text:
Nancy M. Bivens, MD, PhD
Assistant Professor of Psychiatry at CUMC
Dr. Nancy Bivens completed Adult Psychiatry residency and Schizophrenia Research Fellowship training at Columbia University and New York State Psychiatric Institute. She is now an Assistant Professor of Clinical Psychiatry in the Division of Developmental Neuroscience, working on animal models of psychosis, as well as the intersection of human genetics with animal modeling of brain disorders.
She is the recipient of an NIMH funded Career Development Award, the NARSAD Brain Behavior Research Fund Young Investigator Award, and is a Leon Levy Investigator in Neuroscience Research.
Neurological Institute of New York
710 West 168th Street
Floor: 12 New York, NY 10032
Phone: (212) 305-9758
Fax: (650) 646-3120
Public Transportation: Yes
Disabled Access: Yes


text:
Sesame Seed Oil Crust
|Whole wheat flour
|1 1⁄4 Cup (20 tbs), measured scoop and level
|White sesame seeds
|1⁄4 Cup (4 tbs) (Hulled)
|1⁄2 Teaspoon
|5 Tablespoon
|Ice water
|8 Tablespoon (6 Plus 2 Tablespoon)
|Egg white
|1 , lightly beaten
1. Pre-heat the oven to 450°F.
2. Using either your hands or a food processor, mix the sesame seeds whole wheat flour and salt.
3. In a bowl, mix oil and ice water.
4. Slowly add the liquid and continue mixing.
5. Add more water if needed and knead or process the dough into a ball.
6. Flatten it into and 8-inch circle, dust with flour and place between two wax paper sheets.
7. With a rolling pin, roll from the center outward, carefully peeling off the wax paper to release dough and replacing paper. Repeat with the other side. Repeat the process till the dough is 11 inches in diameter.
8. Take a 9-inch quiche pan with removable bottom or a 9-inch greased pie plate and place the dough on it, trimming the excess by rolling to pin on the top of the pan in case of the quiche pan , and trim the edge, leaving a 1-inch overhang, which you should then fold under and flute, in case of a pie plate..
9. Make holes on the surface with a fork.
10. In case of quiche pan, brush with lightly beaten egg white and refrigerate for one hour to dry. In case of pie plate, refrigerate for 10-15 minutes.
11. In the oven, bake for 15 minutes at 400°F if using a pie plate. And you may cook for 2-3 minutes to set the glaze in case of a quiche pan.
12. Cool on a wire rack before filling.
13. Serve with a filling of choice.
You can also bake the crust in a square pan. Take 1/3rd of the dough out and place the remaining in a layer at the bottom.
Divide the 1/3rd dough into four parts and make 6-6 1/2 inch strips of each, to press into the sides with the seams joined and bake.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 1447 Calories from Fat 858
% Daily Value*
Total Fat 97 g149%
Saturated Fat 10.3 g51.4%
Trans Fat 0 g
Cholesterol 0 mg
Sodium 1047.6 mg43.6%
Total Carbohydrates 124 g41.5%
Dietary Fiber 19.8 g79.3%
Sugars 0.9 g
Protein 32 g63.3%
Vitamin A 0.3% Vitamin C
Calcium 6.1% Iron 35.2%
*Based on a 2000 Calorie diet

subset: commoncrawl


text:
The 1905 Cumberland County Courthouse is built of Indiana limestone, a local sandstone known for its distinctive color. The town square is a quaint collection of restored buildings and new businesses "" a representative mix of the history and energy of the town today.
Tennessee Vacation eGuide
The 2016 eGuide gives you instant access to Tennessee's irresistible attractions and destinations. Peruse venues online, then put in your order to get a free guide delivered to your doorstep.


text:
- Players
- 1
- Genre
- Import
- Release Date
Wii Shop
- 21st Jul 2008, 900 points (USA)
- 16th May 2008, 900 points (UK/EU)
- 26th Feb 2008, 600 points (JPN)
About This Game
In the year 2025, a war breaks out between humans and a hostile alien race, and 16-year-old Lucia and the rest of the starfighter pilots in the Earth Federation must join the battle. When Lucia's father is kidnapped by the aliens, she decides to take action and hijacks a brand new prototype starfighter, the Gley Lancer, in order to try to save him.
In this side-scrolling shooter, step into the cockpit of the Gley Lancer as Lucia, rescue your father and destroy the evil aliens that are bent on destroying the human race.


text:
We make Chiropractic FUN for kids. We designate the first Thursday of every month KIDS DAY. Each Kids Day has a different theme. We aim to make the next generation the healthiest generation! Call for details.
The first stress placed on a child's spinal column is during the birthing process. Chiropractic care is a safe and effective treatment. Pediatric chiropractic information brought to you by Integrity Doctors.


