text:
About Sinovac
Sinovac Biotech Ltd. is a China-based biopharmaceutical company that focuses on the research, development, manufacture and commercialization of vaccines that protect against human infectious diseases. Sinovac's vaccine products include Healive(R) (hepatitis A), Bilive(R) (combined hepatitis A and B), and Anflu(R) (influenza). Panflu(R) and PANFLU.1(TM), Sinovac's pandemic influenza vaccine (H5N1) and H1N1 vaccine, have already been approved for government stockpiling. Sinovac is developing vaccines for enterovirus 71, universal pandemic influenza, Japanese encephalitis, and human rabies. Its wholly owned subsidiary, Tangshan Yian, is conducting field trials for independently developed inactivated animal rabies vaccine.
Safe Harbor Statement
This announcement contains forward-looking statements. These statements
are made under the "safe harbor" provisions of the U.S. Private Securities
Litigation Reform Act of 1995. These forward-looking statements can be
identified by words or phrases such as "will," "expects," "anticipates,"
"future," "intends," "plans," "believes," "estimates" and similar statements.
Among other things, the business outlook and quotations from management in
this press release contain forward-looking statements. Statements that are not
historical facts, including statements about Sinovac's beliefs and
expectations, are forward-looking statements. Forward-looking statements
involve inherent risks and uncertainties. A number of
|SOURCE Sinovac Biotech Ltd.
Copyright©2009 PR Newswire.
All rights reserved


text:
EDC Minutes for August 31, 2000, 9D-5, 11:00 – 12:30 p.m.
Members attending
Dwight Ayle
Terri Beam
Julie Bradley
Carol Craig
Bob Farris
Mark Fernandez
Tom McFarland
Dick Ryerson
Linda Small
Kerry Stern
Glenn Vice
Bruce Williams
Vic Zamora
: none
Discussion / Comments / Outcome
New member, Glen Vice, was welcomed to the Educational Delivery Committee.
Meetings and Members
EDC was reorganized during Spring 2000 semester and changed from the Distance Learning Committee. The EDC will be the forum for the following programs to report to: Online Learning, TV, Weekend Business College, Study Abroad, Honors, Work Experience, Service Learning, After School College, Summer Bridge, Freshman Experience, 2 + 2, Langford, and Supplemental Instruction. Meetings are held on the 2nd and 4th Thursday of each month, and representatives from these programs will come to the meeting on the 4th Thursday. A letter was sent inviting these representatives to the meeting on September 28, and a reminder letter will also be sent.
EDC members who are teaching an online class shared how their first two weeks of the fall semester went. Tom McFarland noted that he has observed a distinct difference in his hybrid class between the on-campus and online students.
Terri Beam will be co-chair of the EDC and faculty liaison representative for the 2000-01 academic year.
Academic Senate Report
Terri will be giving EDC reports at each Academic Senate meeting.
To address the Technology Support Resolution 00-09 which was approved by the Academic Senate on 4/13/00, Terri proposed a task list for the following areas:
EDC report which replaces Program Review
Assessment and/or evaluation of existing online courses
Training/mentoring programs
Academic Senate liaisons.
Online Assessment
Kerry Stern, Virginia Bohr, and Terri met regarding assessment of online teaching for the 1999 Summer Sessions and Fall Semester, and 2000 Spring Semester and Summer Sessions.
The four topics comprising the
assessment will be:
comparison of online vs. on-campus teaching.
oes it make a difference what time of day courses are offered?
re hybrid (online-supported) courses doing better than online?
n regards to success and retention, is one term for students more successful than any other term?
Online Courses Update
The EDC will pursue the idea of an Online Fair or Open House with the goal of increasing interest in online teaching.
Open Hourse
Much discussion. Dwight will design an electronic form for faculty. Open Haunted House will be Monday - Thursday, October 30 - November 2, from 1:00 - 4:00 p.m. A sign-up sheet will be distributed at the next EDC meeting. Glen Vice suggested using the words "technology" and "support" in advertising.
Course Approval
Bonnie Jeffers reviewed online course, BUSO 5, Business English.
Course was approved.
In her absence, Kerry reviewed for Andrea Diem the online course, PHIL 5, World Religions.
Course was approved.

subset: commoncrawl


text:
Rutherford et al 2005 highlights
A remarkably close similarity is observed (Figure 3) between the RegEM and Mann et al. (1998) NH annual mean surface temperature reconstructions. The two reconstructions are indistinguishable well within their 2-sigma uncertainties. The RegEM NH reconstruction using all available individual proxy records (rather than replacing spatially dense tree-ring networks with their leading principal components as in the MBH98 Multiproxy/PC network) again yields nearly indistinguishable estimates (Figure 2). The close reproducibility of the MBH98 reconstruction based on both (a) the use of an independent CFR method and (b) the use of the individual proxies used by MBH98 rather than the Multiproxy/PC representation used by MBH98, discredits the arguments put forth by McIntyre and McKitrick (2003) in support of their putative correction to the MBH98 reconstruction.
Figure 1. From Fig. 2, Rutherford et al (2005). Comparisons of stepwise non-hybrid and hybrid-20 reconstructions for annual mean using the multiproxy/PC network and the hybrid-20 reconstruction using the full multiproxy network. Also shown is the instrumental record consisting of the few available data from 1750-1856 (Mann, 2002a) and the Jones et al. (1999) data for 1856-2000.
Figure 2. From Fig. 3, Rutherford et al (2005). Comparison between the RegEM-based hybrid-20 annual mean reconstruction (using the Mann et al. multiproxy/PC network) and Mann et al. (1998) showing overlapping uncertainties in both estimates. Instrumental record is shown for comparison (see Figure 1 for details).
Page 2 of 2 | Previous page


text:
We have over 12,000 students, from over 100 countries, within one of the safest campuses in the UK
97% of Lancaster students go into work or further study within six months of graduating
Research output: Contribution to journal › Journal article
|Journal publication date
|Acta Physica Polonica A
|Number of pages
|Original language
Quantum dots have been identified as potential sources of entangled photon pairs, essential for many applications in quantum information. However, in practice structural properties of the dots result in polarisation splitting of the emission lines, which has prevented the realisation of such a source. Here, we present two techniques developed to control the polarisation splitting in a quantum dot both irreversibly and reversibly. We find that the splitting is strongly dependent upon the emission energy, or lateral confinement, of the quantum dot. Thus by precise control of the InAs deposition thickness, it is possible to produce ensembles from which dots with zero polarisation splitting can be easily selected. Additionally, we demonstrate that by using modest in-plane magnetic fields to partially mix the bright and dark exciton states, the polarisation splitting can be reversibly tuned to zero for most dots of a certain type. Finally, we demonstrate the emission of triggered entangled photon pairs from a quantum dot with approximately zero splitting with fidelity > 70%.


text:
Grand Chifley Adelaide
Grand Chifley Adelaide offers a variety of room types with 138 guest rooms and suites that blend contemporary design with comfort and include video on demand, wireless broadband and personal balcony.
Whether you are planning to conduct business, enjoy South Australia's fabulous food and wine, visit friends and relatives, or just relax and take it easy you are guaranteed to enjoy your stay at Grand Chifley Adelaide.
Grand Chifley Adelaide is a non-smoking hotel.


text:
Size Guide
Sold out
Sheer chiffon blouse. Gathers and ties at neck. decorative pleat at front, and 3/4-length sleeves. Sewn inner top with narrow, adjustable shoulder straps.
100% polyester.
Machine wash warm
H&M uses cookies to give you the best possible experience when visiting our website. By continuing to use our services we assume that you accept our use of cookies.
Read more about cookies


text:
How to Take Care of Your Lips
Ever wondered how Angelina Jolie got those plump lips? Ever wished for that cutie you're crushing on to grab and kiss you? Then read this.
1Fold a tissue in half and run it under water.Ad
2Squeeze it so that it is not soaking, but moist/damp.
3Run it over your lips. This will moisten them.
4Apply Vaseline or petroleum jelly. This will moisten and soften your lips.
5Press your lips together for 15-30 seconds. Make sure the Vaseline/petroleum jelly is well absorbed.
6Apply a thin coat of lipstick (it's usually best to stick with neutral tones, but if bright reds and such work on you, go for it!)
7Blot out by kissing off the excess lipstick on the back of your hand. DO NOT use a tissue for this, as little bits could stick to your lips.
8Apply a thicker coat of lip gloss. This will give it shine.
9Pucker up! (see tips)
10Practice your smile.
11Get some moisturizer (the thicker the better). After you've brushed your teeth at night, put the moisturizer on your lips, rubbing it on them and a little bit around your mouth until you can't see it. Then, get some more and dab a thick layer onto your lips so your lips look white (the color of the moisturizer). Leave it on overnight, and during the night it will soak into your lips and dry, leaving them smooth, soft and perfect. Also, if you don't want to use moisturizer, you can apply a thick layer of Vaseline/petroleum jelly to your lips before you go to sleep.
Give us 3 minutes of knowledge!
- Breathing through your nose, instead of your mouth (when possible) makes a huge difference in the overall appearance of your lips. They dry up and crack when mouth-breathing.
- Remember not to add too much lip gloss on your lips, instead of being shiny it will make it goopy and gross!
- Don't forget to wring out the tissue. Otherwise your Vaseline and everything else will dribble onto your chin.
- Make sure that when you are applying Vaseline you use your fingers instead of a brush.
- if you don't have any lip gloss, you can just apply vaseline/petroleum jelly as this will also add shine and if you are wearing red or a dark lipstick this will take some of it off while you are putting it on.
- Don't stick out your lips when puckering. Adopt a sultry, sexy look that will compliment your great new lips.
- No red colors (for your lipstick). It's best to stick to colors like salmon pink and the peach. Although, if you want your teeth to look whiter, apply some red lipstick. Then over that, apply some bluish toned lip gloss, which makes the white in your teeth stand out.
- Alternatively, instead of tissue, use an old toothbrush, wet it, and lightly brush over your lips. This is great for removing dead skin. Just don't do it too hard, or they'll feel sore afterwards. Use a circular motion, and after you are done, rinse your lips with water and apply Vaseline and/or some other hydrating lip product to your lips. Make sure to coat your lips well. This works best at night, but for during the day, you can apply some lip gloss as the last step. also remember to use a baby toothbrush to brush your lips.
- Make sure your teeth are clean. Your smile will look horrible if your lips are shiny and your teeth are yellow. Gross. There's also no point in having a great smile if you have bad breath. Try using breath spray/mouth wash. Try searching up how to keep your teeth clean or how to brush your teeth properly. As well as how to get your teeth white.
- Do not apply lip gloss all over your lips. Keep it in the middle area and not onto the edges, as this can sometimes make it look messy or look like you've put on too much. Alternatively, too much lip gloss can push someone away, especially if it is sticky. Nothing is more unattractive than sticky lip product.
Things You'll Need
- Lips
- Tissue
- Water
- Vaseline/petroleum jelly
- Neutral toned lipstick
- Lip gloss
- Toothbrush [baby or a soft toothbrush]
Article Info
Categories: Chapped Lips | Facial Preparations
In other languages:
Italiano: Prendersi Cura delle Proprie Labbra, Español: cuidar de tus labios, Deutsch: Lippen pflegen, Português: Cuidar de Seus Lábios, Русский: ухаживать за губами, Français: prendre soin de vos lèvres
Thanks to all authors for creating a page that has been read 344,836 times.


