinagar, Uttarakhand on the Uttarakhand flood in Hindi.
The apocalyptic flood in the Alaknanda showed its destructive dance in Srinagar (Uttarakhand). A large part of the town is completely damaged. The flood has ruined the SSB Academy area and ITI area. About a hundred families living in some 7 residential huildings are now homeless. These residential buildings have 10-12 feet of debris in them. None ofhold goods have remained usable. Dr. Umashankar Thapliyal, a senior journalist who had reported the Belakuchi lood in 1970 is of the opinion that the flood at that time was of similar intensity, but the damage was not as much.
The Srinagar Hydro Electric power project is responsible for the terrible destruction in Srinagar. GVK company developing the project has dumped lakhs of tons of mud on the banks of the Alaknanda from the dam site at Koteshwar to Kikileshwar. The width of the river was reduced due to this. The flood on the 16th and 17th started eating away at this mud, due to which the level of the water rose by up to two meters. At the Seema Surakasha Bal Academy area, where the flood has wreaked destruction, the river curves like the English letter "N" and the width of the river suddenly decreases ahead of that. It is due to this that when the flood came on that day and it brought with the the lakhs of tons of soil from the Srinagar Hydro Electric Project, it could not flow ahead from the area in the quantities it reached. Consequently this tremendous amount of mud started settling back in the SSB Academy, ITI and Vijay Nagar areas. When the flood waters receded, people were in disbelief. 70 residential homes, half the SSB Academy campus, the IT and Shakti Vihar areas, grain storage, gas storage, silk farms had up to 12 feet of mud accumulated! Many residential buildings are buried in the debris to their roofs. The life's earnings of the people living in these homes has been leveled.
One of the affected, Vinod Uniyal says that this flood has deposited eight feet of debris inside his home. Household goods and life's earnings stand finished. He is not able to understand how he will survive in the future. Santosh Chandra Naitiyal, who made a living running the ration shop in Shakti Vihar has been twice devastated. His home and shop both lie buried in the debris. Now he faces a crisis of income as well.
S. Bandopadhyay of the SSB Academy estimates that the damage in the Academy itself amounts to 100 crores. With this as a gague, it seems that the damage in the Srinagar area will could up to 300 crores.
Apart from this, in Devprayag as well, many government and residential buildings stand mired in debris. The lakhs of tons of soil illegally dumped in the river by the Srinagar Hydro Electric Project has now vanished and the river bank is looking like it used to, before the soil was dumped. Srikot too has several buildings next to the river, but it escaped the destruction because most of the soil had been dumped a little below the village from Chouras Jhula up to Kikileshwar. This saved Srikot from being destroyed as well.
Geologist Dr. D. P. Sati estimates that the hydro Electric projects in the valley have a major role in the destruction wreaked.
5 lakh metric tons of soil was dumped in the Alaknanda riverbank for the Srinagar Hydro Electric project. When the flood came, this mud merged with the water under pressure and the volume of the water increased dramatically. Because this mud was placed directly in front of Srinagar, the lower parts of Srinagar accumulated it as it settled and became the cause of ruin.
(The Srinagar Hydro Electric project for 330MW of electricity, at about 30 kilometers from Devprayag on the Alaknanda has been constructed in blatant violations of all environmental norms and solely on political clout. We can provide more information if needed. ~ Vimalbhai)
One Comment
It has become quite fashionable to blame hydro sector for all calamities in hills, – whether it is due to earthquakes or floods. Public is free to make up its mind after considering following facts.
1. All the deposits left in Srinagar town, after the floods receeded comprise of sand. Srinagar Project did not generate any sand during the excvation required for Project structures. So, whatever was left behind in the town did not come from the Project.It travelled from areas upstream over the Dam.
2. It is alleged that, all the 5.0 lakh tonne muck dumped by the Project at the Forest Department approved dump sites found its way into Srinagar town.This is questionable. What is not questionable that, Srinagar Dam has retained and prevented more than twenty million tonnes of muck from flowing past the Dam and depositing in areas downstream.
3. All this can be verified even now, i.e. two months after the flood. It would be more dignified, if allegations are levelled against Hydro Sector not on heresay or with bias.


