text:
Facebook Reportedly Working on Healthcare Features and Apps
Facebook Inc already knows who your friends are and the kind of things that grab your attention. Soon, it could also know the state of your health. ...Read more ›
Facebook Inc already knows who your friends are and the kind of things that grab your attention. Soon, it could also know the state of your health. ...Read more ›
Edison International, the parent company of Southern California Edison (SCE), announces the application period for its 2014-15 Edison Scholars Program ...Read more ›
A survey released by the National Education Association, in conjunction with the American Association of University Women and Tufts University, describes the numerous ...Read more ›
The Heartbeat Music Academy, located in San Diego, California, was founded by entrepreneur and humanitarian, Tyra K. Hawthorne. Ms. Hawthorne ...Read more ›
Thirty-six of California Lutheran University's top undergraduates will present their research findings at the 11th Annual Student Research Symposium ...Read more ›
If you have to ask, then you haven't been paying attention to the seismic shift going on when it comes to marketing to women. For a while now, we've been told that females ...Read more ›
If you were going on an important job interview, you'd dress to impress. That's exactly what you want to do with your home, too–dress or, rather, stage to impress and sell! ...Read more ›
In 2014, there still isn't a cure for glaucoma but researchers in Chicago are one step closer. The research team at Northwestern University Feinberg School of Medicine ...Read more ›
Some of the most serious injuries among older adults, age 65 and older, are caused by falling. More than 1.6 million older Americans end up in the emergency room ...Read more ›
The report suggests that the first end-of-life conversation could coincide with a cherished American milestone: getting a driver's license at 16, the first time a person ...Read more ›
Site Designed By NoRegretMedia.com


text: POPSUGAR Celebrity Inception Photos of Leo by Celebrity 10/05/09 0 Shares Like us on Facebook Previous Photo 1 of 11 View on One Page ADVERTISEMENT ( more slides) Next Share This Link Copy 1 Photos of Leo Previous Photo 1 of 11 View on One Page ADVERTISEMENT ( more slides) Next Read More InceptionEllen PageLeonardo DiCaprio Leonardo DiCaprio Make Leonardo DiCaprio's Billionaire's Beach Home Your Summer Hangout 170+ Celebrities You Should Be Following on Instagram! 36 Celebrity Selfies You Didn't See Coming Spend the Night at Leonardo DiCaprio's Palm Springs Pad Ellen Page 12 Stars Who Came Out in 2014 Heidi Klum Will Inspire You to Start Your Fitness Resolution Now Lena Dunham Goes Topless to Encourage Female Voters Ellen Page and Kate Mara's True Detective Spoof Is Truly Hilarious Get the scoop with our daily newsletter Enjoy the latest news on the celebs you love. A Listers Movies Young Hollywood TV Sign up with Link Love Ashton and Mila aren't afraid to hang with the neighbors — Us Weekly Is this why Hilary Duff dyed her hair blue? — HuffPost Celebrity Katy Perry might have chopped off all her hair — Lainey Gossip Rihanna addresses those Leonardo DiCaprio rumors — Dlisted Is Monica Lewinsky heading to The View? — ET Scott Eastwood continues to make the world swoon — JustJared Emma Roberts shows some serious leg on set — Hollywood Tuna Gear up for Michael Keaton on Saturday Night Live — Pink Is the New Blog You'll never guess who's feuding with Howard Stern — The Superficial Joe Giudice describes Teresa's prison life — I'm Not Obsessed Latest Celebrity

subset: commoncrawl


text:
Fuji X-Pro 1 … Andre' & Garry hit the Streets | Streetshooter
Well, at the last minute I switched bags from the TurnStyle 5 to the Retrospective 5. It just is a little easier to swap cameras from the Retrospective 5. I don't need any Enter & Exit Bag Procedure Intrusions. On the street I adjusted the dividers a little and now everything just works great. So, Andre' has the 23mm on and Garry has the 50mm on and the extra pocket holds the EF-X20 Flash & the 18mm. I'll never use all this stuff out on the street but….if I don't use both cameras, one will go and I know it. So, I need to learn again to depend on using 2 cameras. Besides, what better way to change lenses eh? It was very convenient to swap cameras when I felt like changing Field of Views. It's not hard to take, that I'll say…….
See on streetshooter.net
US-Fotograf Jeff Seltzer: "Die Welt muss menschenleer sein" |
Jeff Seltzer, 43, sucht leere schöne Räume in überfüllten Großstädten. Der US-Amerikaner zeigt auf seinen Fotografien auch gerne Überbleibsel, die Menschen auf Straßen oder Flughafen-Wartehallen hinterlassen haben: Zigarettenstummel, Bremsspuren oder Kritzeleien. Vor allem aber hat er es auf Parkplätze abgesehen. Seltzer wurde in Los Angeles geboren und studierte dann Kommunikation und Rhetorik in San Diego. Er lebt und arbeitet als Fotograf in L. A.
See on www.spiegel.de
Interview with Tom Lazar – Street Photographer | The Fujifilm Blog
Photographer Tomasz Lazar finds that the Fujifilm X100S is the best choice for his brand of street photography. Tomasz Lazar is a photographer from Poland and a graduate of the West Pomeranian University of Technology. His black & white documentary images of street culture have won numerous accolades, including placing in the Sony World Photography Awards and the International Photography Awards. "I also took second place in 'People in the News' category of the 2012 World Press Photo contest and received Picture of the Year at BZ WBK Press Foto 2012," he tells X Magazine……
See on fujifilmblog.wordpress.com
The Streets of San Francisco | Valérie Jardin
From Haight & Ashbury to Chinatown to North Beach, I truly enjoyed doing street photography in San Francisco last weekend. It was a family vacation, not a photography trip. Although I had my Fuji x100s with me at all times, photography was not the focus of this trip (no pun intended). I still managed to grab a few shots here and there. I'll be back and will most likely lead a photo workshop there in 2015…….
See on valeriejardin.wordpress.com
Streets of Dublin | Gabor Nagy
Street photography.. Hm.. I always wanted to do it, but like many of you I was too afraid of it. I was really nervous to get close to strangers and take a candid shot but the last couple of months things have changed. I'm still not a street shooter/warrior, but I feel more confident and step by step I overcome my fear of street photography. I also realized that people usually don't give a damn about somebody photographing them and most of them don't even notice me. Quite often I chose a spot what I find interesting and I wait for someone to walk into my frame. There is still room to improve, I still have to get closer to my subjects but I feel I'm on the right track and I'm really enjoying this whole new experience.
All images were taken with the Fuji X100s and the X-Pro1 with the 35mm lens.
See on gaborimages.blogspot.de
Barcelona, perfect place for street photography | Nicole Struppert
My year has just started from the photography point of view. After my shoulder surgery I am ready to travel again and explore new places. I thought it would be nice to share some insights about the places I travel. I try to give u the best tips for photography spots and other insights. So here is my first review on Barcelona. It was my second visit to Barcelona – time was pretty tight, cause I spent just the weekend there but I tried to make the best out of it. I don't want to get into details about sightseeing places – I am sure this will be on your list when u travel anyway. I highly recommend the beach area of Barcelona. Best time is in the morning – then the place is not too crowded and there is already much to see. Young and older people are coming together for a chat, to bike, skate or just stroll around. This area is fantastic for street photographers – the light is incredible and a perfect spot to catch some nice shots or just have a drink and enjoy the surrounding. Don't miss to walk around the little streets next to the beach area. This area is incredible beautiful and reminded me a bit of Napoli in Italy…….
Review of the Fujifilm X-T1 for Street Photography | Eric Kim
When I was here in Dubai for Gulf Photo Plus, the guys from Fujifilm were generous enough to give me a new Fujifilm X-T1, the Fujifilm 23mm f/1.4 (35mm full-frame equivalent), as well as the Fujifilm 27mm f/2.8 Lens (~40mm full-frame equivalent). I shot with it everyday for around a week. Overall I like the camera a lot and would highly recommend it (superb image quality, great form factor, and responsive). Some downsides are that the AF isn't as accurate and quick as other cameras (like the Olympus OMD)– although it is a huge improvement from the X-Pro 1 and x100s. Hope this improves with future firmware updates…..
See on erickimphotography.com
Barcelona, Saturday Morning at the Beach… | Nicole Struppert
The past weekend I was in Barcelona. It was my first trip after my shoulder surgery and I was very excited how I can handle to photograph again. First goal was to travel light. I left my Nikon D800e at home and took just my Fuji x100s. The x100s was the perfect companion, light, fast and nearly perfect for all situations. I love this little camera more and more… I got some really nice shots Saturday morning at the beach. The mediterranean mood, the sun and the easy beach life felt so good after the long grey wintertime……
Berlin with the X-Pro1 | Peter Evans
I'd never been to Berlin before, but I'd seen a few films set there. Forefront in my mind was 'Wings of Desire', Wim Wender's atmospheric tale of an angel (literally) falling for a trapeze artist in 1980's Berlin. It stars Bruno Ganz, Nick Cave and Columbo and it's all shot in the most gorgeous black and white. And that's how I thought I was going to shoot Berlin. I mean, this is surely a monochrome city, right? Battle scarred buildings, grim brutalist Eastern-bloc architecture, lowering skies, great concrete walls – one amazing oppressive movie set. That's how my mind imagined it. Turns out I'm an idiot. Berlin is tremendously colourful. I arrived on the train from Schoenfeld airport greeted by some beautiful magic hour light. I figured, "What the hell, let's capture a little colour at least," and set the camera to Pro-Neg Hi colour film simulation. I never left that setting. Whenever I confronted a crumbling ruin of a building, a concrete monolith of a towerblock, it always seemed painted in lovely warm light and splashed with the most vibrant graffiti. 'Poor but sexy' indeed and as splashed with colour as a heaping of ketchuppy Currywurst……
See on petetakespictures.com
Fujifilm X-T1 on the street | Lars Øivind Authen
Unfortuneatly I was not able to go to the boxing matches this weekend and test the responsiveness of the X-T1. So, I got out on the streets instead. Together with the Fujifilm X-T1 I used the XF 23mm f/1.4 and XF 35mm f/1.4 lenses. The pictures are shot in raw, converted to Tiff's in Adobe Camera Raw using the new Fuji X camera profile Astia/Soft which pretty much like exactly the same as the in camera Astia Jpegs. The Tiffs were then imported into Lightroom and a bit of contrast and sharpness were added……
See on www.larsauthen.com


text:
Be there and be square (dancing)
Square dancers will do-si-do and area merchants hope to see more dough this weekend as an estimated 800 to 900 square dancers sashay into Brainerd Friday through Sunday for the 60th annual State Square and Round Dance Convention at Brainerd High School.
Get the full story
Subscribe or Log In
Are you a newspaper subscriber but you don't have a Digital Access account yet? Activate your account.
You will need your subscription account number and phone number. Not sure if you have an account? Call 218-855-5897 and we can help you.