text:
Category information Beanies
You can find your new, cool beanie in our shop for sure! No matter if you're looking for a knitted beanie, visor beanie, snowboard beanie, bobble beanie, wool beanie or headbands for girls and boys by brands like DC, Neff, Volcom, SK8DLX, Cleptomanicx, Burton, Element or Billabong, because we have everything. The products impress with their color, two-ply material and different fits like slouch and fold-up. Whether uni-colored, striped, funny, with logo stitch, patch or flag, you will definitely find your new favorite here.


text:
Includes 1.59 Index Single Vision Lenses
Via Spiga Arezzo Black eyeglasses have a classic style that looks great day and night. These glasses are made of black acetate and have a Havana brown interior. Via Spiga displays their logo on the left temple.About the Brand
Named after a street in one of Milan's renowned shopping districts, Via Spiga Eyewear delivers sleek and architecturally inspired styles, bringing luxury fashion within reach. Draw all eyes to yours in these sophisticated and polished Via Spiga eyeglasses.


text: After a day outdoors, pamper yourself and your partner with a one hour rejuvenating massage each (total of 2 hours), a bottle of Cavabrut (or sparkling juice) and nibble on some chocolate covered fruit…all in the privacy of your room or suite. This 2 night package includes sumptuous breakfast each morning and all our special amenities to make this a special romantic getaway. Prices starting at $578.00 for a party of 2.


text:
The Human Form in Clay (Sculpture - 50 Famous Artist Profiles) by: Jane Waller
Click thumbnails for expanded view.
- $49.99
- Quantity Available: 2
Collector Bookstore is a retailer of new books located in Leavenworth, Kansas. We specialize in price guides and reference books for the antiques and collectibles industry.
Table of Contents
This book is a celebration of 50 contemporary artists from around the world who have chosen to create the human form in clay. There is a detailed account of each artist's work, including biographical details and precise descriptions of their working methods. In addition, each artist has been asked to name a major source of inspiration--whether another artist or key work, in any medium and from any culture, explaining the reasons for their choice. The resulting depictions--ranging from the "Willendorf Venus" to a Michelangelo "Pieta"--combine with stunning photos of the artists' work to make this a book that will inform and inspire.
Jane Waller has a Masters in Ceramics from the Royal College of Art; she has been making Millefiori and Raku ceramics for nearly 30 years. Among her previous books are "Hand-Built Ceramics" and "Colour in Clay."
Collector Book Store
- Leavenworth , Kansas
- View Storefront
- View Seller Website
- Contact Seller About This Item
- View Store Policies
Dealer Accepts:
Item Details:
- Reference #
- 2869
- Quantity
- 2
- Category
- Toys, Dolls, Games & Puzzles
- Department
- Antiques
- Year
- 2001
- Country of Origin
- Dimensions
- Width: 8.75 inches
- Height: 11.25 inches
- Depth: 0 inch
- Weight: 2.88 pounds
- Condition
- New
- Material
- Hard Cover with dust jacket


text:
Enemo J: Burton rockers' track set to appear in major new film Shooting Clerk
Posted: July 30, 2014|
A HEAVY metal band are celebrating after one of their songs was picked to feature on the soundtrack of a new film.
Craig Hartwell, frontman of Burton metal band Enemo J, revealed that he and his bandmates could not believe their luck when they heard that a track from their new album would be featured in new movie Shooting Clerks.
The film is based around how cult movie Clerks came to be made and is being put together by a group of young Scottish film-makers and actors.
The buzz around the movie has been so great that big-hitting Hollywood director Kevin Smith, who was the director of Clerks and the person the film is based on, has backed the flick.
Dreadlocked singer Craig, who is a major fan of the director, spoke to the Mail about the news.
He said: "I am a massive Kevin Smith fan and could not believe it when I heard that they were going to be creating a biopic on him.
"I then found out that the film was being made in Scotland.
"So we got in touch with the film-makers to see if we could help them out in any way.
"We helped them raise some money for parts of the film to be shot in America and then they asked if they could use one of our songs in the film itself.
"They have not decided which track it will be yet but they have our entire new album and will pick a song from that.
"It will be a great feeling to hear our song blasting out of the speakers at the cinema when the film is released.
"It will be a very proud day for me and the entire band."
The news comes only weeks after the band rubbed shoulders with rock gods at the at the UK's number one rock music festival Download, which was held at Donington Park in North West Leicestershire.
The band, which formed at the dawn of the new millennium, will be looking to top off 'their best year yet' by supporting Marilyn Manson at Alt Fest in August in Kettering.
More information about the band's performance date and tickets is available online at www.alt-fest.com/2014
Anyone interested can learn more about Enemo J by logging on to Facebook and visiting www.facebook.com/enemoj


text:
The herd is healthy and bulking up for the winter months ahead. It seems strange not to have a baby reindeer to train this summer. Copper has no little brother or sister tagging along. He's a big reindeer but seems to be more immature than his brothers were at this age. Usually a reindeer calf stops barking by its first birthday. Copper continues to greet us with a bark at mealtime. He also sticks pretty close to mom and is still a bit shy with people. Nevertheless, he wants to go for walks with the rest of the reindeer searching for the mushrooms that grow in the fall rainy season.
Summer brings lots and lots of green treats for the reindeer. Dandelions are by far the favorite. As the dandelions phase out, clover is the next best thing.
The reindeer like spring. As the snow disappears there is plenty of young moss and grasses to nibble on. In the lower winter pen, their favorite ice skating area quickly develops a shallow lake on top of the ice. They love to run through the water skating on the ice as they go.
The reindeer spend their days digging through the snow searching for moss and vegetation. They follow close behind whenever we walk through the pens checking fences and picking up any plastic bags that may have blown into the pens. Every once in a while Crystal and Silver get affectionate, "reindeer style."
Now that we're done breeding Crystal and Silver it is fairly quiet in the reindeer pen this winter. For the most part, the herd gets along pretty well. They stick together and most of the time they nap along side each other. Copper and Crystal are usually in the middle.


text:
Flip.to launches Operation Bark-away-from-the-trophies
Duncan, a nearly two-year-old American Foxhound Beagle mix, has joined the team to diligently guard the awards Flip.to has won.
Duncan came highly recommended by his housemate, Richard Dunbar, who also recently joined the Flip.to team. They're both confident that working in the same office will not interfere with their home life.


text:
MLB locals: BC grad Resop nominated for Roberto Clemente Award
NAPLES - Pittsburgh Pirates right-hander and Barron Collier graduate Chris Resop is the Pittsburgh Pirates' nominee for the Roberto Clemente Award.
The Clemente Award is given annually to the player who best respresents the game through positive contributions on and off the field.
Clemente died in a 1972 plane crash while attempting to deliver supplies to earthquake victims in Nicaragua.
During the offseason, Resop and his wife, Kara, raised more than $10,000 for All Children's Hospital in St. Petersburg and Children's Hospital of Pittsburgh, as well as $12,000 for the Pediatric Cancer Foundation.
Kara Resop ran the Naples Daily News Half Marathon in January with donations in her name going to the Pediatric Cancer Foundation.
During the 2012 season, Resop was a co-host for a "Bowling with the Bucs" charity event that raised more than $30,000 for Pirates Charities, and has been a big supporter of other teammates' charitable endeavors.
Last month, Respop was nominated for the Branch Rickey Award for community service. New York Mets pitcher R.A. Dickey won the award.
Subscribe to the Naples Daily News
Need Help? Call us at 1-888-878-6842.
Monday-Friday: 6am-6pm / Saturday: 7am-3pm / Sunday: 7am-2pm