text:
Healthline Blogs
Guess What the McRib Has in Common With Your Shoes?
Never once have I understood the appeal of the McRib, McDonald's elusive and strangely popular BBQ-ish sandwich.
In essence, between two pieces of bread are some onions, a pickle, and drenched in BBQ sauce is a patty of rib meat reconstituted to look like ribs. The point is to give you the taste of some ribs without all of those pesky bones.
That's what you can see in photos and in commercials, but when given the microscope treatment—or when you simply look at the ingredients on the company's website—you'll find ingredients you wouldn't put in there if you were making your own sandwich.
The smart folks over at Time Magazine reported that within the 70 ingredients in a McRib, there are some interesting components that make a McRib sound more like a chemistry project than food. Some of them, according to Time, include:
- Polysorbate 80—a common food additive that makes ice cream smoother, and used in vaccines in Europe and Canada. (If you have Crohn's disease, you don't want this.)
- Ammonium sulfate—a salt whose main purpose is typically for fertilizing dirt, but is also used as a flame retardant. It's been banned in many countries because of certain militants' affinity for using it to blow things up.
- Azodicarbonamide—this flour-bleaching agent is mainly used in making foam plastics, like the insulation on your windows, the soles of your shoes, and your yoga mat.
Yeah, yoga is good for you, but eating your mat might not be the best way of reaching your chakras, or shorten your time on the toilet.
Then again, all of the ingredients in the McRib have been proven safe for human consumption by studies and the U.S. Food and Drug Administration.
Then again, azodicarbonamide has been banned in Europe and Australia, as Time reported, because it can contribute to the development of (or worsen) asthma if you're exposed to it at work. But hey, that's only two out of the seven continents, so it's no big deal.
Then again, the fact that McDonald's food isn't the healthiest is no real surprise. They make mass-produced food that is manufactured for a long shelf life for the least amount of money. There are fillers, preservatives, and—as we've already illustrated—chemicals that are hard to produce.
Real food—the kind that grows on trees, in fields, and doesn't have to be pressed into shapes that look like food—costs money and can spoil, so while it makes tons of nutritional sense, it makes for bad business, if your business is to feed millions globally from a Dollar Menu.
You can either eat healthy like seasonal produce, or you can eat seasonal fast food like the McRib, which happens to be seasoned with things that the FDA says is safe, but other continents do not.
The choice is yours.
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Hotel All Suites Le Teich, Le Teich
- Free WiFi and free parking
Main amenities
- 49 smoke-free guestrooms
- Outdoor pool
- Breakfast available
- Fitness center
- Terrace
- Air conditioning
- Daily housekeeping
- Luggage storage
- Limited hour front desk
For families
- Free cribs/infant beds
- Connecting/adjoining rooms available
- Private bathroom
- Free toiletries
- Terrace
- Daily housekeeping
What's around
- In Arcachon Bay
- Parc Ornithologique du Teich (2.8 mi / 4.6 km)
- Gujan Golf (3.7 mi / 5.9 km)
- Parc de la Coccinelle (4.5 mi / 7.3 km)
- KID PARC Ile d'Aventures (4.5 mi / 7.3 km)
- Maison de l'Huître (4.6 mi / 7.4 km)
More great choices
At a glance
Key facts
Hotel size
- This hotel has 49 rooms
- This hotel is arranged over 2 floors
- Check-in time 3 PM-9 PM
- Check-out time is noon
- Express check-in
- January to April and October to December
- Monday - Saturday: 7 AM - 9 PM
- Sunday: 7 AM - 2 PM
- Check-in is available 3 PM to 9 PM Monday - Saturday.
- May, June, and September
- Daily: 7 AM - 9 PM
- Check-in is available 3 PM to 9 PM daily.
- July and August
- Monday - Saturday: 7 AM - 12:30 AM
- Sunday: 7 AM - 9 PM
- Check-in is available 3 PM to 12:30 AM Monday - Saturday and 3 PM to 9 PM Sundays.
Guests arriving outside of these hours must contact the hotel in advance to arrange check-in. Contact details are provided in the booking confirmation.
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
- Minimum check-in age is 18
Travelling with others
- Pets allowed *
- Free WiFi in public areas
- Free WiFi in rooms
- Free self parking
Other information
- Smoke-free property
Nearby Three Star Le Teich hotels
In the hotel
- Buffet breakfast daily (surcharge)
- Outdoor pool
- Fitness facilities
- Sauna
- Bicycle rentals nearby
- Golfing nearby
- Hiking/biking trails nearby
- Mountain biking nearby
- Sailing nearby
- Water skiing nearby
- Windsurfing nearby
- Front desk (limited hours)
- Luggage storage
- Number of buildings/towers - 1
- Elevator/lift
- Terrace
- Accessible bathroom
- In-room accessibility
- Roll-in shower
In the room
- Air conditioning
- Iron/ironing board (on request)
- Blackout drapes/curtains
- Private bathroom
- Shower only
- Free toiletries
- Hair dryer
- Flat-screen TV
- Satellite TV channels
- Desk
- Free WiFi
- Phone
- Daily housekeeping
- Connecting/adjoining rooms available
Top Le Teich hotels
Recently reviewed hotels in Le Teich
- Kyriad Prestige Bordeaux Ouest MérignacFrom
"Comfy room, great staff and the breakfast was good."Nov 8, 2016
- Hôtel Le RelaisFrom
"A very nice small hotel in a resort area. We were there in the off season. Nice reception, comfy room."Oct 14, 2016
- ibis Styles Bordeaux AeroportFrom
"With a 6 am flight back to the USA the hotel was most accomodating -- securing a taxi for us at 5 am and a complete buffet breakfast at this early hour."Oct 10, 2016
- Best Western Bordeaux AeroportFrom
"We had a 6 a.m. flight. Their shuttle left at 5 on the dot and we were at the airport at 5:05. You are a captive audience for their restaurant and the food prices are a little…"Oct 4, 2016
Nearby hotels
Also popular with travelers from the USA
Special features
On Site
- Fitness facilities
- Sauna
- Bicycle rentals nearby
- Golfing nearby
- Hiking/biking trails nearby
- Mountain biking nearby
- Sailing nearby
- Water skiing nearby
- Windsurfing nearby
Hotel All Suites Le Teich, Le Teich's small print
Also known as
- All Suites Le Teich
- Hotel All Suites Le Teich
- Hotel Teich
- Hotel All Suites Teich
- Hotel All Suites
- All Suites Teich
The property has connecting/adjoining rooms, which are subject to availability and can be requested by contacting the property using the number on the booking confirmation.
Mandatory fees
You'll be asked to pay the following charges at check in or check out:
- A tax is imposed by the city: EUR 0.80 per person, per night. This tax does not apply to children under 13 years of age.
Optional extras
Buffet breakfast is offered for an extra charge of EUR 9.90 per person (approximately)
Pets are allowed for an extra charge of EUR 9.50 per pet, per day
We have included all charges provided to us by this hotel. However, charges can vary, for example, based on length of stay or the room you book.
Recent reviews
Good 3.8 /5 from 135 reviews
quiet hotel, perfect to stop for rest. didn't try the breakfast. room comfortable and clean. accepts dogs and charges 9.90 euros per dog per night, but doesn't provide anything for them, so... what is the charge really for? shops are close by car.
Beds are very hard and low to the ground, it was very warm so wanted the aircon on we were told its only available in high season not april even though it was the warmest it had been for many years.
Very nice. Good service.
Beds very hard with a thin mattress, no aircon allowed until may but the rooms were very warm in April so had to have the windows open so that means mozzys coming in at night. The shower was way to small even for my skinny daughter we had to shower with the door open
Stayed at this hotel twice before when visiting Arcachon and the surrounding area. Location might not be everyone's cup of tea, but we find it convenient and quiet. Housekeeping on this visit wasn't as good as it has been on past visits. After 2 nights, hope the very large cobweb hanging from bathroom ceiling has been removed! The only other criticism is the charge for our 2 small dogs. It's far too high.
Is the description of this hotel not correct? Tell us


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MetLife's Hauppauge HQ on the market
One of Suffolk County's signature buildings, the MetLife building in Hauppauge, is on the market for what industry experts suggest is about $19 million.


text:
Faountainhead Fortune Cookie
Fairfax Station, VA
Sat 10/05/2013
12 miles, Mountain Bike Race
300 Participants
|Sat 10/05/2013
|Mountain Bike Race (12 miles)
|12 miles
REV3 Adventure is excited to bring you The Fountainhead Fortune Cookie ? Not only do we bring you good rides but also good vibes with our all new XC mountain bike race at Fountainhead Regional Park in Northern Virginia. The REV3 Fortune Cookie makes your day of riding just that little bit more lucky. Hit those corners a little harder and ride the guys wheel in front of you a little closer knowing that the luck of the cookie is on your side. This mountain bike race is located in one of Northern Virginia?s premo mountain biking spots, Fountainhead Regional Park. Come and race around these sweet single track trails and show your buddies what you have been working so hard on all season. Skills, fitness and stamina. Newbies will ride around one lap of the Green and Blue trails, Weekend Warriors will do two and the hard core Trail Experts will tear it up for three laps. The Black loop will be out of commission for rehab so you get to ride the fast new trails at the front of the park. Once you have finished riding, compare your luck, eat your cookie and reflect on how awesome your day has been.
- Name:Mark Harris
- Email:email@example.com
- Phone:703-828-5990
- Address:Fairfax Station, VA
Fairfax Station, Virginia 22039