text:
Happy Birthday, Jane ;o)
Jane, here's wishing you a great day and many more to come,
Luv, Karen
Thank you Ladies...
Had a nice day...I only wish the weather were nicer.
Tonight DH & DD surprised me with a Pizza Party & Cake
with my Dad & the GrKids.
Nana...Thanks for asking...I getting around a little better now.
I'm on my third leg cast. This one looks like something out of
'Star Wars' which impressed the Gkids..lol
Two more weeks and then hopefully, I graduate to a foot cast..Yippee!
OA...DH took me to GW a few days ago and I got some nice
Blue Dishes & a nice print for over the Fireplace.
I'm itiching to play with my dishes again.
Thanks again everyone..
Thanks Jeanne...and I do LOVE the card!
I am doing better which is a relief and it was fun
doing some GW shopping. I got another pretty framed print
for the Spring/Summer months to put over the fireplace.
It's called 'The Goose Girl'.
Thought of you when I got it.
Not that you're a goose girl...lol - but that it's the same type of prints we like.
Guess I"m the one to have to say belated HAPPY BIRTHDAY. I thought I posted on this last night, but my computer froze up when I tried to send it. I thought it went, but I guess it is out there for you to find in outerspace( maybe you Starwars cast will help find it,Ha. I am so glad that you had a good day and that you are feeling better. Recovery takes time. My DGD found a white turkey gravy boat the other day atGW and bought it for me. It tickled me that she knew Granny would like it. I think DD and DGD are both developing a affection for dishes. That makes it a lot more fun for me. Take care, Jane and keep improving. A Big BD hug, even if it is late! Janet
Thank you Janet...I was going to post a Happy Birthday
to you too, but read that Karen made a 'blooper' !!
(sorry karen)...lol
You can be sure Janet that we'll ALL be ready
for your Birthday come October ...lol
Karen...I'll post the Star Wars cast on a seperate 'OT'
post....just for fun.


text:
Launcher Update
Get the Desktop App for Battle.net Now
- All your games in 1 place
- Log in once
- Automatic game updates
Posts: 4View profile
I searched for off-handed wepon issues, but I did not see this anywhere. If there is a specific placce for this, let me know, and I'll re-post it there.
The problem I am seeing, is when I am equipping an item in the off-hand, the numbers are off by a factor of ten to a hundred, depending on the item.
For example: Demon Hunter, dual-wielding quill-shots and nail spitter respectively. I equip a quiver, then hilight over the replaced nail spitter. It says it has a -366 damage, but when equipped, the damage is about 36 points of damage off.
Here's where it gets interesting. I then hilight over that same quiver I just unloaded, and IT is now saying it has -366 damage, when in reality, it has a positive damage rating over that nail spitter.
I have seen odd numbers like this with the Barbarian, but I don't have a recent example, since the demon hunter is my most current character. I think someone needs to go in and verify that all the decimal places and addition/subtraction calculations are accurate, at least for these characters...
Threats of violence. We take these seriously and will alert the proper authorities.
Posts containing personal information about other players. This includes physical addresses, e-mail addresses, phone numbers, and inappropriate photos and/or videos.
Harassing or discriminatory language. This will not be tolerated.


text:
Inn on Thistle Hill Bed and Breakfast
No availability found through our partners. Please contact the business directly or check some of our recommended alternatives.
Photos of Inn on Thistle Hill Bed and Breakfast
Travelers also recommend:
TripAdvisor Reviews Inn on Thistle Hill Bed and Breakfast Boston
Travel Blogs from Boston
Wine (without the women and song)
... to notice, better than the ones down our way, which are cheaper and offer cheaper tastings. Nevertheless: we had a nice day. The weather was gorgeous—yet again (this might be the nicest week in a Virginia summer since we moved here)—and the scenery is gorgeous. I've posted lots of photos with descriptions, so have a look.
We ate dinner at the restaurant in the Blackthorne Inn (not to be confused with ...
The Potter Wedding
We have known Kevin Potter for years now and when he said he had bought a ring we set our travel plans because we would not miss it for the world!
Kevin and Ginny Lynn got married on a beautiful Saturday afternoon at her families church tucked outside of Washington DC. They held their reception in her grandfather's back ...
Brightwood Day 3
Woke up again to the rooster! Thanks buddy! The thing with the rooster is... It's not like a normal alarm clock. You can't press a "snooze" button on it! Which is good cause it means you have to wake up! Fed the donkeys, sheep and ducks this morning. The poor duck that was injured was still not looking good so we have put her in a safe little nest in the barn near the donkeys and Fiona the cow. My new daily chore is to check on ...
Mission Reflections and Final Blog Entry
... 8220;T"s crossed and so I never needed to know in any sort of detail what was happening next. The failing for me was that I could never communicate that very effectively to the team.
The lens I view the people I come in contact with was shaped in 1969. The summer of that year I traveled to the then Soviet Union during the height of the Cold War with an organization called the Citizens Exchange Group. Their mission was to help people ...
Mason - Dixon. Day 895.
... answered as they had slavering Rottweilers at the window.
A couple of more houses were empty but then I saw an old woman walking to her garage. I waved and walked toward her trying to look as friendly as possible. Not easy when you're dressed in black motorcycle gear.
Once she realized I was a friendly we chatted about the marker. She said it was right were we parked the bikes but got stolen 3 years ago. F**ckers !
I kicked the tennis ball for ...


text:
The coffee and food served here are consistently good. Most dishes are served with a tasty dressed salad but I always make room for one of the delicious cakes.
Tasty imaginative yet simple Italian dishes using top-quality ingredients. Staff always friendly and attentive. You can't go wrong with anything on the menu but my personal favourite is the Napoli pasta.
Excellent choice of pastas, salads, paninis and wraps - hard to make a choice! And lovely desserts to finish off with. Been wanting to try here for a number of months and so pleased it lived up to expectation. The seating area is much larger than expected as well. Brilliant food, great portions and I would say better than Dolce...
A favourite place to go for a light lunch. Tasty fresh food served by friendly and efficient staff who always make you feel welcome. I wish the seating wasn't quite so cramped though.
It is the perfect place for a nice breakfast or lunch...the atmosphere is great, food is always good and the staff always makes you feel at home!!!!
Smalll basement bistro. Friendly staff and really tasty food. Good size portions, well presented. Value for money.
Basement deli in the centre of town. Clean with Italian atmosphere - great music. I had a very hard toasted mozzarella panini. Difficult to eat. Latte was good. Slow service.
As we shop regularly in Cheltenham we have tried many of the copious amounts of eateries , some we have visited more than once, others do not get a second visit. We have popped into la Scala for coffee on a number of occasions, today we were running late and it was after 2.30 by the time we were looking...
One of the best in Cheltenham for eat in or take away. Quality, freshly prepared food and great coffee, nice for a cheeky red wine too. Great service, wonderful selection on menu and very reasonabley priced. Love it!
I've been coming here for years, it's really a lovely place for lunch or coffee and cake. Gorgeous food and reasonably priced.
Register now for free — and start getting automatic notification of new reviews, responding to traveler feedback, adding new photos to your listing and much more.