text:
Vascular smooth muscle cell migration and accumulation in response to growth factors extensively contribute to the development of intimal thickening within the vessel wall. Cumulative evidence has shown that actin cytoskeleton polymerization and rearrangement are critical steps during cellular spreading and migration. Integrin-linked kinase, an intracellular serine/threonine kinase, is a cytoplasmic interactor of integrin beta-1 and beta-3 receptors regulating cell-cell and/or cell-extracellular matrix interaction, cell contraction, extracellular matrix modification, and cell spreading and migration in response to various stimuli. However, the regulatory role of ILK during vascular smooth muscle cell migration and the importance of integrin signaling in occlusive vascular diseases are not yet fully elucidated.
In the present study, we report that integrin-linked kinase controls mouse aortic smooth muscle cell migration in response to platelet-derived growth factor. We have also identified p38 mitogen activated protein kinase as a downstream signaling pathway of the integrin-linked kinase that regulates platelet-derived growth factor-induced actin polymerization and smooth muscle cell migration.
This study will provide new insights into the potential therapeutic value of modulating integrin signaling in an attempt to block or delay smooth muscle cell migration and the progression of vascular diseases.
The initiation and progression of intimal thickening in arterial walls is largely due to migration and subsequent proliferation of smooth muscle cells (SMCs) in the sub-intimal space in response to various stimuli including oxidized low density lipoprotein (oxLDL), circulating growth factors, and inflammatory cytokines such as platelet-derived growth factor (PDGF), tumor necrosis factor (TNF)-α, and interleukin-1 (IL-1) [1,2]. Of these, PDGF, a growth factor released by vascular SMC, endothelial cells, and platelets, has been reported as the most potent inducer of SMC migration within the injured area of the vascular wall.
PDGF is a heparin-binding growth factor composed of polypeptide chains that can be assembled into homodimers (PDGF-AA, -BB) or heterodimer (PDGF-AB) structures and bind to two related cell-surface receptors with tyrosine kinase activity, PDGF receptor α and β [3-5]. In the last few years two additional homodimers PDGF-CC and PDGF-DD were also discovered [6-9]. PDGF binding to its cognate receptors results in dimerization, and subsequent auto-phosphorylation of specific tyrosine residues outside the kinase domain, creating a docking site for SH2 domain-containing signaling proteins. A large number of SH2 domain containing proteins including the phsophatidylinositol-3 kinase (PI3K), phospholipase C-γ (PLC-γ), the tyrosine phosphatase SHP-2, Ras GTPase activating protein (Ras-GAP), Grb2, Grb7, Nck, and the Src family of tyrosine kinases have been shown to bind to cytoplasmic tails of PDGF receptors activating various downstream signaling proteins involved in cell growth, proliferation, survival, and migration [10].
Cellular migration is regulated via a complex interaction between growth factors that attach to their cognate receptors, transmembrane integrins that bind to the components of extracellular matrix, and mechanical stress, all of which cooperatively induce polymerization and reorganization of the actin cytoskeleton. These events are coordinated by mechanisms involved in assembly or disassembly of local adhesion sites, transient changes in actin filaments dynamics, and formation of discrete structures such as stress actin fibers, membrane ruffles, lamellipodia, and filopodia [11-13]. Integrin-linked kinase (ILK), a serine-threonine protein kinase containing a catalytic domain at C terminus, a central pleckstrin homology (PH)-like domain, and four ankyrin-like repeats at the N terminus, is an important component of focal adhesion complex anchoring actin filaments to integrin receptors and the cell membrane [14]. Previous studies have demonstrated that ILK regulates fibroblast migration through the phosphatidylinositol-3 kinase (PI3K) [15], osteosarcoma cell spreading and motility via Rho-associated kinase (ROCK) [16], and mammary epithelial cell migration through the guanine nucleotide exchange factor α-PIX [17].
There is also growing evidence on the cooperation between PDGF receptor and integrins in regulating cellular survival and adhesion [18-20]. However, little is known about the existence of cross-talk between ILK and PDGF signaling in vascular smooth muscle cells and the probable regulatory role of ILK during PDGF-induced SMC migration. In the present study, we have examined the potential function for ILK as an upstream protein regulating the migratory response to PDGF in a primary mouse aortic SMC culture. We have also characterized one downstream pathway that mediates the regulatory role of ILK in modulation actin polymerization and SMC migration.
PDGF activates integrin-linked kinase in mouse aortic SMCs
To study the effect of PDGF treatment on ILK activation, mouse aortic SMCs were treated with 25 ng/ml of PDGF-BB for various timepoints and cell lysates were collected and subjected to the kinase activity assay. As shown, PDGF treatment increased ILK kinase activity around 30 minutes post treatment in the SMC culture without having any significant effect on total ILK protein expression. PDGF-induced ILK kinase activity declined around 60 minutes after treatment (figure 1A). To ensure the specificity of the test and as a negative control, a group of SMCs culture was transfected with specific ILK siRNA for 96 hours. Such treatment completely abolished ILK protein expression in SMCs (figure 1B). As shown, very little kinase activity was detected in ILK-siRNA group confirming that the results of the kinase activity assay is specific. It is noteworthy that we carefully monitored the morphology of SMCs transfected with ILK siRNA for the incubation period prior to PDGF treatment (96 hours) to assure that siRNA treatment had no cytotoxic effect on mouse SMC (data not shown).
Figure 1. PDGF treatment increases integrin-linked kinase activity in mouse aortic smooth muscle cell culture. A) Primary mouse aortic SMCs were serum-starved overnight and then treated with 25 ng/ml of PDGF-BB. ILK kinase activity was measured at various timepoints following treatment. PDGF-BB increased ILK kinase activity (two fold increase). SMC culture transfected with ILK siRNA was used as negative control in the assay. As shown in first lane of the blot, ILK expression was completely abolished in negative control group that led to a significant decrease in ILK kinase activity in this group. B) Transfection of mouse aortic SMCs with ILK siRNA for 96 hours completely blocked ILK protein expression. Data presents three independent experiments and is presented as mean ± STDEV (n = 3), where P < 0.05 was considered significant.
PDGF induces SMC migration through an ILK-dependent pathway
To investigate the potential function of ILK in regulation of PDGF-induced SMC migration, cells were transfected with specific ILK siRNA, and then treated with 25 ng/ml of PDGF-BB. The migratory response was measured using both modified Transwell Boyden chamber and wound healing assays. Inhibition of ILK protein expression markedly decreased SMC migration indicating that the presence of ILK was required for transducing the migratory signal stimulated by PDGF-BB (figures 2A & 2B).
Figure 2. PDGF induces SMC migration through anILK-dependent pathway. A) Inhibition of ILK protein expression with 20 nM ILK siRNA markedly decreased SMC migration in response to PDGF-BB. Data represents three independent experiments (n = 3 culture plates in each replicate) and the value is presented as mean ± STDEV where P < 0.05 was considered significant. B) ILK inhibition decreased the number of migrated SMCs into the site of injury in response to PDGF-BB treatment as compared to the control group (contorl siRNA). Original magnification, ×40. Data present one of three independent experiments.
PDGF is considered as a very potent mitogen. The observed effect of PDGF-BB on wound closure in figure 2B could be due to the effect of this growth factor on SMC proliferation and/or migration. Thus, to determine whether the proliferatory effect of PDGF-BB had contributed to the observed increase in cell migration., cells were treated with increasing doses of PDGF-BB for 24 hours (the maximum time required for wound closure), and cell proliferation and migration were measured using the modified Transwell Boyden chamber assay and MTS cell proliferation assay, respectively. Treatment of primary mouse SMCs with 25 ng/ml of PDGF-BB significantly increased cell migration without having any noticeable effect on SMC proliferation. To assure the accuracy of both assays and as a positive control, mouse SMC culture were also treated with 10% serum. These findings emphasized that the observed increase in cell migration in both assays used in this study (24 hours post injury or following loading onto Transwell Boyden Chambers) was due to the pro-migratory, not proliferatory, effect of PDGF-BB on mouse aortic SMC (additional file 1, figure S1).
Additional file 1. Figure 1S. Effect of PDGF treatment on mouse aortic SMCs migration and proliferation. Mouse SMCs were treated with increasing doses of PDGF-BB for 24 hours and cell migration and proliferation were measured. Cells treated with 10% serum were used as the positive control. As shown, 25 ng/ml of PDGF-BB significantly increased SMCs migration (3 fold increase) with no effect on cell proliferation indicating that the observed increase in cell migration is not due to the proliferatory effect of PDGF-BB in SMC culture. Data is representative of three independent experiments (n = 3 of each culture condition).
Format: PDF Size: 10KB Download file
This file can be viewed with: Adobe Acrobat Reader
ILK inhibition abolishes PDGF-induced p38 MAPK activation in mouse aortic SMC culture
Studies in various in vitro models, have shown that members of the family of the mitogen activated protein kinases (MAPKs) become activated and play a crucial role in regulating cell migration in response to PDGF treatment [21-24]. However, the actual mechanisms underlying this activation and the identity of the upstream kinases involved are not fully elucidated. Here, we first characterized the dynamics of MAPKs family activation in response to PDGF-BB treatment in our primary mouse aortic SMC model. In agreement with previous reports in various cell models, PDGF-BB treatment resulted in the phosphorylation of all three members of MAPKs family, extracellular signal-regulated protein kinase 1/2 (Erk1/2), Jun N-terminus kinase (JNK), and p38 MAPK starting around 5 minutes and with a peak activation at 20-30 minutes post-treatment (additional file 2, figure S2).
Additional file 2. Figure 2S. Kinetics of MAPKs activation in mouse aortic SMC culture. PDGF-BB treatment resulted in phosphorylation and activation of all three members of MAPKs family in mouse aortic SMCs. Data represents three independent experiments.
Format: PDF Size: 71KB Download file
This file can be viewed with: Adobe Acrobat Reader
To investigate the role of each member of MAPKs family in regulating PDGF-induced cell migration, SMCs were treated with specific MEK/Erk1/2 inhibitor (20 μM U0126), JNK inhibitor (50 μM SP600125), and p38 MAPK inhibitor (10 μM SB202190) for two hours prior to PDGF-BB treatment and cell migration was measured using the modified Transwell Boyden Chamber assay. Inhibition of all members of MAPKs family markedly reduced mouse SMC migration in response to PDGF-BB treatment (figure 3A).
Figure 3. ILK inhibition abolishes PDGF-induced p38 MAPK activation in mouse aortic SMCs. A) Mouse aortic SMCs were treated with specific MAPKs inhibitors at specified concentrations for 2 hours prior to seeding onto Transwell Boyden Chambers and PDGF-BB treatment (25 ng/ml) and during the incubation period (12 hours), and cell migration was measured at 12 hours post culture. As shown, inhibition of all three members of MAPKs family significantly reduced SMC migration in response to PDGF-BB. Data is the representative of three independent experiments (n = 3 cell culture replicates for each experiments). B) Mouse aortic SMCs were transfected with 20 nM of specific ILK siRNA and then treated with 25 ng/ml of PDGF-BB for specified timepoints. Inhibition of ILK protein expression markedly reduced PDGF-induced p38 MAPK phosphorylation in mouse aortic SMCs. The blot presents one of three independent experiments. The value in the graph is presented as the mean ± STDEV of three independent experiments. C) ILK inhibition by specific siRNA had no detectable effect on Erk1/2 and JNK phosphorylation in response to PDGF-BB treatment in mouse aortic SMC culture. Data presents one of three independent experiments (n = 3 cell cultures for each independent experiments).
Various components of focal adhesion complex have been shown to regulate or cross talk with members of MAPKs family [24]. ILK, as a key constituent of the focal adhesion complex, also plays an important function in transducing inside-out and outside-in signals [25]. To investigate the role of ILK in regulating MAPKs activation in response to PDGF, SMCs were transfected with specific ILK siRNA prior to PDGF-BB treatment. Inhibition of ILK expression with specific siRNA resulted in significant decrease in PDGF-induced p38 MAPK phosphorylation (figure 3B), while having no effect on Erk1/2 and JNK phosphorylation (figure 3C). This observation suggests that ILK presence is required for PDGF-induced phosphorylation of p38 MAPK in mouse SMC.
ILK mediates PDGF-induced mouse aortic SMC migration through a p38 MAPK-dependent pathway
Taking into account that PDGF-BB activates both ILK and p38 MAPK in mouse aortic SMCs (figures 1A & Additional file 1, figure S1), and the observation that p38 MAPK has a crucial role in PDGF-induced cell migration (Figure 3A); we sought to determine whether ILK can also modulate the migratory effect of p38 MAPK activation in response to PDGF-BB. To examine this hypothesis, we utilized a gain and loss of function approach using adenoviral constructs of ILK and/or p38 MAPK.
In order to revalidate the important role of p38 MAPK activation during the migratory response, and also to exploit a more target-specific approach, we first infected primary mouse SMCs with adenoviral constructs expressing either a dominant negative form of p38 MAPK (Ad-Dn-p38), a wild type form of p38 MAPK (Ad-Wt-p38), or a GFP (Ad-GFP) protein. Western blot analysis (measuring protein expression) and fluorescent microscopy (visualizing GFP expression) were used to evaluate the transfection efficiency at 36 hours post transfection (Figures 4A & 4B).
Figure 4. Dominant negative form of p38 MAPK significantly decreases SMC migration in response to PDGF. A) Photomicrograph of mouse aortic SMCs showing the morphology and GFP expression at 36 hours following transfection. Sub-confluent SMCs were infected with adenoviral vector at the multiplicity of infection of 100. Note the high level of GFP expression as well as the normal phenotype of Ad-GFP-transfected SMCs. Original magnification, ×200. B) Over-expression of a dominant negative form of p38 MAPK markedly increased total p38 MAPK expression while blocking p38 MAPK phosphorylation in response to serum treatment in SMCs (serum is used as a strong activator for p38 MAPK). Also, note the considerable increase in ILK expression level in SMCs transfected with adenoviral ILK constructs. C) For migration assay, transfected SMCs were cultured in Transwell Boyden Chambers in the presence or absence of 25 ng/ml of PDGF-BB and cell migration was measured 12 hours post culture. Dominant negative p38 MAPK significantly decreases SMC migration in response to PDGF-BB treatment while expression of a wild type form of p38 MAPK increases SMC migration in response to PDGF-BB. In addition, over-expression of a wild type mutant of p38 MAPK subverted the inhibitory effect of p38 inhibition on SMC migration. Data represents three independent experiments (n = 3 cell cultures for each independent experiments).
In concurrence with our previous observations, over-expression of a dominant negative mutant of p38 MAPK significantly reduced PDGF-induced SMC migration (Figure 4C). Furthermore, over-expression of a wild type form of p38 rescued SMC migration in response to PDGF-BB, corroborating the specificity of p38 MAPK inhibition in mouse SMC culture transfected with the Ad-Dn-p38 MAPK construct (Figure 4C).
Moreover, a kinase-deficient form of ILK (E359K), which is also incapable of binding to paxillin and parvin and therefore unable to participate in formation of the focal adhesion complex, resulted in a significant decline in PDGF-induced SMC migration, an event that can be reversed by a wild type form of ILK (Figure 5A).
Figure 5. ILK regulates PDGF-induced SMC migration through a p38 MAPK-dependent pathway. A) Kinase deficient mutant of ILK significantly reduced PDGF-induced SMC migration in a Traswell Boyden Chamber migration assay. As shown, expression of a wild type form of ILK rescued SMC migratory response indicating the specificity of ILK inhibition by the adenoviral construct. B) Over-expression of an active form of MKK3/MKK6 (specific upstream activators of p38 MAPK), but not a wild type form of p38 MAPK, rescued SMC migration in response to PDGF-BB indicating that an active form of ILK is required for the initiation of a p38 MAPK-dependent migratory response to PDGF-BB in mouse aortic SMCs. (n = 3 cell cultures for each independent experiments).
Finally, to establish a cause and effect association between inhibition of ILK, a downstream decrease in p38 MAPK phosphorylation, and the subsequent decline in cell migration, we co-transfected cells with a kinase deficient form of ILK (E359K) along with an active form of MKK3/6 (the specific upstream activator of p38 MAPK) or a wild type form of p38 MAPK. As shown, expression of Ad-Ca-MKK3/6 significantly reversed the inhibitory effect of ILK inhibition (Ad-Kd-ILK) in PDGF-treated SMCs (Figure 5B). In contrast, over-expression of the wild type p38 MAPK in SMCs already transfected with Ad-Kd-ILK, did not rescue cell migration, confirming our previous observation that the existence of an active form of ILK was required to facilitate the p38 MAPK-mediated migratory response to PDGF-BB (Figure 5B).
ILK and p38 MAPK regulate PDGF-induced actin cytoskeleton polymerization in vascular SMC
Actin polymerization and re-organization is a critical step in cell motility and migration. To characterize the role of ILK and p38 MAPK in regulating actin polymerization in response to PDGF, cells were transfected with specific ILK siRNA (20 nM) or treated with specific p38 MAPK inhibitor (10 μM SB202190) prior to PDGF treatment. Inhibition of ILK expression and p38 MAPK activity significantly blocked PDGF-induced actin polymerization and re-organization in mouse SMC, indicating a crucial role for ILK and p38 MAPK in regulation actin-mediated cell migration (Figures 6A, B).
Figure 6. ILK and p38 MAPK regulate PDGF-induced actin cytoskeleton polymerization in vascular SMCs. A) Mouse SMCs were treated with either vehicle (DMSO) or with 10 μM of p38 MAPK specific inhibitor SB202190 for 1 hour prior to PDGF treatment. Following treatment, cells were washed and fixed and then stained for actin filaments. Inhibition of p38 MAPK markedly blocks PDGF-induced actin polymerization and reorganization (original magnification ×400). B) Mouse aortic SMCs were transfected with 20 nM of specific ILK siRNA and then treated with 25 ng/ml of PDGF-BB for 20 minutes. Cells were then fixed and stained for actin filaments. Inhibition of ILK expression in SMCs significantly reduced PDGF-induced actin polymerization and reorganization in mouse aortic SMCs. Images are representative image of three independent experiments (original magnification ×400).
Vascular SMC migration contributes to the intimal thickening in occlusive vascular diseases. Following vascular injury, SMC migratory response to various stimuli begins with activation of cell surface receptors followed by remodeling events that involve focal adhesion complex and filament actin remodeling. Various cytokines, peptide growth factors, and components of the extracellular matrix (ECM) have been characterized as pro-migratory molecules.
PDGF is a very potent stimulant for SMC migration through activation of several signaling pathways [26-28]. In vascular SMC, PDGF activates multiple signaling cascades including the PI3K/Akt, protein kinase A (PKA), Src, Ras, and MAPKs signaling. The family of MAPKs comprises three distinct protein kinases, p38 MAPK, JNK, and Erk1/2, all of which have been shown to be involved in vascular diseases and remodeling [23].
On another note, integrin clustering and its downstream signals are necessary for SMC adhesion to ECM, an event that also initiates pro-migratory responses within the vascular wall. Mawatari and colleagues have shown that in human saphenous vein SMCs beta-3 integrin is required for growth factor and cytokine-induced proliferation [29]. It is believed that a coordinated cross-talk between integrins and growth factor tyrosine kinase receptors is necessary for the effective modulation of cell survival, growth, proliferation, and migration [30].
ILK is a key protein of focal adhesion complex connecting β1 and β3 integrins in plasma membrane to the actin cytoskeletal machinery. ILK also interacts with growth factor receptors through the NH2-terminal ankyrin repeat domain and small GTPase signaling molecules such as Nck-2 [14,31]. Previous reports on the effect of PDGF on ILK expression and activity in non-muscle cell cultures were controversial. While Campana et al reported on a PDGF-induced increase in ILK activity in primary rat Schwann cells [32], Janji and colleagues proposed that PDGF had no effect on ILK expression in melanoma HT-144 cells [33], indicating that the outcome of PDGF treatment on ILK signaling is cell specific.
In the present study, we have shown that PDGF-BB treatment results in an increase in ILK kinase activity in mouse aortic SMC culture. It is of importance that the increase in ILK kinase activity in SMCs is associated with a considerable increase in expression of both β1 and β3 integrins (Esfandiarei & van Breemen, unpublished observation). It is well known that the adhesive function of integrin receptors affects cytoskeleton rearrangement and cell motility. Therefore, it is presumable that increase in β1 and β3 integrins expression contributes to the pro-migratory effect of PDGF-BB in SMC culture. However, further investigation is required to evaluate the potential cross-talk between β1 and β3 integrins and PDGF receptors and the significance of such events in vascular SMC migration.
