text:
Jeeja Yanin with Tony Jaa | Action Demo 2014 MP3
A little practice and warm up with some friends.
JeeJa Yanin Fight MP3
This Video footage is owned and copyrighted by {Sahamongkolfilm International}, {Magnolia Pictures} I do not own or did not make any of the footage seen.
THE KICK (Kim Yiroo VS Kyung Suk & JeeJa Yanin) MP3
The Kick (2011) Kim Yi-roo Cut Kim Yi-roo김이루 VS Kyung-suk Kim 김경숙, JeeJa Yanin Cast Yi-roo Kim ... Yi-roo Kyung-suk Kim ... Tae Mi JeeJa Yanin .
Jeeja Yanin | Video of inspiration | Towards 2015 MP3
Words Jeeja Yanin: "Don't let the negative thoughts in your mind stop you from your dreams!"
Jeeja Yanin - Raging Phoenix fight scene MP3
If you haven't heard, Jeeja Yanin is kind of a big deal. Her second movie, "Raging Phoenix," came out recently and it has some awesome fight scenes--here's ...
Heroes of Martial Arts #2 - Jija Yanin (Jeeja Yanin, Chocolate, Raging Phoenix) MP3
Copyright Disclaimer Under Section 107 of the Copyright Act 1976, allowance is made for "fair use" for purposes such as criticism, comment, news reporting, ...
Chocolate - Fight Scene 6 - Dojo MP3
Jeeja Yanin fights several henchmen. Then through the power of autism, learns how to fight a martial artist with more than a few ticks. Choreographed by ...
JeeJa Yanin - Chocolate making fight scene MP3
Chocolate making fight scene.
JeeJa Yanin - The Kick Fights. MP3
Staring K. Kim, Jae-Hyun Cho, Ji-won Ye, JeeJa Yanin, Brahim Achabbakhe & Petchtai Wongkamlao.
JeeJa Yanin - The Kick (더 킥) - Thai/Korean movie my Fanmade Video MP3
My short fanmade music video, I hope you'll like it! http://www.youtube.com/watch?v=kC8NrKkifxs ORIGINAL SONG!!!!
best of JeeJa Yanin MP3
jeeja yanin.
JEEJA YANIN IN TAIWAN (Full) MP3
JEEJA YANIN In Taiwan Promote Chocolate Movie 29/8/08 - 1/9/08 Jeeja went to Taiwan to promote her movie,Chocolate,at Century . Movie will show there at ...
Practice time with Tony Jaa and Jeeja Yanin MP3
A short practice video for a demonstration at the 2014 S.E. Asia IPO Championship next weekend at Fort Kamphaeng Phet Akkharayothin Thailand.
JeeJa Yanin - Chocolate 2008 Trailer MP3
chocolate trailer.
Jeeja Yanin Taekwondo Blackbelt MP3
Jeeja Yanin, star of "Chocolate" and "Raging Phoenix", posing in her South Korean (Kukkiwon) Taekwondo uniform for a photo shoot.
Jija Yanin V.S Jackie Chan 成龍與琴嘉 MP3
成龍與琴嘉的比較 Comparison between Jija Yanin and Jackie Chan.
She Did It On 'Em - Yanin "Jeeja" Vismistananda MP3
This is a tribute video to a Thai action star that has quickly risen up the ranks to be one of my favorite action stars of this generation! Her name is Yanin "Jeeja" ...
Tributo - JeeJa Yanin MP3
JeeJa (chocolate 2008) stunt show on Johjai TV Part 2/2 MP3
JeeJa [Yanin Vismitananda],the new female superstar in martial arts, shows her moves and stunt show from Thai action movie,Chocolate on JohJai ...


text:
For 44 years, local business and community leaders have come together to support Linfield College through the annual Partners in Progress campaign. Since Linfield's chartering in 1858, Yamhill County has played a pivotal role in advancing the college's mission and has helped to provide critical financial support.
Through PIP, Linfield and the community work toward a common goal: a healthy, vibrant and growing community where opportunities for lifelong learning flourish.
Learn more about the campaign below, or make a gift today.
Gifts through PIP support the resources that provide lifelong learning opportunities for students and Yamhill County residents such as:
Your gift to PIP is an investment in Yamhill County's future.
In addition to the many programs that PIP contributions support, your gift also makes a Linfield education possible for students like Jamie.
"Studying in a small department gives me the opportunity to try out what I want to do when I graduate, and the faculty are very caring. When I looked at other schools, scholarship support, especially in the music department, was significantly higher at Linfield, which made it obvious that this is where I should be. My professors have supported me in building the education I want and need."
~ Jamie Bostock '19 music major from Dundee
Kathleen Bernards '76
Kyle Faulk
Erin Stephenson, Chair
Pamela Watts
For more information about PIP events or giving to Linfield, contact the annual giving office at 503-883-2217.

subset: commoncrawl


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Filepicker: Uploading Photos To Filepicker Now Displays Them In Random Order As Opposed To Previous SequentialAsked by markashton on October 16, 2015 at 04:54 AM
The new implementation to filepicker has changed a behaviour ....
Multiple photos uploaded now show in the submission bin and grid reports in a random order rather than the sequence they were uploaded in.
I think having them show in a sequential order as per upload is much more valuable than having them display in any order.
Previously, the first photo uploaded would be displayed first ... then the second ... etc.
This is a chronological benefit.
At this early stage ... could I ask for this to be reinstated please.
Many Thanks
Regards Mark
JotForm Support
Hi markashton,
I am currently checking the FilePicker functionality. Will be shortly. Thanks.
JotForm Support
Just tested it and confirmed that the uploaded files appear in random order in the submissions page. I will escalate this to our development team so they can bring back the sequential ordering of uploaded files. We will inform you in this thread once this is implemented.
JotForm Support
Hello Mark,
This was also restored. Chronological order of uploading - even via edit mode - is observed.
Please check - let us know if you have any further issues.
Agreed. Many Thanks. Close ticket.
Regards Mark


text:
Doctor insights on:
Quadruple Blood Test In Pregnancy
Does any blood test detect pregnancy?
Yes: A serum b (betta) hcg.Get a more detailed answer ›
Pregnancy (Definition)
When your due date arrives, you will be more than ready to have your baby! Most women deliver the baby somewhere between 37 and 42 weeks. According to the American College of Obstetricians and Gynecologists, only 5% of babies arrive on the exact due date. Approximately 7% of babies are not delivered by 42 weeks, and when that happens, it is referred to ...Read more
How does a pregnancy blood test work?
HCG level: The embryo, after implantation in the uterus, makes human chorionic gonadotrophic hormone (HCG). Both blood and urine pregnancy tests depend on detecting HCG, a marker of pregnancy ...Read more
When can a blood test detect pregnancy?
10 days.: Blood test HCG quantitative test can show pregnancy as early as 10 days of conception. ...Read more
What blood test should I get for pregnancy?
A beta-HCG test.: A serum or urine beta-human Chorionic Gonadotropin test is/are best for determination of pregnancy. There are home pregnancy tests which mostly test urine, and serum/blood tests, which are done in the clinical lab. It would like;y be best to wait a week, and then go see your doctor to get a serum test in the lab as an outpatient. ...Read more
How soon can a blood test detect pregnancy?
Soon: You can get a blood test with same day results and they can be reliable 1 to 2 days before your missed menses. I do not like to do pregnancy tests too early. Several women have biochemical pregnancies that do not grow and get a normal period 1 to 2 days later. So, my suggestion is that you do your blood test when you miss your period. ...Read more
What causes a blood test to determine pregnancy?
See your Physician: He will order HCG quantitave pregnancy test which is very accurate, . ...Read more
Is blood test the most accurate to detect pregnancy?
Yes: In early pregnancy, the blood test is more accurate than the urine test. The accuracy of doing one or two (second one a week later) urine tests is good enough for most people to use. ...Read more
What does a satisfactory pregnancy blood test mean ?
Not Medical: That term is not one used in medicine nor is it in any way explanatory. Perhaps you should ask this specific question of the professional who used it. ...Read more
What are the chances of a negative blood test in early pregnancy?
Very low: It depends on how "early" you are referring to, but generally, the blood test for pregnancy is very accurate with a very low false negative rate. ...Read moreSee 1 more doctor answer
Do I need to take blood test with fasting to confirm my pregnancy?
NO: Fasting is not required to check for pregnancy hormones in your blood. ...Read more
- Talk to a doctor live online for free
- Types of blood test for pregnancy
- Chemical pregnancy blood test
- Bhcg blood test pregnancy
- Ask a doctor a question free online
- At home blood test for pregnancy
- Qualitative blood test for pregnancy
- Blood pregnancy test
- Pregnancy test blood
- Talk to a pathologist online for free


text:
Join JSB to raise money for Movember – all donations go directly to charity
November 29th, 2012
On Friday the 30th of November 2012, the Melbourne JSB Lighting team are putting on a party for a great cause: Movember and its associated charities.
$10 donations on the door will be gratefully received.
18.30 til… Late!
65 Southbank boulevard, Southbank, Melbourne.
INDESIGN is on instagram
Follow @indesignlive
The internet never sleeps! Here's the stuff you might have missed


text:
Have you had trouble getting a job or keeping a job?
SSWAP is a year-long program to help adults like you (over 18) get employed and stay employed.
You will set your own goals and make your own Action Plan, with the help of an instructor/ counsellor.
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Contact us today, or read more about the program.
Watch this video about SSWAP, which was made by one of our participants!