text:
The present invention relates to the measurement of estradiol using competitive immunoassay methods. The inventors unexpectedly discovered that estrone and its derivatives conjugated to a label is a particularly effective tracer when used in conjunction with estradiol specific antibodies to determine estradiol levels in fluid samples. The present invention also utilizes 5α-dihydrotestosterone to enhance the assay performance.
1. A method of determining the amount of estradiol in a blood sample comprising:
a. incubating a mixture of a blood sample suspected of containing estradiol, a solid phase coupled to an antibody specific for estradiol, and a conjugate of the formula ##STR4## wherein Q is a label, to form estradiol/antibody complexes and conjugate/antibody complexes on said solid phase;
b. separating said solid phase from said mixture;
c. measuring the amount of label present in said mixture or on said solid phase; and
d. determining the amount of estradiol in said sample from the amount of label.
2. The method of claim 1, wherein the label is selected from the group consisting of: catalysts, fluorescent compounds, chemiluminiscent compounds, enzymes, and enzyme substrates.
3. A kit for performing an estradiol competitive enzyme immunoassay comprising:
a solid phase coupled to an antibody specific for estradiol and
a conjugate of the formula ##STR5## wherein Q is a label.
4. The kit of claim 3, wherein the label is selected from the group consisting of: catalysts, fluorescent compounds, chemiluminiscent compounds, enzymes, and enzyme substrates.
5. A method of determining the amount of estradiol in a blood sample comprising:
a. incubating a mixture of a blood sample suspected of containing estradiol, a solid phase coupled to an antibody specific for estradiol, and a conjugate of the formula ##STR6## wherein Q is an enzyme, to form estradiol/antibody complexes and conjugate/antibody complexes on said solid phase;
b. separating said solid phase from said mixture;
c. measuring the amount of label present in said mixture or on said solid phase;
d. determining the amount of estradiol in said sample from the amount of label.
6. The method of claim 5 wherein said solid phase is a microparticle and said separation step comprises passing the mixture through a porous matrix.
7. In a method of determining the amount of estradiol in a fluid sample, the improvement comprising:
a. pretreating the sample by incubating the sample suspected of containing estradiol with 5α-dihydrotestosterone at a concentration within the range of about 1 μg/mL to about 5 μg/mL in a buffer at a pH within the range of from about 4.5 to about 6.7; and
b. measuring the amount of estradiol in the mixture.
8. The method of claim 7 wherein said buffer is a combination of glycine and citric acid.
9. A kit for performing an estradiol competitive enzyme immunoassay comprising:
a solid phase coupled to an antibody specific for estradiol;
a conjugate of the formula ##STR7## wherein Q is an enzyme; and an enzyme substrate.
10. The kit of claim 9 wherein said enzyme is a peroxidase or phosphatase.
11. The kit of claim 9 further comprising a sample pretreatment solution comprising 5α-dihydrotestosterone at a concentration within the range of about 1 μg/mL to about 5 μg/mL.
Estradiol (1,3,5(10)-Estratrien-3,17a-diol) is secreted by the ovary and placenta. It is synthesized by the aromatization of androgens in the thecal and granulosa cells of the ovary and placenta. The aromatization is stimulated by follitropin (FSH). Estradiol synthesis in turn stimulates production of Lutropin (LH) receptors necessary for the synthesis of androgen precursors.
Estradiol is important for female sexual differentiation during gestation, sexual development at the onset of puberty, and regulation of the menstrual cycle. The menstrual cycle is the result of a precise coordination of the functional characteristics of the Central Nervous System, the hypothalamus, the pituitary, the ovary, and the endometrium which regulate the cyclic release of Gonadotropin Releasing Hormone (GnRH), LH and FSH, and ovarian steroids (Estradiol and Progesterone). Estradiol is involved in both the stimulation and inhibition of the release of the gonadotropins, exerting both a positive and a negative feedback. Early in the follicular phase, ovarian secretion of estradiol from the thecal and granulosa cells is modest. During the follicular phase, estradiol stimulates endometrial growth (repairing the endometrium after menses). Toward mid-cycle, LH production increases and results in the release of the ovum by the rupture of the developed follicle. After ovulation, estradiol secretion declines slightly. During the luteal phase, estradiol along with progesterone are secreted by the corpus luteum stimulating further endometrial growth. If the ovum is not fertilized, there is a further drop in estradiol and progesterone. This drop in estradiol and progesterone initiates menses.
The measurement of estradiol is important for the evaluation of normal sexual development (menarche), causes of infertility (anovulation, amenorrhea, dysmenorrhea), and menopause. Normal estradiol levels are lowest at menses and during the early follicular phase (25-75 pg/ml). The levels rise in the late follicular phase to a peak of 200-600 pg/mL just before the LH surge initiates ovulation. As LH peaks, estradiol begins to decrease before rising again during the luteal phase (100-300 pg/mL). If conception does not take place, estradiol falls further to its lowest levels, thus initiating menses. If conception occurs, estradiol levels continue to rise, reaching levels of 1-5 ng/ml during the first trimester, 5-15 ng/ml during the second trimester, and 10-40 ng/ml during the third trimester. During menopause, estradiol levels remain low.
There are various methods for measuring estradial levels in serum. However, each of these methods utilize radioactive elements as labels and suffer from several disadvantages. The methods require several different steps including manual steps, several vessels, and are either nonautomated or only semiautomated. Radiolabels are problematic because the expense involved in such assays are much greater than in non-radiolabeled assay formats. For example, more highly skilled laboratory personnel are required and waste disposal is more tightly controlled and regulated. The following are commercially available radioimmunoassay methods for determining estradiol level in serum samples.
One assay for estradiol (commercially available from Pantex) is a direct, non-extraction competitive assay. Radiolabeled estradiol tracer, anti-estradiol antibody and sample are mixed and incubated for two hours. Anti-antibody antibody is added and the mixture is incubated for 15 minutes to form a precipitate. The mixture is centrifuged to pellet the precipitate formed. The supernatant is decanted and the radioactivity of the precipitant is measured. The radioactivity measured is compared to a radioactivity vs. estradiol concentration plot to determine the estradiol concentration in the sample. The following compounds are known to cross-react in the assay: α-Estradiol (1.4%) and Danazol (0.6%).
Another assay for estradiol (Coat-A-Count Radioimmunoassay (RIA) for Estradiol commercially available from Diagnostic Products Corporation) is a direct, non-extraction antibody coated tube competitive assay. Each tube is coated with anti-estradiol antibody. Radiolabeled estradiol tracer and sample are incubated in the tube for three (3) hours. After decanting the mixture and washing the tube, the radioactivity of the tube is measured. The radioactivity measured is compared to a radioactivity vs. estradiol concentration plot to determine the estradiol concentration in the sample. The following compounds are known to cross-react in the assay: Ethinyl Estradiol (1.8%); Estrone (1.1%); Estradiol-3β-D-glucuronide (0.7%); Estradiol-3-Sulfate (0.3%); and 19-Nortestosterone (0.25%).
Yet another assay for estradiol (Estradiol MAIA commercially available from Serono)is also a direct, non-extraction competitive assay. Radiolabeled estradiol tracer, anti-estradiol antibody and the sample are incubated for one to three hours (depending on the assay sensitivity and precision desired) to permit the formation of antibody-estradiol complexes. The complexes are separated from the sample by incubation of the reaction mixture with anti-antibody antibody coated magnetic particles followed by sedimentation of the magnetic particles through the application of a magnetic field. The separated particles are then washed. The measured radioactivity level of the particles is compared to a radioactivity vs. estradiol concentration plot to determine the estradiol concentration in the sample. The following compounds are known to cross-react in the assay: Estrone (2.6%); EstradioI-Dipropionate (0.3%); Estradiol-3β-D-glucuronide (0.2%); and Estriol (0.2%).
Some non-radiolabeled immunoassays have been reported, but do not have the sensitivity or specificity needed to accurately and precisely measure estradiol levels. Dawson et al. (Steroids, 31:357-366, 1978) tested rabbit polyclonal anti-estradiol antibodies using horseradish peroxidase coupled to estrone at carbon 11. The cross-reactivities of the antibodies to estrone ranged from 5% to 120%. Such cross-reactivities would not be acceptable in an estradiol specific assay. Thus, an enzyme immunoassay that utilizes a conjugate where the enzyme is coupled to position 11 of estrone would not likely produce a satisfactory assay.
Pandey et al. (Clinica Chimica Acta, 190:175-184, 1990) reported an enzyme-linked immunosorbant assay (ELISA) using the conjugate estradiol-6-(O-carboxymethyl)oxime linked to penicillinase with anti-estradiol antibody coated wells of a microtiter plate. The assay had a lower sensitivity limit of 25 pg/mL, but required a two (2) hour incubation of the reaction mixture at 37° C. and after washing, the enzyme substrate/enzyme reaction required incubation at 37° C. for one (1) hour. Thus the overall assay time was in excess of three (3) hours. Maurel et al. (J. Immunolog. Methods, 102:165-172, 1987) also reported an ELISA using an estradiol-6-(O-carboxymethyl)oxime conjugated to β-galactosidase, which had improved sensitivity over Pandey et al.'s assay. However, as in Pandey et al.'s method, Maurel et al.'s method requires extensive incubation times. The anti-estradiol antibody coated on microtiter wells are incubated with the sample for ninety (90) minutes at 37° C., then incubated with conjugate for ninety (90) minutes, and then after washing the enzyme activity was determined after two (2 ) hours at 42° C. Thus, the assay time was well in excess of three (3) hours.
De Boever et al. (Clin. Chem., 32:1895-1900, 1986) reported a chemiluminescence immunoassay for estradiol with a sensitivity limit of about 49 pg/mL and an assay time of in excess of ninety (90) minutes. Roda et al. (Anal. Blochem., 156:267-273, 1986) reported a luminescent enzyme immunoassay for estradiol which required over four (4) hours to perform.
De Lauzon et al. (J. Immunoassay, 10:339-357, 1989) reported a competitive enzyme immunoassay for estradiol using microtiter plate wells coated with estradiol coupled to bovine serum albumin (BSA). The sample and peroxidase labeled anti-estradiol antibody were incubated in the wells for two (2) hours at room temperature. Alternatively, biotinylated anti-estradiol antibodies were utilized folilowed by a second incubation of three (3) hours with avidin coupled to peroxidase.
Clearly, there is a need for an estradiol non-radiolabel immunoassay that is rapid, accurate, sensitive, easy to perform, free from interferences and relatively insensitive to experimental variables such as pH and temperature. An object of the present invention is to develop an assay method and reagents to perform estradiol measurements accurately and with precision without the need of a radiolabeled tracer. Because of the very low concentrations of estradiol present in biological fluid samples (0.025-40 ng/mL), any alternative method must be very sensitive. The existing methods utilize long incubation times to overcome this sensitivity requirement. Another object of this invention is to eliminate the long incubation times required in non-radiolabeled immunoassays.
The present invention relates to the measurement of estradiol using competitive enzyme immunoassay methods involving the use of estrone conjugated to a label at the 6 position. The inventors unexpectedly discovered that estrone and its derivatives conjugated to a label at the 6 position may be used in conjunction with estradiol specific antibodies to determine estradiol levels in fluid samples. The present invention also utilizes 5α-dihydrotestosterone to enhance the assay performance.
The present invention relates to methods and reagents useful in the measurement of estradiol in fluid samples. The inventors unexpectedly discovered that estrone and its derivatives conjugated to a label can be used in conjunction with estradiol specific binding members to measure estradiol levels in fluid samples, such as blood, plasma, whole blood, cerebral spinal fluid, saliva, urine, and the like. The present invention uniquely involves the measurement of estradiol concentrations in fluid samples using such estrone label conjugates. In addition, estradiol is known to bind to Sex Hormone Binding Globulin (SHBG) (Philip et al., Steroids, 47:373-379, 1986) which is present in fluid samples, such as serum, plasma, whole blood and the like. In order to measure the total estradiol in a sample containing SHBG, the estradiol bound to SHBG must be released. The inventors discovered that the use of a buffered solution of 5α-dihydrotestosterone will release substantially all of the estradiol bound to SHBG.
Testosterone (Philip et al., J. Steroid Biochem., 32:865-872, 1989) and its derivatives are known to displace estradiol from SHBG at pH 8.0 (De Boever et al., Clin. Chem., 32:1895-1900, 1986). As De Boever et al. reported and the inventors observed, not all of the estradiol is substantially displaced from SHBG at pH 7.5-8.0 by 5-dihydrotestosterone. Substantially all of the the estradiol is displaced from SHBG by 5α-dihydrotestosterone at a concentration within the range of about 1 μg/mL to about 5 μg/mL, preferably within the range of about 1 μg/mL to about 3 μg/mL most preferably at a concentration of about 2 μg/mL, at a pH within the range of about 4.5 to about 6.7, preferably within the range of about 5.0 to about 6.0, most preferably at a pH of about 5.7. Preferably, the buffer is a combination of glycine at a concentration within the range of about 0.25M to about 1M, more preferably within the range of about 0.4M to about 0.6M, and citric acid at a concentration within the range of about 0.2M to about 0.5M, more preferably within the range of about 0.2M to about 0.3M, though other buffers or buffer combinations which are capable of buffering a solution within the above effective reaction range are acceptable. The buffer also preferably contains saponin at a concentration of at least about 0.5% (w/v), more preferably within the range of about 0.5% (w/v) to about 1.25% (w/v), and most preferably at a concentration of about 0.75% (w/v). The presence of red blood cells in a serum sample tended to cause the measured estradiol levels to be elevated. The addition of saponin substantially reduced the effect of the red blood cells.
Specific binding members specific for estradiol include estradiol specific binding proteins, such as monoclonal and polyclonal antibodies and other estradiol specific synthetic or recombinant proteins that specifically bind lipoprotein cholesterol particles. For example, it is well known by those skilled in the art that monoclonal and polyclonal antibodies can be produced that specifically bind to steroids such as estradiol. When an immunogen comprising estradiol or a derivative of estradiol coupled, typically through a covalent bond, to a carrier protein, such as albumin and the like, is injected into an animal, the animal's immune system will produce polyclonal antibodies that specifically bind to estradiol. General methods for the preparation of monoclonal antibodies to analytes using mice or rats are well known to those skilled in the art. More recently the preparation of analyte specific synthetic and recombinant proteins have been reported and the same methods can be readily adapted to the preparation of estradiol specific binding synthetic and recombinant proteins useful in this invention.
The inventors discovered that a compound, estrone, which has a reported very low cross-reactivity (less than 5% and in some cases less than 1% cross-reactivity) with estradiol specific antibody, surprisingly acts as an exceptional labeled reagent for detecting exceedingly low levels of estradiol in a very rapid assay format. The term "conjugate", as used herein, refers to any substance comprising estrone or a derivative of estrone coupled to a label. The coupling is preferably covalent. A preferred conjugate for use in this invention comprises ##STR1## wherein Q is a label.
The term "label", as used herein, refers to any substance which can be attached to estrone or its derivatives and which is capable of producing a signal that is detectable by visual or instrumental means. Various suitable labels for use in the present invention can include catalysts, enzymes, liposomes or other vesicles containing signal producing substances such as chromogens, catalysts, fluorescent compounds, chemiluminescent compounds, enzymes and enzyme substrates, and the like. A large number of enzymes suitable for use as labels are disclosed in U.S. Pat. No. 4,275,149 incorporated herein by reference. Such enzymes include glucosidases, galactosidases, phosphatases and peroxidases, such as alkaline phosphatase and horseradish peroxidase which are used in conjunction with enzyme substrates, such as fluorescein di(galactopyranoside), nitro blue tetrazolium, 3,5',5,5'-tetranitrobenzidine, 4-methoxy-l-naphthol, 4-chloro-l-naphthol, 4-methylumbelliferyl phosphate, 5-bromo-4-chloro-3-indolyl phosphate, chemiluminescent enzyme substrates such as the dioxetanes described in WO 88100694 and EP 0-254-051-A2 and derivatives and analogs thereof. Preferably, the label is an enzyme and most preferably the enzyme is alkaline phosphatase.
The conjugate is preferably prepared by coupling the label to the conjugate precursor (which is commercially available from Sigma Chemical Company) ##STR2## using a carbodiimide compound, such as 1,ethyl-3-(3-dimethylaminopropyl)carbodiimide, or by first forming an active ester, such as an N-hydroxysuccinimide active ester and then reacting the active ester with the label. Other coupling methods and conditions well known to those skilled in the art may also be used to prepare the conjugate.
Preferably, estradiol is measured according to the invention using a solid phase having a specific binding member specific for estradiol bound thereto. The solid phase and the sample are separated so that the amount of estrone enzyme conjugate bound to the solid phase or the amount of estrone enzyme conjugate remaining in solution can be determined. The amount of estrone enzyme conjugate on the solid phase or in solution can be correlated to the concentration of estradiol in the sample using a enzyme activity vs. estradiol concentration plot, typically referred to as a standard curve. A standard curve is prepared by performing the assay using calibrators such as those in Table 1 (below). Controls are used to verify that the curve is viable. When an sample with an unknown estradiol level is assayed, the measured assay signal is compared to the standard curve and the estradiol level corresponding to that signal is the estradiol level of the sample.
The specific binding member may be bound to the solid phase by physical or chemical means, preferably by means of a direct covalent bond. The specific binding member should be bound to the solid phase in such a way that substantially none of the specific binding members detach during the subsequent reactions and wash steps. Regardless of the specific binding member and the coupling method selected, the specific binding member must be able to bind to estradiol and the estrone enzyme conjugate after being coupled to the solid phase.
A solid phase according to the present invention may be a mixture of polymeric microparticles with chemically or physically bound specific binding members specific for estradiol. Microparticles that can be used include polystyrene, carboxylated polystyrene, polymethylacrylate or similar particles with a radius ranging from about 0.1 μm to about 0.25 inches. A preferred separation method for these particles is the use of microparticle capture on a porous matrix such as glass fiber (see below).
Other solid phases that can be used include a mixture of magnetizable polymeric microparticles with chemically or physically bound specific binding members specific for estradiol. Magnetizable microparticles that can be used preferably have ferric oxide or chromium oxide cores and a polystyrene, carboxylated polystyrene or polymethylacrylate coating. Yet other solid supports are known to those in the art and include the walls of wells of a reaction tray, tubes, polystyrene beads, nitrocellulose strips, membranes and the like. Natural, synthetic, or naturally occurring materials that are synthetically modified, can be used as a solid phase material including polysaccharides, e.g., cellulose materials such as paper and cellulose derivatives such as cellulose acetate and nitrocellulose; silica; inorganic materials such as deactivated alumina, diatomaceous earth, MgSO4, or other inorganic finely divided material uniformly dispersed in a porous polymer matrix, with polymers such as vinyl chloride, vinyl chloride-propylene copolymer, and vinyl chloride-vinyl acetate copolymer; cloth, both naturally occurring (e.g., cotton) and synthetic (e.g., nylon); porous gels such as silica gel, agarose, dextran, and gelatin; polymeric films such as polyacrylamide; and the like. In any case, the solid phase material should have reasonable strength or strength can be provided by means of a support, and should not interfere with the production of a detectable signal.
An alternative preferred separation method is described in co-pending U.S. patent application Ser. Nos. 150,278abandoned, filed Jan. 20, 1988 and 375,029, abandoned, filed Jul. 7, 1989 both of which enjoy common ownership and both of which are incorporated herein by reference. These applications describe the use of ion capture separation, in which the specific binding members used in the assay in question are chemically attached to a first polyionic compound and a porous matrix having bound thereto a second polyionic compound that binds to the first polyionic compound. A specific binding pair is formed and separated from the reaction mixture by an electrostatic interaction between the first and second polyionic compounds. A specific binding member of the specific binding pair is preferably covalently coupled to the first polyionic compound.
Preferably, the first polyionic compound is a polyanionic acid, such as polyaspartic acid, heparin, carboxymethyl amylose, polyglutamic acid or polyacrylic acid, and the second polyionic compound is a cationic polymer, such as GafQuattm which is a polymeric quaternary ammonium compound (GAF Corporation, Wayne, N.J., 07470), diethylaminoethyl-dextran (Sigma Chemical Company, St. Louis, Mo.), water soluble cellulose derivatives such as Celquat™ L-200 and Celquat™ H-100 (National Starch & Chemical Corporation, Bridgewater, N.J., 08807) which are both polymeric quaternary compounds, or Merquat® 100 (commercially available from Calgon Corporation). The porous matrix is treated with the cationic polymer to render the matrix positively charged. The cationic polymer is bound to the matrix by absorption, adsorption, or covalent or ionic coupling. The separation of the reaction products is effected by the electrostatic interaction between the positively charged pad and the negatively charged polyanion complex.
A porous matrix for use in this invention can include any suitable porous material. By "porous" is meant that the material is one through which fluids can flow and can easily pass. In the present invention, the matrix can include a polypropylene, polyethylene, Teflon, fiberglass, cellulose, or nylon pad or other porous material well known to those skilled in the art for use in a pour and flow-through assay device having one or more layers containing one or more of the assay reagents.
Preferred porous materials include a porous fiberglass material, such as a "Whatman 934-AH" filter paper, which has a nominal thickness of 0.33 mm, or the disposable IMx® cartridge and TestPack™ (fiber matrix) devices of Abbott Laboratories (Abbott Park, Ill., 60064). The thickness of such material is not critical, and will be a matter of choice, largely based upon the properties of the sample or analyte being assayed, such as the fluidity of the test sample.
Some enzyme substrates are converted into fluorescent compounds, such as 4-methylumbelliferone, by the appropriate enzyme. The quantity or rate at which the fluorescent compounds are formed over time is an indication of the quantity of enzyme present in the reaction. When the enzyme is a label, the quantity of enzyme present is related to the quantity of estradiol present in the sample. Thus the measurement of fluorescence can be related back to the quantity of estradiol present in the sample. The fluorescence can be measured by any method known to the art. For example, a fluorescence spectrometer is desirable, although the fluorescence spectrum can be observed with a visual spectrometer or photographed with a spectrograph of high light-gathering power.
In a preferred embodiment, the fluorescence is detected using an IMx® (Abbott Laboratories, Inc.) automated bench top analyzer that contains an optical assembly which is a fluorometer that uses a mercury arc lamp as its light source. This instrument is described by Fiore et al (Clin. Chem., 34/9:1726-1732, 1988), the contents of which are incorporated herein by reference. The instrument utilizes an IMx® disposable cartridge (commercially available from Abbott Laboratories, Illinois) which contains a porous matrix to capture microparticles containing anti-estradiol antibodies which have been exposed to a conjugate and the sample. The label used in the conjugate is preferably alkaline phosphatase. The conjugate and the estradiol in the sample compete for available binding sites on the microparticles. The microparticles are separated from the sample and the quantity of conjugate present on the microparticles is determined from the rate at which 4-methylumbelliferyl phosphate is converted into 4-methylumbelliferone. From a standard curve of rate of 4-methylumbelliferone formation versus estradiol concentration, the quantity of estradiol in the sample can be determined.
The standard curve, also known as a calibration curve, of rate of 4-methylumbelliferone formation versus estradiol concentration is generally prepared from calibrator solutions containing known estradiol concentrations. Preferably; six calibrators are used to obtain a calibration curve, though more or less calibrators can be used depending on the accuracy and precision of the result desired. Preferably, the calibrators contain increasing amounts of estradiol. For example, Table 1 illustrates the composition of one set of calibrators (see Example 4). Controls are generally used in conjunction with an assay to confirm the viability of a calibration curve or assay reagents. Preferably, the formulation of the controls are the same as the calibrators with the exception that the estradiol concentration may not be identical with any one of the calibrators. For example, controls having an estradiol concentration of 150, 500 and 1125 pg/mL would be suitable controls for the calibrators in Table 1. One skilled in the art would be capable of devising other calibrator and control formulations.
TABLE 1______________________________________Calibrator Estradiol Concentration (pg/mL)______________________________________F 3000E 1500D 750C 250B 50A 0______________________________________
To maintain aseptic conditions throughout the procedure, it may be desirable to add a small quantity of an antimicrobial agent to the system which may include solvents, antibiotics and poisons.
The following examples are illustrative of the invention and are in no way to be interpreted as limiting the scope of the invention, as defined in the claims. It will be appreciated that one skilled in the art can conceive of many other devices and methods of use to which the present inventive concepts can be applied.
The following assay format was performed on an IMx® disposable cartridge by an IMx® instrument (both commercially available from Abbott Laboratories, Illinois and described in EP-A-288 793 and in Fiore et al., Clin. Chem. 34/9:1726-1732, 1988, both of which are incorporated herein by reference). Seventy-five microliters (75 μL) Of a serum sample were mixed with 35 μL of 5α-dihydrotestosterone buffer (DHT Buffer: 2 μg of 5α-dihydrotestosterone and 0.75% (w/v) saponin per milliliter of 0.5 mM glycine and 0.25 mM citrate buffer at pH 4.5), 50 μL of rabbit anti-estradiol antibody coated microparticles as prepared in Example 2, and 90 μL of IMx® Buffer (commercially available from Abbott Laboratories, Illinois). The mixture was incubated from about 27.5 minutes at 37° C. One-hundred seventy-five microliters(175 μL) of the mixture was transferred to the fiber matrix of an IMx® disposable cartridge (the fiber matrix is in liquid communication with an absorbent pad). The particles were captured by the fiber matrix and the solution was absorbed by the absorbent pad. The particles were then washed with IMx® Buffer. Sixty microliters (60 μL) of conjugate was added to the matrix, incubated for 12 seconds, and then the matrix was washed again with IMx® Buffer. Sixty-five microliters (65 μL) of a 1.2 mM solution of 4-methylumbelliferyl phosphate in 0.1M 2-amino-2-methyl-1-propanol buffer at pH 9 was added to the matrix and the rate of 4-methylumbelliferone formation was measured by fluorescence reflectance. The fluorescence was measured with a fluorometer that uses a mercury arc lamp as its light source (as described by Fiore, M. et al., Clin. Chem 34/9:1726-1732, 1988, the contents of which are incorporated herein by reference). Using the calibrators of Table 1, this assay format on the IMx® instrument was observed to have a mean sensitivity of 13.9±4.3 pg/mL.
Anti-estradiol coated microparticles were prepared as follows:
a. Rabbit anti-estradiol polyclonal antibodies were prepared by injecting rabbits with estradiol 6(O-carboxymethyl)oxime coupled to bovine serum albumin (commercially available from Sigma Chemical Company, MO.)in Freund's complete adjuvant and boosting with Freund's incomplete adjuvant.
b. Polyclonal antibodies were isolated from the rabbit serum following standard procedures for antibody precipitation with 35% ammonium sulfate.
c. Enough latex microparticles (commercially available from Seradyne, Ind.) are measured to produce a final reaction concentration of 1% (w/v) solids were mixed with 50 mM 2-[N-morpholino]ethanesulfonic acid (MES) buffer at pH 4-5, antibody from step b was added to a concentration of 0.9 mg/mL, and then 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) was added to a concentration of 0.5 mg/mL. The mixture was incubated for 30 minutes and then diluted in 0.1M Tris buffer at pH 7.4 containing 0.1% (w/v) Tween 20 and 0.1M sodium chloride. The particles were filtered and re-suspended to about 9 μg/mL antibody which is about 0.01% solids with 0.1M bis[2-hydroxyethyl]iminotris[hydroxymethyl] methane (Bis-Tris) at pH 6.5 containing 0.1M sodium chloride, 13.6% (w/v) sucrose and 0.2 mg/mL normal rabbit IgG.
Estrone 6(O-carboxymethyl)oxime alkaline phosphiatase conjugate was prepared as follows:
a. Ten micrograms (10 μG) of estrone 6(O-carboxymethyl)oxime (commercially available from Sigma Chemical Company, MO.) and 10 μG of N-hydroxysuccinimide were dissolved in 100 μL dimethylformamide (anhydrous). Forty micrograms (40 μg) of EDAC was added.
b. An aliquot of the mixture in step a was added to alkaline phosphatase in carbonate buffer at pH 8 so that the ration of estrone to phosphatase is 10:1. The resulting conjugate was purified on a G25 Sephadex column and the purified conjugate was diluted to a concentration of about 1-10 μg.mL in 0.1M Bis-Tris buffer at pH 6.5 containing 0.5M sodium chloride, 1% (w/v) casein, 1 mM magnesium chloride and 0.1 mM zinc chloride.
The calibrators listed in Table 1 were prepared as follows:
a. Steroid stripped serum was prepared by raising the pH of serum to pH 11 with 6N sodium hydroxide and let the mixture stand for about 18 hours at 2°-8° C. The pH was then adjusted to a pH of about 7.4-8. Fifty grams (50 g) of activated charcoal was added per liter of serum and the mixture was stirred for 2 hours at room temperature. The charcoal was then filtered away from the steroid stripped serum.
b. Enough estradiol was then added to aliquots of steroid stripped serum to make the final concentration of each calibrator listed in Table 1. Controls are prepared in the same manner.
To demonstrate the effectiveness of displacing estradiol bound to SHBG with 5α-dihydrotestosterone (DHT), estradiol was spiked into normal human serum samples and then the samples were assayed according to the format described in Example 1 hereof. Estradiol was added to normal human serum samples so that the estradiol level was increased by 300 pg/mL. Prior to assaying the spiked samples, an aliquot of each sample was treated with 5α-dihydrotestosterone at pH 5.7 and at pH 7.5. Table 2 lists a representative sampling of the samples tested. % Recovery was calculated as follows: ##EQU1## Spiked Value is the measured estradiol concentration in the sample after the sample was spiked with estradiol. Nascent Value is the measured estradiol concentration in the sample before the sample was spiked with estradiol.
TABLE 2______________________________________DHT at pH 5.7 DHT at pH 7.5Sample Nascent Spiked %* Nascent Spiked %*______________________________________1 71 428 119 132 484 1172 52 428 125 93 475 1273 0 307 102 21 338 1064 209 540 111 296 656 1205 1455 1746 97 1845 1969 416 0 282 94 38 328 977 155 485 110 273 654 1278 17 299 94 120 516 1329 6 313 102 72 381 103______________________________________ *Percent Recovery
Cross-reactivity studies were performed using the assay format described in Example 1 hereof. To steroid stripped serum prepared as in Example 4 hereof, potential cross-reacting steroids were added and the samples were assayed. Table 3 lists the steroids tested, the maximum concentration tested and the percent cross-reactivity observed.
TABLE 3______________________________________ % Cross-Steroid Concentration (ng.mL) Reactivity______________________________________Aldosterone 300 0Androstenedione 30,000 0.0009Clomiphene Citrate 300 0Corticosterone 100,000 0Cortisol 100,000 0Danazol 100,000 0.000311-Deoxycortisol 4,000 0DHEA-Sulfate 5,000 0DHEA 4,000 05-DHT 30,000 0.0008Estrone 100 0.30Estriol 300 0.05Ethinyl Estradiol 100 0.42Estradiol-3-Glucuronide 10 2.40Extradiol-17-Sulfate 300 0Progesterone 100,000 0Testosterone 100,000 0.0003______________________________________
Using the assay format of Example 1 hereof and the Coat-A-Count Estradiol assay (commercially available from Diagnostic Products Corporation), 434 individual serum samples were assayed for estradiol and the estradiol levels as measured by each method were correlated as shown in Table 4.
TABLE 4______________________________________Number of Estradiol CorrelationSamples Range (pg/mL) Intercept Slope Coefficient______________________________________340 0-1500 34 1.16 0.97434 0-6400 72 1.06 0.98______________________________________
The invention also encompasses kits for the determination of estradiol in a fluid sample. The kits comprise a solid phase coupled to an antibody specific for estradiol; a conjugate of the formula ##STR3## wherein Q is a label. Preferably, Q is an enzyme and the kit would further include an enzyme substrate. The kits can also include a sample pretreatment solution comprising 5α-dihydrotestosterone at a concentration within the range of about 1 μg/mL to about 5 μg/mL.
The embodiments described and the alternative embodiments presented are intended as examples rather than as limitations. Thus, the description of the invention is not intended to limit the invention to the particular embodiments disclosed, but it is intended to encompass all equivalents and subject matter within the spirit and scope of the invention as described above and as set forth in the following claims.
Patent Citations
Non-Patent Citations
Referenced by
Legal Events
Original Image