Numerous studies have provided a wealth of information on the crucial regulatory role for ILK as a protein kinase and also as a molecular scaffold and adaptor protein during cell spreading and migration in a variety of cancer cells [13,34]. Moreover, A recent study has shown that an increase in ILK expression coincides with higher rate of SMC migration within the atherosclerotic plaque in ApoE -/- mice, a reliable model that mimics the progression of atherosclerosis in humans [35]. On the contrary, Ho et al has shown that following balloon catheter injury of the rat carotid artery, a significant decrease in ILK expression is associated with an elevation in vascular SMC proliferation and migration [36]. Nevertheless, there is no clear understanding as to how ILK contributes to the intimal thickening in response to growth factors and cytokines released during the development of the atherosclerotic plaque, and the mechanisms by which ILK may modulate vascular SMC migration to the sub-intimal layer of the vascular wall.
In this study, we examined the role of ILK in cell migration, by inhibiting (using siRNA) or increasing (using an adenoviral vector expressing wild type ILK) ILK expression. Inhibition of ILK by different means significantly decreased cell migration in response to PDGF-BB, demonstrating that ILK plays an important role in regulating PDGF-induced vascular SMC migration. Expression of a wild type form of ILK increased SMC migration even in the absence of growth factor. It is noteworthy that the effect of wild type ILK in increasing SMC migration is more evident when SMCs were treated with PDGF-BB (figure 6A). These observations are in agreement with previous report by Dwivedi et al [37] that over-expression of a dominant negative form of ILK markedly reduced intimal thickening in human saphenous vein organ culture.
ILK mutation within the catalytic domain (E359K) disrupts the interaction and binding of ILK with two important components of focal adhesion complex, β-parvin and paxillin. This interaction is required for the assembly of a functional focal adhesion complex and subsequent actin polymerization and reorganization [38,39]. Over-expression of the E395K mutant of ILK resulted in significant reduction in PDGF-induced SMC migration. The effect was fully reversed by a wild type form of ILK. These findings further underscore the crucial function of ILK as molecular scaffold during SMC migration.
In an effort to understand how ILK mediates the pro-migratory effect of PDGF-BB in primary mouse SMC using RNA silencing technique, we were able to show that ILK inhibition could lead to a considerable decrease in p38 MAPK phosphorylation, while having no visible effect on Erk1/2 and JNK phosphorylation in response to PDGF-BB treatment. We also demonstrated that over-expression of an active form of MKK3/6 (specific upstream activator of p38 MAPK) dramatically reversed the inhibitory effect of ILK inhibition on SMC migration. Expression of a wild type form of p38 MAPK markedly increased SMC migration in the presence of PDGF-BB; however, it could not restore migratory response in cells over-expressing a kinase deficient form of ILK. It is of importance that the latter observation accentuates the view that the presence of a functional ILK is required for the initiation of a p38 MAPK-dependent migratory response to PDGF-BB in mouse aortic SMCs. Further studies are required to characterize the precise mechanisms underlying the ILK-mediated regulation of p38 MAPK in SMCs.
Actin polymerization and reorganization is a crucial step in cell migration. We have shown that PDGF increases actin polymerization and filament reorganization in SMCs. This event was significantly blocked by both ILK siRNA and p38 MAPK inhibitor, confirming the importance of this pathway in regulating SMC migration. The actual mechanism by which p38 MAPK may regulate cell migration is not well understood. Actin polymerization and reorganization is under rigorous regulation by a group of protein kinases and/or phosphatases that coordinate the constant turnover of actin filaments in a stimulated cell. Among these, cofilin is a conserved actin-binding protein that enhances actin filament reorganization through de-polymerization and severing of preexisting actin filaments [40]. It is well known that phosphorylation of cofilin (serine 3) by LIM kinase (LIMK) and testicular protein kinase 2 (TESK-2) results in deactivation of cofilin and subsequent inhibition of actin reorganization [41,42]. In mouse aortic SMCs, PDGF treatment resulted in rapid de-phosphorylation and activation of cofilin around 5-15 minutes post-treatment (additional file 3, figure S3). PDGF also increased LIMK phosphorylation leading to phosphorylation and deactivation of cofilin (additional file 3, figure S3). The feedback mechanism provides a strict control system for cell responses to migratory stimuli such as PDGF. Our preliminary observation suggests that p38 MAPK may control actin reorganization partly via regulating the de-phosphorylation and consequent activation of cofilin (Esfandiarei & van Breemen, unpublished observation). Further detailed studies are ongoing in our laboratory to characterize the signaling network and downstream pathways mediating such regulatory events.
Additional file 3. Figure 3S. PDGF induces LIMK and cofilin phosphorylation in mouse aortic SMCs. Aortic SMCs were serum starved overnight and then treated with 25 ng/ml of PDGF-BB. Cell lysates were collected in various timepoints and phosphorylation of LIMK and cofilin was measured. Data represents three independent experiments.
Format: PDF Size: 66KB Download file
This file can be viewed with: Adobe Acrobat Reader
In summary, we have provided evidence that ILK is an important regulator of mouse vascular smooth muscle cell migration in response to PDGF-BB by modulating p38 MAPK phosphorylation. To our knowledge, this is the first report of the regulation of p38 MAPK by integrin signaling in mediating platelet-derived growth factor-induced migration in vascular smooth muscles. Future in vivo studies using specific small molecule inhibitors for ILK and/or p38 MAPK as well as developing transgenic animal models will provide additional information on the potential therapeutic value of integrin signaling, in general, and the integrin-linked kinase, in particular, in controlling vascular smooth muscle cell migration during the progression of occlusive vascular diseases.
Preparation of Primary SMC Culture
All surgical procedures and animal care were conducted according to the Guidelines for Animal Experiments of Faculty of Medicine, University of British Columbia approved by the University Committee on Ethics of Animal Experiments. Primary mouse aortic SMCs were isolated as described previously [43]. Breifly, aortic segments were isolated, cleaned of excess advential tissue, and digested using collagenase II (0.5 mg/ml). Isolated cells were pelleted, resuspended, and grown in Dulbecco's modified Eagle's medium. Sub-confluent (60-80%) cutlures were used for all experiments. ILK recombinant adenoviral constructs were kind gifts from Dr. Hyo-Soo Kim (Seoul National University Hospital, Korea). Adenoviral constructs for recombinant p38 MAPK and constitutively active forms of MKK3/6 were kindly provided by Dr. Donald R. Menick (Medical University of South Carolina, USA).
Transient Transfection
For transient transfection experiments, cells were infected with adenoviruses encoding dominant negative p38 MAPK (Ad-Dn-p38), wild type of p38 MAPK (Ad-Wt-p38), constitutively active MKK3/6 (Ad-Ca-MKK3/6), kinase-deficient ILK (Ad-Kd-ILK), or wild type ILK (Ad-Wt-ILK) with multiplicity of infection of 100 (MOI = 100). Following overnight incubation at 37°C, SMCs were replenished with fresh medium. Transfection efficiency was measured by fluorescence microscopy and Western blot analysis at 36 hours post transfection. For all experiments, transfected SMCs were used at 36 hours post transfection. For ILK RNA inhibition experiments, SMCs were transfected with 20 nM of a 21-base pair double-stranded small interfering RNA (siRNA) molecule targeting the PH domain of ILK or a control nonspecific siRNA. ILK siRNA was introduced to cells using 2.5 μl of siLentFect Transfection Reagent according to the manufacturer's protocol (Bio-Rad Laboratories, Mississauga, ON, Canada). Twenty four hours post-transfection cells were replenished with fresh serum-containing medium, incubated for 96 hours, and then used for various experiments. ILK silencing was determined by Western blot of transfected lysates with an anti-ILK antibody.
Western Blot Analysis
Cells either untreated or treated with various experimental reagents were washed twice with ice-cold PBS, and kept on ice for 15 min in lysis buffer containing 50 mM pyrophosphate, 50 mM, NaF, 50 mM NaCl, 5 mM EDTA, 5 mM EGTA, 100 μM Na3VO4, 10 mM HEPES (pH 7.4), 0.1% Triton X-100, 10 μg/ml leupeptin, and 1 mM phenylmethylsulfonyl fluoride. Cell lysates were collected by scraping and protein concentration was determined using Bradford assay. Extracted protein (40-80 μg) was fractionated by electrophoresis in 7% to 9% sodium dodecyl sulfate-polyacrylamide gels, transferred to nitrocellulose membranes, and blocked with PBS containing 0.1% Tween-20 and 5% non-fat dry milk for 1 hour. Afterward, the membrane was incubated with specific primary antibody overnight at 4°C, followed by secondary antibody for one hour at room temperature. Immunoblots were visualized with an enhanced chemiluminescence detection system according to the protocol of the manufacturer (Pierce Biotechnology, Rockford, IL, USA). Densitometry analysis was performed by using the National Institutes of Health ImageJ software (version 1.27z). Density values for proteins were normalized to the level for control groups (arbitrarily set to 1.0-fold).
ILK Kinase Assay
Kinase assay was performed using 250 μg of protein lysates immunoprecipitated with 5 μg ILK antibody and protein A Sepharose beads overnight at 4°C, while shaking. The immunocomplex was washed twice with high salt NP-40 lysis buffer (containing 750 mM NaCl), and then three times with wash buffer containing 50 mM HEPES pH 7.5, 85 mM KCl, 10 mM ethyleneglycol tetraacetate (EGTA), 0.1% Tween 80, 1 mM Na3VO4, 10 mM Mg2Cl. The kinase assay was performed in the reaction buffer containing 0.5 μg ATP (250 μM ATP and 5 μCi [γ -32P] ATP) and 5 μg of myelin basic protein (MBP) as substrate at 30°C for 20 minutes in a shaker-incubator. Samples were fractionated by electrophoresis in a 12% sodium dodecyl sulfate-polyacrylamide gel, and phosphorylation of substrate by ILK was measured by a scintillation counter autoradiography.
Cell Migration Assay
Cell migration was measured using QCM™ Transwell Colorimetric Cell Migration Assay according to the manufacturer's protocol (Chemicon International, Temecula, CA, USA). Briefly, 1 × 105 serum-starved SMCs were loaded onto the upper well of the chamber in the presence or absence of inhibitors while lower wells were filled with culture medium with or without PDGF-BB (25 ng/ml). Following 12 hours incubation, non-migrating cells on the upper side of membranes were removed by wiping and rinsing, and migrated cells were counted using colorimetric assay. Data are represented as fold change in cell migration where migratory rates for control groups are arbitrarily set to 1.0 fold.
Wound Healing Assay
Sub-confluent transfected or non-transfected SMCs were grown on glass coverslips. Following overnight serum starvation, cell cultures were scratched with a sterile pipette tip to form a wound, washed with pre-warmed sterile PBS, and incubated with medium in the presence or absence of inhibitors for 18-24 hours. At desired timepoints post injury cells were fixed and subjected to imaging using Nikon inverted microscope and Spot digital camera.
Cell Viability Assay
Sub-confluent SMCs were plated in a 24-well culture plate and treated with various doses of PDGF for 24 hours. The CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) was used to measure cell viability according to the manufacturer's protocol (Promega, Madison, WI).
Actin Cytoskeleton Staining
Cells were grown on glass slides and subjected to various treatments. At desired timepoints cells were fixed (3.7% formaldehyde for 15 min & 70% ethanol for 2 min). Following permeabilization in 0.1% Triton X-100, cells were incubated with AlexaFluor 488-labelled phalloidin (Molecular Probes, Invitrogen Detection Technologies) for 1 hour in room temparature. Glass slides were mounted and sealed using ProLong® Gold anti-fade reagent (Molecular Probes, Invitrogen Detection Technologies), and then imaged with Olmpus inverted microscope and Spot digital camera.
Statistical Analysis
Two-way analysis of variance (ANOVA) with multiple comparisons andpaired Student's t tests were performed. Values shown are the mean ± standard deviation. A value of P < 0.05 was considered significant.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
The scientific idea and experimental design were developed by ME and CVB. Experiments were performed by ME, SAY. ILK kinase assay was performed by VG. Manuscript was written by ME and was revised by CVB and SD. All authors read and approved the final manuscript.
This study was supported by a grant-in-aid from the Canadian Institutes of Health Research (CV); and fellowships from the Heart & Stroke Foundation of Canada/AstraZeneca Canada (ME), Michael Smith Foundation for Health Research (ME), and Canadian Diabetes Association (ME).
Schwartz SM: Smooth muscle migration in atherosclerosis and restenosis.
Journal of Clinical Investigation 1997, 100(11 Suppl):S87-89. PubMed Abstract
Kher N, Marsh JD: Pathobiology of atherosclerosis--a brief review.
Majesky MW, Reidy MA, Bowen-Pope DF, Hart CE, Wilcox JN, Schwartz SM: PDGF ligand and receptor gene expression during repair of arterial injury.
Journal of Cell Biology 1990, 111(5 Pt 1):2149-2158. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Ross R: The pathogenesis of atherosclerosis: a perspective for the 1990s.
Nature 1993, 362(6423):801-809. PubMed Abstract | Publisher Full Text
Chen Y, Xu H, Liu J, Zhang C, Leutz A, Mo X: The c-Myb functions as a downstream target of PDGF-mediated survival signal in vascular smooth muscle cells.
Biochemical & Biophysical Research Communications 2007, 360(2):433-436.
Li X, Ponten A, Aase K, Karlsson L, Abramsson A, Uutela M, Backstrom G, Hellstrom M, Bostrom H, Li H, Soriano P, Betsholtz C, Heldin CH, Alitalo K, Ostman A, Eriksson U: PDGF-C is a new protease-activated ligand for the PDGF alpha-receptor[see comment].
Nature Cell Biology 2000, 2(5):302-309. PubMed Abstract | Publisher Full Text
LaRochelle WJ, Jeffers M, McDonald WF, Chillakuru RA, Giese NA, Lokker NA, Sullivan C, Boldog FL, Yang M, Vernet C, Burgess CE, Fernandes E, Deegler LL, Rittman B, Shimkets J, Shimkets RA, Rothberg JM, Lichenstein HS: PDGF-D, a new protease-activated growth factor.
Nature Cell Biology 2001, 3(5):517-521. PubMed Abstract | Publisher Full Text
Gilbertson DG, Duff ME, West JW, Kelly JD, Sheppard PO, Hofstrand PD, Gao Z, Shoemaker K, Bukowski TR, Moore M, Feldhaus AL, Humes JM, Palmer TE, Hart CE: Platelet-derived growth factor C (PDGF-C), a novel growth factor that binds to PDGF alpha and beta receptor.
Journal of Biological Chemistry 2001, 276(29):27406-27414. PubMed Abstract | Publisher Full Text
Bergsten E, Uutela M, Li X, Pietras K, Ostman A, Heldin CH, Alitalo K, Eriksson U: PDGF-D is a specific, protease-activated ligand for the PDGF beta-receptor.
Nature Cell Biology 2001, 3(5):512-516. PubMed Abstract | Publisher Full Text
Heldin CH, Westermark B: Mechanism of action and in vivo role of platelet-derived growth factor.
Physiological Reviews 1999, 79(4):1283-1316. PubMed Abstract | Publisher Full Text
Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR: Cell migration: integrating signals from front to back.
Science 2003, 302(5651):1704-1709. PubMed Abstract | Publisher Full Text
Hall A: Rho GTPases and the actin cytoskeleton.
Science 1998, 279(5350):509-514. PubMed Abstract | Publisher Full Text
Mitra SK, Hanson DA, Schlaepfer DD: Focal adhesion kinase: in command and control of cell motility.
Nature Reviews Molecular Cell Biology 2005, 6(1):56-68. PubMed Abstract | Publisher Full Text
Hannigan GE, Leung-Hagesteijn C, Fitz-Gibbon L, Coppolino MG, Radeva G, Filmus J, Bell JC, Dedhar S: Regulation of cell adhesion and anchorage-dependent growth by a new beta 1-integrin-linked protein kinase.
Nature 1996, 379(6560):91-96. PubMed Abstract | Publisher Full Text
Qian Y, Zhong X, Flynn DC, Zheng JZ, Qiao M, Wu C, Dedhar S, Shi X, Jiang BH: ILK mediates actin filament rearrangements and cell migration and invasion through PI3K/Akt/Rac1 signaling.
Oncogene 2005, 24(19):3154-3165. PubMed Abstract | Publisher Full Text
Khyrul WA, LaLonde DP, Brown MC, Levinson H, Turner CE: The integrin-linked kinase regulates cell morphology and motility in a rho-associated kinase-dependent manner.
Journal of Biological Chemistry 2004, 279(52):54131-54139. PubMed Abstract | Publisher Full Text
Filipenko NR, Attwell S, Roskelley C, Dedhar S: Integrin-linked kinase activity regulates Rac- and Cdc42-mediated actin cytoskeleton reorganization via alpha-PIX.
Oncogene 2005, 24(38):5837-5849. PubMed Abstract | Publisher Full Text
Ahlen K, Rubin K: Platelet-derived growth factor-BB stimulates synthesis of the integrin alpha 2-subunit in human diploid fibroblasts.
Experimental Cell Research 1994, 215(2):347-353. PubMed Abstract | Publisher Full Text
Kirchberg K, Lange TS, Klein EC, Jungtaubl H, Heinen G, Meyer-Ingold W, Scharffetter-Kochanek K: Induction of beta 1 integrin synthesis by recombinant platelet-derived growth factor (PDGF-AB) correlates with an enhanced migratory response of human dermal fibroblasts to various extracellular matrix proteins.
Experimental Cell Research 1995, 220(1):29-35. PubMed Abstract | Publisher Full Text
Sundberg C, Rubin K: Stimulation of beta1 integrins on fibroblasts induces PDGF independent tyrosine phosphorylation of PDGF beta-receptors.
Journal of Cell Biology 1996, 132(4):741-752. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Lundberg MS, Curto KA, Bilato C, Monticone RE, Crow MT: Regulation of vascular smooth muscle migration by mitogen-activated protein kinase and calcium/calmodulin-dependent protein kinase II signaling pathways.
Journal of Molecular & Cellular Cardiology 1998, 30(11):2377-2389.
Matsumoto T, Yokote K, Tamura K, Takemoto M, Ueno H, Saito Y, Mori S: Platelet-derived growth factor activates p38 mitogen-activated protein kinase through a Ras-dependent pathway that is important for actin reorganization and cell migration.
Journal of Biological Chemistry 1999, 274(20):13954-13960. PubMed Abstract | Publisher Full Text
Zhan Y, Kim S, Izumi Y, Izumiya Y, Nakao T, Miyazaki H, Iwao H: Role of JNK, p38, and ERK in platelet-derived growth factor-induced vascular proliferation, migration, and gene expression.
Arteriosclerosis, Thrombosis & Vascular Biology 2003, 23(5):795-801.
Huang C, Jacobson K, Schaller MD: MAP kinases and cell migration.
Journal of Cell Science 2004, 117(Pt 20):4619-4628. PubMed Abstract | Publisher Full Text
Legate KR, Montanez E, Kudlacek O, Fassler R: ILK, PINCH and parvin: the tIPP of integrin signalling.
Nature Reviews Molecular Cell Biology 2006, 7(1):20-31. PubMed Abstract | Publisher Full Text
Koyama N, Morisaki N, Saito Y, Yoshida S: Regulatory effects of platelet-derived growth factor-AA homodimer on migration of vascular smooth muscle cells.
Journal of Biological Chemistry 1992, 267(32):22806-22812. PubMed Abstract | Publisher Full Text
Jawien A, Bowen-Pope DF, Lindner V, Schwartz SM, Clowes AW: Platelet-derived growth factor promotes smooth muscle migration and intimal thickening in a rat model of balloon angioplasty.
Journal of Clinical Investigation 1992, 89(2):507-511. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Buetow BS, Tappan KA, Crosby JR, Seifert RA, Bowen-Pope DF: Chimera analysis supports a predominant role of PDGFRbeta in promoting smooth-muscle cell chemotaxis after arterial injury.
American Journal of Pathology 2003, 163(3):979-984. PubMed Abstract | PubMed Central Full Text
Mawatari K, Liu B, Kent KC: Activation of integrin receptors is required for growth factor-induced smooth muscle cell dysfunction.
Journal of Vascular Surgery 2000, 31(2):375-381. PubMed Abstract | Publisher Full Text
Comoglio PM, Boccaccio C, Trusolino L: Interactions between growth factor receptors and adhesion molecules: breaking the rules.
Current Opinion in Cell Biology 2003, 15(5):565-571. PubMed Abstract | Publisher Full Text
Tu Y, Li F, Goicoechea S, Wu C: The LIM-only protein PINCH directly interacts with integrin-linked kinase and is recruited to integrin-rich sites in spreading cells.
Campana WM, Myers RR, Rearden A: Identification of PINCH in Schwann cells and DRG neurons: shuttling and signaling after nerve injury.
GLIA 2003, 41(3):213-223. PubMed Abstract | Publisher Full Text
Janji B, Melchior C, Vallar L, Kieffer N: Cloning of an isoform of integrin-linked kinase (ILK) that is upregulated in HT-144 melanoma cells following TGF-beta1 stimulation.
Oncogene 2000, 19(27):3069-3077. PubMed Abstract | Publisher Full Text
Hannigan G, Troussard AA, Dedhar S: Integrin-linked kinase: a cancer therapeutic target unique among its ILK.
Nature Reviews Cancer 2005, 5(1):51-63. PubMed Abstract | Publisher Full Text
Friedrich EB, Clever YP, Wassmann S, Werner N, Bohm M, Nickenig G: Role of integrin-linked kinase in vascular smooth muscle cells: regulation by statins and angiotensin II.
Biochemical & Biophysical Research Communications 2006, 349(3):883-889.
Ho B, Hou G, Pickering JG, Hannigan G, Langille BL, Bendeck MP: Integrin-linked kinase in the vascular smooth muscle cell response to injury.
American Journal of Pathology 2008, 173(1):278-288. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Dwivedi A, Sala-Newby GB, George SJ: Regualtion of cell-matrix contacts and beta-catenin signaling in VSMC by integrin-linked kinase: implications for intimal thickening.
Basic Res Cardiol 2008, 103(3):244-256. PubMed Abstract | Publisher Full Text
Yamaji S, Suzuki A, Sugiyama Y, Koide Y, Yoshida M, Kanamori H, Mohri H, Ohno S, Ishigatsubo Y: A novel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction.
Journal of Cell Biology 2001, 153(6):1251-1264. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Nikolopoulos SN, Turner CE: Integrin-linked kinase (ILK) binding to paxillin LD1 motif regulates ILK localization to focal adhesions.
Journal of Biological Chemistry 2001, 276(26):23499-23505. PubMed Abstract | Publisher Full Text
Condeelis J: How is actin polymerization nucleated in vivo?
Trends in Cell Biology 2001, 11(7):288-293. PubMed Abstract | Publisher Full Text
Toshima J, Toshima JY, Takeuchi K, Mori R, Mizuno K: Cofilin phosphorylation and actin reorganization activities of testicular protein kinase 2 and its predominant expression in testicular Sertoli cells.
Journal of Biological Chemistry 2001, 276(33):31449-31458. PubMed Abstract | Publisher Full Text
Arber S, Barbayannis FA, Hanser H, Schneider C, Stanyon CA, Bernard O, Caroni P: Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase[see comment].
Nature 1998, 393(6687):805-809. PubMed Abstract | Publisher Full Text
Troussard AA, McDonald PC, Wederell ED, Mawji NM, Filipenko NR, Gelmon KA, Kucab JE, Dunn SE, Emerman JT, Bally M, Dedhar S: Preferential dependence of breast cancer cells versus normal cells on integrin-linked kinase for protein kinase B/Akt activation and cell survival.
Cancer Research 2006, 66(1):393-403. PubMed Abstract | Publisher Full Text