text:
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text:
Talk:Analytical hierarchy
|WikiProject Mathematics
|(Rated B-class, Mid-importance)
boldface distinction[edit]
Problem here -- the boldface/lightface distinction is not clearly made --Trovatore 7 July 2005 19:31 (UTC)
The term "analytical" refers exclusively to the lightface concept, whereas "projective" is a boldface notion. Therefore "projective set" should not redirect here. I'm planning a pointclass page where it might redirect instead. Also "analytic" and "co-analytic", which are boldface notions, should be removed from this page. --Trovatore 7 July 2005 20:35 (UTC)
The other major problem with this page is the conflation of formulas and sets. Consider for example the following passage:
- A formula is a formula of the form , where X is now a predicate and , while a set is a set of the form
- ,
- where S is Borel and R is a relation.
First, the definition is incorrect, because no restriction is placed on the definability of R. But the problem that more exemplifies the difficulty with the page as a whole is that nothing is said about the underlying Polish space. --Trovatore 7 July 2005 20:48 (UTC)
major problems seem to be fixed[edit]
Accuracy tag removed (thanks to Ben Standeven). Tech tag removed; subject is inherently technical. --Trovatore 02:18, 20 September 2005 (UTC)
edit on 2006-6-13[edit]
The main changes are:
- Carefully separate the cases of sets of numbers and sets of reals. This is an experiment that may be useful on arithmetical hierarchy as well.
- Emphasize the fact that this is lightface (maybe more emphasis needed?)
- I rephrased the introduction. The anlytical hierarchy is not really about second order logic; it is about higher type languages such as Z_2 or type theory in first order logic. Every use I know of is in the context of first order ZFC or first order Z_2.
- Add a reference to Rogers' book. I plan to add more references.
I've seen redlinks to 'hyperarithmetical heirachy' many places, and just recently noticed hyperarithmetical theory so I've created a redirect for now. It's possible that one or the other will need to be renamed in the future, but I figured this was a cheap way to get rid of a lot of dead links at least for now. --- all assuming that 'hyperarithmetical theory' is talking about the same thing... here's hoping! Zero sharp 20:46, 30 August 2007 (UTC)
Removed sentence[edit]
"Note that it rarely makes sense to speak of a formula; the first quantifier of a formula is either existential or universal."
Well, not really, the first quantifier of the normalized form, but not of the formula. As we just demonstrated any formula in or is in for all m>n. Rich Farmbrough, 16:23, 14 December 2009 (UTC).
MAJOR PROBLEM here -- the description of the the abstract levels of the hierarchy do not adequately reflect the alternating nature of the quantifiers. In fact, no where on the page is the word "alternating" found. See the planet math article for a correct description. —Preceding unsigned comment added by 22.214.171.124 (talk) 18:44, 24 June 2010 (UTC)
- It is true that the page doesn't use the word "alternating", but I think the definition here is correct. Is there something in particular wrong with it? — Carl (CBM · talk) 00:26, 25 June 2010 (UTC)
"Indicies of Computable ordinals"[edit]
It is unclear what the indicies of computable ordinals are. See the discussion at http://math.stackexchange.com/questions/72826/complexity-of-the-set-of-computable-ordinals. It may refer to recursive well-orderings of or to Kleene's O notation. If it is Kleene's O, there should be a link to http://en.wikipedia.org/wiki/Kleene%27s_O. Either way (or if it is both and these are equivalent statements), this should be clarified — Preceding unsigned comment added by 184.108.40.206 (talk) 20:40, 2 November 2011 (UTC)


text:
Update – 7/4/11:
According to Mashable, Google's agreement with Twitter that gave the search engine full access to public Twitter feeds has expired. Because of this development, Google has taken down their realtime search feature.
Original Post- 2/17/11:
Google just made search a little more personal by integrating your social accounts into your results. At first blush, it seems pretty cool.
Wired has a pretty good rundown of the social search changes. So, what do you think? Useful?


text:
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text:
PDAC is a group of key stakeholders from a wide variety of child related industries who work to advance the field of early care and education, school-age, youth development and family support.
The PDAC section is under construction and work is being completed to update the committee membership, scheduled meetings, minutes and agendas. Check back soon for updated information.
It also provides standards, guidelines, resources and supports to help you make sensible changes that lead to better quality outcomes for children.


text:
News and Events
Recent Releases
07/20/2017 - 3:00pm
Does every cancer need immediate treatment? The case for active surveillance for low-risk cancersMichigan Medicine experts discuss whether active surveillance could reduce overtreatment for some thyroid, prostate and breast cancers
07/13/2017 - 3:30pm
Appropriate Distress Screening and Follow Up Leads to Fewer ER Visits and Hospitalizations in Patients with Cancer, Study FindsFollowing a cancer diagnosis, all patients experience some level of distress -- regardless of disease stage. When severe and left untreated, distress can have a significant impact on health outcomes, lead to greater mortality and morbidity, affect immune function, and result in higher health care expenditures.
06/30/2017 - 10:30am
Immunotherapy, Gene Therapy Combination Shows Promise Against GlioblastomaNew research out of the University of Michigan supports combining two approaches to fight back against gliomas: attacking the tumor with gene therapy while enhancing the immune system's ability to fight it, too.
06/29/2017 - 11:30am
Researchers propose new approach to identify genetic mutations in men with prostate cancer
In a new study, researchers used a patient's own cancer history rather than family history to identify genetic mutations that might influence cancer treatment and risk for family members.
06/27/2017 - 1:30pm
Grant establishes proteogenomics center at U-M
Researchers at the University of Michigan will lead one of five nationally funded centers dedicated to accelerating research into understanding the molecular basis of cancer and sharing resources with the scientific community.


text:
Watertown City Council and mayoral candidates will face questions on Tuesday in what traditionally is a sparsely attended debate. Topics generally center on specific issues, when they should zero in on leadership qualities.
Those with leadership skills are more capable of responding correctly to a large number of issues.
Unfortunately, council leadership has been lacking in the past four years and voters can vote for more of the same, or take a chance on some of the new candidates.
Leadership is something that takes time and work to develop, and it was noteworthy that none of the current council or mayoral candidates attended a recent day-long leadership forum sponsored by the Prairie Lakes Health Care Foundation. Two council challengers, Beth Mantey and Don Roby, did attend.
Well over 150 local residents were inspired that day by a slate of national speakers who coached people across the country to take stronger roles in their community.
Effective leadership, they stressed, requires a combination of character and competency. Many leaders fail in one of those areas.
Character means that a person has the integrity and the intent to do what is right. Competency means they have the capability to achieve results.
Character is relatively self-explanatory and is obvious in a person's current and past conduct. Competency is based on experience, a person's beliefs and their ability to act.
With that in mind, candidates should be asked questions that give us insight into why we should trust them with the city's future.
The incumbents are well known by anyone who has observed the council. We've seen many of their actions. Incumbent candidates Dennis Solberg, Mike Christians, and Dana Jones, regularly gathered with a fourth member, Al Meisenheimer, who is up for re-election in two years, for questionable Saturday morning breakfast meetings. They almost always voted as a block at council meetings. During much of that time, mayoral candidate Steve Thorson presided as council president over a group that many believe was the most ineffective council in a long time. Never did he publicly challenge this practice.
So the questions to these men are, "How will your style change in the next four years? What do you intend to do to make Watertown a better place to live? What is your guiding force in wanting to be on the Watertown City Council, or in Thorson's case, to be mayor? Why will the next four years be any different than the past four?
The question for incumbent Bill Rieffenberger is a bit different. He consistently has shown leadership and an ability to achieve results through his attention on improving the Uptown District and Watertown Transit, among others. For him, the question is, "What will you spend the most time working to accomplish during the next four years."
And for Mayor Gary Williams, the question is, "How would you change your leadership during the next four years to become more inclusive in engaging council members and the public in moving Watertown forward?"
For the council challengers, Mantey, Roby, Dan Albertsen and Blake Dahle, the questions are simple. "What experience do you bring that will benefit Watertown? What beliefs do you have about the city's future and what motivates you? What action will you take to improve Watertown?"
Oh yes, one last thing, the Watertown Chamber of Commerce's Government and Local Affairs Committee, which is sponsoring the 5:30 p.m. forum on Tuesday at the Watertown Event Center's Heritage theater, should arrange for it to be professionally videotaped for replay on our local Gov-TV channel. A much larger audience deserves to hear the debate.
Brad Johnson is a Watertown businessman and journalist who is active in state and local affairs.
(Editor's note: The city council/mayor forum will be livestreamed Tuesday on P/O-TV for those unable to attend. Go to www.thepublicopinion.com and click on the P/O-TV banner.)