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Discussion in 'General MMA discussion' started by Fight Genome, Jul 31, 2012.
VERY good post
Joe "Steven King is a literary GIANT" Rogan is really getting on me nerves these days.
He really is.
We'll, in terms of sales, popularity, & volume of books, he's not wrong. Especially the influence of the horror genre.
In terms of actual art, probably not.
Mark DellaGrotte on now ( 30 min in )
Really good so far.
They say Rogan get's like 30 to 40k for every episode. What in the world, can that be true.
One of the most popular podcasts, + sponsorship, + youtube... I reckon it's true
I heard rumours of more like 80-85k. But yeah 30-40 is definitely a reasonable estimate. From the youtube revenue alone, he'd be at about $10k/episode but he does 10 minutes of advertising before each audio recorded podcast which is where he probably generates the majority of the revenue. 2-5 million downloads per episode would surely have advertisers paying big money.
JRE MMA Show #38 with Gaston Bolanos & Kirian Fitzgibbons
I don't like the way Kirian Fitzgibbons comes off in this. He seems like a good guy, good mind... but then he veers into pompousness and absurdity. He really loves to focus (several times throughout the talk) about what he does for his fighters monetarily. It's cool to hear once but after like the third time he seems to be tooting his own horn. He has a pretty apparent hate on for Bang Ludwig. WHen he brought up the incident (for no apparent reason, other than Ludwig's name came up) that he had with him, it seemed force. Joe handled it nicely though ("yeah, I'm friends with Duane; he's crazy" chuckle and moved Fitzgibbons away from it).
Then Fitzgibbons going on about "I am convinced that Cody is a much better boxer than TJ. And I'm not sure why it didn't come through in those fights. I really can't figure it out." Uhhh, could it be that it's an MMA fight and not a boxing match?
Gaston Bolanos seems very cool, though. Joe loves that spinning elbow (he's talked about it on past podcast as a pure piece of art...fun to listen to him talk about). I will be excited to see his next Bellator fight.
Cerrone feels slighted as f--- and for good reason. Cerrone is pissed, this will most certainly make headlines.
Yeah it will this is f---ing good right off the bat, there was little warming up into this.
Man, this is the Jones/Evans situation all over again but worse. In all honesty given what Cowboy just said it makes me think differently about Jones's more recent issues.
s--- Cowboys story on his nearly dying on the cave diving thing is intense.
I had to step out for work so I missed a good chunk, I'll catch it later. I knew he was going to talk about it though, was looking forward to it.
Just putting yourself in his shoes as he is relating to what happened and f--- it gets intense.
From Joe's reaction it sounded like the situation there wasn't known. It's kinda f---ed that Perry has got Cowboy's long time gym behind him.
Separate names with a comma.


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Pictured (left to right) are participants: Honigman Partner Khalilah Spencer, who served as moderator; Miller Canfield Senior Counsel Saul Green; Western Michigan University Cooley Law School Dean James McGrath; University of Michigan Law School Dean Mark West; Wayne State University Law School Dean Richard Bierschbach; Michigan State University College of Law Dean Lawrence Ponoroff; and Bodman Partner and Diversity and Inclusion Committee Co-Chair Damali Sahu.
On September 10, Bodman PLC hosted the monthly meeting of the African-American Partners' Collaborative at its Detroit office at Ford Field. The meeting featured a panel discussion on diversity in the legal profession and in legal education, and included the deans of all five of Michigan's law schools and members of their senior staffs.
Also participating but not pictured was University of Detroit Mercy School of Law Dean Phyllis Crocker.
The African-American Partners' Collaborative is an organization comprising partners, principals, members, and other senior attorneys at large metropolitan Detroit law firms who seek to increase the number of African-American attorneys at firms, as well as improve opportunities for them to succeed professionally and participate in firm management.


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Throughout the city, decorations for the upcoming season were already in full force. As I have always wanted to visit Quebec City at Christmas time, I was thrilled. Most were tasteful and pretty; all that was missing was the snow! Here's a selection:
Archives for posts with tag: Holiday Decorations