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Your phone will ring when the dealer answers.
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text:
Hello again Kitty,
I have three plants I traded for last summer now in the garden.
All are
variations on a theme... each one is larger than the other in size,
appearance. Amount of arch to the leaf, rippling along the mid-rid or
margin, coloring, etc. I am considering making a few more trades for
some of
the more exotic cultivars ... read much more expensive plants.
By themselves the plants are not all that jump up in your face
beautiful. Just different and one of those plants that fits a difficult
niche in the shade garden. Just so happens that it goes very well with
others. Great companion plant or not bad on its own. Can't ask much
than that.
I did order a couple of the more expensive cultivars, but they
were sold
out when my turn came at shipping.
Gene E. Bush
Munchkin Nursery & Gardens, llc
Zone 6/5 Southern Indiana
To sign-off this list, send email to email@example.com with the
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text:
Cotton Embossed Tunic Dress
by Area
$121 $604
Product details
Area Cotton Tunic Dress
- Cotton popover dress with button sleeve closure
- Dry clean only
- Made in New York City
* PLEASE NOTE: Each garment is embossed by hand resulting in a one of a kind piece: imperfections are part of the process. Embossing is meant to wear in with time. *


text:
Novi Vinodolskiのベッドルーム1部屋のアパートメント
お部屋の大きさ55 m²
Floor No.
- キッチン
- 鍵つきの部屋
- ペット可
- 朝食
- ジム
- ベランダ/テラス
- ベビーベッド
- 利用可能な駐車場
Accommodation summary
Largest wellness resort in Europe with 10.000 m2 of indoor area, 750 m2 of indoor and outdoor swimming pools, Kids' town (4.000 m2) with pools, playgrounds, Mini club and animation. Teen disco and gaming zone. 8 restaurants, 8 bars. Family friendly, modern, luxurious resort "NOVI SPA HOTELS AND RESORT". 1 km from the centre of Novi Vinodolski, in a quiet, sunny, elevated position, directly by the sea, directly by the beach. For shared use: grounds 31 ha (fenced) with plants, 7 swimming pools, indoor pool (27.03.-31.10.) with salt water and built-in steps. Paddling pool, tennis (3 x hard court, 2 x clay court) (extra), basketball, children's playground. In the complex: reception, restaurant, bar, fitness room, massage (extra). Spa area, games room, shops, boutique, laundry (extra). Linen change weekly. Towel change daily. Breakfast and half-board possible on request (extra). Parking, communal covered parking (extra). Sandy beach "NOVI SPA RESORT" pebble beach "NOVI SPA RESORT". Please note: car-free holiday complex. Entertainment in high season. Suitable for families, baby equipment on request, child minding available in high season (extra). Loading and unloading of baggage is possible at the holiday house. The complex has night security. The photograph is a typical example.
|Check-in before
The people were very friendly, the studio offered everything you needed, the bed was comfy and the location was good, just 350 m to the beach. You had your own terrace and parking lot, which was great. It is a very quiet location, good to calm down and relax. For two persons it is very big, I was scared, but of course most of people would enjoy that luxury of lots of space. The accommodation definitely is a great choice for that price!
The location of the apartment was great and the size was excellant plus with a private parking space in France unbelievable. Downside was cleaning fee per person when the floors were SO dirty that you wouldn'd want to walk on them without shoes on.
Danielle was very helpful. The house, next to her family's home was very clean and comfortable. The view of the mountains was wonderful. we highly recommend Landhaus Wegscheider to anyone interested in traveling to the Innsbruck area.
Fab apartment, fab views. All amenities on our doorstep, would highly recommend this apartment, will definitely be booking again. (unfortunately there are no English TV channels so bring a book)
Beautiful view of the Zugspitz from a private patio. Pleanty of room for two couples with 2 bedrooms. Beds very comfortable. Clean with friendly service.
A group of five girls stayed here and it was fantastic! The views were spectacular the location was spot on couldn't fault the apartment it was spotless and the finishing touch of the towels made up as swans on each bed with small toiletries was a lovely touch. We picked up the keys early from the interhome office the lady was extremely helpful. Would highly recommend we will be using this apartment again on our next visit to benidorm. Thank you
perfect!! the host were so accomodating...nice flat with a private pool...100% coming back!
A little romantic house near lake como. It's a historic stone house, which has been restored beautifully. Perfect for two persons, but also possible with two kids.
Beautiful, comfortable flat with a magnificent view over plage du sillon
Very nice and cosy little appartment. It's small but has everything one needs, the hosts are very friendly, and the location is both quiet and beautiful. Even the dog loved the stay!
- すべてのお部屋はチェック済み
- どのホストともお電話で直接お話いただけます


text:
Easy Cornbread
|Yellow cornmeal
|2 Cup (32 tbs)
|All purpose flour
|1 Cup (16 tbs)
|Baking powder
|1 Tablespoon
|Savory seasonings garden blend
|1⁄2 Teaspoon
|1⁄2 Cup (8 tbs)
|1 1⁄2 Cup (24 tbs)
|Whole kernel corn
|1 Cup (16 tbs)
|2 Large, beaten
Preheat oven to 350°.
Lightly-grease 9-inch square baking pan.
Combine cornmeal, flour, baking powder and barden blend in large bowl.
Set aside.
Melt butter in small saucepan over medium heat.
Add buttermilk and cook until warm; stir to combine.
Add buttermilk mixture, corn and eggs to diy ingredients; stir until moistened.
Place cornbread mixture into prepared pan.
Bake 30 minutes or until wooden pick inserted in center comes out clean.
Cool completely in pan on wire rack.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 2754 Calories from Fat 1031
% Daily Value*
Total Fat 116 g177.8%
Saturated Fat 61.1 g305.4%
Trans Fat 0 g
Cholesterol 665.9 mg222%
Sodium 1502.8 mg62.6%
Total Carbohydrates 364 g121.4%
Dietary Fiber 25.2 g100.8%
Sugars 2.5 g
Protein 62 g124.6%
Vitamin A 66% Vitamin C 2.1%
Calcium 120.3% Iron 110.7%
*Based on a 2000 Calorie diet