text:
Domain Name: SMAN11MKS.COM
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text: Washington (ANI): Katie Holmes rep has shot down reports that the actress will soon be trying her hand at designing a childrens clothing line. OK! Magazine had reported that Holmes was so inspired by her daughter Suri, 2, that she had decided to team up with Giorgio Armani for a line for kids. However, her rep has now shot down the claims, stating that the reports are not true. Katie Holmes is not designing a childrens clothing line for Giorgio Armani, Usmagazine.com quoted the rep, as saying. However, that doesnt mean that Armani would mind teaming up with Holmes. The designer, who created the actress wedding dress, has previously said: Everything about Katie Holmes is refreshing.


text:
MIAMI (AP) Ryan Howard's unfamiliar with the Hall of Famer he surpassed by reaching a home run milestone Thursday night.
The Philadelphia Phillies slugger hit his 200th home run, achieving the total in fewer games than any player in major league history. Ralph Kiner, who played in the 1940s and '50s, previously held the record.
What does Howard know about Kiner?
"Uh, he's the guy whose record I broke," Howard said with a smile. "Not to be disrespectful or anything, but he was before my time."
Howard achieved the feat in his 658th game, hitting his 23rd homer of the season in the sixth inning of a 4-0 win over Florida. Kiner hit No. 200 in his 706th game.
"It's a nice feat," Howard said. "It's a nice record to have. I'll take it and run with it."
Howard is the eighth Phillies player to hit 200 homers.
Copyright 2009 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.
Guidelines: You share in the USA TODAY community, so please keep your comments smart and civil. Don't attack other readers personally, and keep your language decent. Use the "Report Abuse" button to make a difference. Read more.


text:
|2012-07-21, 09:51
|Link #34445
Titans Test Team Pilot
Join Date: Feb 2008
Location: The Elysium
Should prolly post it come Valentines next year...
|2012-07-21, 10:02
|Link #34446
I am no one
Join Date: Apr 2010
Location: Inside your head
@ Demi
I don't really know, honest. The closest character currently I write to fit that is Saruyama, which is annoying as hell but overall a decent and loved character. However, he is a sidekick, so such reception is expected.
@ Stratos
Fr some reason, I'm not keen writing about 'special day' chapter as separate, only if it's related to plot.
|2012-07-21, 10:31
|Link #34449
I am no one
Join Date: Apr 2010
Location: Inside your head
Its really depend on the purpose of flying at that speed, look how Cecilia casually talk with Ichika. As Eras put it, even talk about weather is appropriate if you're not in emergency. if you are, then just talk about on topic things.
|2012-07-21, 10:36
|Link #34450
That one guy
Join Date: Nov 2011
It's during the race. Ichika just barely kept himself from smashing into the sidelines thanks to the intervention of Charlotte. Charlotte Makes small talk as she comes up beside him. Ichika does a doubletake at her presence. She goes angel of death at his "almost unhappy" reception of her. Ichika somehow notices and amends quickly. Charlotte cools down to normal. Then that's where I left off.
|2012-07-21, 10:39
|Link #34451
I am no one
Join Date: Apr 2010
Location: Inside your head
Just continue with them talking little about the race condition, then something else. For example, what if they work together to get rid of other rivals. Ichika is frightened, but he agreed so long as it won't be too dangerous.
Relationship EXP + 3?
|2012-07-21, 11:13
|Link #34454
That one guy
Join Date: Nov 2011
No... I can make Ichika likable to me. But that makes him run parallel to Gentarou in so many ways that I don't want. Gentarou is the hot blooded hero. Ichika is supposed to be the rational one. I want to portray him as someone who grows into confidence. But sometimes I feel my portrayal pushes him into recklessness @_@
also, if someone dominates a personal IS pilot with only a regular Uchigane by taking advantage of the rules. Would that be bad-ass normal?
|2012-07-21, 11:28
|Link #34455
I am no one
Join Date: Apr 2010
Location: Inside your head
Then that makes you like Gentarou archetype more. This normal BTW, I like Rito more than I like Ichika. Though both of them has been changed form canon portrayal, Ichika is downright OOC sometimes.
All I said, never sacrifice his role because you can't just ditch the protagonist. You can change him as you wish. (I regularly omit major character besides the protagonist though)
Yes, it's downright insane, considering their portrayal in canon, default configuration Uchigane is a piece of shit for a pro pilot (shown in my fic as well). You can adjust their parameter but you can only go so far before the need of parts customization. The most important would be spare armaments.
|2012-07-21, 12:04
|Link #34456
That one guy
Join Date: Nov 2011
I'm trying to do something like improving Ichika's attittude about his IS rather than improving his IS. I'm sure Izuru is trying to steer away from the standard archetype of how the machine is a blitz oriented fighter but... that's about all I can see it as.
Spoiler for to not be a bother:
opinion please XD
|2012-07-21, 18:36
|Link #34457
I am no one
Join Date: Apr 2010
Location: Inside your head
Ichika, FAIL.
Best freudian slip ever.
In TLR Stratos, Ichika is going so far as rehauling the machine (incomplete ByakuShiki) himself before Tabane can get a hand on it. It become a lightning bruiser close combat unit, at the cost of endurance (due to compact design = smaller battery). Simply put, it never had Yukihira or Reiryaku Byakuya.
|fanfic ideas, fanfiction, ff.net, harem, is fanfic, warning fanfic spoilers, wincest
|Thread Tools


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I was totally in the mood this weekend to spend time in my kitchen, as you already know from my post about Cherry Crisp. [2]
I was craving a healthy-ish dip for pretzels we happened to have (leftover party food [3]- I don't usually buy them because they're so dry), and hoping to find a recipe for something beyond honey mustard.
I found this Pretzel Mustard Dip recipe on allrecipes.com [4], and I found it to be perfect. I even adjusted it a bit to make it even healthier, and it's really good. We even tried it on hot dogs and it was perfect.
Source: tasteofhome.com [6] via Meg [7] on Pinterest [8]
Pretzel Mustard Dip
1/4 cup mayonnaise (I used low-fat)
1/4 cup prepared yellow or Dijon mustard (I actually used a bit more mustard than this, and a little less mayo. It makes the dip sharper, but that's OK for us, as Rob loves mustard)
2 tablespoons finely chopped green onion (you can use regular onion but I used green onion because it's milder)
1 tablespoon ranch salad dressing mix
2 1/4 teaspoons prepared horseradish
In a large bowl, combine mayonnaise, mustard, onion, salad dressing mix and horseradish. Cover and refrigerate for at least 30 minutes. Serve with pretzels.
Even though this is an easy recipe, it prompted me to find another for homemade ranch dressing mix. I don't buy the packaged ranch dip mix because it somehow contains buttermilk and I'm lactose intolerant. Also, salt and MSG are the first two ingredients.
Luckily, I found this quick, salt-free recipe for ranch dip mix [9].
I didn't change it at all, so the entire credit for this recipe goes to Budget Savy Diva. You can find her if you click the recipe link above.
One-Minute Homemade Ranch Seasoning Mix Recipe
2 tablespoons dried parsley
1 teaspoon dried dill
1 teaspoon garlic powder
1 teaspoon onion flakes
1/2 teaspoon dried basil
1/2 teaspoon pepper
Place all ingredients in a bowl. Mix. Store in an airtight continer.
To make ranch dressing, add a tablespoon of this mix to 1/3 cup mayonnaise or nonfat Greek yogurt, 1/4 cup of milk and salt to taste.
To make ranch dip, add 1 tablespoons mix to to 1 1/2 cups of sour cream. You can adjust the amount of mix you add to taste, and add salt to taste, as well.
No chocolate and pretzels? I've had some chocolate chips recently without lactose that made me really happy. Lactose is everywhere. And I've been looking for a ranch dressing mix. I'll try it with the new lactose-free yogurt, vanilla flavor since they don't have a plain, and add a tablespoon or two to balance the sweetness out. Good timing.


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text: the lawman - Unregistered September 27, 2009 9:20 AM Some of you guys need help: Buckland, Quentin snap out of it. Someone mentions Hamilton's name or shows they're not in favor of Hamilton and some you you get right out of shape...I'm surprised your moms let you stay up and watch the damn race...fairdinkum. 8 agree 0 disagree