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American teenager Cori 'Coco' Gauff claimed the first Top 10 win of her fledgling career over top seed Kiki Bertens as she moved into her maiden WTA semi-final at the Upper Austria Ladies Linz on Friday.
Sometimes I'm motivated and then I have moments when I don't want to do it [homework] at all, Cori Gauff
The 15-year-old from Delray Beach, Florida, who has attended school online since 3rd Grade, is starting to write records as well as essays after upsetting the Dutchwoman, 7-6(1) 6-4.
In so doing Gauff became the youngest woman to reach a WTA semi-final since Nicole Vaidisova won the Tashkent title in 2004, and will take on Andrea Petkovic on Saturday for a spot in the final.
The 75th-ranked Petkovic booked her first semi in a year, defeating Viktoria Kuzmova of Slovakia, 6-4 6-1.
Gauff's strategy sessions for her first semi were put on hold for a few hours due to high-school homework that needed to be tackled after her late-afternoon win, with lessons beckoning in her hotel room.
"I just got off the court, so I've not thought about the semi-final," she said. "I'm thinking of the homework I have to do tonight. I don't know what it is yet.
"I'm studying the typical stuff: math, English, history… I don't know what I have to do – but I know it's something."
Gauff is not bothered by juggling her tennis with school commitments: "Sometimes I'm motivated and then I have moments when I don't want to do it [homework] at all," she admitted.
The 15-year-old has experienced a huge change of fortune during the week, going from losing in the final round of qualifying to entering the main draw as a lucky loser and attaining the first Top 10 victory of her career with her 97-minute victory over top-seeded Bertens
The teenager earned her place in the field as a lucky loser from qualifying when Greek Maria Sakkari withdrew injured at the start of the week.
The Floridian, still under WTA age eligibility restrictions, is playing her fifth Tour-level main draw of 2019 after reaching the Wimbledon fourth round and making the third round at the US Open.
The Linz week has been a breakthrough for the World No 110, who will crack the Top 100 on Monday, when she travels to Luxembourg for the last event of her season, and she is already assured of a spot in the Australian Open field in January with her rise in the rankings.
Gauff maintained control throughout over the 8th-ranked Bertens.
"I'm glad I was able to come out on top. I've really gotten a lot of support from the crowd," she told a sellout audience in this upper Austrian regional capital.
"There have been a lot of firsts this week. I'm really happy and proud of how I'm playing.
"It was definitely crazy," Gauff later told the press, after her massive victory. "Today was my first Top 10 win, so I'm super proud of myself for that.
"It was a good match, Kiki played great, and obviously I couldn't ask for anything better so far in this tournament."
"I didn't have any nerves until the match points came along. Kiki was playing well and there was just a difference of one or two points."
Very little separated the two players, with a single break of serve in the encounter determining the victor.
Gauff staved off both of the break points she faced in the match, won three-quarters of her first-service points, and claimed half of the points off of Bertens's second serve.
She was never threatened by her more experienced opponent, who won titles this season at St Petersburg and Madrid, and reached the Beijing semi-finals last week before returning to Europe.
Gauff stepped up to win the opener in a tie-break and broke for 2-1 in the second.
The match ended on Gauff's second match point as Bertens sent a return wide.
"I guess [Linz is] my little lucky place," Gauff added. "I didn't know what to expect coming here, and I'm glad I was able to take the best of the opportunity.
"I got in as a lucky loser and I'm in the semis, so I hope I can keep this going."
Gauff exhibited exceptional prowess in the forecourt as the match commenced, punching victorious volleys and delivering divine drop-shots, but the thunderous power game of Bertens kept her on level footing as she blasted her way through any moments of peril in the early going.
Bertens had to grit her way through a tough service game to hold for 4-4, as she double faulted to give Gauff the first break point of the match.
A strong serve by the Dutchwoman erased that chance, and she continued to find robust deliveries which pulled her through the game, as the combatants continued to stay deadlocked throughout the opening frame.
Despite an evenly-matched run to 6-6 with zero breaks of serve, Gauff quickly turned the pivotal first-set tiebreak fully in her favour.
After opening the breaker by winning a scintillating rally with a drop volley winner, the American stayed steely as a run of unforced errors came from the racquet of Bertens.
The Dutchwoman hit her third double fault of the set to cede a 5-1 lead to Gauff at the changeover, and after another wide forehand by Bertens, the teenager reached 6-1 and held 5 set points.
Gauff only needed one, ending the set with a forehand winner and putting her on the precipice of a signature upset.
In the second set, Bertens had an early opportunity to pull ahead, firing a forehand winner to queue up a break point for a 2-0 lead.
Gauff, however, had other ideas, as she executed a drop volley to force an error and pull herself out of danger, eventually moving to a hold for 1-1.
Gauff was immediately rewarded in the next game as Bertens misfired on forehands, as the American suddenly attained the first, and what would prove to be the only, service break of the match.
Bertens never faltered on serve after that, but the Dutchwoman got no further looks on Gauff's delivery, and the teenager would go on to close out the victory on her second match point.
Gauff now takes on former Top 10 player Andrea Petkovic of Germany for a spot in her first-ever WTA Tour singles final.
Petkovic dispatched Slovakia's Viktoria Kuzmova, 6-4 6-1, to book her place in her first semi-final of the 2019 season.
The other semi-final will see 8th seed Ekaterina Alexandrova take on Jelana Ostapenko from Latvia.
Russian Alexandrova beat Kristina Mladenovic from France, 6-2 6-4, while former French Open champion Ostapenko scored a 7-5 6-1 win over Elena Rybakina from Kazakstan.


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Okay, I have a confession to make....I'm a BIG fan of the Teenage Mutant Ninja Turtles! Ever since my local comic book store suggested I pick up a copy of the book and check it out, I was instantly hooked. Although I collected a lot of comics in high school, I wasn't a huge fan of B&W comics back then, so I would have completely skipped it otherwise. But I gave it a shot and I loved it. Over the years I have amassed quite a collection of TMNT comics, books, and, of course, toys!
The Turtles were a constant source of inspiration in many of my art projects in college, and I have all kinds of drawings, painting, sculptures featuring the "Heroes in a Halfshell", and I was even hired to paint the cast of the TV show on the walls of a birthday party room in a local arcade that I worked at during those years!
Action Figures
So obviously, I collected the TMNT action figures (why else would I be writing this, right?), and although Playmates created hundreds of action figures over the years, my focus stayed on the main team and any of the supporting cast that I liked. I am not a completist by ANY means for this line (which still continues to this day, thanks to last year's CGI movie!), but the figures I did get have always been some of my favorite toys ever!
Following the animated series' lead, the Turtles wear color-coded (a brilliant move, BTW) so that each one had a unique colored outfit. This helped to tell them apart, because previously, they all wore red.
Now that I had the main TMNT cast in my collection, it was a tough call to determine which additional figures to add to it. April didn't interest me, and neither did the villains. While I knew Shredder & the Foot Soldiers from the comics, I didn't like the action figures versions, so I just passed on those. As you know from previous columns, I got pretty picky with teh supporting cast members, and I only wanted to get the characters that were in the comics. This is a rule I would break of course, but hey, I was on a college student budget!
So out of the hundreds of TMNT figures that came out, here are the ones I did pick up.
So there you have it! The first few years of the TEENAGE MUTANT NINJA TURTLES line had a lot of interesting figures in it, but these were the only ones I collected. Characters like Ace Duck, Napolean Bonafrog, Krang, Slash, and Mondo Gecko were fun for kids, but I didn't get into them. There WAS another related figure that spun out of the TMNT line, which I'll feature as a surprise over the weekend.
See you next week!


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Kelli Fox was born in 1964 in Vancouver, British Columbia, Canada. She is an actress, known for Timecop (1994), Double, Double Toil and Trouble (1993) and Moonlight and Valentino (1995).
Fox has been a member of the Shaw Festival acting company for several seasons. She is now appearing at the Stratford Shakespeare Festival in Stratford, Ontario.
Kelli Fox is the younger sister of actor Michael J. Fox and sister-in-law of actress Tracy Pollan.
Community content is available under CC-BY-SA unless otherwise noted.