text:
Mirk/Dyrk1B mediates G0/G1 to S phase cell cycle progression and cell survival involving MAPK/ERK signaling in human cancer cells
- Jingchun Gao†1Email author,
- Yi Zhao†2,
- Yunyi Lv1,
- Yamin Chen2,
- Bing Wei1,
- Jianxin Tian1,
- Zhihai Yang1,
- Fandou Kong1,
- Jian Pang1,
- Jiwei Liu2 and
- Hong Shi1Email author
© Gao et al.; licensee BioMed Central Ltd. 2013
Received: 2 October 2012
Accepted: 8 January 2013
Published: 11 January 2013
Mirk/Dyrk1B contributes to G0 arrest by destabilization of cyclin D1 and stabilization of p27kip1 to maintain the viability of quiescent human cancer cells, and it could be negatively regulated by mitogenic-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling. This study was performed to investigate the effect of Mirk/Dyrk1B on cell cycle and survival of human cancer cells involving MAPK/ERK signaling.
The correlations between Mirk/Dyrk1B expression and active ERK1/2 detected by western blot in both ovarian cancer and non-small cell lung cancer (NSCLC) cells were analyzed by simple regression. Mirk/Dyrk1B unique phosphopeptides with sites associated with Mirk/Dyrk1B protein were isolated and quantitated by liquid chromatography coupled to tandem mass/mass spectrometry (LC-MS/MS) proteomics analysis. The human cancer cells were treated with small interfering RNAs (siRNAs) and/or U0126, an inhibitor of MEK for indicated duration, followed by investigating the alterations of cell cycle and apoptosis as well as related proteins examined by flow cytometry and Western blot, respectively.
Our study demonstrated the widely expressed Mirk/Dyrk1B proteins in the human cancer cells were positively correlated with the levels of activated ERK1/2. Moreover, Mirk/Dyrk1B protein expressions consistent with the tyrosine autophosphorylated levels in the human cancer cells were increased by U0126 or growth factor-depleted culture. Conversely, knockdown of Mirk/Dyrk1B by siRNA led to up-regulated activation of c-Raf-MEK-ERK1/2 pathway and subsequent changes in cell cycle proteins (cyclin D1, p27kip1), accompanied by increased growth rate and cells from G0/G1 into S of cell cycle which could be blocked by U0126 in a dose-dependent manner, indicating Mirk/Dyrk1B may sequester MAPK/ERK pathway, and vice versa. Whereas, combined Mirk siRNA and U0126 induced cell apoptosis in the human cancer cells.
These data together show that Mirk/Dyrk1B mediates cell cycle and survival via interacting with MAPK/ERK signals and simultaneous inhibition of both pathways may be a novel therapeutic target for human cancer.
KeywordsMirk/Dyrk1B MAPK/ERK siRNA Cell cycle Cancer cells
The serine/threonine kinase, Mirk/Dyrk1B is expressed in few normal tissues, but in skeletal muscle and many types of human cancers [1]. Mirk/Dyrk1B has the ability to auto-phosphorylate on tyrosine activating itself and then phosphorylate other substrates on serine and threonine; therefore, it has been categorized as a dual function kinase. One role of Mirk/Dyrk1B in skeletal muscle differentiation after a stress signal of serum deprivation is to block cycling myoblasts in the G0 quiescent state [2] by phosphorylation of the cell cycle regulators cyclin D1 and CDK inhibitor p27kip1 [3, 4]. Specificially, phosphorylation by Mirk/Dyrk1B at a conserved ubiquitination site Thr288 initiates proteolysis of cyclin D1, while p27kip1 was stabilized following phosphorylation by Mirk/Dyrk1B at Ser10. As normal cells in quiescence activate pathways that protect them from metabolic stress, the subpopulation of tumor cells is likely to utilize similar pathways to survive within the tumor microenvironment. Recently, it has been reported that Mirk/Dyrk1B functions independently and additively to regulate the exit of cancer cells from quiescence through regulating cyclin D turnover and p27kip1 stabilization in colon, pancreatic and ovarian cancer cells shown by Mirk-depletion studies [5–7]. As a result, the quiescent cancer cells depleted of Mirk/Dyrk1B out of G0 entering into the cell cycle may enhance cancer cell kill by chemotherapeutic drugs or radiation, while having less effect on normal tissues in which Mirk/Dyrk1B levels are quite low.
Currently, ovarian cancer and NSCLC are among the leading causes of cancer-related mortality in the world [8]. The presence of drug-resistant, higher proportion of quiescent cancer cells with high clonogenic capacity and tumorigenicity is known to increase recurrence of human cancer and decrease patient survival. Our recent studies have found Mirk/Dyrk1B is overexpressed in a wide spectrum of cell lines and tumor specimens of ovarian and lung cancers [9, 10]. Furthermore, knockdown Mirk/Dyrk1B by small interfering RNA (siRNA) induced cell apoptosis and increased sensitivity of human cancer cells to conventional chemotherapeutics in vitro[9–12]. More recently, Mirk/Dyrk1B has been found to contribute G0 arrest and maintain viability of the quiescent cancer cells via mediating cyclin D1 and p27kip1 which is associated with reduction of reactive oxygen species (ROS) [5, 13], therefore, we hypothesize that Mirk/Dyrk1B pathway may be a novel target for overcoming the drug-resistance and recurrence of various human cancers.
As well as Mirk/Dyrk1B regulating cell cycle-related proteins cyclin D1 and p27kip1, it has been demonstrated that blocking mitogenic-activated protein kinase kinae (MEK) - extracellular signal-regulated kinase (ERK) signaling pathway increases Mirk abundance by up-regulating Mirk/Dyrk1B transcription in either myoblast or colon cancer cells [2], suggesting the possible involvement of mitogenic-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling in Mirk/Dyrk1B functions. However, to date, insufficient data regarding the interaction between Mirk/Dyrk1B and MAPK/ERK in human cancer cells are available, and the mechanisms involved need to be elucidated. In this study, we have identified that the expressed Mirk/Dyrk1B in both ovarian cancer and NSCLC cells is positively correlated with expression of activated ERK1/2. Mirk/Dyrk1B mediates G0/G1 to S of cell cycle and cell survival involving MAPK/ERK pathway in the human cancer cells. It may be a novel target via inhibiting both Mirk/Dryk1B and MAPK/ERK signals for the treatment of human cancer.
Materials and methods
The rabbit polyclonal Mirk/Dyrk1B antibody (C-term, AP7538b) was purchased from Abgent (San Diego, CA, USA). Anti-p27kip1, anti-cyclin D1, anti-ERK1/2, and goat anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-poly (ADP-ribose) polymerase (PARP) and anti-phosphotyrosine (pY) purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-C-Raf and -phosphorylated C-Raf (P-C-Raf), anti-phosphorylated ERK1/2 Threonine 202/Tyrosine 204 (P-ERK1/2) were purchased from BD Biosciences PharMingen (San Diego, CA, USA). Anti-β-actin and donkey anti-rabbit IgG HRP-conjugated secondary antibody were purchased from Sigma (St. Louis, MO, USA) and Amersham Biosciences (Piscataway, NJ, USA), respectively.
Cell lines and cell culture
Human ovarian cancer cell lines used were OV2008, OVCAR3, OVCAR5, SKOV3, MDAH2774, OVCAR10, OV1063, and OVCAR8. Of eight cell lines, SKOV3 and OVCAR3 were purchased from American Type Culture Collection (Manassas, VA, USA); others and all NSCLC cell lines used in this study, such as HCC827, PC-9, H1975, H292, H358, H441, A549, and H1299 were gifts from H. Lee Moffitt Cancer Center and Research Institute, USA. All lines were maintained in DMEM supplemented with 10% heat-inactivated (56°C, 30 minutes) fetal bovine serum (FBS; Invitrogen, Grand Island, NY, USA). Monolayer cultures were incubated at 37°C in a 95% humidified atmosphere air containing 5% CO2.
Small interfering RNA treatment
Cells were reverse transfected with small interfering RNAs (siRNAs) using lipofectamine 2000 transfection reagent (Invitrogen) according to the manufacturer's instructions. The Mirk/Dyrk1B siRNA duplexes as well as the corresponding nonspecific control siRNA duplexes as described [10] were supplied by Dharmacon (Pittsburgh, PA, USA). For combined treatment, cells were pretreated with U0126, an inhibitor of MEK purchased from CalBiochem-NovaBiochem Corporation (La Jolla, CA, USA) at dose escalation for 1 h followed by combination with a constant 20nM dose of siRNAs. Through indicated duration of each treatment, cells treated were harvested and saved for the following experiments.
Cell proliferation assay
Cells were plated in 96-well plates, and siRNA transfection was performed for 72 hours as described above. Cellular proliferation was measured by [3-(4,5)-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) analysis [14]. Briefly, after cells were washed with PBS, they were incubated in MTT solution for 4 hours and then supplemented with 100 μl of dissolving solution (10% SDS in 0.01 M HCl). The absorbance (optical density units) was measured with a microplate spectrophotometer (Bio-Rad Laboratories, Hercules, CA, USA) with Microplate Manager 5.1 software at wavelengths of 590 nm and 660 nm. Each assay was performed in quadruplicate.
Flow cytometry analysis
After 72-hour treatment with siRNAs, cells were subjected to flow cytometry analyses of apoptosis. Apoptosis was assayed using Pharmingen PE-conjugated monoclonal active caspase-3 antibody apoptosis kit without modification as described previously [10]. We determined the percentage of cells in G1, S, and G2/M by propidium iodide staining as described previously [15]. A total of 10,000 cells per experimental condition were used for processing and analysis of fluorescence on Becton-Dickinson FACScan (BD, Franklin Lakes, NJ, USA) using CellQuest software.
Western blot analysis
Cells were washed twice with cold PBS and lysed with buffer A [10 mM Tris–HCl (pH 7.4), 1% Triton X-100, 0.1% SDS, 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol, 0.5 mM phenylmethylsulfonyl fluoride, 10 μg/ml leupeptin, 5 μg/ml aprotinin]. After incubation for 30 minutes on ice, the suspensions were centrifuged (15,000 g for 30 minutes). The supernatants were removed and stored at −80°C until analysis using gel electrophoresis. The protein concentration was determined by Bio-Rad protein estimation assay according to the manufacturer's instructions. For Western blot analysis, ~60-100 μg of whole cell proteins were separated using 10% or 12% SDS-PAGE and transferred to nitrocellulose membranes. After blocking of the membranes with 10 mM Tris–HCl (pH 7.4), 150 mM NaCl, and 0.1% Tween 20 containing 5% nonfat dry milk at room temperature for 60 minutes, the membranes were incubated with indicated antibodies at 4°C overnight and then with the HRP-conjugated secondary anti-rabbit or anti-mouse antibodies at room temperature for 60 minutes. Each protein was detected using the enhanced chemiluminescence (Amersham Biosciences, Piscataway, NJ, USA) system. β-actin was used as an internal control.
Immunoprecipitations were performed with 500 μg of whole cell protein lysates, using Protein A-agarose (Roche, Indianapolis, IN, USA). Briefly, equal amount of protein lysates were incubated with Mirk/Dyrk1B antibody and normal rabbit IgG used as negative control. After incubation for overnight at 4°C, the immune complexes were precipitated with Protein A-agarose. The immunoprecipitates were washed with lysis buffer according to the manufacturer's instructions, then separated by SDS-PAGE, and transferred to nitrocellulose membranes followed by incubation of pY or Mirk antibodies for western blot analysis as described above.
Phosphopeptide immunoprecipation and analysis by liquid chromatography coupled to tandem mass/mass spectrometry (LC-MS/MS)