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Could anyone write a routine that would convert a string to a list and vice-versa? I would like it to use 1 as A and 2 as B and so on…
Text to numbers
Text to numbers
mrmprog 08 Dec 2010 01:22
Unfold Text to numbers by mrmprog, 08 Dec 2010 01:22
Re: Text to numbers
Timothy Foster 08 Dec 2010 04:05
Might any of these routines help?
For turning a string into a number, you can use the expr( command.
Timothy Foster - @tfAuroratide
Auroratide.com - Go here if you're nerdy like me
Unfold Re: Text to numbers by Timothy Foster, 08 Dec 2010 04:05
Re: Text to numbers
Xeda Elnara 08 Dec 2010 04:13
Hey! Would you like an assembly program/opcode? I have made a program that does something very similar and would only need 4 bytes changed to make it work. Who knows, the original you might find better.
(I am looking for it…)
Found it:
The opcode is found in the Source.txt document.
47%? Take a look and try to imagine how cool 100% will be. This has won zContest 2011 and made news on TICalc. This compromise between Assembly and BASIC parses like BASIC and is fast like assembly. Grammer 2
Last edited on 08 Dec 2010 04:18 by Xeda Elnara Show more
Unfold Re: Text to numbers by Xeda Elnara, 08 Dec 2010 04:13
Re: Text to numbers
mrmprog 11 Dec 2010 01:52
O.K. thanks, both of those look good.
Unfold Re: Text to numbers by mrmprog, 11 Dec 2010 01:52
Re: Text to numbers
Adeadbody 15 Dec 2010 00:28
Unfold Re: Text to numbers by Adeadbody, 15 Dec 2010 00:28


text:
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You'll be entertained in unforgettable and unusual ways at the Louis Braille School Benefit Auction on Saturday afternoon, March 21st, from 1:00 to 3:00 pm, in the South County Senior Center along the waterfront in downtown Edmonds at 220 Railroad Avenue.
The SeaSharps, one of Seattle's finest barbershop quartets, will sing tight harmonies and beautiful melodies that appeal to all ages.
We're bringing a hand-cranked ice cream maker to the auction for you to take a turn at creating a fresh dessert and eating it.
Puppies in training for Guide Dogs for the Blind will put on a demonstration. Pet them and watch how they begin to get a foundation as working dogs.
Inhale the fragrance of tomatoes, olive oil, and pizza crust topped with delicacies, both of the meat and vegetarian kinds, from Pagliacci Pizza. And try what many top food critics consider America's finest gourmet sodas, supplied by the Dry Soda Company in downtown Seattle, made from the finest natural fruits, herbs, and flowers.
Enjoy all the activities while supporting the work of the Louis Braille School. It's a wonderful arrangement when your money goes to the education of blind and partially sighted children while you can purchase a gift or practical item for yourself or a loved one.
With your chance to obtain something you want or need at a better-than-usual price, our auction items are especially attractive in the current economy.
If you'd like to get away for a trip, we have lodgings at the Inn at Cannon Beach (Oregon), weekend admission and camping passes for the Darrington Bluegrass Festival (in July), and a ride on the Chehalis Steam Engine Train.
Restaurant gift certificates for American, Greek, Japanese, French, health food, Thai, and other cuisines.
Premium compost and mulch, delivered to your home. Those aren't the only delivery opportunities. How about hormone-free, home-delivered milk, cheese, and eggs?
Artworks. Cultural events. Clothing. Beauty and body care products. Jams and jellies. Husky football tickets. And a ride on an Edmonds fire truck.
Join us. You won't forget the entertainment, you'll help make homemade ice cream, pet puppies, and take home something you're happy with. And you'll know you're supporting good, important work that makes a difference in the lives of blind and partially sighted kids.
Visit Auction Donations Online to preview items available for bidding. Questions? Call Eric 425 778-2384.


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The specialty pharmaceuticals universe encompasses a variety of technologies, among them innovative drug delivery systems and unique pain management and orphan disease treatments. In this interview with The Life Sciences Report, Scott Henry of ROTH Capital Markets touts the value of "sound business models" in the specialty pharma space, and names companies with interesting prospects and/or stock-moving catalysts on the horizon.
The Life Sciences Report: Scott, is right now a good time to be investing in biotech?
Scott Henry: It depends on what you invest in. Picking sentiment is a fool's game, in my opinion. Rather, I try to find good companies with sound business models at reasonable prices. This sector is highly volatile and investors have to develop intrinsic valuation models for companies. That way they are best positioned to take advantage of volatility, as opposed to being a victim of it.
TLSR: Let's talk about some of the biotech sectors in your coverage universe. What do you see going on in inhalable medicines?
SH: I look at the inhalable category on a product-by-product basis, because one must demonstrate material incremental advantages to warrant the increased premium pricing over cheaper alternatives. One company I cover that I believe demonstrates these advantages is Alexza Pharmaceuticals Inc. (NASDAQ:ALXA).
Alexza has a soon-to-be-launched product in Adasuve [Staccato loxapine], for agitation in patients with bipolar disorder and schizophrenia. The product is approved in the U.S. and Europe and the company has marketing partners lined up-Teva Pharmaceutical Industries Ltd. (NYSE:TEVA) in the U.S. and Grupo Ferrer Internacional S.A. in Europe. Adasuve is an easily inhalable version of the antipsychotic loxapine, and could replace slow-acting pills or painful needles. I would take an inhaler over a needle any day.
For investors, I believe that Alexza offers opportunities because expectations are so low. The market cap of the company is ~$90 million [$90M]. By comparison, Teva paid $40M alone for rights to market Adasuve in the U.S., and Teva will supplement this payment with ongoing royalties. My target price on Alexza is $10/share, which is based on a sum-of-the-parts valuation, including $5.50/share for U.S. Adasuve royalties, and $2/share for ex-U.S. rights to Adasuve.
TLSR: Let's look at pain management and oncology support. What products are in the pipelines in that space? Any recent trial results that investors should take a peek at?
SH: Again, I like to find specific investment opportunities as opposed to focusing on a category. That said, pain management is a sector that I know well.
TLSR: One of my relatives recently had a terrible case of shingles. Are there any new treatment products for this condition coming our way? And do companies in this space have other products that treat central nervous system disorders and inflammations?
SH: First, I am sorry to hear about your relative, but perhaps we can find him or her some good investments to help relieve the pain. Depomed Inc. (NASDAQ:DEPO) may be one of those. The company has a high-reward, low-risk grouping of assets. The shares have performed well this year, as this proposition has become better known to investors. Shares now approach $7.50/share-up 50% from the lows. Our price target is $9/share.
The company just added ~$240M to its balance sheet by converting assets into cash. We believe that putting this cash to work could drive upside to our price target. Importantly, management has been shrewd with cash lately-including the recent purchase of the approved pain treatment Lazanda [fentanyl nasal spray]. Also, the company's shingles drug, Gralise [gabapentine], continues to grow inline and ahead of our targets. Depomed could be an emerging specialty pharma company looking to make the shift to the elite world of profitable healthcare companies. The risk here is the redeployment of cash into assets that produce high returns. We believe the current management track record could bode well for investors.
TLSR: Let's look at generic topicals. Any picks in this space?
SH: Generic topicals have been a very favorable environment, because the space is under the radar for most big players. There are also clinical barriers to entry, such that supply tends to be in check with demand. In the past few years, this has led to strong upward pricing power, which has raised all boats higher.
One of my favorite names in this space is IGI Laboratories Inc. (IG). This small, generic company has many potential positive inflection points.
First, the company could turn earnings-per-share [EPS] profitable this year, which tends to be a good time to buy generic stocks. Second, the company has 13 proprietary abbreviated new drug applications [ANDAs] at the FDA. The first proprietary approval could come in the next six months. That could validate the whole business model and send shares significantly higher.
Our valuation on IGI Labs is $3.50/share, relative to the current price of ~$2. We base this valuation on a 20x multiple of forecasted 2016 EPS. The risk here is that the approvals fail to materialize.
TLSR: What about molecular diagnostics? I understand you have a target price on PDI Inc. (PDII), which has recently entered in to a collaboration agreement. Can you talk about the prospects of this firm?
SH: PDI is a really neat story; it just expanded into molecular diagnostics. The company is a contract sales organization [CSO], which means it has a "sales force for hire" for other pharmaceutical companies. That's been a tough business model the past few years, but we expect it to improve the next two years. That alone makes PDI stock interesting to us, given that it trades just below $5, with ~$3.50/share in net cash.
The kicker is that PDI is putting small amounts of cash to work that could have large payouts. This includes two recent asset acquisitions for small up-front payments. The company can market these products with its excess CSO capacity such that it mitigates the marketing risk. The payouts from these recent deals could start to be apparent in H2/14. We think now may be the time to invest.
Our price target on PDI shares is $9, which is derived from cash [~$3.50/share] and the base business [~$5/share based on 20x EPS potential in 2015].
TLSR: Do economic stresses in the international market [including North America] impact the prospects of the firms we have talked about today?
SH: Fundamentally, economic circumstances do not reduce demand, as people always get sick. Economic stress may hurt healthcare coverage and reimbursement, but not in a material way. The one thing that does come into play is financing. In a low-risk investment environment, investors will demand a higher premium to invest in money-losing companies. Companies with greater financing flexibility will tend to outperform in tougher economic times.
TLSR: Thanks for your time, Scott.
SH: Thank you.
This interview was conducted by Peter Byrne of The Life Sciences Report and can be read in its entirety here.
Scott Henry is the head of healthcare research and a senior research analyst with ROTH Capital Partners, covering the specialty pharmaceutical sector. He came to ROTH Capital from OppenheimerFunds, where he led coverage of the U.S. pharmaceutical sector. He has also been affiliated with Thomas Weisel Partners, ABN AMRO and Leerink Swann & Co. His coverage universe has included specialty pharmaceuticals, large-cap pharmaceuticals, biotech and selective medical device names. He has 12 years of sellside experience. Henry was ranked #2 in The Wall Street Journal's "Best on the Street" stock-picking survey in 2010, #3 on the Forbes/Zacks Investment Research "Best Analysts" list for the Drugs category in 2010 and #3 in the pharmaceutical sector by Forbes.com/StarMine for earnings estimate accuracy in 2007. His investment views have been cited in The Wall Street Journal and The New York Times and he has made frequent appearances on CNBC, CBS Marketwatch, and Bloomberg. Henry attended the University of Rhode Island and received a master's degree in business administration with distinction from Cornell University.
1) Peter Byrme conducted this interview for The Life Sciences Report and provides services to The Life Sciences Report as an independent contractor. He or his family own shares of the following companies mentioned in this interview: None.
2) The following companies mentioned in the interview are sponsors of Streetwise Reports: None. Streetwise Reports does not accept stock in exchange for its services.
3) Scott Henry: I own, or my family owns, shares of the following companies mentioned in this interview: Alexza Pharmaceuticals Inc., Depomed Inc., PDI, Inc., IGI Laboratories Inc. I personally am, or my family is, paid by the following companies mentioned in this interview: None. My company has a financial relationship with the following companies mentioned in this interview: None. I was not paid by Streetwise Reports for participating in this interview. Comments and opinions expressed are my own comments and opinions. I had the opportunity to review the interview for accuracy as of the date of the interview and am responsible for the content of the interview.
4) Interviews are edited for clarity. Streetwise Reports does not make editorial comments or change experts' statements without their consent.
5) The interview does not constitute investment advice. Each reader is encouraged to consult with his or her individual financial professional and any action a reader takes as a result of information presented here is his or her own responsibility. By opening this page, each reader accepts and agrees to Streetwise Reports' terms of use and full legal disclaimer.
6) From time to time, Streetwise Reports LLC and its directors, officers, employees or members of their families, as well as persons interviewed for articles and interviews on the site, may have a long or short position in securities mentioned and may make purchases and/or sales of those securities in the open market or otherwise.
Disclosure: I have no positions in any stocks mentioned, and no plans to initiate any positions within the next 72 hours. I wrote this article myself, and it expresses my own opinions. I am not receiving compensation for it. I have no business relationship with any company whose stock is mentioned in this article.