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Employer Summary
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Poor bonus scheme which doesn't reflect value.
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Everything from the pension to the maternity pay is the bare minimum legal amounts required by law
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Being able to buy additional holidays and health benefits can also apply for loan for travel expenses
List based on reports from current and former employees. It may not be complete.


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A week packed full of announcements
A stack of information published this week should ensure some volatility for markets around the world.
The EUR/USD pair had a mainly stable night of trading, weakening in the lead up to the US Non-Farm Payroll figure. It hit session lows of $1.22719 just before the NFP figure, but began to strengthen upon release. It then reached session highs in the two hours following the release, peaking at $1.23337. It is now currently trading at $1.23091. The GBP/USD was very different. It hit session lows of $1.37869 early before steadily climbing. It reached session highs of $1.38883 but then gave up some of those gains to now be trading at $1.38532.
The Dow had a solid night of trading. After a flat start to the session, dropping to session lows of 24,835.40, the Dow took off when the NFP figure was released. It rallied over 400 points to end the session at 25,333.26. It has had a leg-up this morning, currently trading at 25,413.00. The Dax posted some solid gains as well in the last trading session. After reaching session lows of 12,281.53 in early trading, it followed the Dow in climbing upwards when the NFP data was released. It got to session highs of 12,411.35, then gave a chunk of gains before recovering again to where it is currently trading at 12,372.10.
Plenty of vital data set for release this week. The data released today and the most significant data published this week include:
Today – Japanese BSI Manufacturing Index & Prelim Machine Tool Orders, Eurogroup Meetings, US Federal Budget Balance, NZ FPI
Tuesday – UK Annual Budget Balance, US CPI & Core CPI, BoC Governor Stephen Poloz speaks
Wednesday – China Industrial Production, US Retail & Core Retail Sales, US Crude Oil Inventories & PPI, ECB President Mario Draghi speaks, NZ GDP
Thursday – US Import Prices, Unemployment Claims & Empire State Manufacturing Index
Friday – US Building Permits & Housing Starts, European Final CPI
Disclaimer: The information in this website is of a general nature only and the advice has been prepared without taking account of your objectives, financial situation or needs. Accordingly, before acting on the advice, you should consider the appropriateness of the advice having regard to your objectives, financial situation and needs. We recommend you seek independent advice if necessary.


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Glade Springs
Web Development
The Glade Springs Village Property Owners Association (POA) was incorporated in 2001 as a domestic, non-profit corporation in West Virginia. The original developer, Cooper Land Development, Inc., an Arkansas corporation has filed a Declaration of Covenants and Restrictions for Glade Springs Village in order to impose restrictions under a general plan of improvement for the benefit of all the property owners and to establish a method of maintenance, preservation, use and enjoyment of the property. The Declaration is recorded with the Raleigh County, West Virginia Recorder at Book 5004, Page 6485. A copy of the Declaration is available on our owner information page. Justice Holdings, LLC purchased all interest in the POA from Cooper Land Development in October 2010.


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Trio drives to Louisiana to help animals rescued from Hurricane Katrina
September 12, 2005
By Doyle Murphy
Imagine an arena like Island Grove Regional Park, and imagine it's filled with animals, said area veterinarian Colin Combs.
That's the reality Combs, his wife, also a veterinarian, and brother-in-law found when they reached the Lamar-Dixon Expo Center Monday morning near Gonzales, La. About 3,000 animals rescued from Hurricane Katrina have ended up at the Expo Center.
Combs, 39, and his wife, Rebecca Hubert, 38, of Greeley took time off from West Ridge Animal Hospital, 6525 28th St. in Greeley, to help care for the animals. Hubert's brother, Alan, 44, of Evans is a member of the Air National Guard and former volunteer firefighter. The trio drove down Sunday.
A second center in Baton Rouge, La., handles most of the pets affected by the hurricane. At the Expo Center, Combs said, the majority of the animals are strays. Some arrive after two weeks spent in polluted waters.
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"I think that's been one of the huge heartbreaks, is everyone seeing all the animals on TV," Combs said.
Rescue workers deliver more animals, mostly dogs, every day. Combs said it's a myth police officers shoot animals rather than rescue them. A hundred animals arrived on Monday morning. Combs expected at least 30 more in the evening.
If needed, the three planned to stay a week. Until then, they'll count on co-workers to take care of the animals in Weld County.
Evergreen Colorado Kennel Club, which hosted dog shows at Island Grove on Saturday and Sunday,
collected money and matched the amount donated. It will send $784 to the AKC/CAR Canine Support and Relief Fund and $1,984 to Take the Lead, a nonprofit organization supported by professional dog handlers and judges which provides financial support for human dog fanciers affected by catastrophic events. Greeley Kennel Club will also donate $2,000 to the AKC/CAR.