Phosphopeptide immunoprecipitation for eight NSCLC cell lines: HCC827, PC9, H1975, H292, H358, H441, A549, and H1299 was performed using phosphoscan kit (P-Tyr-100, Cell Signaling) according to the manufacturer's instructions. Using an immunoaffinity peptide profiling technique, Mirk/Dyrk1B unique phosphopeptides with sites associated with Mirk/Dyrk1B protein were isolated and quantitated by LC-MS/MS proteomics analysis as described previously [16]. Results were subjected to sequest-IPI database searching according to criteria specified by molecular and cellular proteomics/cell signaling technology (MCP/CST). Duplicate samples for each cell line each with two technical runs were then filtered according to a 80% peptide identification probability and a 50% protein identification probability.
Statistical analysis
Each experiment was repeated three times. Data are presented as mean ± SD. Statview 5.0 software was used for statistical analyses. Statistical comparison among the groups was performed using one-way analysis of variance (ANOVA), followed by the Fisher least significant difference test. The correlations between Mirk/Dyrk1B expression and active ERK1/2 were analyzed by simple regression. Differences were considered to be statistically significant when P was less than 0.05.
Widely expressed Mirk/Dyrk1B in the human cancer cells is positively correlated with activated ERK1/2
Enrichment of autophosphorylated Mirk/Drk1B consistent with the protein expression may be mediated by activated ERK1/2
Mirk/Dyrk1B regulates progression from G0/G1 to S phase of the cell cycle via MAPK/ERK signaling
Mirk modulates cell survival associated with activation of MAPK/ERK
Mirk/Dyrk1B is a member of a conserved family of serine/threonine kinases that are actived by intramolecular tyrosine phosphorylation which mediate maturation in defferent tissues: Mirk in skeletal muscle, Dyrk1A in brain, etc. [1]. Previous studies show that Dyrk1A activity was increased by binding to 14-3-3 protein [17], and is mediated by autophosphorylation at a C-terminal serine [18], which is not conserved in Mirk. The possible YxY activation domain of Mirk within the conserved kinase region has been known to be intramolecularly phosphorylated only during translation, and the mature members of Mirk family have only serine/threonine kinase activity [19]. Whereas, in this study we utilized a phosphoproteomics screen to identify peptides corresponding to the tyrosine autophosphorylation site of Mirk/Dyrk1B in the human cancer cells and demonstrated a positive correlation between the expression of Mirk protein and the phosphotyrosine abundance of Mirk/Dyrk1B. This suggests that activation of Mirk/Dyrk1B kinase in the deregulated cancer cells may be mediated by tyrosine autophosphorylation, although further study is required.
Mirk/Dyrk1B is also an arginine-directed serine/threonine kinase which has limited expression in normal tissue with highest expression seen in skeletal muscle, heart, tests, and brain. Initially, most of the studies of Mirk/Dyrk1B have been conducted using myogenesis as a model system. It has been known that Mirk/Dyrk1B functions as a transcription factor activator in muscle differentiation. In cultured myoblasts, mitogen deprivation increased Mirk/Dyrk1B protein levels predominantly via transcriptional mechanisms regulated by RhoA and Cdc42, and to a lesser extent by Rac1 [2]. Inhibition of MAPK/ERK activity by removal of serum mitogen [20] or by addition of the MEK inhibitor increased endogenous Mirk/Dyrk1B protein levels and its promoter construct [2], suggesting Mirk/Dyrk1B induction in muscle differentiation requires not only active Rho proteins but also inhibition of the MAPK/ERK signaling pathway. Consistently, the colon cancer cells has been found to utilize similar pathway as myoblasts through which MEK inhibition activates Mirk/Dyrk1B promoter constructs and increases Mirk/Dyrk1B transcription [2]. In this study, we also found Mirk/Dyrk1B could be up-regulated by serum depleted culture or U0126 treatment in both ovarian cancer and NSCLC cells. Therefore, Our study along with others [2, 21] suggests that MAPK/ERK signaling may inhibit Mirk/Dyrk1B transcription and functions in human cancer cells. In addition, our results in the study demonstrate that the widely expressed Mirk/Dyrk1B in both ovarian cancer and NSCLC cells largely correlates the activated ERK1/2. Moreover, we have also found Mirk/Dyrk1B protein levels were increased by inhibition of MEK-ERK1/2, and knockdown of Mirk/Dyrk1B resulted in up-regulation of activated ERK1/2 as well as up/downstream signals. All together indicate the possible interaction between Mirk/Dyrk1B and MAPK/ERK signals in human cancer cells. Namely, Mirk/Dyrk1B may sequester MAPK/ERK, and vice versa in either myogenesis or cancer.
In the past decade, growing evidence has demonstrated that knockdown Mirk/Dyrk1B could induce cell apoptosis and increase sensitivity of various human cancer cells to conventional chemotherapeutics. Furthermore, the study in osteosarcoma demonstrates that the overall survival rate of patients is negatively correlated with the levels of Mirk/Dyrk1B protein expression [12]. Our previous studies have also shown Mirk/Dyrk1B function in an NSCLC orthotopic mouse model [10], and the involvement of FoxO1/3A in the Mirk-mediated ovarian cancer cell survival [9]. Recently, studies have focused on the effect of Mirk/Dyrk1B on various human cancer cells arrested in a reversible quiescent state (G0-G1) to undergo DNA repair or survive suboptimal growth conditions [6, 7, 13]. Moreover, Mirk/Dyrk1B, through regulating cyclin D turnover and p27 stabilization, functions independently and additively to regulate the exit of cancer cells from quiescence G0 or early G1 into S and G2/M of cell cycle [6]. Although it has been reported that Mirk could be negatively regulated by MAPK/ERK in colon cancer, our study is the first to show the possible interaction between Mirk/Dyrk1B and MAPK/ERK signals or sequestering with each other in both ovarian cancer and NSCLC cells. In this study, knockdown of Mirk/Dyrk1B by siRNA in either ovarian cancer cells or NSCLC cells led to up-regulated activation of c-Raf-MEK-ERK1/2 pathway, accompanied by increased growth rate and cells from G0/G1 into S of cell cycle which could be blocked by U0126 in a dose-dependent manner. Furthermore, combined Mirk siRNA and U0126 induced cell apoptosis in the human cancer cells. All of above suggest that Mirk/Dyrk1B may mediate cell cycle and cell survival through interacting with MAPK/ERK pathway in human cancer.
Taken together, the widely expressed Mirk/Dyrk1B in the human cancer cells is positively correlated with the levels of activated ERK1/2. Mirk/Dyrk1B mediates G0/G1 to S of cell cycle and cell survival in both ovarian cancer and NSCLC cells may be associated with MAPK/ERK signaling. Therefore, simultaneous inhibition of Mirk/Dyrk1B and MAPK/ERK may be a novel target for treatment of human cancer.
In Mirk-mediated cancer cell cycle and survival.
We thank Dr. Yuyan Zhu (University of South Florida Medical College, Tampa, FL, USA), for helpful discussions as well as English Editing. This work was supported by the National Natural Science Foundation of China (81172457 to J. G.).
Authors' Affiliations
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- Deng X, Mercer SE, Shah S, Ewton DZ, Friedman E: The cyclin-dependent kinase inhibitor p27Kip1 is stabilized in G(0) by Mirk/dyrk1B kinase. J Biol Chem. 2004, 279: 22498-22504. 10.1074/jbc.M400479200.View ArticlePubMedGoogle Scholar
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- Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ: Cancer statistics, 2008. CA Cancer J Clin. 2008, 58: 71-96. 10.3322/CA.2007.0010.View ArticlePubMedGoogle Scholar
- Gao J, Yang X, Yin P, Hu W, Liao H, Miao Z, Pan C, Li N: The involvement of FoxO in cell survival and chemosensitivity mediated by Mirk/Dyrk1B in ovarian cancer. Int J Oncol. 2012, 40: 1203-1209.PubMed CentralPubMedGoogle Scholar
- Gao J, Zheng Z, Rawal B, Schell MJ, Bepler G, Haura EB: Mirk/Dyrk1B, a novel therapeutic target, mediates cell survival in non-small cell lung cancer cells. Cancer Biol Ther. 2009, 8: 1671-1679. 10.4161/cbt.8.17.9322.PubMed CentralView ArticlePubMedGoogle Scholar
- Deng X, Ewton DZ, Li S, Naqvi A, Mercer SE, Landas S, Friedman E: The kinase Mirk/Dyrk1B mediates cell survival in pancreatic ductal adenocarcinoma. Cancer Res. 2006, 66: 4149-4158. 10.1158/0008-5472.CAN-05-3089.View ArticlePubMedGoogle Scholar
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- Hu J, Friedman E: Depleting mirk kinase increases cisplatin toxicity in ovarian cancer cells. Genes Cancer. 2012, 1: 803-811.View ArticleGoogle Scholar
- Gao J, Niwa K, Sun W, Takemura M, Lian Z, Onogi K, Seishima M, Mori H, Tamaya T: Non-steroidal anti-inflammatory drugs inhibit cellular proliferation and upregulate cyclooxygenase-2 protein expression in endometrial cancer cells. Cancer Sci. 2004, 95: 901-907. 10.1111/j.1349-7006.2004.tb02200.x.View ArticlePubMedGoogle Scholar
- Song L, Turkson J, Karras JG, Jove R, Haura EB: Activation of Stat3 by receptor tyrosine kinases and cytokines regulates survival in human non-small cell carcinoma cells. Oncogene. 2003, 22: 4150-4165. 10.1038/sj.onc.1206479.View ArticlePubMedGoogle Scholar
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- Kim D, Won J, Shin DW, Kang J, Kim YJ, Choi SY, Hwang MK, Jeong BW, Kim GS, Joe CO, Chung SH, Song WJ: Regulation of Dyrk1A kinase activity by 14-3-3. Biochem Biophys Res Commun. 2004, 323: 499-504. 10.1016/j.bbrc.2004.08.102.View ArticlePubMedGoogle Scholar
- Alvarez M, Altafaj X, Aranda S, de la Luna S: DYRK1A autophosphorylation on serine residue 520 modulates its kinase activity via 14-3-3 binding. Mol Biol Cell. 2007, 18: 1167-1178. 10.1091/mbc.E06-08-0668.PubMed CentralView ArticlePubMedGoogle Scholar
- Lochhead PA, Sibbet G, Morrice N, Cleghon V: Activation-loop autophosphorylation is mediated by a novel transitional intermediate form of DYRKs. Cell. 2005, 121: 925-936. 10.1016/j.cell.2005.03.034.View ArticlePubMedGoogle Scholar
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Is it good to have some
Is it good to have some compost mixed in soil while planting bulbs? I live in North Georgia. Is it too late to plant bulbs here?
A closer look at a daffodil.Annette McCarthy
Here are key flower bulb growing tips from The Old Farmer's Almanac.
Is it good to have some compost mixed in soil while planting bulbs? I live in North Georgia. Is it too late to plant bulbs here?
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I Keep planting sunflower seeds and they never come up. What am I doing wrong.
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I live here in Chicago is it too late to plant bulbs for this summer? I planted some canna lilies I got about 2 weeks ago and they are coming up but I wanted to plant more in my girlfriend's yard. Would it be too late?
I want to split my hosta and replant them in different areas, can I do that now (May 12) or do I need to wait until the fall?
Should I try to cover my Daffodils that have just bloomed this past week? We live in central Texas and will probably have a freeze tonight.
Daffodil bulbs can take temperatures up to low 20s and bounce back-- though prolonged freezing temperatures may cause some damage to flowers and foliage. If flowers are in full bloom, the flowers may wilt as the flower stalk thaws. Protection such as garden fabric is a good idea.
I have 3 Amaryllis in containers that have about 6 inches of foliage should I cut them back or will they go ahead and bloom
Only remove the leaves if they turn brown. Your amaryllis will bloom when the bulbs are ready.
Help they are calling for snow and my hyaciths are about 2inchs up out of the ground what should i do!! they are calling for 6-10inchs should i cover them or leave them be!!
Hyacinths are very hardy. They are not going to die and the snow acts like a cozy blanket around your hyacinths.
I planted daffodil bulbs in November. They started coming up in Feb. when we had a warm spell. The beginning of March we had frost which hurt all the blooms. Should I deadhead them or leave them alone?
I have canna bulbs and was told to plant them in the spring.....is that not right?