text:
Mb Motor Group Bradford Vans
Search and browse 0 cars currently available from Mb Motor Group Bradford Vans, a car dealer in Bradford. You can also read reviews of this dealer from customers who have bought cars there, or contact the dealer directly to discuss what you're looking for.
Call dealer nowTel: (01274) 506749
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- 30 day exchange plan
- Minimum 2 year vehicle warranty
- 2 year roadside assistance
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About this dealer
Mb Motor Group Bradford Vans
"We pride ourselves on supplying best quality cars on market at unbeatable prices to you the customer. A family run business with over 50 years motor trade experience. We have 2 large showrooms housing our extensive stock including brand new Mitsubishi's, we will not be beaten on price! "
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text:
Science Department contact information
The Science curriculum enables students to gain scientific knowledge by theory, investigation and experimentation. The laboratory component provides students with the opportunity to link concepts to hands-on experience and develop a deeper awareness of the investigative nature of science.
Course Offerings: • Biology • Honors Biology • Chemistry • Honors Chemistry • Physics • AP Physics I • Human Genetics • Experimental Genetics • Anatomy and Physiology I • Anatomy and Physiology II • AP Biology • AP Chemistry • AP Environmental Science • AP Physics C • Vertebrate Zoology • Invertebrate Zoology
Bishop Eustace Preparatory School
5552 Route 70, Pennsauken, NJ 08109-4798
Phone: 856.662.2160


text:
August 1, 2008, Pantaray Research announces SuperOrca MSI editor
Use this security logo:
Freeware is computer software that is available for use at no cost or for an optional fee.
Tutorial not found. Let know us about any useful video tutorial.
orca, orca msi editor, microsoft installer, windows installer, msi database, mergemodule, validate msi, msi validation, installation, uninstall, uninstallation, installshield, development tools
First Release
You may include this program on any CD compilation.
"SuperOrca" is a freeeware program.
You may use this software for any length of time.
You may not alter this software in any way, including changing help and text files.
"SuperOrca" IS DISTRIBUTED "AS IS". NO WARRANTY OF ANY KIND IS EXPRESSED OR IMPLIED. YOU USE IT AT YOUR OWN RISK. "Pantaray Software Systems" WILL NOT BE LIABLE FOR DATA LOSS, DAMAGES, LOSS OF PROFITS OR ANY OTHER KIND OF LOSS WHILE USING OR MISUSING THIS SOFTWARE.
You are hereby licensed to make any number of backup copies of this shareware and documentation. You can give the copy of the software to anyone or distribute the freeware version of the software providing that no fee is charged and the package files are not modified in any way.
All copyrights to "SuperOrca" are exclusively owned by "Pantaray Research Ltd.".
Installing and using "SuperOrca" means the acceptance of these terms and conditions of the license. If you do not agree with the terms of this license you must remove "SuperOrca" files from your storage devices and cease to use the product.
Copyright © 2002-2008 Pantaray Resarch Ltd., All rights reserved.
Internet: http://www.pantaray.com
Email: firstname.lastname@example.com


text:
It only takes a minute to get started. Fill out our uncomplicated sign up form and login to the site.
This step is elective but will dramatically multiply your chances of getting interest from other over 50s members on the site. Let people know a bit about yourself, your pasttimes and attach a nice photograph of yourself.
Use our website's advanced search to uncover over 50s members who match your needs. Or just look through listings and uncover folks that arouse your interest.
Found someone who ticks all the boxes? Send them a word or two or just a wink and see if they're interested. With a little luck you'll be on your way to a lifetime of happiness.
It only takes a few clicks


text:
Robie Robe - Purple
Product Details
Don't freeze your boots off getting changed this winter!
We all know what a pain getting changed in winter is. Rain, sleet, snow, wind-chill, towels blowing off, hands not working. Well, forget about all that, because St Ives surfers Scrappy and Martin (Tarps) Tarpley have come up with these heavyweight changing robes. They're hooded, really thick and having stood next to them freezing my arse off down at 'Leven last January while they laughed at me, I can tell you they work a treat!


text:
NXP Semiconductors NV, NXPI is in the exchange NASDAQ and its industry is Semiconductor – Broad Line in the sector of Technology. Based in Netherlands, NXP Semiconductors NV, NXPI has a market cap of 26459.05. Since its IPO date on the 8/6/2010, NXP Semiconductors NV, NXPI performance year to date is -6.05%. Today NXP Semiconductors NV, NXPI has gained 1.71%, with a current price of 80.5.
Ownership of the company is 86.40% for insider ownership while institutional ownership is 91.30%. The management of the company have seen the company have a payout ratio of 0.00%. Return of assets are at 7.20%, with return on investment at 10.20%.
In terms of debt levels and profit levels, NXP Semiconductors NV, NXPI is seeing a long-term debt/equity of 0.74. While Total debt/equity is 0.8. With a profit margin of 18.00%, this is combined with a gross margin of 39.30%, and operating margin of 18.20%. NXP Semiconductors NV ability to meet debt levels, with a current ratio of 1.6, while the quick ratio is 1.
For the last year NXP Semiconductors NV, NXPI has seen a EPS growth of 181.20%. A performance for the year of -12.55%. The 52-week high is -19.71%, and the 52-week low is 30.66%. The average volume for NXP Semiconductors NV, NXPI is 2964.
With a target price of 107.86, can NXP Semiconductors NV, NXPI reach this target? Looking at the value indicators of NXP Semiconductors NV, NXPI. NXP Semiconductors NV has a P/E of 15.41 and a forward P/E of 10.63. Perhaps the more useful indicator than P/E, is PEG which has a value of 0.45. NXP Semiconductors NV also has a P/S and a P/B of 3.86 and 2.42 respectively. For P/cash, NXP Semiconductors NV has a value of 17.78, while it is *TBA for P/free cash flow.
At the current price of 80.5, NXP Semiconductors NV has a dividend yield of *TBA. We see a return on equity of 20.40%.
Looking more long-term NXP Semiconductors NV, is projected to get an EPS growth for the next five years of 34.00%. In the short-term an EPS growth of 30.56% in the next year is forecasted. This is after a EPS growth of 181.20% for this year and for the last five years a 36.40% growth has been seen.


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Pleasure SpotFavorites [+]
Pay-Per-Minute Viewing
You can now enjoy this movie on your iPhone®, iPad®, or compatible Smart TV & Android devices.
Purchase Options
Pay-Per-MinutePurchase Minutes
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Available MP4 sizes:SD 480p (1.01 GB)
DRM-free downloads offer content that is not limited by standard Digital Rights Management (DRM) access technologies. These technologies allow hardware manufacturers, publishers, copyright holders and individuals to limit the usage of digital content.
When you purchase DRM-free downloadable content, that content is available for you to download on any computer and save to a disk whenever you like. You can also download to multiple computers. There is no timeframe associated with your purchase and the content will not expire.
Description: Pleasure Spot...everybody scores when they hit it! Pleasure Spot is a story of a woman who never knew what she was not getting out of sex until a wacky transplant operation gave her a whole new attitude! Do you have an organ to donate?
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text:
Sewer manholes are positioned at regular intervals (approximately every 100 metres), or wherever there is a change in grade, direction or materials in the sewerage system. They allow us to:
It is important that manholes are kept free of any obstructions.
Due to movements in the soil around manholes, we sometimes need to raise or lower a manhole to keep it level with the ground.If the manhole is on your property, it is your responsibility to maintain the area around it. To have the height of a manhole adjusted, please complete an Water and Sewerage Supply Services Private Works Application (PDF).
Please note: In line with the Water Supply (Safety and Reliability) Act 2008, it is an offence to interfere with a public sewer or manhole, and penalties may apply.
Approximately one pool in every 20 has a leak! Even a minor leak can result in a major water loss. Under pressure a tiny leak can lose more than 3000 litres a day.


text:
|First Seen In
|Born to Run
|Last Seen In
|The Incident
|Owned By
Kate Austen owned a New Kids On The Block themed lunchbox which she shoplifted when she was young.
A New Kids On The Block lunchbox is shoplifted by young Kate Austen and her friend Tom Brennan. The shop owner catches them and recognizes kate as Diane Austen's daughter. When he threatenes her of calling her mother, Jacob appeares and offeres to pay for the lunchbox.
He hands over the lunchbox to kate and makes her promise that she will never steal again. Jacob blesses her by tapping on her nose telling "Be good Katie".
Time CapsuleEditKate and Tom make a Time Capsule using the lunchbox and burries it underneath an old oak tree.
Years later, Kate approaches Tom at the hospital that he works, and asks him to help her meet his mother who is believed to be dying, in the same hospital. Later at Tom's house, Kate mentions the Time Capsule to him. Tom agrees to retrieve the box after kate explains that it might be the last time they could retreive the box. Kate and Tom arrives at the oak tree that they burried the lunchbox underneath, at night, and retrieves the box together.
They find a red cap, tom's Toy Airplane, and a cassette inside the box. Tom is happy and surprised to see his Toy Airplane.
The CassetteEdit
Tom and Kate listen to the audio cassette together,which contains a conversation between young Kate and Tom.
YOUNG TOM: Is it on? I don't think it's on.
YOUNG KATE: It's on.
YOUNG TOM: Okay, this is Kate Austen and Tom Brennan and this is our dedication for our time capsule, here on August 15th, 1989. Hey, give me that back.
YOUNG KATE: Why are you putting this stupid plane in there?
YOUNG TOM: Because it's cool, Katie. I got it when I flew to Dallas by myself.
YOUNG KATE: Ooh, that is cool, just like this time capsule.
YOUNG TOM: It'll be totally cool when we dig it up in like 20 years.
YOUNG KATE: How do you know we'll be together?
YOUNG TOM: Because we'll be married and you'll be a mom and we'll have 9 kids.
YOUNG KATE: I don't think so. As soon as I get my license we should just get in a car and drive. You know, run away.
YOUNG TOM: You always want to run away, Katie.
YOUNG KATE: Yeah, and you know why.
The audio tape reminds Kate and Tom of their old times and the two end up making love.
- New Kids on the Block is an American boy band that enjoyed success in the late 1980s and early 1990s


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Davy Hite has a special affinity for Louisiana. He won the Classic on the Delta in 1999. And not too long after that, he won a Bassmaster event here on the Red River.
Like every one else he also wants to win this event. He has plans on winning a second Classic this year or the next. His motivation is simple. He explains it in this video clip:


text:
Israelis Do the Wave After Windsurfer Earns Country's First Olympic Gold Medal
Television and radio stations in Israel cut away from their mid-day programming. News Web sites were updated faster than even the nimblest of fingers could press "refresh." It wasn't another terrorist attack in Israel, but some good news for a change: On Wednesday, the Jewish state entered the fraternity of Olympic gold-medal winners. "I felt… More ▸


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Family in West Chester
mainly Saturdays during day, looking for someone to take care of well behaved 6 year old twins. own a catering company I have many meetings during day, need a responsible fun sitter, the girls are well behaved and easy to deal with, they do get bored easily, so need someone that will keep them entertained with either learning activities or simple fun
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Expression of matrix metalloproteinase-12 in aortic dissection
Hypertension and peripheral vascular disease
Aortic dissection(AD) is an acute process of large blood vessels characterized by dangerous pathogenic conditions and high disability and high mortality. The pathogenesis of AD remains debated. Matrix metalloproteinase-12 (MMP-12) participates in many pathological processes such as abdominal aortic aneurysm, atherosclerosis, emphysema and cancer. However, this elastase has rarely been assessed in the presence of AD. The aim of the present study was to investigate the expression of MMP-12 in aortic tissue so as to offer a better understanding of the possible mechanisms of AD.
The protein expression levels of MMP-12 were analyzed and compared in aorta tissue and the blood serum samples by reverse transcription polymerase chain reaction(RT-PCR), Western blotting, immuno-histochemistry, fluorescence resonance energy transfer ( FRET ) activity assay and enzyme-linked immuno sorbent assay ( ELISA ), respectively. Ascending aorta tissue specimens were obtained from 12 patients with an acute Stanford A-dissection at the time of aortic replacement, and from 4 patients with coronary artery disease (CAD) undergoing coronary artery bypass surgery. Meanwhile, serum samples were harvested from 15 patients with an acute Stanford A-dissection and 10 healthy individuals who served as the control group.
MMP-12 activity could be detected in both AD and CAD groups, but the level in the AD group was higher than those in the CAD group (P < 0.05). MMP-12 proteolysis existed in both serum samples of the AD and healthy groups, and the activity level in the AD group was clearly higher than in the healthy group (P < 0.05). For AD patients, MMP-12 activity in serum was higher than in the aorta wall (P < 0.05). MMP-12 activity in the aortic wall tissue can be inhibited by MMP inhibitor v (P < 0.05).
The present study directly demonstrates that MMP-12 proteolytic activity exists within the aorta specimens and blood samples from aortic dissection patients. MMP-12 might be of potential relevance as a clinically diagnostic tool and therapeutic target in vascular injury and repair.
Acute aortic dissection(AD) is the most common aortic catastrophe. The overall in-hospital mortality rate for acute aortic dissection in China is 17.7%, with most of the deaths occurring during the first week
Considering that MMPs play a central role in matrix degradation and aneurysm progression, it seems that these proteinases constitute attractive targets of drug treatment. Over the past few decades , selective inhibitors of MMPs have been developed, but in most cases these compounds have not met the promise and expectations in clinical trials
MMP-12 (matrix metalloproteinase12), also known as macrophage metalloelastase, was first identified as an elastolytic metalloproteinase secreted by inflammatory macrophages 30 years ago
Although there are substantial numbers of reports on the localization of several MMPs in aortic aneurysms, little is known about the roles of MMPs in the pathogenesis of AD. In this study, using immunohistochemical and biochemical methods, the expression of MMP-12 and its roles in AD was examined in the aorta and blood serum samples of patients with AD. The results present new evidence that MMP-12 is involved in the development of AD.
The institutional research ethics committee of Kunming Medical University approved the study, and written informed consent was obtained from each subject. This study was conducted in accordance with the ethical requirements.
Intraoperative aortic samples were obtained from 12 patients with Stanford Type A acute aortic dissection as experimental group. These patients received Bentall operation in the period from April 2010 to June 2011 at the No. 1 Hospital Affiliated to Kunming Medical University. 9 were male and 3 were female (mean age, 44.7 years); 8 patients (66.67%) had a history of hypertension and 7 patients (58.33%) used to smoke. All of them had a color Doppler ultrasonic examination and computed tomography and all were diagnosed as acute aortic dissection Stanford A. Control aortic tissues were obtained from 4 cases of CAD without aorta wall damage who underwent coronary artery bypass surgery; samples were taken by punching a hole on the aortic wall tissue with a hole punch. All patients with CAD were male, aged 65.5±3.3; 2 patients had a history of hypertension and 3 patients had a history of smoking. Both groups were ruled out of the possibility of inflammation of the chest cavity, Marfan syndrome, bicuspid aortic valve abnormalities, cardiomyopathy, chronic pulmonary heart disease and infectious endocarditis possibilities.
Serum samples were obtained from 15 patients with Stanford type A acute aortic dissection as the experimental group. All patients were hospitalized in the period from April 2010 to June 2011 at No. 1 Hospital Affiliated to Kunming Medical University. Ten age-matched healthy people were included as a control group.
Recombinant human MMP-12 (GenBank: AAI43774.1) was obtained in our laboratory. Briefly, the prokaryotic plasmid pET -17×b - MMP-12 was constructed and transformed into E. coli strain BL21 (DE3). The recombinant protein expressed in
Figure 1
The domain structures of human MMP-12
The domain structures of human MMP-12. The latent form of human MMP-12(top), the active form of MMP-12 with molecular weight 45 KDa (middle), and the catalytic domain of MMP-12 with molecular weight 22 KDa (bottom) are illustrated. The precise amino acid residues are known from protein sequencing.
Reverse transcription-polymerase chain reaction
Aortic wall tissues (400 mg) was ground to a fine powder using a mortar and pestle in liquid nitrogen, to which a certain amount of Trizol (Gibco Brl, Rockville MD, USA) was added to extract RNA according to the manufacturer's instructions. Approximately 1.5 μg of total RNA from each sample was used to conduct reverse transcription reaction in a 50 μl volume using the RNA PCR KitVer.3.0 (Takara Biotechnology, Dalian, China). The reaction mixture was incubated at 42°C for 2 h and the reaction was terminated by heating to 99°C for 5 min. The synthesized cDNA was used for PCR amplification or stored at −80°C for further analysis.
PCR primers (GenScript, Nanjing, China) were designed to amplify MMP-12 cDNA. The forward primer 5'-CGATGAGGACGAATTCTGGACTAC-3' is situated in the exon 4, and the reverse primer 5'-GGTTCTGAATTGTCAGGATTTGGC-3' is situated in the exon 6. The primer sequences correspond to residues Asp211 to Pro292 in the catalytic domain of human MMP-12. The PCR reaction was performed in a 50 μl volume containing 0.5 mM of each primer, 5 μl PCR buffer, 0.5U Ex-Taq DNA polymerase (Takara Biotechnology, Dalian, China). Reaction conditions included initial denaturation at 94°C for 2 min, followed by 35 cycles at 94°C for 1 min, annealing at 55°C for 1 min and extension at 72°C for 1 min, followed by a 10 min final extension at 72°C. PCR products were separated on 1.0% agarose gels and visualized by Gelview (Bioteke,Beijing,China) staining.
The quality of the total RNA was determined by RT-PCR for the house-keeping gene GAPDH (glyceraldehyde-3-phosphate dehydrogenase). The primer sequences were as follows: GAPDH sense, 5'-CCCATCACCATCTTCCAGGAGCG-3'; anti-sense, 5'-GGCAGGGATGATGTTCTGGAGAGCC-3' (GenScript, Nanjing, China). The PCR reaction contained 10.0 μl of cDNA, 0.5 μM of each primer, 5 μl PCR buffer, 0.5 U Ex-Taq DNA polymerase in a final volume of 50 μl using the following conditions: 95°C for 5 min, 34 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min, with a 10 min final extension at 72°C.
Immunohistochemical staining
Aorta wall tissues (4 cm)were obtained at the aortic dissection site, The blood was washed from the newly obtained sample with 0.9% sodium chloride, dipped the sample into 10% Formalin solution for 24 h for serial sections of 4 μm thick, then, stained with hematoxylin for observation. The antibody for MMP-12 (Epitomics, Inc, CA, USA) was a rabbit monoclonal antibody generated by immunizing rabbit with the synthetic peptide corresponding to residues on the C-terminus of human MMP-12; After incubating with Biotin-conjugated goat anti-rabbit IgG (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 15 min at room temperature(RT), the streptavidin-alkaline phosphatase-biotin complex solution was applied to all sections for 15 min according to the manufacturer's instructions. Positive results of immunohistochemistry indicated yellowish brown or brown particles presented in the cytoplasm. A random selection 10 fields were viewed under a 400× microscope and calculated the percentage of positive cells was calculated. Rating criteria: no positive expression scored 0 point (negative), positive expression less than 25% scored 1 point (weak positive), positive expression higher than 25% and lower than 50% scored 2 points (positive) and positive expression higher than 50% scored 3 points (strong positive).
Western blotting
Aorta wall tissue (400 mg) was obtained and ground into powder in liquid nitrogen, to which 4 ml Trizol was added and mixed, then it was centrifuged at 11,000 rpm for 15 min. The supernatant was then spun for an additional 10 min and collected. The protein (15 μl) from the tissue samples was immersed in boiled water for 15 min. The total protein was separated on 12% gels. The proteins were transferred to polyvinylidene fluoride (PVDF) membranes. Recombinant human MMP-12 was used as positive control. Briefly, after blocking for 1 h at 37°C in 10 mM Tris–HCl pH 7.5, 100 mM NaCl, triethanolamine-buffered saline (TBS) containing 3% bovine serum albumin and 0.1% Tween 20, the membrane was incubated with an antibody (rabbit anti-human MMP-12 mAb, Epitomics) at 1:1000 dilution for 2 h at RT . After repeated washes, the membrane was incubated with Biotin-conjugated goat anti- rabbit IgG (Santa Cruz, American) at 1:1000 dilution for 1 h at RT , followed by incubation with Streptavidin-alkaline phosphatase (R&D, USA) for 1 h at RT. Finally, the membrane was incubated with BCIP/NBT (Calbiochem, Bad Soden, Germany) in the darkness.
MMP-12 FRET activity assay
The activity of MMP-12 was assessed using a peptide-based FRET assay (EnzoLyte MMP-12 Assay Kit, AnaSpec, Inc, Fremont, CA,USA) in accordance with the manufacturer's protocols and fluorescence intensity was determined using a fluorescence analyzer (Fluoroskan Ascent 2.6, Thermo) at excitation/emission wavelengths of 340 nm/490 nm. Briefly , aorta wall tissues (200 mg ) in assay buffer(0.5 ml component D, AnaSpec) were ground in liquid nitrogen, centrifuged at 12,000 rpm for 10 min at 4°C and the supernatant collected and stored at −70°C until use. The blood samples(5 ml, no anticoagulant added) were centrifuged at 3000 rpm for 20 min at 4°C and the serum collected and stored at −70°C until use. Samples were incubated with APMA (4-aminophenyl- mercuric acetate, in component C, AnaSpec) at a final concentration of 1 mM in the assay buffer for 2 h at 37°C to activate MMP-12 prior to the experiment. After activating MMPs with APMA for 2 h , blood samples were incubated with 1 μM MMP Inhibitor v (Merck KGaA Frankfurter Str. 250 64293 Darmstadt, Germany), an orally active non-peptidyl hydroxamate compound that acts as an effective broad-spectrum inhibitor against MMP-2, -3,-8,-9, -12, -13, for 1h at 37°C. The assay was carried out in black 96-well microplates. Fluorescence intensities were recorded using a Fluoroskan Ascent fluorimeter at an interval of 5 min over a period of 1 h until peak fluorescence was achieved. The total volume of the assay was 100 μl assay buffer (SensoLyte TM 490 mmp-12 Assay Kit, ANA SPEC) containing various optimized sample volumes.
Enzyme-linked immuno sorbent assay
Protein samples were prepared by the methods mentioned above. An ELISA kit for MMP-12 detection is commercially available from EIAab (Wuhan EIAab Science Co.,Ltd,China). This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for MMP-12 had been pre-coated onto a microplate. Standards, samples and the antibody that is bound by horseradish peroxidase were pipetted into the wells, which were then incubate and washed. Peroxidase can catalyze the oxidation of (3,3',5,5'-tetramethylbenzidine)TMB into two colored products. The first product is a blue which, if sufficient acid exists, the blue product will be further oxidized to a yellow diimine, that is stable in acidic conditions and has a maximal absorption wavelength of 450 nm. The depth of the color is positively correlated with the concentration of MMP-12. Samples were diluted with the appropriate amounts of sample dilution buffers provided with the kits. All tests were performed in duplicate for each sample. The optical density OD was determined using a microplate reader set to 450 nm, the average OD was calculated for each set of standards.