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Louisiana Chicken Pasta – Parmesan Crusted Chicken in a Spicy New Orleans Sauce
Prep Time 20 mins
Cook Time 35 mins
Total Time 55 mins
Servings: 6 servings
Calories: 674 kcal
Delicious Parmesan crusted chicken is served over bow tie pasta with mushrooms, green onions, bell peppers and tomatoes in a spicy New Orleans sauce. Very easy, one skillet meal that comes together quickly and is full of flavor!
- 1.5 lb chicken breasts thin (4 to 5 pieces)
- ½ cup Parmesan cheese
- 1 cup bread crumbs
- 1/3 cup flour
- 2 eggs
- ½ tsp salt
- 1 tsp Italian Seasoning
- 1 tbsp olive oil for cooking chicken
Veggie part of the sauce:
- 2 tbsp butter
- 1 tbsp olive oil
- 8 oz cremini mushrooms sliced
- 2 green onions chopped
- ½ large yellow bell pepper chopped
- ½ large red bell pepper chopped
- 3 cloves garlic sliced
- 1 small tomato chopped
- 1 cup heavy cream
- 1 cup chicken stock sodium free (adjust salt if not sodium free)
- 1 tbsp flour
- 1 cup Parmesan cheese shredded
- 1 to 1.5 tsp Cajun Seasoning
- 1 tablespoon salt
- 2 teaspoons garlic powder
- 1 tablespoon paprika
- 1 teaspoon ground black pepper
- 1 1/2 teaspoons dried oregano
- 1 1/2 teaspoons dried thyme
- 1 teaspoon onion powder
- 1 teaspoon cayenne pepper
- ½ tsp paprika
- ½ tsp red pepper flakes
- Salt to taste
- 10 oz bow tie pasta
TO MAKE BATTER FOR CHICKEN: Even though we won't cook chicken in this step, we want to make the flour mixture now ahead of time so that it's right there in front of us when we need to use it. In a shallow dish mix ½ cup Parmesan cheese, 1 cup bread crumbs, 1/3 cup flour, ½ tsp salt and 1 tsp Italian Seasoning and set aside. Add 2 eggs into another dish or a bowl and whisk them until mixed.
TO COOK VEGGIES FOR THE SAUCE: Heat 1 tbsp olive oil and 2 tbsp butter on medium high heat in a large stainless steel skillet until melted. Add 8 oz sliced cremini mushrooms to the hot pan and cook for a total of 3 minutes – 1.5 minute on each side. Remove mushrooms, if you want them to stay more flavorful and intact for the sauce. Otherwise, you can leave them in the same pan. I only like to remove them because they retain flavor and texture better that way – it's not necessary.
Add ½ red bell pepper and ½ yellow bell pepper, chopped to small cubes and 2 chopped green onions and a splash of olive oil. Cook for 4 minutes on medium high until softened. Add 3 garlic cloves and cook for 1 additional minute until garlic is fragrant.
TO MAKE SAUCE: Stir in 1 tbsp flour and then add 1 cup chicken stock, 1 cup heavy cream, 1 to 1.5 tsp Cajun Seasoning, ½ tsp paprika and ½ tsp red pepper flakes and salt to taste. Add 1 small chopped tomato and simmer over low heat for about 5 minutes.
Add 1 cup shredded Parmesan cheese to sauce, stirring to incorporate. Taste the sauce now before adding pasta to make sure it's seasoned to your liking, add a pinch more of Cajun seasoning, red pepper flakes and salt if necessary.
TO COOK PASTA: In the meantime bring a large pot of water to boil, add pasta and cook it according to instructions. Drain the pasta, but do not rinse. For authentic "al dente" pasta, boil your pasta uncovered, stirring occasionally for 12 minutes. For more tender pasta, boil an additional 1 minute.
TO COOK CHICKEN: If your chicken breasts are not thin, butterfly them, or flatten them by pounding with a meat tenderizer. The thinner the chicken breasts the better.
Take a chicken and dip in flour mixture first, then into the egg bowl, then back into the flour mixture. I went from one bowl to the other until I liked how much stuff I had on the chicken. Repeat with each chicken breast and set them aside on a plate.
Heat 1 tbsp olive oil on medium high in the stainless steel skillet (2nd skillet, not the one that has the sauce in it) and cook on medium high for 3 to 4 minutes on each side (total of 6-8 minutes) – just set the timer and flip when it beeps. Check to make sure the chicken is fully cooked.
TO PUT IT ALL TOGETHER: Add pasta to the pan and stir until fully coated in sauce. Add mushrooms (if you removed them earlier) to the sauce now. Season with more salt if necessary. Slice chicken into thin strips and serve on top of pasta. Sprinkle with additional shredded Parmesan and chopped fresh green onions, if desired.