text:
Credits & Info
- Uploaded
- Apr 5, 2011 | 3:57 PM EDT
- Daily 4th Place April 6, 2011
If you liked this, check these out!
Author Comments
Rated 3 / 5 stars
very nice but very small.
Rated 5 / 5 stars
Nice job, Kreid nice artwork keep it up...I mean I'm an artist too but I don't know how animate it!! :L sorry for this comment just trying the help by making people happy u know?Hehehehehe...ohhh.
Rated 5 / 5 stars
i know one thing on a last stand .... no fat chicks
Rated 5 / 5 stars
Don't they know the song?
~Shoot the zombies in the head,and you on't be dead,shoot the zombies,shoot the zombies,yeah!~
Rated 5 / 5 stars
best way to die
If theres ever a zombie apocolypse I wanna be the retard who brings a nerf or water gun
Kreid responds:
Spot's taken bro!


text:
Alright so I moved to Estes park in march and am getting ready for the winter. Golite's 4 season adrenaline bags are on sale for a crazy price right now. Will that bag be enough for the winter up here?
The last few trips in the NM or CO winter high country I've used my Feathered Firends eVent shelled 10 bag. I also have my Stephenson's DAM, I think the pad underneath is almost as important as the bag around you. I think the Golite bag will be fine, but see if you can wear some clothes inside it just in case.
estes is quite the base camp. the 4-season bag should be enough. I'd highly recommend the neoair xtherm for a pad if you can find a deal. so light, so warm. i'm in boulder if you wanna hike or scramble sometime.
The Golite bag will work. If your worried about fit, they have a permanent store on the mall now if you want to lay in it first. I'm also in Boulder if you want to go for a hike or need trip ideas. Enjoy Estes!
You must login to post.
MEMBERSHIP IS REQUIRED TO POST: You must be a Forum, Annual or Lifetime Member to post messages in the backpackinglight.com forums.