text:
Fusion Hotel
No availability found through our partners. Please contact the business directly or check some of our recommended alternatives.
- Shuttle bus service
- Continental Breakfast
- Restaurant
- Beach
- Free High-Speed Internet
- Bar/lounge
Photos of Fusion Hotel
Travelers also recommend:
TripAdvisor Reviews Fusion Hotel Playa del Carmen
Travel Blogs from Playa del Carmen
Mexico (Playa del Carmen)
... heard of him?), Lauren Lane and John Acquaviva, and from researching their bio's I had doubts they even played a set this night as none of them looked anything like them (no female DJ's so I couldn't get that one wrong!) Nonetheless, whoever was playing, played an awesome eclectic blend of house and deep house tunes, which had us bopping along with the rest of the punters. As it is with festivals we soon had made a few friends, and spent ...
Mexico Night 2: Lost in...Tequilalization? pt. 1
... the disco and, like true Canadian tourists, headed straight to the bar. There were all kinds of bottles I had never seen before, and people around were downing drinks of every color you can imagine. Someone nearby was holding one that I'd never seen - the bottom layer was purple and the top layer was bright blue. I asked him what it was.
"A superman!" he said, really enthusiastically.
"Un Spiderman!!" corrected the bartender. Ok, well ...
We love turtles
... bus) to Akumal and got a guide to take us to swim with turtles. It was pretty terrestrial rain but the water was so clear and we saw about 30 turtles just from swimming out to snorkel off the beach. They come up for air every couple of minutes then they float down and eat it's very cute.
Next day we visited another cenote called Cenote Azul, since we got there so early in the morning there we had the place to ourselves which was awesome, the water was cold but so clear. In ...
Day 1 - habla espanol pequeno
... bags and spent the rest of the afternoon by the pool just hanging out doing absolutely NOTHING! A huge change of pace from the usual hustle bustle routine. So we took in some sun, and luke warm pool water, while the remaining parties in our group found us. Went to dinner, bought some groceries from the store across the street and are turning in for what looks to be a great week away from the everyday of design life in Michigan.
Day 324-339- 25th- August-8th Sept MEXICO
... from Ohio, USA a lovely 23year old full of passion and energy. Unfortunately the weather has turned, the sea is choppy and the wind strong, I think Beyers the sailorman could handle it but the sports manager and life guards have called the red flag on the beach, meaning it is too dangerous to go out in the sea. Alas this did not stop us from having a wonderful morning or more food, coffee and then to top it off before Beyers birthday massage a drinking game down by the pool bar. Yes 9am ...