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Bacillus clausii SIN is one of the four strains of B. clausii composing a probiotic administered to humans for the prevention of gastrointestinal side effects due to oral antibiotic therapy. The strain is resistant to kanamycin, tobramycin, and amikacin. A gene conferring aminoglycoside resistance was cloned into Escherichia coli and sequenced. The gene, called aadD2, encoding a putative 246-amino acid protein, shared 47% identity with ant(4′)-Ia from Staphylococcus aureus, which encodes an aminoglycoside 4′-O-nucleotidyltransferase. Phosphocellulose paper-binding assays indicated that the gene product was responsible for nucleotidylation of kanamycin, tobramycin, and amikacin. The aadD2 gene was detected by DNA-DNA hybridization in the three other strains of the probiotic mixture and in the reference strain B. clausii DSM8716, although it did not confer resistance in these strains. Mutations in the sequence of the putative promoter for aadD2 from B. clausii SIN resulted in higher identity with consensus promoter sequences and may account for aminoglycoside resistance in that strain. The aadD2 gene was chromosomally located in all strains and was not transferable by conjugation. These data indicate that chromosomal aadD2 is specific to B. clausii.
Probiotics are administered to humans for the prevention of gastrointestinal side effects due to oral antibiotic therapy. Ingestion of high quantities of spores is thought to restore an intestinal flora following the destruction of the normal flora by antibiotics (17). Enterogermina is a preparation of Bacillus available in Italy (3, 18). It is composed of four antibiotic-resistant strains, OC, NR, T, and SIN (3), initially identified as Bacillus subtilis but recently assigned to the Bacillus clausii species (26). For the prevention of intestinal disorders, the strains of Bacillus used in the Enterogermina preparation were rendered multiantibiotic resistant to survive in the presence of the antibiotics which are coadministered (3, 18). One of the strains, B. clausii SIN, is resistant to kanamycin, tobramycin, and amikacin (3). Resistance to aminoglycosides is generally due to enzymatic modification of the drugs and is widespread in a variety of bacterial pathogens (27). However, aminoglycoside inactivation has rarely been reported for Bacillus spp. An aph(3′)-IVa gene from a butirosin-producing strain of Bacillus circulans was reported and subsequently sequenced (4, 10). This gene encodes an aminoglycoside 3′-O-phosphotransferase type IV which confers resistance to butirosin, gentamicin B, kanamycin, lividomycin, neomycin, paromomycin, and ribostamycin by modifying these aminoglycosides at the 3′-hydroxyl group (10). B. subtilis 168 possesses a chromosomal aadK gene which encodes an aminoglycoside 6-O-adenylyltransferase that confers resistance to streptomycin (23). A kanamycin 4′-O-nucleotidyltransferase encoded by plasmid pTB913 from a thermophilic Bacillus sp. was found to be similar to that encoded by the gene ant(4′)-Ia or aadD borne by plasmid pUB110 from Staphylococcus aureus (16). The aim of this study was to characterize the mechanism of resistance of B. clausii SIN to aminoglycosides and to elucidate the genetic basis of this mechanism.
Bacterial strains.The four B. clausii strains OC, NR, SIN, and T used for the production of Enterogermina were obtained from Sanofi-Synthelabo OTC SpA (Milan, Italy) as separate spore suspensions. B. clausii DSM8716 was used as a reference strain.
Susceptibility to antibiotics.Antibiotic susceptibility was tested by the disk diffusion method, and MICs were determined by agar dilution according to the recommendations of the National Committee for Clinical Laboratory Standards (21). Susceptible, intermediate, and resistant categories were defined according to the recommendations of the National Committee for Clinical Laboratory Standards (22).
Mating experiments. Enterococcus faecalis JH2-2 (11) and Enterococcus faecium HM1070 (1), both resistant to rifampin and fusidic acid, and B. subtilis UCN19, resistant to ciprofloxacin, were used as recipients in mating experiments (5, 11). The latter strain was obtained by stepwise selection of a mutant from B. subtilis 168 (12) on agar plates containing increasing concentrations of ciprofloxacin. In every transfer experiment, E. faecalis BM4110 containing the conjugative plasmid pAMβ1 (14) was used as a control. Agar plates for the selection of transconjugants contained rifampin (50 μg/ml) plus fusidic acid (20 μg/ml) or ciprofloxacin (8 μg/ml) combined with 128 μg of kanamycin per ml for enterococci and 20 μg/ml for B. subtilis. All mating experiments were repeated a minimum of three times.
PCR.The deoxynucleotide primers ANT4-1 (5′ TAAATATGGGGATGATGTTAAGGC 3′) and ANT4-2 (5′ TGGTATGCGTTTTGACACATCCAC 3′), specific for ant(4′)-Ia, were used. S. aureus BM3002 was used as a control (6).
DNA manipulations.Total and plasmid DNA from Bacillus strains was extracted as described previously for enterococci (2). Cloning was carried out by standard techniques (25). Total DNA from B. clausii strains was digested with various restriction enzymes, cloned into plasmid pUC18, and introduced by electrotransformation into Escherichia coli DH10B (2). Recombinant plasmids were selected on agar plates containing ampicillin (200 μg/ml) and kanamycin (20 μg/ml). A DNA fragment conferring kanamycin resistance was then subcloned into the shuttle plasmid pJIM2246 conferring resistance to chloramphenicol (24) and introduced into E. faecalis JH2-2 by electrotransformation (1). Sequencing of DNA was performed on ABI PRISM 377 (Perkin-Elmer Corp., Norwalk, Conn.) with big dye terminator according to the protocol supplied by the manufacturer. Nucleotide and amino acid sequences were analyzed by using the software available online at Pedro's Biomolecular Research Tools website (http://www.up.univ-mrs.fr/∼wabim/pedro/research_tools.html ) and at the National Center for Biotechnology Information website (http://www.ncbi.nlm.nih.gov/ ). Putative promoters were identified by using the Neural Network Promoter Prediction software (www.fruitfly.org/seq_tools/ ). Score predictions of >0.9 were considered to be significant.
Southern hybridization.DNA from B. clausii strains was digested with I-CeuI or SmaI (New England Biolabs, Beverly, Mass.), separated by pulsed-field gel electrophoresis with a technique used for enterococci (2), transferred onto a nylon membrane, and hybridized to a probe specific for aadD2 of B. clausii and consisting of the entire gene amplified by PCR and labeled with digoxigenin (Boehringer Mannheim). DNA was then hybridized to a probe specific for the 16S rRNA. The 16S rRNA probe was obtained by labeling the DNA fragment amplified from B. clausii DSM8716 with deoxynucleotide primers 5′ AACTGGAGGAAGGTGGGGAT 3′ and 5′ AGGAGGTGATCCAACCGCA 3′ (7).
Aminoglycoside-modifying enzyme assays.Bacteria were lysed by ultrasonic disintegration, and the resulting extracts were centrifuged at 100,000 × g for 45 min. Aminoglycoside-modifying activity in the supernatant (S100) was tested by the phosphocellulose paper-binding technique with [α-32P]ATP (specific activity, 30 Ci/mmol) as described previously (8). Aminoglycosides (67 μg/ml, i.e., 1.4 × 10−1 mM kanamycin or tobramycin) were incubated with S100 extracts at 30°C for 30 min in TMND buffer (0.06 M Tris-HCl, 0.04 M MgCl2, 0.4 M NH4Cl, 1.6 × 10−4 M dithiothreitol [pH 7.1]).