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|"Yonik Seeley (JIRA)" <j...@apache.org>
|[jira] Commented: (LUCENE-1187) Things to be done now that Filter is independent from BitSet
|Sun, 24 Feb 2008 22:47:14 GMT
[ https://issues.apache.org/jira/browse/LUCENE-1187?page=com.atlassian.jira.plugin.system.issuetabpanels:comment-tabpanel&focusedCommentId=12571979#action_12571979 ] Yonik Seeley commented on LUCENE-1187: -------------------------------------- {quote}also I think there is a hard_to_detect_small_maybe_performance_bug in ConjuctionScorer {quote} Yup, I introduced that feature + bug. I just committed a fix that makes the code do what the comments say (no correctness implications, just perhaps minor performance). > Things to be done now that Filter is independent from BitSet > ------------------------------------------------------------ > > Key: LUCENE-1187 > URL: https://issues.apache.org/jira/browse/LUCENE-1187 > Project: Lucene - Java > Issue Type: Improvement > Reporter: Paul Elschot > Priority: Minor > > (Aside: where is the documentation on how to mark up text in jira comments?) > The following things are left over after LUCENE-584 : > For Lucene 3.0 Filter.bits() will have to be removed. > There is a CHECKME in IndexSearcher about using ConjunctionScorer to have the boolean behaviour of a Filter. > I have not looked into Filter caching yet, but I suppose there will be some room for improvement there. > Iirc the current core has moved to use OpenBitSetFilter and that is probably what is being cached. > In some cases it might be better to cache a SortedVIntList instead. > Boolean logic on DocIdSetIterator is already available for Scorers (that inherit from DocIdSetIterator) in the search package. This is currently implemented by ConjunctionScorer, DisjunctionSumScorer, > ReqOptSumScorer and ReqExclScorer. > Boolean logic on BitSets is available in contrib/misc and contrib/queries > DisjunctionSumScorer calls score() on its subscorers before the score value actually needed. > This could be a reason to introduce a DisjunctionDocIdSetIterator, perhaps as a superclass of DisjunctionSumScorer. > To fully implement non scoring queries a TermDocIdSetIterator will be needed, perhaps as a superclass of TermScorer. > The javadocs in org.apache.lucene.search using matching vs non-zero score: > I'll investigate this soon, and provide a patch when necessary. > An early version of the patches of LUCENE-584 contained a class Matcher, > that differs from the current DocIdSet in that Matcher has an explain() method. > It remains to be seen whether such a Matcher could be useful between > DocIdSet and Scorer. > The semantics of scorer.skipTo(scorer.doc()) was discussed briefly. > This was also discussed at another issue recently, so perhaps it is wortwhile to open a separate issue for this. > Skipping on a SortedVIntList is done using linear search, this could be improved by adding multilevel skiplist info much like in the Lucene index for documents containing a term. > One comment by me of 3 Dec 2008: > A few complete (test) classes are deprecated, it might be good to add the target release for removal there. -- This message is automatically generated by JIRA. - You can reply to this email to add a comment to the issue online. --------------------------------------------------------------------- To unsubscribe, e-mail: email@example.com For additional commands, e-mail: firstname.lastname@example.com
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- 1(of a law, rule, regulation, etc.) very strict and that must be obeyed stringent air quality regulations Licences are only granted under the most stringent conditions.
- 2(of financial conditions) difficult and very strictly controlled because there is not much money the government's stringent economic policies Word Origin mid 17th cent. (in the sense 'compelling, convincing'): from Latin stringent-
BrE BrE//ˈstrɪndʒənt//; NAmE NAmE//ˈstrɪndʒənt//(formal)
Check pronunciation: stringent