text:
~ George Demeter ~
~ Anonymous ~
Please be sure to visit our
Parks & Recreation Department website at :
www.BSLPRD.com or Click ~ Here
To read the newsletter, please click ~ here
For a full schedule go to the Fitness Class Tab on the
Parks and Recreation Website Click ~ Here
2nd Thursday of each month 6 - 8 pm
Lucky Girls
Call BSL Community Center 363-0018 for details
4th Thursday of each Month 6 – 8 pm
Hired Hands Bluegrass/Country
For more information, please click ~here~
~ Parks and Recreation ~
Help Us Send a Kid to Camp
For more information on how to help send a kid to camp, please click ~here~
Boiling Spring Lakes Board of Commissioners
Have suspended the Citizen Forum Meetings.
The Public Comment time allowance at the
Board of Commissioners regular monthly meetings has
changed from three (3) minutes to five (5) minutes.
When: Saturday, March 28th
Where: BSL Police Department, 9 E. Boiling Spring Road (parking lot)
Time: 10:30 AM until gone
Tickets: Contact the BSL Police Department for tickets 363-0011
During the month of April, Parks and Recreation is offering a yearly special that includes two months FREE!
Join as a yearly member to use the fitness room, fitness on demand, or even a combo now, and your membership lasts for 14 months!
Join the Special Events Committee in their 2nd Annual Spring Bicycle Ride!
Prizes will be awarded for first place and last place. Lunch provided.
Proceeds will benefit The Navy Seabee Veterans of America
When: April 18, 2015
Where: Ride starts at Spring Lake Park on Pine Road
Time: Registration at 9:00 AM
For more information about the event and registration prices, please click - here
Audio of Board of Commissioner Meetings 2014 available ~ Here
Audio of Board of Commissioner Meetings 2015 available ~ Here
The City of Boiling Spring Lakes Invites Construction Proposals
for Paving of Four City Streets, with an
alternate for additional paving.
A Pre-Bid Conference will be conducted by the City Engineer
at City Hall on Wednesday March 18, 2015 ~ 10:00 A.M.
Sealed bids endorsed as "ROADWAY PAVING CONTRACT" will be
received by the City's Public Works Department at:
9 East Boiling Spring Rd. Southport, NC 28461
until 2:00 p.m. Friday March 27, 2015.
Bids may be mailed or hand delivered only. If mailed they must
be received by the city in advance of the bid deadline
or they will NOT be opened
Full instructions, contract and proposal ~ Here
Addendum Number One to the Paving Contract ~ Here
Note ~ E-verify is not required to be submitted for this proposal
~ Note ~
If you are having trouble opening this link please contact
City Hall at 910-363-0025 or email firstname.lastname@example.com
April 13 - April 18, 2015
April 18, 2015
Shallotte Middle School
For more details on these events please
visit Brunswick County's Website ~ Here
Spraying will be done in late April or early May
If you prefer to not have your area sprayed
Please contact Aquatico @ 910-367-6869 or
email @ firstname.lastname@example.com
Feasibility Study available ~ Here
Please be sure to visit us and read our newsletters ~ Here
Join the Library Commission at Barnes and Noble at Mayfair Town Center in Wilmington for their Book Fair event.
All sales will directly benefit the Library Commission.
When: In store from April 24 through April 26
Online from April 24 through May 1
Where: Barnes & Noble, Mayfair Town Center, Wilmington, NC
Please use voucher when making purchases so the Library Commission will receive credit - voucher
Vouchers also available at City Hall
Event hosted by the Library Commission
When: Saturday, April 25, 2015
Time: 8:30 AM to 12:30 PM
Where: Spring Lake Park on Pine Road
For ages 3 to 12. Pre-register your Small Fry at the
BSL Police Department or at the Community Center.
For more information, please contact Allison at 363-0011