Nucleotide sequence accession number.The 1,006-bp fragment containing aadD2 was submitted to GenBank and assigned accession no. AF539790 .
Antibiotic resistance in B. clausii strains.By disk diffusion, all B. clausii strains were found to be resistant to penicillin G, cefalotin, cefotaxime, erythromycin, and lincomycin. Resistance to aminoglycosides was detected only in B. clausii SIN, and the MICs of certain aminoglycosides for the B. clausii strains are shown in Table 1. B. clausii SIN was resistant to kanamycin, tobramycin, and amikacin but susceptible to gentamicin and netilmicin.
Characterization of the aadD2 gene from B. clausii SIN.An 8-kb EcoRI DNA fragment originating from the DNA of B. clausii SIN was cloned into plasmid pUC18. The recombinant plasmid, pUV11, conferred resistance to ampicillin and kanamycin. A 2.2-kb Sau3A fragment that also conferred resistance to kanamycin was subsequently subcloned into pUC18, generating plasmid pUV12, and sequenced. Analysis of the sequence revealed the presence of an open reading frame of 765 bp or an alternative open reading frame starting 6 bp upstream and putatively encoding 254- or 256-amino-acid proteins. Both ATG start codons were preceded at 9 and 10 bp by ribosome-binding site-like sequences, 5′ GAGATGGAAG 3′ and 5′ CAAAAGGAGA 3′, complementary to five and seven bases (underlined), respectively, of the 3′-OH-terminal sequence (5′ UCUUUCCUCC 3′) of B. subtilis 16S rRNA (19).
A search for the presence of motifs in the deduced amino acid sequence detected a nucleotidyltransferase domain between amino acids 14 and 59 that is shared by a large family of nucleotidyltransferases (15). In this family, highest homology (identity, 47%; similarity, 65%) was found with the enzyme encoded by ant(4′)-Ia or aadD (GenBank accession no. V01282 ) of plasmid pUB110 from S. aureus (Fig. 1) (20, 27). As previously mentioned, a gene that is identical, except for a 1-bp substitution, was reported in a thermophilic Bacillus sp., where it was borne by plasmid pTB913 (16). This nucleotidyltransferase modifies kanamycin, tobramycin, and amikacin and confers a resistance phenotype similar to those of B. clausii SIN and the E. coli transformant (Table 1).
The aadD2 gene was subcloned into shuttle vector pJIM2246, and the recombinant plasmid, pUV13, was electrotransformed into E. faecalis JH2-2. For the transformants, the MICs of kanamycin and tobramycin were elevated and that of amikacin was moderately higher but the MICs of gentamicin and netilmicin remained unchanged. The resistance phenotype was consistent with modification of the 4′-hydroxyl group of aminoglycosides.
Analysis, by the phosphocellulose paper-binding assay, of bacterial extracts from E. coli DH10B/pUV12 indicated that aminoglycoside resistance was due to a nucleotidylating activity. Kanamycin A nucleotidylation was defined as 100%, and tobramycin (91%), amikacin (16%), isepamicin (14%), and neomycin B (100%) were modified at the indicated percentages. Gentamicin C1 and netilmicin were not substrates for the enzyme. On the basis of the substrate profile, the enzyme encoded by pUV12 was an aminoglycoside 4′-O-nucleotidyltransferase. Similar results were obtained with B. clausii SIN extracts and, surprisingly, also for B. clausii DSM8716, although the enzyme activity was much weaker in the latter strain. According to the nomenclature proposed by Shaw et al. (27), the kanamycin nucleotidyltransferase of B. clausii was assigned to the Ant(4′)-I enzyme family and the corresponding new gene was tentatively called aadD2.
Distribution and localization of aadD2.After digestion with the I-CeuI enzyme, which recognizes a sequence which is specific for rRNA operons (13), total DNA of the B. clausii strains yielded apparently six fragments, indicating that this species contains a minimum of six rRNA operons (Fig. 2). Southern experiments showed that all fragments hybridized with an rrs probe specific for 16S rRNA. The aadD2 probe hybridized to a single large chromosomal fragment of each strain. B. clausii OC, NR, SIN, and T had indistinguishable SmaI-generated patterns, and hybridization showed that the aadD2-like genes were borne by a fragment approximately 30 kb in size in the probiotic strains and by a larger fragment of ca. 50 kb in B. clausii DSM8716 (data not shown). Thus, aadD2-like genes were present in all the strains of B. clausii studied but were phenotypically expressed only in B. clausii SIN.
Sequence analysis of the 8-kb EcoRI insert of plasmid pUV11 revealed the presence, 3′ downstream from the aadD2 gene, of genes homologous to blaZ, blaI, and blaR from Staphylococcus, responsible for the production and regulation of a penicillinase (our unpublished data), a ytrA-like gene encoding a putative 135-amino-acid protein homologous to tRNA synthetases, the structural gene for an ABC transporter, and a gntR-like gene putatively encoding a transcriptional regulator. The presence of aadD2 adjacent to genes known to be part of the chromosome confirms its chromosomal location. In addition, repeated attempts to transfer resistance to aminoglycosides and to beta-lactams from B. clausii strains to E. faecalis JH2-2, E. faecium HM1070, and B. subtilis UCN19 by conjugation were unsuccessful.
Analysis of the putative promoter of aadD2 from B. clausii SIN.There was a discrepancy between the presence of aadD2 in all strains of B. clausii studied and the various levels of aminoglycoside resistance of the strains. We thus amplified and analyzed a DNA sequence of nearly 200 bp upstream from the aadD2 genes of B. clausii SIN (resistant to aminoglycosides), NR, OC, T, and DSM8716 (susceptible to aminoglycosides). Potential −35 and −10 consensus sequences were identified, one of which may be the recognition site for a DNA-dependent RNA polymerase (Fig. 3). In all strains, one promoter sequence was confidently predicted (prediction score, 0.93). Comparison of the sequences revealed three nucleotide substitutions in the B. clausii SIN sequence. Two of these substitutions were located in putative −35 and −10 regions. A C-to-A mutation rendered the −35 sequence (TTAGCA) identical to the consensus promoter sequences of E. coli and B. subtilis (9, 19), which might result in a stronger promoter. Another mutation was located in the −10 sequence (Fig. 3). This mutation led to the prediction (score, 0.95) of an additional putative promoter. The consequence, if any, of the third substitution remains unknown.
Although the origin of B. clausii SIN is not known, the identity of the SmaI restricted DNA profiles of strains composing Enterogermina strongly supports the notion that SIN is a derivative of one of the other B. clausii probiotic strains or of a common parental strain.
This study was supported in part by a grant from Sanofi-Synthelabo OTC SpA, Milan, Italy.
- Received 26 August 2002.
- Returned for modification 15 November 2002.
- Accepted 18 January 2003.
- Copyright © 2003 American Society for Microbiology