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Reaction with a phenol yields, after hydrolysis, the diaryl carbonate: Angew. Chem. Int. Ed., 10, 573 (1971). For reactions with active methylene compounds, see: Angew. Chem. Int. Ed., 10, 574, 575 (1971). 2-Hydroxy acetophenones lead to 2-dimethylamino chromones: J. Org. Chem., 57, 6502 (1992); 61, 3218 (1996):/n
Dehydrates aldoximes and aldehyde phenylhydrazones to nitriles: Synthesis, 563 (1974), 201 (1985), respectively. N, N-dialkylureas are dehydrated to carbodiimides: Tetrahedron Lett., 37, 7047 (1996)./n
Effects cyanation of electron-rich aromatics and heterocycles: Tetrahedron Lett., 27, 1939 (1986)./n
Converts THP protected alcohols directly to alkyl chlorides in good yield: Tetrahedron Lett., 38, 3517 (1997)./n
For review of reactions, including use as Mannich or Vilsmeier intermediates, see: Angew. Chem. Int. Ed., 12, 806 (1973)./n
See also (Chloromethylene)dimethylammonium chloride, B24172./n
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Game of Thrones and more at C2E2
This past weekend was the 4th annual C2E2 (Chicago Comic and Entertainment Expo for those not familiar with the event). C2E2, which is organized by the same people who put New York Comic Con and PAX, is a convention spanning the latest and greatest from the worlds of comics, movies, television, toys, anime, manga and video games.
This year the event returned to the large hall at McCormick Place and filled every inch of their available space with amazing guests, countless hours of programming, talented artists and an incredible show floor.
The line to get into the panel wrapped along the wall and escalator.
Around this point a fan asked how since the viewership of the show is easily women are we ever going to see more male nudity to make it more balanced and how particular scenes with Littlefinger can get a bit cringeworthy. Natalia went on to say that every time we see breasts on screen that we should see cock and she hopes one day that it's true.
When asked who should sit on the Iron Throne, Kristian said that it has to be a Stark and he's pulling for Arya. Natalia said it should go to Osha.
The lead in to this that I didn't record was the fan referencing the rape scene with Jaime and Cersei and how it completely changed his character (and obviously wasn't what happened in the books).
Kristian Nairn surprises a fan with a video message from C2E2. I was there when he got to find out about Jace's reaction and he was really touched by the whole thing.
One of the more interesting booths this year was for Friends With Scales: Reptile Rescue. They had different rescue snakes that you could see and take pictures with. For a suggested donation of $10 you got to pose with a large yellow python named Lemondrop who resembled this iconic snake (only bigger).
Source: Me and my friend's camera, plus YouTube