text:
Welcome to BADEN - Kunama.Com
The Kunama is a minority
ethnic-group living in the western part of Eritrea.This page exposes the
unjust and discriminatory activities of the Eritrean government. It also
participates in the political dialogues in Eritrea.
G15 Germano Nati
Arabic / Tigringa / Feedback
Adolay Kornelios Museum
The geographical changes of the Eritrean regions Read more
THE NATIVES AND THE INDIGENOUS POPULATIONS OF ERITREA "THE KUNAMA AND THE BARIA /NARA". "Nil-Chary tribes". VKP 2009 (Reviewed, January, 11.20112). Read more
1.- Adolay-Cha-Fanede.Assi tukumu - 2 - ADOLAY MABUSA KUTUMA KUNAME. Assi tukumu 3.- ERKUKODISU ANTATE Assi tukumu . 4.- .ADOLAYKIME TEWELDETTE MIAMA KUNAMA KISHE Assi tukumu. ,.5.- ISASA UKUYYA BAKATIYA KINTI 2003.Assi tukumu
IwanIkka Isha Shetanana.
24 Marso 2015 Facebookla.
WOYNAKKESI KOSONA SOOTA.:- Ide tamana tuffanki kittitasi jarimatata doromunasi arfana gonke ditta kalmana meynammema saneya limu. Kunama tummadala mekona data ellana kosimmema tabaromu.
EREMIAS GOGNE- Oinabu amanana halalanakin degata kun koshimesi hellena sase hellamma ede tummamu alama fardata kalokema. Tamana alala guja koshignan ditta itenni!!!.
KAM=Kunama Agara Mena , Marso 27, 2015 Assi tukumu
Response to Nati Galla: on, "now our patience, in wearing thin for some unbriddled Kunama guys"; :- Nati Galla, we do know who you are, but, what are you, and what is your position, your work, within DMLEK, "Er-ku-ko-di-su?"
Are you one of its leadership-members?
Its active member?
Its bookkeeper?.
Its strongest political and financial supporter, or are you just, Kornelios Adolay Osman Aggar's body-guard, trying to protect him from far-away-Canada, where you now live?
The Voice of the Kunama People, (VKP) Kunama Agara Mena, (KAM) , March 26, 2015. Read more
The jossurney and arrival of Mr. Kornelios Adolay Osman Aggar, in the U. S. A.
According to very well founded and ascertained sources, Mr. Kornelios Adolay Osman Aggar, the chairman of the Democratic Movement for the Liberation of the Eritrean Kunama, (DMLEK); Eritrea Kunama Koibisha Sungada, (Er-Ku-Ko-Di-Su.),
arrived in Waschington, on March 19, 2015, and was received and welcomed, by a Kunama, by the name of Anene Dagu, resident of Mineapolis, (Minnesota), who himself is an operative of the Eritrean PFDJ`s dictatorial regime. Anene Dagu is reported to have accompanied Mr. Kornelios Adolay Osman Aggar to Minniapolis, from where, a certain Kunama, by the name of Sirnay, resident of De-Moine, (IWOA), presumably a relative or a very supporter of Mr. Kornelios Adolay Osman Aggar, took him to De-Moine, and is now hiding him in his own house/appartment. The Voice of the Kunama People, (VKP);
Kunama Agara Mena, (KAM)
March 23, 2015. Read more
The bunker lies on the right hand side of the valley.
Br.General YAKOB SHINGGIRAI (DMLEK)
YAKOB SHINGGIRAI SHIMELBA KAMBA KIKISHIMA KABARA.- Ai tagamong gommo laga ambommola mishima tama agarossi "anda baia mamim mashikema agaratte kanene, sangga-nene okoske, sananga baioabbu ashkarena auiki kambata kolaj gongkema dabbu nadadongki dagang gonumana kotikgoske" attile oina fogadeoaikin KODUSE mikosumme, ena Yakob Shinggirai nokossumia kamba niki kalma ninama kalla agara haddadong gongke. KAM= Kunama Agara Mena, Marso 23, 2015; Read more
O sons of Kunama and Nara
And for that purpose our brothers in Europe, America and Australia took the first initiative and established the first free Kunama civic organization by the name "The voice of Kunama" (Kunama Agara Mena) . Following their foot steps and as a continuation of their noble nationalist efforts, and in unity and coordination with them, we the sons of Kunama and Nara and sons of Gash Setit generally, in the Middle East, Sudan and Ethiopia, held a general meeting from 17 - 19 / 3/2015 and issued the following declaration : -
March 23, 2015 Read more
WHO ARE THESE? These are the Kunama individuals who were execute by DMLEK´s leader Adolay Kornelios Osman .
Published on Wednesday, 11 March 2015 10:40
Amedem, Adolay Kunama kisha artesi, Shabia asamedab kibingki ililiki, jajenaski, komaldala iafaskisso, ame malatta maudamoassina, Adolay iafada tabilebu, tamma Kunama kishelana iafada badam uno, okoiafang gongkemaki malattamoassi, ena "nna kenaebu abagnanga, deres kisha goske/komala kiddikengki" nilattanasi, Adolay ainiana, agara agasala, (Paltalkla), "naiakesuma", ena iammeng gonube, o maida kiakeng gonube. Nna kibishi kaso.
KAM= Kunama Agara Mena ,(Marso 22, 2015) Assi tukumu
بسم الله الرحمن الرحيم
اعلان هام
يا أبناء الكوناما والنارا
ويا ابناء قاش- سيتيت المناضلين الصا
المجد للثورة الأرترية
الخلود لعواتي ورفاقه الأبرار
العار للطائفية والشوفينية والتبعية
تاريخ 19/3/2015م
Read more


text:
Hotel las Leyendas
No availability found through our partners. Please contact the business directly or check some of our recommended alternatives.
Photos of Hotel las Leyendas
Travelers also recommend:
TripAdvisor Reviews Hotel las Leyendas Avila
Travel Blogs from Avila
Palace and El Escorial
... to decorate every wall and floor there is. This palace is only used now for stately functions, wedding etc, while the Spanish royal live a separate residence. I love the grand staircase leading up the rooms , the throne room and the massive dining room with a long dining table that can easily fit over 130 guests . The quarter also houses some of worlds best violins collections by Antonius Strdivarius. the violins are over 300 + years old and produces some incredible music. ...
Luxury on the road
I know most of u r have been missing our blog.
Bnb secure internet - 4* hotel not secure - go figure.
We r in avila now in a supposed paradore! Very nice but has lost it originallity.
Leaving Cagliari, arrived at the airport lined up 4 baggage drp. Madrid is closed - panick - translation - flights 4 for madrid r not open yet!
CATTLE CLASSryanair. We couldn't understand the english, god knows how the none ...
... peaceful enclosed garden spaces. There were lots of cute little kittens sunning themselves at the entrance. MP= "That's because they don't believe in contraception"
We ate well there too. Simple hearty food but we got homemade empa�adas and the local biscuits. On the whole we're finding the Spanish fare much blander than what we expected. Eggs, cheese, potato and many varieties of pig. So ...
Does a walk on a medieval wall negate 32 oz steak?
... price that one. I will only put my neck out to offer to take care of the project management for the fist 30 years, after that I will only say "it is not my problem any more, let the next 10 generations worry about it." Luckily the Roman Catholic Church had more money than the queen of England so fitting the bill was no issue...if we only had clients like that. On our way out of town we spotted an old bull fighting ring, I have no idea how old it was, but we had ...
Castles in Castille
... only vehicle in it! Despite the new car park, parts of the village look almost mediaeval.
Then it's on to the town of Aranda de Duero, where it actually begins to rain for a short time. Here I cross the Rio Duero, another of Spain's longest.
I'm now firmly committed to using the backroads rather than any fast dual carriageways. Apart from going very slowly through the occasional village or town, progress is quite good as there are long ...


text:
Anger That a Herod Show Uses West Bank Objectstags: NYT, Jerusalem, Herod, West Bank, Second Temple
JERUSALEM — In one room sits a sarcophagus of reddish-pink limestone believed to have held the body of King Herod, painstakingly reconstructed after having been smashed to bits centuries ago. In another, there are frescoes from Herod's elaborate underground palace, pieced together like a jigsaw puzzle. Throughout, elaborate animated videos show the king's audacious construction — atop the desert fortress Masada; at his burial place, Herodium; and his most famous work, the Second Temple of Jerusalem.
The Israel Museum on Tuesday opened its most ambitious archaeological exhibition and the world's first devoted to Herod, the lionized and demonized Rome-appointed king of Judea, who reigned from 37 to 4 B.C.E. and is among the most seminal and contentious figures in Jewish history. But the exhibition, which the museum director described as a "massive enterprise" that involved sifting through 30 tons of material from Herodium and reconstructing 250 artifacts, has also brought its own bit of controversy.
The Palestinian Authority says the exhibition is a violation of international law because much of its material was taken from near Bethlehem and Jericho, both in the Israeli-occupied West Bank. An Israeli group of archaeologists and activists complains that the museum, however unwittingly, is helping the Jewish settlement movement advance its contention that the West Bank should be part of Israel and not a Palestinian state....
comments powered by Disqus
- A grandmother's trove of Civil War photos goes to Library of Congress
- Tribes See Name on Oregon Maps as Being Out of Bounds
- Holy Haystacks! Researchers Have Officially Discovered A New Monet
- Joan Baez, Sly Stone, Steve Martin, Ben E. King -- all honored by the Library of Congress
- StoryCorps to Launch Global Expansion With $1M TED Prize
- OAH denounces anti-gay legislation signed by Indiana governor
- Emory's Leslie Harris says we should remember the racist roots of American colleges as we think about what went wrong at OU and other schools
- Stanford historian looks to the U.S. Postal Service to map the boom and bust of 19th-century American West
- U.S. historian denounces Japanese scholars' statement over wartime sexual slavery
- Timothy V Johnson Named Head of Tamiment Library


text:
Radiant Tinted Moisturizer SPF 15 is a lightweight tinted moisturizer with an in-part avobenzone sunscreen. It has a cushiony, moist texture on application, but sets to a soft matte finish with the tiniest hint of shine. You'll achieve soft color and very sheer coverage from each of the shades, all of them outstanding. This formula is best for normal to dry skin. Those with oily or blemish-prone skin will not appreciate the short-lived matte finish, and the thickeners and waxes aren't the best over breakout-prone areas.
Note: This foundation's rating is due to its overall performance rather than its SPF rating. Due to concerns about people not applying sunscreen liberally enough to get the amount of SPF protection stated on the label, it is often recommended to look for SPFs with ratings higher than 15. If you plan to use foundation as your sole source of facial sun protection, consider using one rated SPF 20 or greater. If the foundation with sunscreen you choose is rated less than an SPF 20, we strongly advise applying it over a daytime moisturizer rated SPF 15 or greater and following it with a pressed powder rated SPF 15 or greater. That way, you're ensuring sufficient broad-spectrum protection which is essential for having and maintaining healthy, younger-looking skin at any age.
Active: Octinoxate, Avobenzone; Other: Water, Cetearyl Alcohol, C12-15 Alkyl Ethylhexanoate, Dimethicone, Ethylhexyl Palmitate, Polyglyceryl-4 Isostearate, Cetyl PEG/PPG-10/1 Dimethicone, DEA-Cetyl Phosphate, PEG-40 Castor Oil, Polysorbate 60, Hexyl Laurate, Phenoxyethanol, Sodium Dehydroacetate, Cetyl Dimethicone, Dromiceius Oil,Tocopheryl Acetate, Carbomer, Allantoin, Magnesium Aluminum Silicate, Sodium Ascorbate, Methylparaben, Isopropylparaben, Isobutylparaben, Butylparaben, Propylparaben; May Contain:Titanium Dioxide, Iron Oxide, Iron Oxide, Iron Oxide, Mica
Sonia Kashuk At-A-Glance
Strengths: Affordable and widely available (though exclusive to Target stores); good makeup remover; the makeup is still the star attraction, with impressive options for foundation, powder, blush, lip color, eyeliner and especially makeup brushes.
Weaknesses: The concealers, eye pencil, and brow shapers are a letdown; the makeup palettes may seem convenient, but several of the included products perform poorly.
Perhaps best known for her years of work with supermodel Cindy Crawford, makeup artist Sonia Kashuk's well-chosen selection of makeup has been available exclusively at Target stores since the mid-1990s.
Kashuk's brushes have been a strong point, and now that they've been redone they're better than ever, and still affordable. This is definitely a makeup collection worth visiting, and you'll find some premium options among Kashuk's powder foundation, regular powders, and blush. Several other products, from foundation to lipsticks, are also worthy of attention, though this is not a line that caters to darker skin tones. Ironically, it's the latest cosmetic products that met with the most critical reviews, largely due to poor execution or their gimmicky nature.
For more information about Sonia Kashuk, visit soniakashuk.com or www.target.com.
Note: Sonia Kashuk products are also available at select Bloomingdales locations in New York and California. The company also maintains freestanding stores in New York and New Jersey.
The Beautypedia Research Team is dedicated to helping you find the absolute best products for your skin, using research-based criteria to review beauty products from an honest, balanced perspective. Each member of the team was personally trained by Paula Begoun herself.
I have been using this product for about 3 years and I love it. It has good coverage and lasts until I wash it off at night. I am over 50 and have good skin so I don't need too much coverage. I tried using different tinted moisturizer brands, and none compare to Sonia Kashuk. The others
came out too white or pink and left an unnatural color on my skin.
Terms of Use
Thanks for sharing your experience with this product! Approved reviews appear on our site within 3-5 business days.
We reserve the right to reject or modify reviews without notice. All submitted reviews become the property of Paula's Choice, LLC.
Call us Mon–Fri 6am-6pm PT, Sat–Sun 8am-4:30pm PT
Connect with us:
Our Privacy Policy: We will not sell or share any of the information you provide.
*Promotion expires 9pm PT, April 7, 2015. No cash value. Applies to in stock items only. Cannot be combined with other offers or applied to previous purchases.
**Promotion expires 9pm PT, April 7, 2015. No cash value. Applies to US and Canada only.
***Promotion expires 9pm PT, April 7, 2015. Choose any 3 from 33 available FREE samples on orders of $5 or more.
Please sign in and access your saved member information.
You will have the option to create an account after you have submitted your order.


text:
|Premier Wine & Spirits
|USD 22.99 $22.99 750ml
Dashe Cellars Zinfandel Dry Creek Valley 2009
Winemaker's Notes:
Color: Black-purple, with reddish hues. Aroma: Black raspberry, black cherry, cassis, mineral, chocolate, black pepper, anise. Taste: Black cherry, black raspberry, chocolate, licorice, clove spice, good acidity, wonderful viscous velvety texture on the mid-palate, complex, long sweet finish of black cherry fruit, and peppery spice.
Member Reviews for Dashe Cellars Zinfandel Dry Creek ValleyAdd your review
Musty cranberries on the nose. Dark fruit and a funky dry quick finish.
Liquid rasberries. Pour over haggen daz or slurp out of gothic girlfriends belly button and fulfill her fetish of blood play without risk of HIV and having to look depressed emo for work on Monday.
Classic DCV zin. Very much a medium-bodied style that is full of cranberry and raspberry fruit with dust and spice.
A great wine price right! Geat structure,balance,full of fruit. a must try! Don't sleep on this one!
Four and a half glasses
Dashe was recommended to me while tasting around Healdsburg, CA and it did not disappoint. Dry Creek Valley is arguably the best place for Red Zinfandels in the entire US. This wine is intense but not overpowering and for someone who loves this type of wine, it is one of the best I have ever tasted (and may ever taste). I've had two separate vintages of this wine and both were equal and above nearly everything else. It's not expensive to buy (~$25) which is very nice because this is a rare find in that you get such an amazing wine for an affordable price. Highly, HIGHLY recommended.
Four glasses
External Reviews for Dashe Cellars Zinfandel Dry Creek Valley
With a little Petite Sirah and Carignane blended in for depth and complexity, this is a nice Zinfandel for everyday drinking. It's dry and full-bodied, and while the tannins are strong, they're perfect for busting up rich barbecue. Shows all the fruity spiciness you expect from a lusty Zin.
The 2009 Zinfandel is a very pretty, mid-weight wine laced with sweet red berries, flowers, licorice and leather. This is a decidedly feminine, gracious style, but it works very nicely. The 2009 is a very pleasing wine best enjoyed over the next 2–3 years. Anticipated maturity: 2012–2017.
Rustic and ripe, with dried cherry and licorice aromas and hardy plum, blueberry, roasted sage and cracked pepper flavors. Drink now through 2012.
Somewhat oak-driven, the plum, raspberry, blackberry and herb flavors take a backseat. Still, the wine shows well, with fine tannins framing the flavors, which finish with a chocolatey edge. Long and well balanced.
Made in a totally different style, the 2005 Zinfandel Todd Brothers Vineyard is a deep ruby/purple-tinged offering displaying more chocolatey black fruits, tremendous intensity (it's one of the most concentrated Zinfandels of the vintage), and outstanding balance, richness, and length. Full-bodied with stunning purity and texture, it is a genuine revelation in this vintage. Drink it over the next 2-3 years.
Rich and nuanced, the flavors range from blackberries and blueberries through spiced plums and white chocolate. Best of all is the texture: rich, velvety, supple and flattering. It's a big wine, but wears its size well.
The opaque purple-colored 1999 Zinfandel Todd Brothers Ranch boasts a superb bouquet of pure, jammy black raspberries, cherries, minerals, and subtle notions of earth and wood. Full-bodied, powerful, big, and thick, this dense, blockbuster Zinfandel can be drunk now and over the next 7-8 years. Bravo!
All Prices
Premier Wine & Spirits
Amherst, NY (280 mi)
Dashe Cellars Zinfandel, Dry Creek Valley 2009 / 750 ml. »
$22.99 750ml USD 22.99
Food Pairings for Dashe Cellars Zinfandel Dry Creek Valley
Photos+ Add Image
Best Wine Deals
$22.9515%offChateau Patache d'Aux 2010
$34.9922%offJelu Torrontés Valle de Zonda 2008
Zinfandel Top Lists
Community Activity
Whatcha drinking tonight?Replied
Frisk Prickly Riesling (2013)Listed
Rock View Riesling Columbia ValleyListed
Atwater Estate Vineyards Riesling Bubble (2013)Listed
Jones of Washington Riesling Wahluke Slope (2008)Listed
Nefarious Cellars Riesling Columbia Valley (2011)Listed
Maryhill Riesling (2012)Listed
Kiona Riesling Red Mountain White Riesling (2009)Listed
Zero One Vintners Riesling (2007)Listed
Snoqualmie Eco Organic Riesling (2012)Listed