text:
Oil & Gas news that matters News - Articles
Data analytics is the new frontier in cost reduction
- Category: Technology
- Published: 2015-Sep-29
- Author: Dale Lunan
In the oil and gas business, data is measured in terabytes and petabytes. To understand how much data that is, here is a comparison: a gigabyte is seven minutes of high definition video; a petabyte is 13.3 years of the same video feed.
Smart oil
- Category: Technology
- Published: 2015-Sep-11
- Author: Sebastian Gault
Artificial intelligence is shaping the future of energy, but don't forget the risks
"Data is the new oil." One often hears that saying these days, but what does it really mean? For some people, oil has long been the ultimate symbol of wealth and power, but some like to suggest its role is being replaced by information.
Learning curve
- Category: Technology
- Published: 2015-Sep-11
- Author: R.P. Stastny
Refrac tests hold promise as the industry races to lower its costs
Everyone loves a good deal: Black Friday, Boxing Day and refracs. Refracturing horizontal multistage wells has been touted as a cost-effective way to unlock additional resource-play production because it avoids the expense of drilling. So the promise of refracking is huge in the current low commodity price environment.
Side of veggies
- Category: Technology
- Published: 2015-Sep-11
- Author: Jim Bentein
Plant-based chemical innovations aren't likely to push petrochemicals away from the table
It's unlikely plant-based chemical sources will ever play a larger role than fossil fuel–based petrochemicals in the Canadian economy, says the head of an organization that helped attract a new multi-million dollar biochemical plant to economically depressed Sarnia, Ont., Canada's historic Chemical Valley.
Triple play
- Category: Business
- Published: 2015-Aug-07
- Author: Dale Lunan
A trio of industry stalwarts are poised for induction into the Canadian Petroleum Hall of Fame
A triple play in baseball is a rare occurrence—fewer than 700 have been recorded in the last 139 years of Major League Baseball play—but that has nothing on the rarity of a triple play induction into the Canadian Petroleum Hall of Fame: 2015 marks the first time in 19 years that only three have been named to join the 146 honoured members already enshrined there.
Tread lightly
- Category: Environment
- Published: 2015-Aug-07
- Author: Jim Bentein
Oilsands projects are critical to Canada's economy, but operators need to significantly reduce their footprint
Even an economist with an environmental organization that spends much of its time criticizing Canada's oilsands sector acknowledges that it is a large part of the country's economy and shutting it down would be committing economic suicide.
Pause button
- Category: Business
- Published: 2015-Aug-07
- Author: R.P. Stastny
Slowed oilsands development will still require more pipelines
"Dirty oil" activism over the years held Canadian export pipeline projects hostage in the battle to contain oilsands growth, but $100 oil was the trump card that allowed industry to mostly stay on track in its growth plans. Bitumen-by-rail mushroomed and alternate pipe routes and expansions stepped up to move ever-increasing crude oil volumes to refining markets. In fact, business was so good just a year ago that overheated oilsands construction costs prompted some producers to shelve capital-intensive investments in new projects or expansions.