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Today's Guests:
The Educated Investor with David Cichon of the the Householder Group and Cichon Financial.Today David discusses the importance of the foundation as you build your financial house. Then Allen Stack, Owner of Stack Optical joins us to talk all about the services at Stack Optical. Get your $69 Eye exam booked today! Then it's Your Health Matters brought to you by Integrated Health Systems. Today Jess Sorci, Metabolic Coordinator for IHS talks about building your immune system. Integrated Health Systems offers a free consultation when you mention Haystack Help radio, visit their website at www.IntegratedHealthDenver.com.
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Creative Market 2015 Year in Review
Long scrolling One Pager highlighting 33 awesome moments from Creative Market's 2015 year.
2015 has been all about growth for the Creative Market community. Everything we do is to help independent creators make a living doing what they love – creating beautiful design content for anyone working on a creative project – and in 2015 we've soared past 11,000 independent shops, selling 455,000+ unique products, to our community of more than 1,500,000 members.
Categories Annual Report
- Need Hosting? I'd highly recommend Bluehost
- Broken link? Please Email or Tweet me
- Credits: Creative Market
- Reviewed: 18 Jan 2016 by Rob Hope


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Our students designed some art creations using recycled products and they were worth seeing…
Category: Kırsehir Merkez Cumhuriyet Ortaokulu
Toy houses by Kırşehir students
Our students are working hard for both their lessons and their nature. They are building toy houses for their little brothers and sisters from recycled products and they are enjoying their time.
Orienteering in Kırşehir
Our students had a day out this week.They practiced orienteering. They had compasses and maps in their hands and they were able to find their way to the target destinations. It was fun for bothContinue reading
A trip to Japanese Garden
A garden with every different type of trees to be discovered and observed, Japanese garden will always be a quest in visitors' heart.The same happened for our students,too. We organized a trip to Kaman forContinue reading
A day off
Students and teachers from Kırşehir Cumhuriyet had a day-off together saying goodbye to summer and welcoming the autumn.They danced, played games and observed nature. The volleyball match between the teachers and the students ended inContinue reading


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Past Events › Festival
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March 2019
medienwerk.nrw at META Marathon 2019
"META Marathon" is a New Technology Festival. From 15-17 March 2019, the NRW Forum will be transformed into a Living Lab with a 42-hour non-stop programme. On the 17th at 11am the Office medienwerk.nrw presents a panel with Katrin Hochschuh, Adam Donovan and So Kanno.Find out more »
May 2019
Blue Skies #1 Bodies in Trouble
The growing impact of technology on life as we know it brings changes to our bodies, our communities and the environment – what can we do? How can we understand and shape these changes collectively? These are the questions PACT Zollverein asks in its new reccuring festival "Blue Skies". The first edition is developed together with the Office medienwerk.nrwFind out more »