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How would you like to work in an organization where you have a lifetime appointment? You can be fired only for some extreme violation of ethics. You choose what to work on. If you do not like your assignment, you can refuse it until they give you something you want to do.
Read Whole Story
On Thursday, August 7, busses brought back about half of the families of Kibbutz Kfar Aza who decided to return after the Egyptian-brokered 72-hour ceasefire went into effect.
Twenty-five years ago, in late 1989, I enrolled in an ulpan or Hebrew immersion program at Mishmar Haemek, a kibbutz in the Jezreel Valley. When I met the kibbutzniks as well as the other students in the ulpan, many of them would ask me two questions.
Fifteen seconds will never mean the same to me again.
Even after visiting Israel many times, I still forget how astonishing the establishment of Israel really is. Occasionally, however, a simple intrusion in my life can abruptly focus my amazement on that little state.
Caviar may be the food of kings, but one brand of $4,000-per-jar caviar found in New York City restaurants has its roots in socialism.
Kibbutz Dan...
Israel, on my first visit in 1994, was teeming pitas, tomato-and-cucumber breakfasts and fresh-squeezed Jerusalem juices. Still, I could not find a cornerstone of my culinary upbringing.
Israel's founders understood that Israel cannot treat its minorities the way that Jews were treated throughout history. A world view of how the Jews can realize their national aspirations in a socially just manner is as relevant today as it was then.
I feel pretty amazing most of the time. And I worked hard to get here -- a decade of learning about nutrition, yoga, holistic wellness and my body, while implementing lifestyle adjustments like tons of exercise and fresh air.
This is the tale of Mezer and Meiser, communities that have lived side by side for over 55 years, offering a model of what peaceful coexistence in the region might look like.
JUBA, South Sudan.
The Kibbutz movement is dying? Don't tell that to Emmanuel Logoro. "I have a dream," he says, sitting in his hut near a plastic C...
Marge Piercy's novel He, She and It appeared almost 20 years ago. My review appeared in Tikkun magazine in 1992. Now many aspects of her novel loom even more prophetic than they did before.
To me, Mishmar Ha'emek's Pinat Hagola is an example of what we should strive for in public art built in memory of tragedies. Often, the smallest places can say the most.
Once, long ago, my friend and I bandied the concept of what American Judaism would be like if there were no Israel, and what Israel would be like if there were no American Jews.
A few dozen socialists in the north of Israel met recently to raise glasses of wine and toast their latest business success.
The residents of Kibbutz...
Israel was built in places like Kibbutz Mishmar Ha'emek, a hilly village whose roughly 1,000 residents share a view of the northern Jezreel Valley - a...
Shared consumption and production is a nice idea -- but it's ultimately unrealistic, especially when surrounded by an intensively capitalistic environment, as there is in Israel today.
By Robert L. Smith
Religion News Service
CLEVELAND HEIGHTS, Ohio (RNS) Raychel Bone, Sam Selker, Naima Cohn and Taliesin Haugh sat on their front por...
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Cara Delevingne
Numero September 2013 by Karl Lagerfeld
Designer and photographer Karl Lagerfeld captures the striking cover story of Numero Magazine starring fashion's top models Nadja Bender,...
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DKNY Fall Winter 2013.14 by Mikael Jansson
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Order ielts, toefl, toeic,ESOL,GMAT Without attending the exam. Original passport,Driving License(ieltsassist
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Joad Sportz Supplies
Bicycle Stand, Kick Stand, Kickstands
The BIPOD bicycle kick stand has become a world favourite for all touring bicycles riders in Australia and New Zealand. The PLETCHER made bicycle kick stand is all about quality, refinement and strength. If you need a bicycle kick stand that will last and last then look no further than the BIPOD Pletscher double leg kick stands. The Rolls Royce of bicycle kickstands imported directly from Switzerland by Joad Sportz Supplies Sydney Australia. The clever Audax Bicycle Kickstand. The Pletscher Bipod stands attach just behind your bottom bracket and both legs cleverly fold to the left hand side of the bike. By lifting the front wheel off the ground the stand distributes the bikes weight on it's points of contact and your bike's back wheel. Availiable from Joad Sportz Supplies at www.joadsportzsupplies.com.au
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Professional grade long haul bicycle mud flaps from Widek. Modern sturdy mud flaps that are fast and easy assembly, no tools required. Can be easily fitted to the front mud guard, (no tools required) but can also be riveted into place. Made in Holland
Your future cycling safety is important and you need to be safer whilst riding your bicycle. This is made possible by applying PROVIZ to your everyday cycling trip.
Proviz High visability cycling wear from is now avaliable in Australia from Joad Sportz Supplies. Cycle with the safety in mind of knowing you can been seen on the busy road ways. The unique triviz illuminated cycling wear is a world first and has won the 2013 Red Dot awards. You can now be seen on your bicycle at dusk at night or during the day. Dont leave home on your bicycle without Proviz on your body to show others your on the road.
The original foldable rear fender, that is only there when you need it. It's the fendor bendor mud flap, mud guard.
In investment owned brands, the first design-oriented brand "JYE-SEN" introduced "DRIVELINE"─ a highly regarded innovative, R & D team that is always listening to user's experience" and "Real time cyclists riding demands". The primary consideration according to the different demands of end user and preferences are the crank leg shape, tooth plate in their design creating the best cranks money can buy with their highly customized range. The "DRIVELINE" R & D team further explores the innovation
Bicycle Cranks by DRIVELINE now avaliable from Joad Sportz Sydney Australia
Uusus Wheels, whose factory is in Rosá, northeast of Vicenza, Italy, not only makes wheels, it supplies hubs to other wheel makers. They currently supply wheels to the Farnese Vini pro team.
Imported from Italy URSUS Bike Wheels
Fully imported Ursus Wheels are created and produced through thorough research, allowing the cyclist to optimize the power of their pedaling, using the least amount of expended energy, whilst not giving up quality or reliability.
Purveyors of fine european made cycling parts and accessories.
Pletscher Kickstands to suit modern day bicycles
Kickstands to suit TREK bicycles from Pletscher switzerland
SUP surfers all require a theft security system for their stand up paddle boards, with a locking system that gives them piece of mind as they leave their SUP board locked up secure from theft by applying a Docks Locks stand up paddle board locking system.
Lock up one or multiple boards using DocksLocks surf lock. Then relax, knowing your equipment is safe and you'll catch tomorrow's waves.
DocksLocks surf lock has the answer: Lock your surfboards with DocksLocks Surfboard Security System.
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