text:
Apple has kicked off a new countdown promotion, this time for the 50 billionth app to be downloaded from its iTunes App Store. As always, the winner will nab a $10,000 / €10,000 gift card.
The company announces on iTunes, "The App Store is about to hit 50 billion downloads. And you could win a $10,000 Appe Store Gift Card."
"As we approach 50 billion app downloads, we'll be awarding a $10,000 App Store Gift Card to the lucky customer who downloads the 50 billionth app," the company elaborates.
Unlike previous years, this time around Apple will also be giving out $500 / €500 gift cards to each of the next 50 people who download an app.
On the announcement page, Apple posts the most popular iOS apps, paid and free, "each one having played a big role in achieving this historic benchmark."
Go ahead and check out the promotion at this link.
The promo is open to entrants who are at least 13 years old at the time of entry. You must also be located in a participating country.
The download limit is 25 titles per day, and you can limit your downloads to free apps only – no need for any purchases.
Apple clarifies that the contest ends after the download of the 50,000,000,050th app.
According to the application charts, the all-time favorite paid apps include Angry Birds, Fruit Ninja, Doodle Jump, Cut the Rope, WhatsApp Messenger, Camera+, and lots of other titles (mostly games).
In the free-apps department, things are a little bit different. The all-time favorite is (unsurprisingly) the Facebook iOS client, followed by Pandora Radio, Instagram, YouTube, Skype, The Weather Channel, Temple Run, Google Search, Netflix, Shazam, and others.
On the iPad, things aren't much different, though it's worth noting that the #1 paid app is actually Apple's Pages productivity app.


text:
Goodfellow Inc.
Flooring Adhesive Stauf PUM 950 Urethane Adhesive - 3 Gallon Pail
Model: 711040005 | Store Special Order SKU: 1000744826
Stauf 950 is a high performance Urethane adhesive that spreads easily and quickly. It can be used with any species, width, thickness or length of flooring. It has no VOC's and most importantly will not etch floor finish during installation.
- Urethane Adhesive
- No etch to floor finish
- Radiant Heat Approved
- No VOC
Warranty Information
For more information please contact 1-800-628-0525.
|Assembled Depth (In Inches)
|Assembled Height (In Inches)
|Assembled Weight (In LBS)
|38 Lbs.
|Assembled Width (In Inches)
|Country of Origin
|Item Depth
|12 In.
|Item Height
|10.5 In.
|Item Weight
|38 Lbs.
|Item Width
|12 In.
|Store SKU
Price and availability may vary by store.
*Valid only on date printed
User Reviews
Get the inside scoop from users like you!
Thank you!
Your quick rate has been submitted.
Please note it may take up to 8 hours for your quick rating to appear.
Sorry we are currently experiencing technical difficulties. Please try and resubmit your Quick Rating.
The Home Depot offers more than 2,000 Eco Options products online and in store that can help you save money and are better choices for the environment.
Energy Star
The Home Depot offers close to 1,000 ENERGY STAR® qualified products, which meet or exceed the highest energy efficiency standards without compromising performance.
Shipping calculator
Shipping estimate is based on total order weight and destination. Actual shipping charges will be confirmed at checkout.
Remote areas may exceed this range.
Flat Rate Shipping
Some exceptions apply. Shipping rate is applied on a per product basis. Orders/deliveries to select remote locations will need to be placed through Customer Support where shipping rates will be provided prior to order confirmation/completion.
Shipping calculator
Shipping estimate is based on total order weight and destination. Actual shipping charges will be confirmed at checkout.
Remote areas may exceed this range.
We offer 250,000+ products through our in-store special order program. Please visit your nearest Home Depot location for details.


text:
The release of the final build for Opera 12.10 did not come as a surprise this week, as there were clear signs that the developer managed to overcome some of the most nagging difficulties.
Touted as a security update, Opera 12.10 incorporates repairs as well as new features. Among the most significant are support for SPDY, the full screen API, compatibility with the latest operating systems from Apple and Microsoft (Mountain Lion and Windows 8), as well as page load improvements.
On the latest Windows operating system, the browser features inertia scrolling and pinch-to-zoom action. Pages should also look better, as the new Opera has support for ICC profile v4.
On Mac systems, there is support for Retina displays, thus making everything look much better. There is also integration with the Notification Center.
The current release integrates DNS prefetching, a feature that enables the browser to do a DNS look-up when the user merely hovers the mouse over a web link. The result of this action is that the page loads faster if the user decides to visit it.


text:
Team Deployment
On September 15, 2008, the ASPCA deployed members of our Field Investigations and Response Team to assist Texas in dealing with the aftermath of Hurricane Ike. Members included veterinarians, veterinary technicians, disaster responders and sheltering professionals, along with an ASPCA disaster response trailer. The group was stationed at the Area Command Center at the Texas Animal Health Commission in Austin and provided assessments in Liberty County, just north of Galveston, one of the hardest-hit areas.
"We were pleased to lend our assistance again, both in terms of human resources and equipment, in relief efforts for Hurricane Ike, especially so closely on the heels of Hurricane Gustav," said ASPCA President and CEO Ed Sayres. "By using our resources strategically and making key connections at the local levels, the ASPCA's Field Investigations and Response Team is able to effectively respond to shelters and animals in need."
Financial Support
In addition to deploying the disaster team, the ASPCA dispersed and pledged more than $75,000 in grant funding to organizations in both Texas and Louisiana following Hurricanes Ike and Gustav. The grant funding covered recovery costs, as well as transportation assistance, distribution of food and supplies, and other needs. Grant recipients included:
- Animal Aid of Vermillion Parish, Louisiana, for recovery and supplies
- Walter Ernst Foundation of the Louisiana Veterinary Medical Association, for equine hay drops in southwest Louisiana
- Habitat for Horses Inc. in Hitchcock, Texas, for emergency hay after destruction of a new barn and sheds, and for 60 horses on site and another 300 in foster care across several states
- Hopeful Haven Equine Rescue Organization in Shreveport, Louisiana, for assistance with removal of hoof stock from flooded areas in Vermillion Parish, as well as hay
"Thanks to the kindness and generosity of our supporters, the ASPCA is able to offer grants and emergency funds to assist organization affected by both Hurricanes Ike and Gustav," said Sayres. "Over the years, these organizations have shown exceptional commitment and dedication the animals in their care, and we are happy to be able to provide assistance during their time of need."
For vital information on creating disaster plans that include your pets, visit our Disaster Preparedness Tips.


text:
Kristin Davis, Aaron Sorkin Kiss on Red Carpet, Confirm Romance!
New love in the city for Kristin Davis!
After weeks of speculation, the Sex and the City star, 47, and famed screenwriter Aaron Sorkin confirmed their rumored romance -- with a passionate kiss on the lips on the red carpet at the Los Angeles premiere of HBO series The Newsroom Wednesday night.
PHOTOS: Hottest Hollywood hookups
With plenty of cameras present to capture the moment, 51-year-old Sorkin (creator and executive producer of Newsroom) grabbed Davis and gave a long, lingering kiss. Davis, pretty in an A-line aquamarine dress and strappy stilettos, grabbed her new man by the shoulders.
"They are really happy," a source told E! News in late May. "They have actually known each other a while through mutual friends and would sometimes bump into one another at industry events, but it has only just turned into something more."
PHOTOS: Kristin and other 90s stars then and now
The Oscar-winning screenwriter divorced Julia Bingham back in 2005; they share daughter Roxy, 12. Never-married Davis adopted a baby girl, Gemma, last year.


text:
Dell Streak 2 M02M spotted in testing: Opus One or Looking Glass?
Dell's second-gen Streak tablet has seemingly passed through WiFi certification, with the Dell M02M spotted getting a test certificate. Though the Streak name isn't mentioned, the M02M product code is one digit away from the original Streak's M01M, and it is described as a "mobile internet device" with WiFi b/g/n.


text:
RayMartell Moore was born in Frankfurt am Main, Germany (military brat). Throughout his childhood he traveled all over Europe and America. Finally, his family settled down in Tampa, Florida where he completed middle & high school. After graduating Brandon High he received a scholarship to continue his education in New York City at the New York Conservatory for Dramatic Arts (SFT).
Upon graduation he was cast the lead role, in the independent feature film "Finding Me". The entire process of "Finding Me" gave him his sense of vocation, experiencing the lives he's touch with such a role. His focus with every role is to tell it truthfully & honestly so that someone, somewhere can see their story whether it be on stage or on the big screen.
RayMartell Moore has also studied with the O'Neill Studio and became an honorary Independent Artist, having the great opportunity to work with the multiple Tony Award winning actress, Marian Seldes.
As he continues to sharpen his craft through film, television, theatre and print, RayMartell also has a plethora of stories of his own to tell with the screenplays he's currently writing. Thrilled with all that has come, is coming and is yet to come, RayMartell currently resided in Upper Manhattan, ready for the world.