Statistical analysis
Analysis was performed by using SPSS version20.0. Descriptive statistics analysis was done. An independent sample
Reverse transcription-polymerase chain reaction
mRNA were isolated from human aorta and then cDNA were synthesized by reverse transcription using the first strand cDNA as templates. The target DNA sequence was amplified by PCR (polymerase chain reaction) with the primers that primed amplification of an 250-base-pair (bp) segment of the MMP-12 gene and a 410-base-pair (bp) segment of the GAPDH gene. The authenticity of amplified PCR products was confirmed by nucleotide sequence analysis. MMP-12 and GAPDH mRNA had been detected by RT-PCR in ascending aorta specimen from AD and CAD patients (Figure
Figure 2
Agarose gel electrophoresis analysis of PCR fragment obtained from human aorta
Agarose gel electrophoresis analysis of PCR fragment obtained from human aorta. MMP-12 gene fragments were amplified from human aorta wall by RT-PCR. PCR amplifications were then analyzed by electrophoresis using 1% agarose gel stained with Gelview. The gel was viewed on a Tanon 3500R Gel Documentation System and recorded. (A) cDNA corresponding to MMP-12 in human aorta was PCR-amplified using primers for MMP-12. (B) cDNA corresponding to GAPDH in human aorta was PCR-amplified using primers for GAPDH. Lanes 1–2: Aorta wall of coronary heart disease patient; Lanes 3–10: AD; Lane M: DNA ladder; Lane 11: negative control.
Immunohistochemical staining
The culprit lesion was obtained from a typical patient who underwent aortic replacement. Representative micrograph of MMP-12 immunostained specimens are shown in Figure
Figure 3
Immunohistochemical staining
Immunohistochemical staining. Ascending aorta specimen in the entry site of the dissection from a AD patients were sectioned and labeled with MMP-12 antibodies as indicated. Sections were developed with alkaline phosphatase anti-alkaline phosphatase (APAAP) techniques and counterstained with hematoxylin. Total cells and positive cells were counted under the microscope. Arrows indicate examples of MMP-12 positive cells, A (magnification×200), B, C, D (magnification×400).
Western blotting
Recombinant MMP-12 with molecular weights of 45 KDa and 22 KDa could been identified by Western blotting (Figure
Figure 4
MMP-12 expression in the human aorta wall
MMP-12 expression in the human aorta wall. Lysates from tissues samples collected from AD and non-AD patients were separated on 12% SDS-PAGE gels and were analyzed by immunoblotting using an antihuman MMP-12 monoclonal antibody. Recombinant MMP-12 expressed in
Enzymatic assay of MMP-12 activity using the FRET peptide
Comparing with recombinant MMP-12 which exhibited the highest activity in 5 mM CaCl2 (Figure
Figure 5
Effects of different concentration of Ca2+ ions on MMP-12 activity
Effects of different concentration of Ca2+ ions on MMP-12 activity. The MMP-12 assay was based on the MMP-12 hydrolysis of FRET substrate ( AnaSpec, Inc, Fremont, CA,USA) .A total of 100 μl reaction mixture consisting of 1 μl (100 ng) MMP-12, different concentrations of Ca2+ ions (0-40 mM) and 50 μl of buffer ( AnaSpec, Inc, Fremont, CA,USA)were incubated at 37°C for 10 min. Fluorescence was read at Ex/Em wavelengths = 340 nm/490 nm for 60 min. RFU: Relative Fluorescence Unit.
Figure 6
Continuous assay for MMP-12
Continuous assay for MMP-12. Continuous fluorometric assay of MMP-12 in (A) aortic specimen from AD(n=12) and CAD(n=4), (B) blood serum sample from AD (n=15) and healthy(n=10) , (C) aortic specimens and sera from AD, (D) aortic specime n from AD incubated with MMP Inhibitor v. Assays were performed at RT in a total of 100 μl assay buffer containing various optimized sample volumes, by activating MMPs with 1mM APMA , inhibiting MMPs with 1 μM MMP Inhibitor v, or 100 ng of recombinant MMP-12 served as a control. Fluorescence was read at Ex/Em wavelengths = 340 nm/490 nm for 60 min. RFU: Relative Fluorescence Unit; AD: aortic dissection; CAD: coronary artery disease.
Enzyme-linked immuno sorbent assay result
MMP-12 concentrations measured by ELISA were used to assess the levels in ascending aorta specimen from 12 AD, 4 CAD, and serum samples from 15 AD and 10 healthy patients. Higher MMP-12 concentrations were detected in aorta specimen and serum samples of the AD group compared with aorta specimens of the CAD group and serum samples of healthy controls (3.7±0.9, 0.6±0.16 vs. no detectable quantity of MMP-12, 0.05±0.03ng/ml, P<0.05, P<0.05, respectively) (Figure
Figure 7
MMP-12 levels in the aortic specimens and sera. MMP-12 levels were measured using ELISA as described
MMP-12 levels in the aortic specimens and sera. MMP-12 levels were measured using ELISA as described. (A) Level of MMP −12 in the aorta wall. Cases 1–10 are the aorta tissues of AD, and Cases 11–16 are aorta tissues of CAD. (B) level of MMP-12 in sera. Cases 1–15 are the sera of AD, and cases 16–25 are sera of healthy patients.
Acute aortic dissection is the most common catastrophic aortic condition but lacks an effective pharmaceutical therapy. The aortic wall consists of 3 layers: the intima (inner layer made of endothelial cells), media (containing muscular elastic fibers), and adventitia (outer connective tissue). In the human aorta, the elastin-to-collagen ratio decreases in progressively distal regions making the aorta most flexible at proximal regions
Western blotting confirmed that the 22 kDa MMP-12 was present in the lysate of damaged aorta walls. This is, however, not surprising because MMP-12 is secreted as a pro-enzyme and undergoes self-activation through autolytic processing
Clinical and basic studies have implicated changes in MMPs activity in aortic Dissection, but the key effectors and mechanism of action is presently unknown.
Kurihara and colleagues
Accordingly, through the combined use of a fluorogenic substrate and ELISA, the present study demonstrated that a continuous fluorescent signal and the level of MMP-12 could be detected from serum samples of AD. These results indicate that an ambient level of MMP-12 proteolytic activity exists within the blood circulation. With the occurrence of AD, serum MMP-12 activity was transiently increased. Thus, this study demonstrated for the first time that MMP-12 activity exists in the serum of AD. We also identified the presence and activation of MMP-12 within the entry site of aortic dissection, so, the results of the present study set the stage for future mechanistic studies especially in MMP-12 induction and activation on the development of AD.
These data reported here demonstrated that MMP-12 activity exists in the serum and aorta wall of AD. This new information adds to our understanding of the pathophysiology of the dissection. Measurement of serum MMP-12 levels with a peptide-based FRET assay or ELISA in AD is a simple and relatively rapid laboratory test, that can be used as a biochemical indicator of aortic disease. Further studies are required in order to draw precise conclusions on the clinical significance of serum levels of MMP-12 in diagnosis and prognostic applications in the treatment of this deadly disease.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
YS and YHX contributed equally to this work. ZHM, RWM , YHX, and YS conceived and designed the study and wrote the manuscript. ZHM and YHX designed the primers and constructed MMP-12 expressive plasmid . RWM , YS, FL and QL collected tissues. RWM, YHX, FL, CZ carried out the immunoassays. CZ, RY, SB, QFY, JXW implemented purifcation and characterization of human MMP-12. FL and HBW performed reverse transcription-polymerase chain reaction. FL, CZ and RY carried out FRET assay . RWM, YHX, QL and YS participated in the experimental data collection and analysis. YS, YHX and FL performed the statistical analysis. All authors read and approved the final manuscript.
This work was supported in part by grants (30860278 and 81160025) from the National Natural Sciences Foundation of China, Leading Talent Program of Yunnan Province of China (L-201203), the grants (2011CI057 and 2006C0071M) from the Department of Science and Technology of Yunnan Province, and the grants (2007C0001R and 2012FB005)from the Joint Special Funds for the Department of Science and Technology of Yunnan Province-Kunming Medical University, the grant (ZD2010007) from the Department of Education of Yunnan Province and the grant(2011JY38)from Kunming Medical University.
Acute aortic dissection in China
Centers for Disease Control and Prevention. United States Department of Health and Human Services
Deaths: final data for 2005
American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines; American Association for Thoracic Surgery; American College of Radiology; American Stroke Association; Society of Cardiovascular Anesthesiologists; Society for Cardiovascular Angiography and Interventions; Society of Interventional Radiology; Society of Thoracic Surgeons; Society for Vascular Medicine: guidelines for the management of patients with thoracic aortic disease
Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function and biochemistry
Matrix metalloproteinases in peripheral vascular disease
Patterns of expression of fibrinolytic genes and matrix metalloproteinase-9 in dissecting aortic aneurysms
Collagen and elastin degradation by matrix metalloproteinases and tissue inhibitors of matrix metalloproteinase in aortic dissection
Up-regulation of matrix metalloproteinase-2 and membrane-type 1-matrix metalloproteinase were coupled with that of type I procollagen in granulation tissue response after the onset of aortic dissection
Matrix metalloproteinases and cardiovascular disease
Evidence for altered balance between matrix metalloproteinases and their inhibitors in human aortic diseases
Matrix metalloproteinases in vascular remodeling and atherogenesis: the good, the bad, and the ugly
Decreased tissue inhibitor of metalloproteinase-2/matrix metalloproteinase ratio in the acute phase of aortic dissection
Matrix metalloproteinase inhibitors: new challenges in the era of post broad-spectrum inhibitors
Progress in the development of matrix metalloproteinase inhibitors
Synthetic active site-directed inhibitors of metzincins: achievement and perspectives
Prolonged administration of doxycycline in patients with small asymptomatic abdominal aortic aneurysms: report of a prospective (Phase II) multicenter study
Mouse macrophage elastase. Purification and characterization as a metalloproteinase
Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages
Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase
Expression and localization of macrophage elastase (matrix metalloproteinase- 12) in abdominal aortic aneurysms
Carotid atherosclerotic plaque matrix metalloproteinase-12-positive macrophage subpopulation predicts adverse outcome after endarterectomy
MMP12, lung function, and COPD in high-risk populations
T: Localization of aortic disease is associated with intrinsic differences in aortic structure
Extracellular matrix remodeling and matrix metalloproteinases in the vascular wall during aging and in pathological conditions
Atherosclerosis of the aorta in patients with acute thoracic aortic dissection
Pathway analysis of differentially expressed genes in patients with acute aortic dissection
Gene expression in acute Stanford type A dissection: a comparative microarray study
Differences in matrix metalloproteinase-1 and matrix metalloproteinase-12 transcript levels among carotid atherosclerotic plaques with different histopathological characteristics
Carotid atherosclerotic plaque matrix metalloproteinase-12-positive macrophage subpopulation predicts adverse outcome after endarterectomy
Crystal structure of human macrophage elastase (MMP-12) in complex with a hydroxamic acid inhibitor
Macrophage metalloelastase, MMP-12, cleaves human apolipoprotein(a) in the linker region between kringles IV-4 and IV-5. Potential relevance to lipoprotein(a) biology
Neutrophil-derived matrix metalloproteinase 9 triggers acute aortic dissection
The prominent expression of plasma matrix metalloproteinase-8 in acute thoracic aortic dissection
Association of smooth muscle cell phenotypes with extracellular matrix disorders in thoracic aortic dissection
Rule out of acute aortic dissection with plasma matrix metalloproteinase 8 in the emergency department
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These tutorials were made using the Playstation 3 version of PES 2012/2013/2014Basic Dribbling TutorialLearn the fundamentals of Dribbling in Pro...Free
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