text:
The system works completely automatically, and does not offer any way for the driver to lock the torque distribution. Taking the Crosstour up a gravelly slope, we found the system worked as advertised with the grind of one of the front wheels spinning free being quickly replaced by continued ascent as the rear wheels dug in.
The Crosstour rides higher than the Accord sedan, with 6 inches of ground clearance. That may not be river-fording height, but it will keep the Crosstour from getting bogged down in snow and let it navigate rutted roads.
On normal roads, the Crosstour shows an easy drivability. The steering uses a variable ratio, but the wheel feels loose, offering a comfortable amount of play. It doesn't require much input during long freeway cruises, but also doesn't feel like precision steering. In dense urban areas, the turning radius seemed a little wide, requiring more than one instance of backing and filling to exit a parking space.
Taking a turn at speed, the Crosstour shows some good stability, with no excessive body roll. But at the beginning of a turn the car feels like it wants to neatly rotate. Then something weird happens--possibly the stability electronics taking over--and resets the car's balance. That behavior underlines the fact that the Crosstour is not a sports car.
Although the Accord sedan can be had with a four- or six-cylinder engine, Honda makes the Crosstour with a V-6 only. This 3.5-liter V-6 appears in many different Honda vehicles, and uses Honda's VTEC variable valve timing to improve efficiency. Output is 271 horsepower and 254 pound-feet of torque--not the most spectacular numbers when compared with newer, direct-injection engines.
But power feels more than adequate from the driver's seat. The Crosstour responds well to accelerator input. It moves fast enough for comfortable freeway merges and jumps off the line with a satisfying push.
The automatic transmission's lack of a sixth gear puts it in the Stone Age category. Rather than manual gear selection, Honda gives it three low ranges, which works for the car's mission but will bother drivers who want precise gear control.
Fuel economy for the Crosstour with all-wheel drive is EPA rated at 18 mpg city and 26 mpg highway. CNET's review car showed midteens in city driving, but ended with an average of 18.6 mpg over a mixed course of city, freeway, and mountain driving.
Honda employs the trick of active cylinder management to get the Crosstour's highway number. The engine shuts down up to half of its cylinders when cruising at steady speed, seamlessly reactivating them when the driver hits the gas. The system runs transparently, without intruding on the driving experience.
The car offers some advice on increasing mileage in the form of a green Eco light in the instrument cluster. But keeping that light shining restricts you to only the lightest pressure on the accelerator. Any call for even a measure of power will extinguish it.
In sum
Honda has not been pushing the tech envelope lately, a fact the 2011 Accord Crosstour EX-L demonstrates. The navigation system is at least six years old, but it is nice to see iPod integration and a decent Bluetooth phone system. The stereo's sound quality stands out as the car's high point for cabin tech.
The onscreen cabin tech interface uses an easy-to-understand paradigm, but its design is chunky and ugly. The dual voice command systems show an overall lack of integration when it comes to cabin tech. The hatchback cargo area offers good practicality, although the obstructed rear view is troublesome. As for the car's exterior design, it will certainly have its detractors, but it does have a unique appearance.
The only real highlight of the Crosstour's performance tech is the engine's cylinder management. All-wheel drive is a nice addition, but this system offers few features. The transmission is low-tech by today's standards.
|Tech specs
|2011 Honda Accord Crosstour
|Power train
|3.5-liter V-6, 5-speed automatic transmission
|EPA fuel economy
|18 mpg city/26 mpg highway
|Observed fuel economy
|18.6 mpg
|Bluetooth phone support
|Disc player
|MP3-compatible 6-disc changer
|MP3 player support
|iPod integration
|Other digital audio
|USB drive, auxiliary input, satellite radio
|Audio system
|Seven-speaker 360-watt system
|Driver aids
|Rearview camera
|Base price
|Price as tested


text:
Juniper acquires virtualization security firm
Juniper Networks Monday announced it has acquired Altor Networks, a provider of virtualization security for data centers, for $95 million.
Juniper Networks Monday announced it has acquired Altor Networks, a provider of virtualization security for data centers, for $95 million.
F5 is making file virtualization more cloud friendly with the introduction of software that translates storage protocols, making it possible to store files in public or private cloud networks using a range of technologies.
IBM's research division is working on several virtualization projects that could boost security of cloud computing networks, reduce data center power costs, and improve the ability to run multiple hypervisors and operating systems, including Linux an...
When you consider technologies that make corporate IT more efficient by improving utilization of computing resources, <a href="http://www.networkworld.com/news/2010/060710-tech-argument-citrix-vmware-microsoft.html?source=NWWNLE_nlt_daily_am_2010-06-...
Business agility is slowly replacing cost as the factor driving most companies down the path of desktop virtualisation. According to a survey from Molten Technologies, companies are also unsure whether the drive to desktop virtualisation would be a w...
A new tool from Citrix has been designed to help XenDesktop users gather more detailed information about the hardware that is being used.
In an era of technology industry consolidation, the questions raised by Novell's sale to Attachmate for $2.2 billion ought to be familiar by now. Analysts can only speculate -- and users can only wonder -- what may happen to Novell's deep and extensi...
Microsoft doesn't earn billions of dollars every month by giving away its software. But even Microsoft is no stranger to offering freebies in a bid to increase customer loyalty.
Cisco today announced two new virtual desktop devices, more efficient virtualization software and more affordable videoconferencing endpoints, building on its line of video collaboration tools and capabilities for companies.
Companies looking to adopt cloud computing could cut their chances of being compromised by using a statistical framework to assess risk according to Runaware. Concerns about security are the main inhibitors for companies looking to adopt cloud techno...
Don't look now, but the data center is transforming. Seeking greater efficiencies, enterprises of all sizes have steadily evolved from application-based silos to virtualized environments. Now this evolution is taking the next leap forward as innovati...
Oracle has reportedly changed its position on support for its RAC (Real Application Clusters) software running in VMware virtualized environments, saying it will provide it under certain circumstances.
In an effort to ease the complexity of system administration, Oracle has expanded its Enterprise Manager software to include servers, operating systems, networking gear and virtual environments, the company announced Tuesday.
At the Computerworld Premier 100 IT Leaders conference in March, one CIO stood up to express his unease about the security of a virtual infrastructure that has subsumed more than half of his company's production servers. Two other IT executives chime...
Microsoft has partnered with six major hardware vendors to provide reference architectures that should help enterprises build private IaaS (Infrastructure-as-a-Service)-styled clouds, the company announced Monday.
The latest business news, reviews, features and whitepapers
Visit our shop for the best deals on hot products
Watch our video news and reviews from around the world
Comprehensive buying guides, features, and step-by-step articles
First impression on unpacking the Q702 test unit was the solid feel and clean, minimalist styling.
For work use, Microsoft Word and Excel programs pre-installed on the device are adequate for preparing short documents.
The Fujitsu LifeBook UH574 allowed for great mobility without being obnoxiously heavy or clunky. Its twelve hours of battery life did not disappoint.
The screen was particularly good. It is bright and visible from most angles, however heat is an issue, particularly around the Windows button on the front, and on the back where the battery housing is located.
My first impression after unboxing the Q702 is that it is a nice looking unit. Styling is somewhat minimalist but very effective. The tablet part, once detached, has a nice weight, and no buttons or switches are located in awkward or intrusive positions.


text:
Foxgloves in containers
I love foxgloves but, living in an apartment, don't have any wet, shady areas. Has anyone successfully grown foxgloves in containers on their balcony?
Although my deck is south-facing, I can create some shade and in fact have a large conifer covering a portion of the deck, which gives some filtered sunlight, at best, to that area.
I live in Vancouver, so it isn't too hot here at the best of times.
I'd appreciate any advice; thanks,
My containered foxglove is putting on a show right now!
My balcony faces south as well; the plant is growing in what passes for full sun out there - sunrise to about 11:00 A.M. - and part sun/part shade for the rest of the day.
My only regret is that I don't have enough space to have a whole stand of them.
Check out the link below to see ...
Here is a link that might be useful: My foxglove story


text:
The Next System Project, which launches today, offers systemic solutions to the systemic climate, political, economic, social and cultural crises the United States is experiencing.
A construction accident illustrates the differential risks still facing Latino workers, often employed by a tangled web of contractors and subcontractors.
Lesley Clark works for The Miami Herald and The Bradenton (Florida) Herald. She's worked for The Miami Herald in Washington since February 2011, and before that in Miami and Tallahassee, covering state government and politics. She's a graduate of the University of Massachusetts at Amherst.


text: Maine is a beautiful place of the country with offerings of nature, outdoor activities, great music and arts and the ocean nearby. After living in large cities for about 10 years prior to moving here, we have found the gentleness and innate beauty of Maine to be a welcoming respite. We look forward to meeting those of you who wish to visit and experience the splendor of Maine.


text:
Origami News
News on Origami continually updated from thousands of sources around the net.
Daly Projects celebrates divergent roster with new show
Wednesday | Planet Jackson Hole | Comment?
Origami crane dresses are one highlight of successful M-A fashion show
Wednesday Mar 25 | InMenlo.com | Comment?
Single-molecule chemical reactions on DNA origami
Wednesday Mar 25 | CiteULike | Comment?
There's A 5,000 on the streets of London today and it's yours if you find it
Wednesday Mar 25 | Metro UK News | Comment?
This Collection Is Like Origami For Your Body
Tuesday Mar 24 | Refinery 29 | Comment?
Upcoming Drone Game Lets You Feel the Horror of Killing Middle Eastern Civilians
Tuesday Mar 24 | The New York Observer | Comment?
30 Celebrities Who Have Twin Children
Did The Pope Perform a Miracle in Naples?
topix 297
Word Search - All Around Topix
topix 74
Design Principles for Single-Stranded RNA Origami Structures
Monday Mar 23 | CiteULike | Comment?
ChangeThis: Jeremy Gutsche's Better and Faster Manifesto Explores How ...
Thursday Mar 19 | Trend Hunter Magazine | Comment?
Bookniture is Furniture Hidden in a Book
Tuesday Mar 17 | Small Business Trends | Comment?
The epigenetic mechanisms that underlie health and disease
Tuesday Mar 17 | Immunology and Cell Biology | Comment?
Croll to Receive Air Force Young Investigator Award
Monday Mar 16 | Newswise | Comment?
REDCAT gala honors Gabriel Orozco and Michael and Jamie Lynton
Monday Mar 16 | Los Angeles Times | Comment?
Pupils show their skills n in Origami at Holbrook's Japanese dayPics Trevor Porter
Monday Mar 16 | Wiltshire Times | Comment?
Celebration of Us at Digby Elementary School
Monday Mar 16 | Nova News Now | Comment?
An Origami Yoda Activity Book
Sunday Mar 15 | Examiner.com | Comment?
Student programs DNA to form Script Ohio
Sunday Mar 15 | The Columbus Dispatch | Comment?
Sponsored: Celebrating Eileen Fisher with Your Favorite Fashion Bloggers
Thursday Mar 12 | New York Magazine | Comment?
Trending Now
Updated: Thu Mar 26, 2015 12:42 pm
Copyright © 2015 Topix LLC


text:
Website on a budget required
Closed - This job posting has been filled and work has been completed.
Job Description
I need someone to turn my website design into html and java:
I would also like input on the colour schemes, a blog i can easily update, key words for google search found in the programming, and a contact page which goes directly to my email. Its pretty basic.
I can send all photos.
Skills: design


text: Select your motorcycle to guarantee fitment. Custom Dynamics HD SMART Triple Play Brake / Running Light and Turn Signal Conversion Kit J&P Part: AI632-165 Regular Price: $89.99 Scratch & Dent: $62.99 Scratch & Dent Item Product Description: Manufacturer Part Number: GEN-SMART-TPU-HD More Info In Stock, usually ships same business day. More Info


text:
OP Junior Member
Thanks Meter: 0
5 posts
Join Date:Joined: May 2010
AT&T has tried to issue me unlock codes twice, but neither works. I'm using an old t-mobile SIM to attempt the unlock.
Does anyone know if maybe updating the CID impacts the ability to use a carrier unlock code?