text:
Urban Decay Surreal Skin Cream to Powder Foundation Review
I suspect many of you may be excited about the brand new Urban Decay Surreal Skin Cream to Powder Foundation. Upon first seeing it I was pretty excited myself. My first foundation and the foundation that remained with me for many years prior to my experimenting with different formulas was in fact a cream. Becoming deeply attached to my cream foundation over the years it was of course very exciting to see one of my favorite brands releasing a cream compact foundation.
Jump ahead for a peep at the new Urban Decay Surreal Skin Cream to Powder Foundation.
I will in fact express my disappointment this isn't cream foundation but a cream to powder. I love cream foundation however I'll begrudgingly admit, in all my years of using it, it doesn't wear as long as I'd like. I'll also add that one thing about cream is the fact it doesn't oxidize on my skin.
Anyway….cream to powder doesn't hold too much appeal for a dry skin girl like myself as powder always holds a great danger of flaking up on my skin, aging me, and just making me look washed up.
Urban Decay Surreal Skin Cream to Powder Foundation impressed. That's say alot considering how I normally don't like this style of foundation or powder in general for that matter.
The foundation comes housed in the slick translucent purple compact that UD has embraced lately with many of it's new compact releases such as their blotting powder. Goth fans shall rejoice at the purple color and graphic designs of which include a small dagger, skulls, and spiders. For those of you that aren't in touch with your goth side you'll still enjoy whipping this out of your purse for touch ups on the fly. The compact is a chunky dual chamber compartment compact which houses the foundation and a mirror on top and a brush applicator on the bottom.
Mmm yes I said brush. Surprised? I was. I expected a sponge applicator but a brush it is kids. The brush is a half moon and large in size. Bristles are slightly scratchy and I'm unsure if I'm ever going to grow accustom to brushing on a cream foundation. You see…I'm normally a finger girl and apply foundation as such. It's rare I use a sponge or brush for foundation application, to tell me otherwise is like trying to break a dog into trying new tricks. I will say the idea of applying a cream to powder is logical as the brush sweeps the cream into place and blends it nicely but I do think it needs a little pat and rub here and there from my finger to get it in place.
Now the formula….
The formula is possibly the most interesting one I've come across in regards to cream that converts to powder. I've tried a few and have always either had too powdery a finish or not a powder finish at all. This almost instantly, upon contact with your skin, turns quickly to a powder. It's buttery to the touch at first and quickly turns into a velvety smooth finish. So far I quite like it. I admit I've ditched the brush and have been applying with a sponge as I feel like I get a more even finish.
Some things you should know…..
I think it should work quite nicely with oily skin. Do prep alot prior to use. If your skin isn't properly prepped you'll feel like you have a mask on and the foundation will settle on your skin in a most undesirable way, believe me I know as I tried it on unprepped skin and proceeded to look terrible. Be sure your skin is nicely moisturized prior to use also apply primer as it creates a nice working canvas for the makeup.
The foundation wears for around 4 hours on me before I need a touch up but I can stretch this wear with proper priming to around 5 hours. One thing that delights me is how easy it is to retouch up as it's as simple as applying powder. Sweep on with the brush or smooth on with your sponge, 1, 2, 3 retouch! I really loved how the foundation managed to stay fresh as well without oxidizing or turning gray, ashy on my skin.
Overall, Urban Decay Surreal Skin Cream to Powder should appeal to anyone that's a fan of cream to powder finishes. The foundation has the potential to become a great product for Urban Decay as much of their Surreal Skin collection is quite good. The range of shades available should appeal as their appears to be a shade that suits everyone.
Urban Decay Surreal Skin Cream to Powder Foundation is available now from Urban Decay and later this month at Sephora.
For your chance to win an Urban Decay Surreal Skin Compact for yourself please see this link!
Anyone indulging?
Indulged already?
What do you think?
9/24/09 7:57 Kate:
This foundation is my holy grail of foundations. i've been wearing it since august, and i dont think i'll ever wear another foundation. Urban Decay is my favorite brand of makeup, and this doesn't disappoint. I, however, get a full day's wear with this. The only time I touch up is if i have to go somewhere after work, and even then i don't always have to.
1/5/10 9:49 Kay:
Thanks for the review, this foundation looks like just what I want. But still no UK release! Gah!
Love your blog, I consult before all my UD purchases 😛
1/14/10 8:12 Kay:
As was I! But they seem to randomly omit things for release over here. For example, I have yet to find the Universal Mineral Powder. There are 6 lipsticks that have never appeared over here, Frigid shadow is nowhere to be seen, and we only got one of winter's 24/7 sets! Maybe they think we're not ready 😉 Either that or I'm not looking hard enough!
Can't wait to see the review, loving the look of everything! Loving the sparrrrrkle!
1/14/10 15:26 Kay:
I am on MUA yeah, so maybe I should go for that approach. There's only so long you can wait before you have to give up on getting stuff here! 😛
I'm not on twitter actually (maybe I should be XD) but I have seen bits and pieces of the sparkle and quite a lot on the Alice palette! Looks gorgeous but I want to find out what the shades are, since they're renames of existing shadows. At least I know we're getting her over here!
1/14/10 16:08 Kay:
Thanks so much, at least I'll have a month and a half to contemplate getting it after the US release 😛 Wooo! *goes to log on to MUA*
1/14/10 16:14 Kay:
Aw, thank you so much! I'm sure there will be more nice people out there to help a poor UD starved Englishwoman 😛
1/21/10 12:55 Shel:
Do you have any advice for "setting" this foundation? It is nice and creamy but it stays a bit too creamy and moist for me. I was hoping it would turn to a nice powder finish that I didn't have to constantly touch up. Do you use the UD powder over this?
5/1/10 12:50 Minttu:
I am dying to now what is your favourite cream foundation..(S)?:)
So far I am happy with my Arden flawless finish sponge-on and Blasco.I finally got my MUFE face&body and I LOVE IT! I am also sucker for cream to powder and powder compacts allthough I have quite dry skin.
So,miss Beautiful,would you PLEASE tell me ALL your favourites..
5/4/10 6:47 Minttu:
When can I find it? Weird,never heard.
Anythin more common in Europe Or USA?
What is your favourite mousse make up,that quality of foundations never tried but think they don`t cover much
My Guerlain parure aqua compact is coming,think,
Any BBcreams for europian markets..?
5/8/10 13:10 Minttu:
Hi pretty!
I have A stupid question..how do you use Macs mineralize skinfinish?
My friend has it for brunettes and she might sell it to me.She also has those in few another colours.
I found a great mousse foundation; Isadoras 16h active moist make-up and spf 30!
What do you think is Macs best foundation for normal/dry skin for at least medium coverage? Thinking about maybe studio fix or stick.
Bobbi Brown also has moisturizing cream compact,might like that.
But my next order is absolutely MUFEs HD..
I'm glad I'm not the only one who shelved the brush. I tried a foundation brush and it was ok. I'll be trying a sponge instead! ^_^
12/6/10 12:36 passerby:
Hi there. May I know what shade is your UD Surreal cream to powder foundation in? I'm a mac NC25, what shade do u recommend?
12/20/10 22:37 Whit:
I am having some problems with dry spots with this foundation. For the most part I have pretty oily skin. I always moisturize and prime my face before application. Perhaps it is because the winters are very dry in the desert? Should I wait until summer to try this again?


text:
- Northern New York Newspapers
- Watertown
- Ogdensburg
- Massena-Potsdam
- Lowville
- Carthage
- Malone
- NNY Business
- NNY Living
- NNY Ads
Below is my story on SLU coach Joe Marsh and his decision to take a medical leave from the team, the length of which he does not know. I will have more later in the week, including excerpts from my 40-minute interview, additional interviews and player reactions.
CANTON — St. Lawrence University men's hockey head coach Joe Marsh will not be behind the bench to start the 2011-12 campaign.
He plans to take an indefinite medical leave and has applied to do so through the university.
Marsh, now in his 27th year as head coach at SLU, missed several games and countless practices last season while battling a chronic arthritic condition he's had for several years. It became the enduring narrative of the season's final three months.
He is confident, though, that he will regain control and return to coaching this year.
"What I'm looking to avoid is a continuance of how last year went — was I in or was I out?," Marsh said. "Who knows? Maybe I'll come back soon, maybe I'm not coaching again, I don't know. Whatever happens, I want it to be my decision and do it the right way and leave it the right way. I've done this for half my life, so obviously what's left is pretty important to me."
The pain associated with his condition makes it impossible for Marsh to wear a skate or shoe, stand behind the bench or coach the team the way he always has — with vitality.
"You can't be in and out," he said. "You have to be in it, day after day and feel it, smell it, taste it and be in it. I've always coached from a more guttural, emotional level. You have to have a finger on the pulse. I'm not there, yet."
Newly appointed associate head coach Mike Hurlbut and assistant Greg Carvel will handle the day-to-day duties. Each has extensive professional coaching or playing experience. Carvel spent 15 years coaching, most-recently with the NHL's Ottawa Senators.
Marsh also hopes to bring in former Saint Jared Keller to help with recruiting efforts.
"I think we could be pretty good," Marsh said. "I want the attention on that. (Hurlbut and Carvel) are not novices. Carvel coached in the Stanley Cup Finals. We've got a very experienced coaching staff."
Last season, Hurlbut, then an assistant, was in a similar position next to then-associate head coach Bob Prier. The duo controlled practices and coached the four games Marsh missed.
"We're very confident in our abilities and I know Joe is confident in our abilities," Hurlbut said. "We're focused and the (players) understand the situation. With my playing experience and Greg (Carvel's) coaching experience, I think it's a good blend here. We're very comfortable with the state of the team and the program."
St. Lawrence senior captain Jacob Drewiske approached Marsh shortly after he privately announced his plans to the team last week. Drewiski assured him the team was as strong as if Marsh was an everyday presence.
It is a team that, despite Marsh's sporadic appearances late last year, upset Princeton in the ECAC Hockey Tournament and pushed top-seeded Yale to a decisive third game.
"The team's in good hands and that starts first and foremost with the two assistants that we have," Drewiske said. "We have a lot of faith and trust in those guys. We just want (Marsh) to be healthy, and at the same time it inspires us to play harder as a team. Maybe what we do on the ice can help him heal faster somehow, some way."
Ultimately, Marsh wants his program to focus on what it needs to do to be the best it can be and not get distracted by his off-ice dilemmas. For those reasons, he will spend at least the start of the season on the periphery.
"I want the emphasis on them and what we have here," Marsh said. "Not the fact that I'm sitting around on the shelf. I don't want to mess it up by mistake by being half-in, half-out. It's better for everybody if I just sort of step back."
Thousands of readers like you have made Northern New York Newspapers the number one ranked website* for local news and sports coverage in the north country.
You can continue on to this article in a few seconds, or you can visit one of our websites: