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"Soul Asylum's 1998 Professional Undoing Candy from a Stranger: Is it Their Best Album?"
I can no longer contain myself… this Soul Asylum album from '98 is GREAT. It's everything '90's rock was at its best — it's powerful, gritty and honest, it wields trenchant tension pitted by one scale against another (major/minor), it's got snuff juice in its joints, limbs swirlin' and whirlin'.
And as many know, and as, yes, I've past detailed on this site, it did get them dropped from their label (Columbia). [1] Now, in order to avoid actually knowing the label's reasoning behind this, [2] I've abstained from doing too much research on it, but I have a feeling their offices are like one big beer-bonging frat party… they probably wanted bands like Lit's "The car is in the front yard" juvenility or Eve 6's concentrated confrontation. [3]
So all of these cultural accoutrements, with rap/rock and boy bands to follow in addition to Eminem's much-excused outright weirdness, clearly occlude what was happening in rock and roll, a baton I think would be picked up most worthily down the line by the Asylum's Midwest brethren The White Stripes, with deliberate rondos like "I Want to Be the Boy" and then the raucous party "Girl, You Have No Faith in Medicine." The Stripes, that is to say, were not a THROWBACK the way The Strokes and definitely Franz Ferdinand were (The Libertines were fine but somewhat ephemeral) — it struck the listener as music irreplaceable that when absent, should have been there, to fit in as a piece of a larger puzzle, one possibly likewise contributed to by forerunners from Zeppelin to the Meat Puppets and Sebadoh.
But songs in the mainstream in the late '90's were almost like little mini-reality shows: even Everclear, who I have to say rock with ear-splitting abandon, definitely had a penchant for melodrama, taking pot shots frequently at kissups and hypocrites, or people THEY SAW as hypocrites. [4] What's an old sh**-kicker with nothin' to lose to do these days? Soul Asylum seemed destined for indie rock all the way, though I guess this didn't work out, the way it maybe did for Jawbox's J. Robbins who would go on to not only start his own label (De Soto), but belt out some timeless post-rock in the indie sphere with newly assumed outfit Burning Airlines.
Soul Asylum always had a knack for what I'd like to call creative dissection — copying, in other words, ripping off a riff from another song and using it in their song (the guitar intro for Rush's "Fly by Night" for their own intro in the still excellent "Black Gold" being a case in point), and I detect a hint of this also on Candy's second track "I Will Still Be Laughing" which mind you made it worthily onto the band's closely subsequent greatest hits. That is, I think I hear a strain from the song by Seal "Prayer for the Dying," but again, the effect is positive and altogether uplifting — that is to say, no listener would assume the position of denial toward Dave Pirner's worthiness of the position. It'd be like disrupting an old person who's playing bingo, or watching his or her grandson or granddaughter playing in the sandbox, and really having a good time. I mean, he seems to be a genuine muse relaying a celestial cluster family of rock vocal, and the whole band is locked in a tight groove throughout this whole album, with kinetic energy breaking and crumbling out of their very joints like phosphorous. What I'm saying is, let's not overestimate how cool the '90's were. This muse had to come from a certain snowballing of feeling, a certain introspection to create the fermented stew of outpour.
[1] In my opinion, it's all the fault of those organized crime goons Smash Mouth and all their intimidating boorishness.
[2] For anyone who doesn't remember, the late '90's actually kinda sucked, I mean Celebrity Deathmatch was one of the most popular shows, for Christ's sake. Also there was that whole "swing" revival thing.
[3] I once saw Eve 6 in Chicago and to my eye Max Collins was deliberately attempting to appear homosexual, traipsing around in a tight white shirt and red leather pants and asking something like "Does this outfit showcase my figure well?" Anyway, they definitely make for more DRAMA, with the unsettled semantic specificity of "Tongue Tied" and "Leech."
[4] As Nirvana had already proved, bashing public figures in high-profile interviews is a readily demonstrated Northwesterner's habit.


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A 3 Day Route In Austria
Every time I go home to Austria, I love discovering new parts of the country. The last time I visited, my brother and his wife suggested
News for nerds
Every time I go home to Austria, I love discovering new parts of the country. The last time I visited, my brother and his wife suggested
MLBPA executive director Tony Clark told Bob Nightengale of USA Today that he is "optimistic that the possibility of playing still exists" in 2020. So
Switzerland announced on Thursday a three-stage easing of the restrictions imposed to suppress the COVID-19 pandemic, with some shops and services allowed to reopen from
"As shutdowns continue, job losses will likely extend into other areas of the labor market," warned one analyst. The Dow Jones Industrial Average ticked upwards
If you're wondering where to buy an Xbox One, you're not alone, with stock starting to slip there are only a few bundle deals still
A Michigan mother is accusing her local school district in a lawsuit of not protecting her 15-year-old son from months of anti-gay bullying and abuse
Six years ago today, the Giants released pitcher Dontrelle Willis from his minor league contract. He would have just one more attempt to reach the
According to a clinical study conducted by the School of Chinese Medicine (SCM) at Hong Kong Baptist University (HKBU), nearly 90% of the patients who
Comcast's new streaming service, Peacock, could debut earlier than planned, the company said Tuesday, citing the "unprecedented and historic turn of events brought about by
Mobile World Congress (MWC) will continue to be held in the city of Barcelona until 2024 as per the terms of a new agreement. The

subset: commoncrawl


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This year both the High Carbon Stock Approach (HCSA) and Palm Oil Innovation Group (POIG) hosted booths at the 16th Annual Roundtable Conference on Sustainable Palm Oil (RT16), staffed by the Helikonia team. This year marks a pivotal point for the Roundtable on Sustainable Palm Oil (RSPO) and the sustainable palm oil community as the updated Principles and Criteria were adopted at the General Assembly, which reflects new requirements to halt deforestation and to improvements in human rights protection. Both the HCSA Steering Group and POIG members have diligently worked to contribute to the Principles and Criteria consultation process over the past one and half years.
RT16 was held at The Magellan Sutera Resort, Kota Kinabalu, Malaysia from 12-15 November 2018, with over 850 representatives from the global palm oil industry in attendance, including corporate leaders in sustainability, financial institutions, policymakers, academics, and social and environmental NGOs from around the world.


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Security as a Priority
September 28th, 2016 by admin
Looking for the best surveillance system that uses leading technology to reliably secure your facility? BTI can help you manage your business more efficiently by making sure you and your employees are safe. When safety is our priority, you can be assured that you will get the reliable, foremost technical quality and most secure, user friendly platform out there.
Our Axis systems are market leaders in the digital surveillance business. Our access control systems can help you minimize the cost for locks and doors via integration of electronic readers. Additionally, our fire and alarm systems can protect your business from many threats. Find out more about our surveillance and surveillance products at: https://www.btigroup.com/solutions/security.aspx
Posted in: BTI, Industry Solutions


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Pastoral Care Training
For 2020 the diocese is once more commending an accredited Pastoral Carers Training Course which has been well received by our churches in previous years.
The course will enable participants to develop their understanding of pastoral care from a Christian perspective and to become more effective in relating to and caring for the people and communities around them. It is run by ACC and Pastoral Care NI.
Who is the course for?
- those who see themselves helping others informally in their daily relationships and who desire to learn more about pastoral care generally without being part of a pastoral care team.
- those who may be members of a pastoral care team and who desire to learn more about pastoral care in order to help others both informally and formally.
- those in church leadership or who have specific church roles and who desire to develop their awareness of pastoral care in order to support those for whom they have responsibility.
- those who wish to help others in specific community and workplace settings and desire to learn more about pastoral care in outreach and mission.
- those who desire to explore a call to pastoral care.
Starting date: Tuesday 11 February 2020
Venue: Christian Fellowship Church, 10 Belmont Road, Belfast BT4 2AN
Cost: £90 (non–refundable)
How do I apply?
To find out more and to apply please download the leaflet here. Note that you should discuss your application with your rector as they will need to recommend you by letter or email.
The diocese offers Down and Dromore participants subsidies from £90–£60 (please notify the tutor on application).


text: Choose Options Quick view Crown 2 Wire 1/2" CROWN PREMIUM Rope Light - 206CP $42.75 - $73.50 Crown Premium 1/2" 2 Wire Rope Light NOTE: Above shipping charge is per roll. Cost is based on shipping UPS Ground within the continental United States. If faster delivery is needed, or shipping outside the US, the additional charges will be... $42.75 - $73.50 Choose Options Quick view


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Recent Posts
- Catalysis Today: 'Prof. Dragomir Bukur' Special issue, edited by prof. Elbashir is published!
- A new news article published in an international news outlet about the group research activities
- News from our group
- CaRE-Fuel Lab researchers develop natural gas processing technology that could reduce Qatar's carbon footprint
- National and International newspapers highlight the research work of Prof. Nimir Elbashir group and TEES Gas and Fuels Research Center


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A Community-Centered Project
Bringing Fiber-Based Internet Service to Lyme, New Hampshire
What is LymeFiber?
LymeFiber is a community-centered project offering fiber-optic Internet data service to all residences, businesses, and public organizations in Lyme. Levels of service range from 25 Mbps up to 800 Mbps. All service is symmetrical - same speed upload and download - and are free of data caps.
Monthly rates begin at $72. Residential and business phone lines will be available as an option starting at $25 per month.
We will begin to offer services in Fall 2020. For more information, see our Frequently Asked Questions page.
- Universal Coverage. We aim to provide high-speed, fiber-optic-based Internet service to every reachable business and residence in Lyme.
- Net Neutrality. LymeFiber will not throttle or otherwise interfere with your data.
- A Future-Proof Design. Fiber-optic cable provides almost unlimited capacity for whatever new applications come along.
- Competitive Pricing. The cost of our service will be competitive with similar service elsewhere in the region.
- Community Focus. Bringing a needed service to Lyme - our community - is our primary goal.
LymeFiber News
LymeFiber Trucks on the Job
In January, people started to see LymeFiber trucks working in Lyme. This photo shows Klee Smith, a contractor for ValleyNet in front of Lyme's horse sheds. Klee is conducting a pole survey to create an inventory of all the poles along with their geographic coordinates.
Over 200 signed up!
As of January 2019, the number of signups crossed 200! We're expecting that between 300 and 400 people will ultimately sign up by the time we begin to provide service in Summer 2020. There's no obligation, but if you would like to indicate your interest in a high-speed fiber connection, please fill out the form …
Signups for LymeFiber now open
We opened the signup process for LymeFiber in mid-December 2018 to allow Lyme residents to express an interest in becoming a customer of our universal high-speed internet service. After just two weeks, right before Christmas, over 160 people had already signed up. There is no obligation – we will get back to you to confirm …


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Yep, it's real. The Canon 7D is an 18MP semi-pro DSLR that shoots 1080p video in 24—or 30—glorious frames per second for $US1899, kicking Nikon's previously unchallenged D300s where it hurts. But it's also an odd little beast.
The 7D feels like Canon took the results of a survey they handed out to people about what they wanted in a camera and crammed 'em all into one product targeted at semi-pros. Full HD video with manual exposure in 24, 25 or 30 frames per second, check. More rugged, weatherproof body than 5D, check. Customisable buttons, including a new multifunction button, check. A dedicated button for switching to RAW+JPEG mode. Um, check. Electronic axis level? Also check. It has dual DIGIC IV image processors, the first model outside of the pro 1Ds line with dual image processors for fast burst shooting: 8FPS with 94-shot JPEG bursts (124 with UDMA card) or 15 RAW shots, all at full resolution with 14-bit A/D conversion.
But, it's not full-frame: They've crammed 18 megapixels into an APS-C-sized sensor (like in the Rebel series or 50D, versus full-frame in the 5D) with an ISO range from 100-6400, and a Hi setting of 12,800. Canon says they've shortened the distance between the photodiodes in the sensor, which decreases light falloff, supposedly translating into better high ISO performance.
We got to shoot with a beta pre-production model for a little while in midtown using a couple of Canon's new EF-S lenses—a 15-85mm ($US800, coming in October) that's replacing the old 17-85mm, and a 100mm macro lens ($US1050, out in September)—so you can see some of the low-light results, along with other samples, below. (Again, Canon would like us to reiterate that the pictures are from a pre-production sample, i.e. not final product.)
Here's a close-up on those guitars comparing the different ISO levels. At ISO1600, it's not so bad, even if it's not 5D level, but it starts getting kinda gruesome at ISO3200 (click to make bigger):
It might just be the best video on a DSLR yet: Manual exposure from the get-go, and oh yeah, the ability to shoot 1080p video at 30, 24 or 25 frames per second. If you cut it to 720p, you can shoot up to 60FPS. Also, video is encoded using H.264 now. There's a dedicated switch to flip from shooting stills to video that encircles a start/stop button for recording, so it's much easier to get right to shooting video than the 5D. You can see some sample videos below:
Everything Else
Okay, so what else? Focusing system has 19 cross-type points, with a new way to focus in addition to the usual full-blast autofocus vs. single-point—zones, which are clusters of points. You can also lock different default focus points for horizontal and vertical orientations, so you can turn the camera back and worth without having to repick your focus point. (Setting it up is a little confusing—even the Canon rep took a minute to make it happen.) AI servo focus is supposedly smarter, so it tracks moving objects better, and it works with a new light-source detection system that adjusts for flickering light. (We didn't get a chance to check that out.)
The viewfinder provides 100 per cent coverage, like the 1Ds, and it uses a new polymer LCD network with a different graphics overlay than the 5D—that's a lot like what Nikon's got in the D300, actually—so it can do things like display the neat new 3D-axis electronic level in the viewfinder (which feels like an iPhone tilt game from hell if you're holding the camera by hand trying to level it on both axes). In a first for Canon, the built-in, wide-coverage flash can command other wireless flash modules.
Oh, did I mention a ton of new buttons? Seriously: A dedicated RAW+JPEG button that'll shoot your next shot in RAW+JPEG if you're only shooting one at the moment. A new start/stop live view button with a control ring to quickly switch from live view to video mode. There's also a new multi-function button on the top right. Aaand another for the custom controls menu, where you can assign different features to different buttons if you want.
Honestly, a lot of the deep customisation and new interface commands were confusing, at least in the short time I spent with the camera—like setting different autofocus points for whether you're holding the camera vertically or horizontally. There was some other shortcut that required pressing and holding two buttons at once on the camera. And I never quite understood how to use the multifunction button or pick what graphical display showed up in my viewfinder. I think you would learn a lot of that stuff in time, though. In terms of overall handfeel, I slightly the ergonomics and weight of the D300, FWIW.
Final Thoughts
While we can't give any final judgements on this in some ways odd camera until we get a review unit, one thing seems certain: The value proposition for Nikon's $US1800 D300s—which might've already seemed dicey, given that it's the guts of a two-year-old camera with a dash of limited 720p video—just got a lot less enticing, simply on account of the 7D's fairly wide video powers. That's before you even take anything else into consideration, like more pixels for more crop potential (if the pixels are good ones, that is). Obviously, we'll be head-to-heading these two as soon as we can. Which should be shortly, since the 7D is due by the end of next month, at $US1900 for the kit with an 18-135mm f/3.5-5.6 lens, or $US1700 body only.
AU: Local pricing is still TBC for the camera and the new lenses, unfortunately. –NB


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Give Them the VIP Treatment with Crystal Awards & Gifts
Recognizing customers "just because" they are great customers helps them feel connected and drawn to your company. By acknowledging your customers, you show them just how important they are to you. Going one step further and treating a customer like a VIP can help them intuitively become a brand ambassador for you. When you focus on customer appreciation, it can make a huge difference to your company and your brand.
The best rewards come in the form of acknowledgment and appreciation – offered in both words and symbolic items. It is worth spending a little extra on customers that are engaged and care about your business.
Consider coupling a handwritten note of appreciation with a small crystal keychain, crystal diamond, or crystal rectangle. These gifts, and the others pictured below, are made of flawless optical crystal and support a variety of imprint process options. This simple gift comes in beautiful black packaging that adds to its value. Gifts like these can be sent out on the spot to your best customers by your sales or customer relations team.
This appreciation idea, along with an article, can be downloaded by clicking here.
Keychain Lighted Rectangle
Keychain Lighted Square
Elena Bowl
Denton Diamond
White Swirl Award
Alliance Goal-Setter
Triumph Trophy Vase
Clipped Globe
Denton Rectangle
Like these ideas and need additional options?


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Construction Clean Up Las Vegas
Construction Clean Up Las Vegas
No matter the job, construction waste can be cumbersome and time-consuming to remove. But at A Track Out Solution, construction waste is no problem at all. We specialize in fast, efficient, professional waste removal in Las Vegas, NV. When you call us, your construction waste is off your property—and off your mind—in no time.
When you need debris, junk or commercial waste gone in a hurry, A Track Out Solution is the waste removal service you can count on to provide quick, hassle-free service. Call A Track Out Solution in Las Vegas, NV today to learn more about our pricing and services.
Residential and Commercial Clean Up
At A Track Out Solution, we take quality and customer service very seriously, which means you can expect nothing less than the best when you choose to work with us. We know that prompt, reliable Construction waste removal is important to the continued success of your business. That's why we work hard to ensure your continued and complete satisfaction with our service, exceeding your expectations every chance we get.
Construction Clean-up Services
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Our team considers your total satisfaction with our service to be our highest priority. We go the extra mile to exceed your expectations at every turn. As our client, you can expect attentive service and outstanding results. We simply wouldn't have it any other way.
At A Track Out Solution, we pride ourselves on being the best in the business. We've worked hard to build a reputation of excellence, and we're committed to doing everything possible to maintain it. So if you're stuck with piles of construction waste, one call to A Track Out Solution is all you need to make the problem go away. Contact us in Las Vegas, NV to schedule a pick-up or request an estimate.
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Sun: Closed


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How Invention Ideas and New Technology are Helping Businesses
They feel that that necessity is a new mother associated all innovations. Nowadays, the boom in technology ensures and enables the distribution of new inventions toward interested part in people should. Social media networks and other samtale sites also help returning to spread the exact word more or less inventions and make the type of people concern to do new things.
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Nowadays, items are most of the time based in new advancement. A lot of organisations and businesses depend on new technology to be sure the profitability of their enterprises and to particular that his / her processes can be efficient as customer well-behaved.
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While Hilda and Paulina enjoy their raunchy nylon licking they both break out hard plastic dildos to work themselves into frenzy. These two nylon licking lesbians plow into their hot moist pussies and lick up the spent hot pussy juices that stream down their nylon covered legs. Now their face to crotch mashing can gobble up the juices and have them cumming in waves.
View Gallery :: Brought to you by LickNylons.com @ FerroNetwork
Check Official Reviews to learn more about FerroNetwork sites


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Montreal, 1951. Rebecca Wiseman, 18 years old, from a Catholic-Jewish family, lives with her parents in Outremont. At a local dance, she briefly meets a handsome young man, but has little hope of seeing him again. When Sol Gottesman tracks her down and asks her on a date, her joy mingles with disbelief when she learns he is the son of a wealthy Westmount businessman.
When Sol takes her in a chauffeured Rolls-Royce to the most expensive restaurant in the city, Rebecca enters a world of upper-class wealth and privilege unknown in her working-class family. Despite the usual bumps in any relationship, Rebecca believes her life is perfect.
She soon learns that despite Sol's outward charm, he lacks self-confidence. On a visit to Mount Royal overlooking the city, Sol reveals the simmering conflicts in his family and his fears that his brother plans to drive him out of the family business. Rebecca's wants to protect Sol, but helping him stand up to his family, puts her squarely in the midst of it all.
Class, religion, family conflict and sexual secrets test their love and Rebecca struggles to control events. And then, a late night telephone call changes her life forever.
Echo from Mount Royal
Cover in category: 'Fiction' > 'Romance'
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Rapid Endothelialization of Off-the-Shelf Small Diameter Silk Vascular Grafts
Author + information
- Received June 30, 2017
- Revision received December 13, 2017
- Accepted December 15, 2017
- Published online February 26, 2018.
Author Information
- Elysse C. Filipe, MSca,b,
- Miguel Santos, MSca,b,
- Juichien Hung, MSca,
- Bob S.L. Lee, MSca,b,
- Nianji Yang, MPhila,b,
- Alex H.P. Chan, BSca,b,
- Martin K.C. Ng, MD, PhDa,c,
- Jelena Rnjak-Kovacina, PhDd,∗∗ (j.rnjak-kovacina{at}unsw.edu.au) and
- Steven G. Wise, PhDa,b,∗ (steve.wise{at}hri.org.au)
- aApplied Materials Group, The Heart Research Institute, Sydney, New South Wales, Australia
- bSydney Medical School, University of Sydney, Sydney, New South Wales, Australia
- cDepartment of Cardiology, Royal Prince Alfred Hospital, Camperdown, New South Wales, Australia
- dGraduate School of Biomedical Engineering, University of New South Wales–Sydney, Sydney, New South Wales, Australia
Visual Abstract
Electrospinning of silk to create nanofibers, which deposit onto a rotating collector. This results in the formation of a pure silk conduit of 1.5 mm internal diameter. These conduits are then implanted into the descending abdominal aorta of Sprague Dawley Rats with end-to-end suturing, and left for 3, 6, 12, and 24 weeks. Endpoint histologic analysis of the explanted grafts demonstrate hyperplasia stabilization, complete endothelialization and excellent blood compatibility.
• Bombyx mori silk was electrospun to engineer mechanically robust vascular grafts that support endothelial cell attachment and monolayer formation while resisting blood adhesion and fibrin assembly in vitro.
• Silk grafts were evaluated in a rat aortic interposition grafting model at 3, 6, 12, and 24 weeks. Graft survival for silk grafts (95%) was superior to control ePTFE (73%) at 24 weeks.
• Comprehensive histologic analysis of the grafts demonstrated near complete endothelialization by 6 weeks, which occurred concomitantly with neointima regression, characterized by a loss in cellularity, a phenotypic switch in SMC and an increased ECM expression.
• Silk grafts appear to undergo positive remodeling over time, with temporal reductions in apoptotic and inflammatory cells, adventitial granulation tissue, and the deposition of new ECM proteins.
Synthetic vascular grafts for small diameter revascularization are lacking. Clinically available conduits expanded polytetrafluorethylene and Dacron fail acutely due to thrombosis and in the longer term from neointimal hyperplasia. We report the bioengineering of a cell-free, silk-based vascular graft. In vitro we demonstrate strong, elastic silk conduits that support rapid endothelial cell attachment and spreading while simultaneously resisting blood clot and fibrin network formation. In vivo rat studies show complete graft patency at all time points, rapid endothelialization, and stabilization and contraction of neointimal hyperplasia. These studies show the potential of silk as an off-the-shelf small diameter vascular graft.
In the absence of appropriate autologous grafts, revascularization of ischemic tissue is reliant on the availability of effective synthetic conduits. In applications in the coronaries and lower limbs, where diameters are <6 mm, current synthetic graft materials uniformly fail, driving significant research in bioengineering, aiming to generate viable alternatives (1,2).
However, more than 2 decades of research has failed to provide the ideal platform for engineering of small diameter synthetic conduits for clinical use. A significant proportion of preclinical research has used vascular cells as a tool to produce a native scaffold, rich in extracellular matrix (ECM) components. This is frequently in conjunction with lengthy maturation phases ex vivo (1,2). The most successful have demonstrated feasibility in large animal models (3–9). However, the use of cell-containing scaffolds or cell by-product scaffolds poses a significant limitation to the clinical translatability of such approaches, both logistically and financially. The most promising approach to bioengineered conduits are decellularized before use to reduce immunogenicity and improve translation. A recent phase II human trial has demonstrated the clinical feasibility of this approach, but additional studies of efficacy are still required (9). Similarly, traditional polymeric scaffolds such as polyurethane have also failed to improve clinical outcomes due to unexpected degradation in vivo, pro-inflammatory by-products, and poor interactions with vascular cells (1,10,11). Therefore, there remains a significant unmet need for new materials for synthetic vascular conduits that combine favorable mechanical properties and compatibility with vascular cells and blood, which can be readily translated to the clinic.
Bombyx mori (silkworm) silk fibroin (referred to as "silk") is a naturally occurring protein polymer with unique mechanical and physical characteristics, which make it a widely used biomaterial. It has a long history in applications as diverse as skin repair, retina replacement, and clinically as sutures (12). Recent methodological advances in silk processing have significantly improved silk fibroin purity reducing the inflammation that has been considered a weakness of silk in the past (13). In addition, advancements in silk processing and manufacturing methods have made the production of mechanically robust conduits feasible, leading us to revisit a well-established biomaterial for vascular repair applications (12,14). In this work, we demonstrate the engineering of a silk conduit, electrospun from water and cross-linked with water vapor, thus eliminating the use of potentially toxic chemical precursors. Silk conduits manufactured in this way have tunable physical properties and high vascular compatibility in vitro and were assessed in a rat model of abdominal aortic replacement. Our conduits demonstrate excellent surgical handling and in vivo patency for up to 6 months with near-complete endothelialization as early as 6 weeks with concomitant stabilization and contraction of neointimal hyperplasia. Silk grafts appear to undergo positive remodeling over time, with temporal reductions in apoptotic and inflammatory cells, adventitial granulation tissue, and the deposition of new ECM proteins. Overall, this work demonstrates the robust in vivo performance of a cell-free silk scaffold for use as a small diameter vascular graft, a simple, cost-effective platform designed to be readily translated for broader clinical use.
Full methods are included in the Supplemental Appendix.
In vivo rat model of aortic replacement
Study approval was obtained from the Sydney Local Health District Animal Welfare Committee (protocol number 2014/028). Experiments were conducted in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purpose. Briefly, electrospun silk grafts were prepared and disinfected in 70% ethanol with exposure to ultraviolet light for 30 min. Samples were stored in air until the time of surgery. Expanded polytetrafluoroethylene (ePTFE) was purchased from Bard Scientific (Covington, Georgia) and stored in air until the time of surgery. Forty-three 8-week-old male rats (Sprague Dawley) were purchased from Laboratory Animal Service (New South Wales, Sydney Australia). Rats were given a single intramuscular injection of ketamine (75 mg/kg) and medetomidine (0.5 mg/kg) to induce anesthesia. The abdomen was shaved and a midline laparotomy incision was performed. The infrarenal abdominal aorta was exposed and clamped proximally, just below the renal arteries, and distally, just above the aortoiliac bifurcation. The abdominal aorta was resected and a 0.8-cm section of silk or ePTFE (n = 21 and 22 for silk and ePTFE, respectively) was implanted with 8 interrupted 9-0 silk sutures at each anastomosis. After re-establishment of blood flow, the abdominal cavity and overlying skin were sutured closed. At the end of surgery, rats were given atipamezole (in equal volume to medetomidine; intramuscularly) to reverse the anesthesia. For pain management, temgesic (buprenorphine 2 mg/kg body weight) was provided once a day for 24 to 48 h. Animals were kept on normal food and water ad libitum for the duration of the study and no ongoing anticoagulants were given. Rats were followed up for 3, 6, 12, and 24 weeks (n = 5 per time point) at which time, animals were anesthetized (as described) and the graft excised for scanning electron microscopy and histologic analysis.
Histologic staining and immunohistochemistry
On explantation, all samples were fixed in 4% paraformaldehyde, overnight. Samples were then processed for scanning electron microscopy imaging or histologic analysis by paraffin processing and embedding (Leica Microsystems, Wetzlar, Germany). Graft cross sections, from distal to proximal were cut. For staining, slides from 6 equidistant points throughout the graft were deparaffinized and immediately stained with hematoxylin and eosin, Milligan trichrome for collagen deposition, orcein for detection of elastin, or alcian blue for detection of proteoglycans. For immunohistochemistry, slides were deparaffinized and subject to heat-mediated antigen retrieval prior to permeabilization and blocking with 10% goat serum. Primary antibodies (rabbit α–von Willebrand factor [vWF] [Dako, Agilent, Santa Clara, California]; rabbit α–proliferating cell nuclear antigen [PCNA] [Abcam, Cambridge, United Kingdom]; rabbit α-CD34 [Abcam]; rabbit α-CD68 [Abcam]; rabbit α-vimentin [Abcam]; rabbit α-caspase [Cell Signaling Technology, Danvers, Massachusetts]; mouse α–smooth muscle cell actin [SMC-α] fluorescein isothiocyanate [Sigma-Aldrich, St. Louis, Missour]) were incubated at a dilution of 1:250 in antibody diluent (Dako). Secondary antibody (goat α-rabbit Alexa 594 [Abcam]) was incubated at 1:250 in antibody diluent, after which NucBlue (Invitrogen, Thermo Fisher Scientific, Waltham, Massachusetts) was used to detect cell nuclei. Slides were coverslipped in anti-fading medium (Dako) and imaged in the Zeiss Axioscan (Jena, Germany) at 20× magnification.
Image analysis
All image analyses were performed in Image Pro Premier (version 9.2, Media Cybernetics, Inc., Rockville, Maryland), where a minimum of 3 sections were analyzed per region (distal to proximal) per graft for each stain. For endothelialization analysis, vWF and CD34-positive portions of the luminal cross sections were measured and are represented as percentage of endothelialization relative to the total circumference of the lumen of the same section. For hyperplasia and granulation tissue analysis, the neointimal and adventitial tissue areas were calculated and are represented as a percentage relative to the total lumen size of the graft in each section. For quantification of CD68+ cells, the amount of red staining was quantified in the granulation tissue and is represented as an absolute pixel value. Collagen analysis was performed by calculating the area of collagen (aqua/blue) and cells (purple) and is represented as a ratio of collagen to cells in each section. Elastin analysis was performed by calculating the total area of positive elastin staining (dark purple) and is represented as a percentage elastin relative to the total hyperplasia region of each section. Quantification of proteoglycans was performed by calculating the amount of positive (aqua/blue) staining, relative to the total hyperplasia region of each section. The SMC-α/PCNA double stain was analyzed by quantifying the total green and red areas of the hyperplasia, respectively, and data are represented as percentage of positive green or red staining relative to the total stained area in the hyperplasia per section.
Statistical analysis
Data are expressed as mean ± SEM and statistical analysis was performed using GraphPad Prism 7 version 7.02 for Windows (GraphPad Software Inc., San Diego, California). The normal distribution of our data sets was confirmed using the Kolmogorov-Smirnov test and parametric analysis was performed. Where appropriate, data were analyzed by unpaired Student's t-test when comparing 2 samples and analysis of variance when comparing multiple samples. Post hoc comparisons were analyzed with Tukey multiple comparisons test. For analysis of Figures 5E, 6C, 6D, 7C, 7D and Supplemental Figures 4B, 6B, and 7B, 1-way analysis of variance was performed on the group averages (represented as a dashed line for each time point). For group averages, all points throughout the graft were averaged for each animal (n = 4 animals). The survival of grafts over time was assessed using Kaplan-Meier survival curve analysis with a Mantel-Cox test. We considered p values <0.05 to be significant.
Silk scaffolds provide appropriate mechanical properties for use as a vascular graft material
Electrospun silk scaffolds (Figure 1A) demonstrated an average fiber diameter of 355 ± 5.0 nm and a relative porosity of 23.3 ± 1.2% (Figure 1B). The overall β-sheet crystallinity of the silk fibroin was unaffected by the electrospinning process and was increased on cross-linking, from 17.58% to 36.08%, inducing water insolubility (Figures 1C and 1D). The mechanical properties of the electrospun silk scaffolds were evaluated in comparison to commercial ePTFE and native rat aorta. The elasticity of the scaffolds was determined by the slope of the linear region of the stress/strain curve of each replicate sample. Electrospun silk scaffolds revealed a Youngs modulus of 4.2 ± 0.5 MPa, not significantly higher than the native rat aorta (2.1 ± 1.0 MPa), and 7.6-fold more elastic than ePTFE (31.9 ± 1.3 MPa) (Figure 1E). Similarly, the ultimate tensile strength of the silk scaffolds (1.0 ± 0.03 MPa) were more similar to rat aorta (2.4 ± 1.87 MPa) than ePTFE (9.5 ± 0.47 MPa) (Figure 1F). Finally, elongation data found that the silk scaffolds tolerated an increased stretch before breakage (163 ± 11.7%) when compared with the ePTFE grafts (105 ± 5.5%), albeit not significantly different to the native rat aorta (134 ± 13.2%) (Figure 1G). Silk scaffolds were then engineered into small diameter (1.5-mm) grafts (Figure 1H) for analysis of graft-specific mechanical properties. Burst pressure analysis further demonstrated the strength of the silk grafts that burst at an average of 849 ± 59 mm Hg. Although the values obtained were less than those for the native rat aorta (1,896 ± 260 mm Hg), they were in large excess of physiological blood pressure (Figure 1I). Suture retention studies showed that the silk grafts withstood an average 0.86 ± 0.1 N of force before tearing (Figure 1J). These values were below those of the native rat aorta (1.71 ± 0.3 N), but superior to previously published data for other synthetic graft materials (15).
Endothelial cells form a monolayer on silk scaffolds in vitro with excellent blood compatibility
We next sought to evaluate both endothelial cell and blood compatibility on electrospun silk scaffolds. Endothelial cells rapidly attached to and spread on the surface of the silk scaffolds, after just 3 h in culture (Figure 2A). The same cells on ePTFE remained poorly attached and did not spread on the surface (Figure 2B). Longer time points of 1, 3, and 6 days showed an increase in cell density on the silk scaffolds, with cells maintaining a spread-out morphology (Figure 2C). In contrast, the endothelial cells on ePTFE remained poorly attached even at 6 days post-seeding (Figure 2D). Endothelial cell seeding and subsequent cross-sectioning of the silk scaffolds demonstrated the formation of a monolayer on the surface, without penetrating the silk scaffold (Figure 2E). Further vascular endothelial cadherin immunostaining of confluent endothelial cells confirmed the formation of tight junctions between the seeded cells, indicative of a nonpermeable endothelial barrier (Figure 2F). Conversely, cells on ePTFE were unable to form these junctions (Figure 2G).
Blood compatibility assays also demonstrated the favorable blood compatibility of electrospun silk scaffolds. Polycaprolactone (PCL) was chosen as a control for electrospun scaffolds, as this polymer yields a material with similar fibrous morphology to the electrospun silk and which has been well validated as a synthetic graft material (16,17). Contact with platelet-rich plasma enriched with fluorescent fibrinogen demonstrated negligible fibrin network formation on the surface of the electrospun silk, while both the ePTFE and electrospun PCL control scaffolds displayed extensive fibrin networks at the same time point (Figure 2H). Quantification of the fluorescence intensity confirmed 6.6- and 7.8-fold less staining on electrospun silk when compared with that of ePTFE and PCL, respectively. Contact with whole blood further validated these results, with macroscopic images demonstrating the complete absence of clot formation on silk, while significant clot formation was visible on both control scaffolds. Scanning electron microscopy confirmed this, with almost no red blood cells on the surface of silk. The ePTFE and PCL scaffolds displayed extensive red blood cell deposition (Figure 2I, black arrow) with clear fibrin network formation in the latter (Figure 2I, white arrow).
Silk grafts display high patency at 6 months in a rat
Having confirmed appropriate mechanical and biological compatibilities of the silk scaffolds, small diameter silk grafts were implanted into the abdominal aorta of Sprague Dawley rats (Figures 3A and 3B). Control animals were implanted with commercial ePTFE of similar diameter (Figure 3C). Silk scaffolds demonstrated excellent handling and were successfully implanted in all animals, with explantations performed at 3, 6, 12, and 24 weeks. Graft survival was 95% (20 of 21) for the silk grafts, with only 1 death within 24 h of the implantation (Figure 3D). However, ePTFE failure was higher at 27% (6 of 22) with graft failures relating to deaths occurring post-operatively (n = 2) or, alternatively, at the explantation time point of 3 (n = 2) and 6 weeks (n = 2) due to complete occlusion (Supplemental Figure 1). These results demonstrate a high degree of variability in ePTFE survival, which is consistent with observations in humans (18).
Rapid endothelialization, complete by 12 weeks
The rate of endothelialization following injury is a critical determinant of vascular lesion formation and areas of injury that rapidly endothelialize have significantly less intimal thickening and stenosis, while also deterring thrombus formation (19). To evaluate this, we first performed scanning electron microscopy of the luminal side of the grafts at each explantation time point. The images reveal that silk grafts have complete cell coverage as early as 3 weeks, with no underlying silk visible (Figure 4A), a feature that is maintained at all later time points. Higher magnification further showed a rough cobblestone cell morphology at 3 weeks, changing to a more elongated morphology by 6 weeks (Figure 4B). The resolution of the images was not sufficient to allow more detailed morphological analysis of the cells. Conversely, the scanning electron micrographs of the ePTFE grafts revealed a bare surface, with the underlying material morphology clearly visible at 3 and 6 weeks (Figures 4C and 4D). At later time points of 12 and 24 weeks, focal points of cell coverage were observed; however, large areas of uncovered ePTFE remained. Immunohistochemistry of graft cross sections with vWF was performed to quantify endothelial cell presence, which is consistent with multiple previous studies (3,4,8,15,20). Silk graft cross sections showed endothelial cells present as early as 3 weeks, with complete coverage of anastomotic regions by 6 weeks and full endothelialization by 12 weeks (Figures 5A and 5B). The vWF staining of the ePTFE graft cross sections was confounded by significant non-endothelial cell–specific staining, particularly at earlier time points. This may be due to physical collection in the porous graft wall, as previously reported (21) (Supplemental Figure 2). Additional staining for CD34+ endothelial progenitor cells was also performed to establish the contribution of these cells to endothelialization (17). Results show CD34+ staining throughout the length of silk grafts as early as 3 weeks (Supplemental Figure 3A). This was shown to increase at later time points, consistent with the vWF data (Supplemental Figure 3B). Overall, the degree of CD34+ staining was lower than for vWF, suggesting that although these cells contribute to endothelialization, additional mechanisms, including transmigration are also playing a role (22).
SMC-driven neointimal hyperplasia stabilizes and contracts by 6 weeks in vivo
The major driver of long-term graft failure is neointimal hyperplasia within the graft lumen. We extensively characterized this by evaluating the extent of neointimal formation over time, while also investigating the phenotype of the SMC present. Hematoxylin and eosin staining of the graft cross sections demonstrated the characteristic bell-shaped neointima distribution from distal to proximal throughout the graft, with greater amounts observed at the anastomotic regions (Figures 6A and 6C). At 6 weeks, however, hyperplasia was equal throughout the graft length at an average 40.9 ± 3.8%. Interestingly, hyperplasia at later time points was reduced to 27.9 ± 1.9% and 27.0 ± 0.3% at 12 and 24 weeks, respectively. These data suggest that hyperplasia stabilizes by 6 weeks, corresponding to when endothelialization is also almost complete and potentially contributing to the reduced proliferation of SMC. This phenomenon has been called neointimal retraction and is similarly observed in cases of human stent implantation (23,24), generally at later time points of >18 months.
To better understand the change in hyperplasia composition over time, we evaluated ECM deposition (collagen, elastin, and proteoglycans) and SMC phenotype. Milligan trichrome staining of the graft cross sections revealed a loosely packed collagen matrix with abundant cell presence at 3 weeks (Supplemental Figure 4). This abundance of cells was lost at later time points, with extensive and compact collagen detected instead. Staining of caspase-3+ apoptotic cells, evident at early time points but reducing over time (Supplemental Figure 5A), demonstrates cell death consistent with this overall loss. The progressive loss of cells is similarly observed in stented human samples where a decrease in SMC is accompanied by a regression in percentage stenosis, the point at which some investigators suggest healing is complete (23,24). Orcein staining of the graft cross sections showed increasing levels of elastin secretion in the hyperplasia, with greater amounts detected at longer time points (Supplemental Figure 6). Elastin secretion by SMC has been attributed to a contractile SMC phenotype (23,25), suggesting an increased presence of mature SMC in the hyperplasia layer at these later time points. Alcian blue staining also demonstrated increasing levels of proteoglycans in the hyperplasia, particularly in the mid-section of the graft, where levels increase by more than 5-fold from 3 to 24 weeks. Similarly to elastin, proteoglycans are recognized as important regulators of SMC phenotype, promoting maintenance of contractile phenotype and reducing SMC proliferation (Supplemental Figure 7) (26). Finally, a double-stain immunohistochemistry for PCNA, a proliferation marker, and SMC-α, a marker for mature SMC, was performed to evaluate the proportion of proliferative/stable SMC in the hyperplasia layer. The results clearly demonstrate a phenotypic switch in the cells that at early time points are mainly PCNA+, but at later time points (from 12 weeks) reveal a dominance of SMC-α+ cells (Figures 6B and 6D). These results indicate the increasing stability of the neointima that initiates as early as week 6 post-implantation.
Granulation tissue and inflammation significantly reduces over time
The formation of granulation tissue is an expected outcome for any implanted material. What dictates implant survival, however, is the way this tissue resolves and the extent of ECM deposition and vascularization in the long term (27,28). Quantification of the granulation tissue area surrounding the silk grafts confirmed formation of a substantial adventitial layer at 3 weeks, which is reduced at later time points where only a thin layer of cells and matrix is visible (Figures 7A and 7C). Caspase-3 immunostaining confirmed apoptotic cells within the granulation tissue, particularly at early time points (Supplemental Figure 5B), also reducing over time. Staining of CD68+ macrophages further demonstrated an initial inflammatory response at 3 weeks (Figures 7B and 7D), significantly reduced at later time points, suggesting resolution of the acute inflammatory response. Immunostaining for vimentin, an established marker for fibroblasts, showed an abundance of fibroblasts in the granulation tissue (Supplemental Figure 8). Staining for collagen, elastin, and proteoglycans confirmed the presence of collagen alone in the granulation tissue, with no elastin or proteoglycan visible, which is suggestive of a late-stage fibroblast-mediated matrix deposition (28). CD34, a marker for progenitor endothelial cells, staining also revealed the extent of neovessel formation in the granulation tissue surrounding the silk grafts, as neovessel formation is propagated by circulating endothelial progenitor cells (29). The fluorescence images clearly demonstrate extensive neovessel staining as early as 3 weeks, which is maintained at all later time points (Supplemental Figure 3C). Collectively, these results show an initial foreign body response to the silk graft at early time points; however, positive remodeling and resolution of the acute inflammatory response suggest that the grafts are extremely well tolerated in the mid-term.
Over the past decades, there have been various approaches to the fabrication of an artificial blood vessel, particularly for small diameter applications where current surgical intervention relies entirely on autologous grafts. Several groups have reported unique bioengineering techniques that demonstrate potential in in vitro assays and preclinical animal models with modest results in human trials of hemodialysis access. Although these approaches are promising, they nevertheless fail to fulfill all the requirements of a synthetic conduit—good mechanical properties and blood compatibility while simultaneously enhancing endothelial cell migration and proliferation. Furthermore, for widespread translation to the clinic, a synthetic vascular graft should ideally be cost-effective, cell-free, with an easy and scalable manufacturing technique, and available in an off-the-shelf manner.
Our work is the first to describe the comprehensive in vivo performance of an electrospun small diameter silk vascular graft generated from and cross-linked using only water. Previous studies demonstrating the potential of silk as a graft material have used fiber winding, braiding, and gel-spinning techniques, leading to suboptimal mechanical properties (13). Long-term evaluation of a combination graft made from plaited silk fibers and wound cocoon filaments showed 85% patency at 1-year and replacement of the graft material with collagen over time (30). Together, these previous studies demonstrate the promising biological and physical properties of silk, manufactured in a way that precludes ready clinical translation. Herein, we demonstrate that electrospun silk can be engineered to have appropriate mechanical properties in a scalable, chemical-free process, while demonstrating favorable endothelial cell and blood compatibilities. In vivo performance of electrospun silk grafts was assessed in a rat model of abdominal aortic replacement at 3, 6, 12, and 24 weeks, showing high graft survival, near complete endothelial cell coverage by 6 weeks, and concomitant smooth muscle cell stabilization and contraction. Comprehensive assessment of inflammatory and remodeling cell types, and newly deposited ECM proteins, further suggests positive remodeling of the silk grafts over time and demonstrates a high degree of in vivo compatibility.
The use of silk, a natural polymer, as a biomaterial for vascular graft engineering has implicit advantages over many competing synthetic polymer-based approaches. Although synthetic polymers impart strength to a biomaterial, they generally have poor cellular compatibility, with many of the degradation products eliciting adverse immune responses in vivo (1,17). Of the natural polymers, silk is of particular interest due to its well-established strength, low immune response, controlled degradability in vivo, and versatility of manufacture (12). For these reasons, the utility of electrospun silk as a biomaterial has been well studied. Previous work on electrospun silk has established some of the important production parameters that control the mechanical properties of this material and demonstrated favorable endothelial cell attachment and growth (31–36). However, no comprehensive preclinical assessment of highly purified electrospun silk conduits has yet been carried out.
In this work, we fully characterized the mechanical properties of the electrospun silk, showing it to be significantly more elastic than the ePTFE. Other mechanical endpoints such as ultimate tensile strength demonstrated that although silk is not as strong as ePTFE, it is more closely matched to the native rat aorta, substantially reducing problematic compliance mismatch. Graft-specific properties, such as burst pressure, revealed that the silk grafts could withstand internal pressures of 849 mm Hg, which is well above physiological values and outperforms other electrospun silk tubes previously described (31,32,34). Furthermore, the silk grafts withstood a suture pull-out strength of 0.86 N, which is within the range of previously published results for small-diameter polymer-based poly(glycerol sebacate) and polycaprolactone grafts that were also implanted into the rat abdominal aorta position (15).
Biological compatibility of the silk materials was shown here to be favorable for use as a vascular graft material. Specifically, endothelial cells interacted well with the electrospun silk, with cells attaching and proliferating well on the surface of the scaffolds, forming a complete and functional monolayer evident by the expression of vascular endothelial cadherin. Another crucial factor for graft patency is the blood compatibility of the material, an aspect commonly overlooked. In this work, we describe the striking blood compatibility of electrospun silk scaffolds. We demonstrate a significantly reduced fibrin network formation on silk, when compared with other electrospun scaffolds (PCL) and the clinical control ePTFE. Furthermore, whole blood contact displayed less clot formation on the silk scaffolds with negligible red blood cell attachment visible under scanning electron microscopy.
In vivo implantation of an electrospun silk graft has not before been reported, with previous in vivo studies being performed with woven silk and gel spun silk tubes. Gel-spun small-diameter grafts were manufactured by winding silk fibers onto a cylindrical mandrel to create highly porous conduits for short-term studies in rats. Initial findings demonstrated significant levels of SMC hyperplasia, leading some investigators to suggest further optimization of mechanical properties, and subsequent modification of the silk may be necessary to limit SMC proliferation (37,38). A separate approach utilized unmodified, knitted silk to engineer small-diameter grafts; however, this manufacturing approach resulted in material strength and compliance insufficient for in vivo evaluation in pre-clinical models (30,39–41).
In a rat model of abdominal aortic replacement, our electrospun silk graft enabled rapid coverage of the graft lumen, contrary to ePTFE, which remained largely uncovered, even at 24 weeks. Immunohistochemistry confirmed the presence of endothelial cells as early as 3 weeks, with an almost compete monolayer forming by 6 weeks. Further histologic analysis of the neointima formation and composition over time suggests a stabilization of neointima from 6 weeks, with regression of total neointima accompanied by the increased collagen, elastin, and proteoglycan production. A phenotypic switch of the cells in the hyperplasia to less proliferative, SMC-α expressing cells further supports this. Furthermore, positive remodeling of the silk graft was observed, with the formation of a thin, collagen rich capsule with extensive neovessel formation. The influence of a healthy endothelium in blood vessels is widely recognized, with past studies establishing the importance of endothelial-derived molecules such as nitric oxide in the regulation of SMC proliferation and maintenance of vessel patency (42). Our results similarly suggest the benefit of a rapidly endothelialized lumen, which may responsible for the observed stabilization of hyperplasia progression observed from 6 weeks.
In this work, we evaluate the mechanical and biological properties of an electrospun silk graft, demonstrating the feasibility of engineering an entirely cell-free, synthetic scaffold with mechanical properties similar to native vessel tissue. Furthermore, the ability to favorably interact with endothelial cells in vitro while resisting clot formation further supports the use of this material as a vascular graft. We show, for the first time, the in vivo performance of an electrospun silk scaffold in a rat model of abdominal aortic replacement for up to 6 months. Our results show early endothelial cell interaction and near complete luminal coverage by 6 weeks post-implantation. Further analysis into the matrix and cellular composition of the hyperplasia and adventitial tissues reveals clinically relevant regression from 6 weeks onward, suggesting the stabilization of the neointima and positive remodeling of the silk graft. Collectively, our results illustrate the promise for electrospun silk conduits to be used as small diameter vessels, representing a cell-free and cost-effective option for small diameter synthetic grafts in clinical use. These results justify further exploration in larger preclinical animal models to evaluate long-term graft survival and stability.
COMPETENCY IN MEDICAL KNOWLEDGE: Clinical applications for small diameter (<6 mm) arterial bypass grafting include coronary artery bypass graft and peripheral vascular applications. The 2 clinically available synthetic graft materials, polyethylene terephthalate (Dacron) and ePTFE produce an unfavorable immune response and are highly thrombogenic, making them innately unsuitable for vascular implantation. New synthetic graft materials with improved efficacy have long been required. This study demonstrates the potential of synthetic silk conduits as off-the-shelf small-diameter grafts.
TRANSLATIONAL OUTLOOK: There is currently a large unmet need for synthetic small-diameter vascular grafts that are effective in small-diameter applications. Small-diameter, cell-free, electrospun silk grafts were shown here to remain patent for up to 6 months in a preclinical rat model of aortic replacement, with results suggesting almost complete endothelialization and neointimal hyperplasia stabilization as early as 6 weeks. Positive remodeling over time also suggests the potential for improved biointegration. Further in vivo studies in larger animal models would be necessary to validate the use of electrospun silk scaffolds in models that also evaluate other aspects important to graft patency in humans such as long-term graft survival.
The authors acknowledge the facilities as well as scientific and technical assistance at the Australian Center for Microscopy and Microanalysis, The University of Sydney.
This work was supported by the National Health and Medical Research Council (APP1066174; Dr. Ng) and an Australian Research Council grant (DP150104242; Dr. Rnjak-Kovacina). Ms. Filipe is a recipient of an Australian Postgraduate Scholarship. Mr. Santos has received funding support from The Heart Research Institute. Financial support was received from E. Brackenreg. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose.
All authors attest they are in compliance with human studies committees and animal welfare regulations of the authors' institutions and Food and Drug Administration guidelines, including patient consent where appropriate. For more information, visit the JACC: Basic to Translational Science author instructions page.
- Abbreviations and Acronyms
- extracellular matrix
- ePTFE
- expanded polytetrafluoroethylene
- polycaprolactone
- proliferating cell nuclear antigen
- SMC-α
- smooth muscle cell actin
- vWF
- von Willebrand factor
- Received June 30, 2017.
- Revision received December 13, 2017.
- Accepted December 15, 2017.
- 2018 The Authors
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Why People Think Vacations Are A Good Idea
By : Jane -
Tips To Have A Great Vacation Enjoying your own vacation is something that can be done in different ways. It's a fact that most people tend to spend their vacation by staying in a hotel, but house rental is also a great idea. Getting the house rental option is also something that's considered a great choice especially when you are in Comporta. Of course, you will still need to find the right rental house that you need. So if you want your family to feel at home while having the vacation that you want in Comporta, then this choice is what you need. Also, you should know that in Comporta, this kind of choice is the most economical one. In any event, this article will help you know more about your choice for the house rental. You should be aware that the type of accommodation can change depending on certain factors. Choosing the best rental that you can find You should know that there are some factors that you need to consider first before you decide to have your own rental. If you want to get the quiet vacation that you want in Comporta, then you will need to find a quiet place to rent. Being able to rent the right cottage in Comporta beach is also something that you should plan for. However, you will need to know the things that you have to do for the reservations of such places. With the help of the online network, you should have no trouble making the reservations that you need in such place. Also, going to Comporta means that you will be able to have a good time enjoying white sands and local culture.
How I Became An Expert on Traveling
You will also want to think about the kind of holiday that you'll be having in this place. Doing this will help you greatly when it comes to having the accommodation that you require for your vacation. Also, being prepared for a group vacation is something that you have to focus on. Also, before you decide to bring your pet to vacation, you'll need to know which of the holiday rentals allow pets in the first place. If you want to find the right holiday rental that you need, you'll want to read some reviews about them first. If you're in need of certain tools, you should also check the facts about the holiday rental before you decide to get it.
Learning The "Secrets" of Vacations
Asking the rental service a few questions is something that's also necessary. In addition to those things, your budget is something that's very important to consider before you decide to get the holiday rental that you need.


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DAN LEGER: Australian bushfires hold lessons for the whole world
Aussie PM Morrison denies link between climate policies, devastating fires
Australia is so vast that even Canadians struggle to comprehend the scale of the disaster unfolding there now. Think Fort McMurray in 2016, only monstrously larger and with no end in sight.
The Fort McMurray fires burned 589,000 hectares and caused $11 billion in damage with an epic firefighting battle that transfixed the country. Australia's bushfires are on another scale entirely.
Fires are burning in every state, already destroying 5.9 million hectares or 15 million acres. As of this writing, 19 people have died and 1,400 homes destroyed, with more of both expected. In New South Wales, Australia's most populous state with Toronto-sized Sydney in its centre, 3.6 million hectares have burned and a state of emergency is in force.
The fires are so big and widespread that smoke dims the sun over much of the country. It's estimated that 450 million wild animals have perished. Mandatory mass evacuations of communities have been ordered in many areas and the Australian navy is rescuing people from burning shorelines.
Bushfires aren't new in Australia. In 2009, they killed 175 people and caused massive property damage. The current fires are worse by almost every measure.
There hasn't been proper rain anywhere in Australia since 2017 and none is expected for months. Rivers are drying up and the grasslands are parched. With summer temperatures at all-time highs, the entire continent is a tinderbox.
But Australia's problem isn't the weather. It's the climate, which is changing for the worse. The ongoing heat wave has been relentless and record setting. Even the ocean around Australia is warmer than has ever been recorded.
Inevitably, Aussies are asking whether their government has done enough, or actually anything, to prevent the fires, beyond fighting them after the fact.
Prime Minister Scott Morrison, a conservative who ignited public fury by vacationing in verdant Hawaii while fires burned at home, is no fan of climate science. He calls the fires routine and refuses to see any link between his government's pro-coal policies and the continent burning around it.
Australia is one of the world's largest coal producers, with $61 billion in annual exports. That means jobs, votes and corporate cash for Morrison's party.
He argues that because Australia only produces one per cent of global greenhouse gas emissions, there's no point developing policies to reduce them. His government axed some preliminary climate programs.
If Morrison's arguments sound familiar, that's because they're precisely those of the Canadian denial caucus: climate change isn't real but if it is real, it's not human-made. Even if it is real and caused by humans, they say, Canada is too small to reverse it and therefore we should do nothing.
Morrison, an anti-immigration populist, is an unabashed fan of the world's foremost climate science denier, Donald Trump. The U.S. president has lavished praise on Morrison, hosted him at a rare state dinner at the White House and even asked him for help digging up dirt on Joe Biden.
Now Morrison, re-elected only last May, finds himself in political hot water at home. Cameras captured angry people in fire-struck rural areas refusing to shake the prime minister's hand and hurling ripe Aussie profanities as he scarpered to his luxury SUV.
Still, Morrison can't make the connection between coal and fire, claiming that no "single policy, climate or otherwise" could have prevented the disaster.
This is deceptive and dangerous. No one believes any "single policy" will stave off catastrophic changes in the global climate. Fighting climate change requires a whole range of actions from governments, corporations and the global public.
When it doesn't rain for years amid persistent record high temperatures, that's not weather. It's climate. And while you can't draw a direct line, yet, linking coal, greenhouse emissions and the Australian fires, ignoring the possibility amounts to criminal negligence.
It's time for climate change deniers, like Morrison and too many Canadians, finally to wake up and smell the smoke.


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-What's happening, guys? Oh, you know, the usual. Just f--king a motherf--king f--kity f--k. But who doesn't do that? Yeah well, you know what most people don't do? Bowl as well as this guy. That dude is so good, he's apparently at the trials for Team USA. That dude is so good that even when he f--ks up, he wins. Now that got almost 600,000 views in a week and don't you just love that? Dude is so good he's gonna go ahead and knock down his opponent's pins too like, "Don't mess with me. I'll knock down every pin up in this motherf--ker." No but I'm guessing one of his fingers got caught in the ball and that's why the ball slipped away like that. And the dude even celebrates later with a little guido fist bump like he meant to do it. -Whoo! Go me! -I don't know, it made for a good video though. And to be honest, I bowl exactly like that. But I bowl drunk as s--t though. I'm always happy to review a type of video that I've never reviewed before. And yesterday, you guys sent me one. It was uploaded to YouTube about a year ago, but it just went viral in the last two days getting 1.3 million views. Geezus! And the woman in the video explains what's going on pretty well. -What are you doing, Sam? -So what I've done is boiled water in the kettle so it's boiling water and I pour it in here and I throw it in the air and it's gonna evaporate. Ready? -Oh, wow! That's amazing. -Usually if it were warm out, it would go kerplunk in the snow but... -Yeah, that is pretty cool isn't it? I guess they shot this video up in Northwest Canada which is where all the caribou but yet away from all the French people, I guess. Anyway, I'm not really sure why I'm asking, but did you see that modern act of witchcraft there? I can't even believe that happened. It must be really cold for that to happen. It must be colder than a prostitute's heart up there. Man, it's so cold up there, you put things in the fridge to heat them up. It's so cold up there the fire department recommends that you set your house on fire. It's so cold up there, well, you get the point. It's f--king cold. Yet she is using that awesome trick to make even more snow. And clearly what they need in this picture is more snow. Yes, see, that's interesting. I always wondered what they do for fun up in Canada. Now this last video should be the big video of the week but it's not. I thought it was really funny, but you know, who the f--k am I? As of now, it's only got 400,000 views it was featured on Attack of the Show and I think ESPN. It's this NBA game right. Now notice the Utah Jazz's mascot, the Jazz Bear get into a little argument with this Cleveland Cavaliers fan. Fake, Cleveland Cavaliers don't have fans. Calm down, troll. Wow, I haven't heard from you in a while troll. Go f--k yourself. All righty then. Anyway, I guess the security starts to escort this dude out and then the mascot like smarts off to him. [crowd cheers] It's funny 'cause he just got his ass whipped by a giant teddy bear. Now I don't usually review videos of people fighting but, one, they weren't really fighting as much as one dude just got knocked on his ass. And two, if I had to make a judgement call on this one, I'd say the video's fake. And by fake, I mean the Utah Jazz staged it to liven up the crowd. It's just that the guy broke away from security a little too easy and the dude in the forty pound bear costume just dominated that fight. I don't know, stuff in real life doesn't usually go down like that. I mean, he even stages like a little crane move at the end like he's Karate Kid. Yeah, I'm definitely going on fake on this one. And I don't know if the camera guy is in on this one too, but he didn't a conveniently good job filming that on his iPhone. I'm just sayin' Mr. Cameraman. You happened to be at the right place at the right time, I'm suspicious. You know, it's probably a damn conspiracy. The whole stadium's probably in on it. And now that I think of it, I'm pretty sure that bear helped stage the moon landing too. In fact, I bet all of Canada is in on that too. But you know who's probably not in on it? The comment question of the day which comes from a user named, bam!, and they said... -'Sup forum? Our comment question of the day is what is Lady Gaga really hiding in her pants? -All right, five naked dudes asking the comment question of the day. Good times. So, what's Lady gaga really hiding in her pants? Leave your interesting or creative responses in the comments section below or on Facebook or Twitter. But thanks for watching today's episode of =3. I'm Ray William Johnson and I approve this message. So tell me guys, what's your flavor? [Stalkin' Your Mom playing] Captioned by SpongeSebastian
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Duration: 5 minutes and 18 seconds
Country: United States
Language: English
Genre: Viral Video
Producer: Ray William Johnson
Director: Ray William Johnson
Views: 124
Posted by: spongesebastian on Jan 23, 2011
Equals Three with RayWilliamJohnson.
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Thursdays Mean Music in Devou Park
Behringer-Crawford Museum 1600 Montague Road, Covington, Kentucky 41011
The Music@BCM series on Thursdays, May through August with some exceptions for holidays. The schedule includes:
May 16: Nicole Zuraitis
May 30: Phil Degreg & Brubeck Tribute - Jazz
June 6: Leroy Ellington's Sacred Hearts - Roots
June 13: Mt. Auburn Brass - Brass
June 20: Mambo Combo - Latin
June 27: Comet Bluegrass All Stars - Bluegrass
July 11: Jake Speed - Family-oriented
July 18: Ben Levin & Heaters - Blues
July 25: Bam Powell and the Troublemakers - Northern Kentucky Rock
August 1: The Bluebirds - Blues
August 8: Robin Lacy and DeZydeco - Cajun
August 15: The Company with Ramona Blaine - Old-school show band
Weather permitting, concerts are held in BCM's outdoor amphitheater at 1600 Montague Road - Devou Park, Covington KY 41011. Concert-goers are invited to bring folding chairs or blankets for seating. In case of inclement weather, the events will move indoors. Guests are encouraged to check the museum's website, www.bcmuseum.org , or call 859-491-4003 to confirm the schedule.
Admission is $5 for adults and free for children ages 3-12, who can participate in a new Kids' Music Club offering free children's activities during the concerts. Concert-goers who ride their bikes to Music@BCM get free admission, courtesy of the Devou Good Project. Parking is free. Adult beverages are available for purchase. Concert-goers can also purchase food by Colonial Cottage of Erlanger, KY, supporting local community organizations.
About Behringer-Crawford Museum:
From the ridiculous (BCM's infamous two-headed calf) to the sublime (our fascinating collections), this family-friendly museum celebrates the unique arts, heritage and culture of Northern Kentucky as part of the Ohio Valley. Examine a mammoth fossil, take a ride through time with the beautifully restored 1892 streetcar "Kentucky," activate the engines, lights and music of a miniature, mid-century community in the Faragher G-gauge train display and even watch a drive-in movie from the seat of a 1959 Buick Electra convertible. View works by prominent national and international artists Mary Bruce Sharon, Wolfgang Ritschel, Harlan Hubbard and others from our region. Music@BCM concerts are "the place to be on Thursday nights" during the summer, while the annual Holiday Toy Trains display is a child's delight in winter.
For more information call (859) 491-4003 or visit bcmuseum.org


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PHOTOSET 244 | Red Adidas nylon shorts and white t-shirt
191 photos in this set + 6:09 min full HD video
Clothes used in this photoset
Red adidas "chile" shorts with white and red soccer t-shirt
Photoset Size: 73 MB Format: ZIP
Video available to watch online in High Definition.
Available for download in 4 sizes. You can download the original full hd video with a resolution of 1920x1080px.
Duration: 6:09 min.
"Recorded in high resolution, each frame is a great photo!"


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I have been greatly interested recently by pronouncements by certain individuals in the press attributing our recent weather disasters to homosexuality, gay marriage, Mitt Romney, Obama, and/or loose morals in New Orleans. The capacity of the human mind to find causal relationships between totally unrelated things or concepts is fascinating.
According to Wikipedia "In statistics, a spurious correlation[1][2] is a mathematical relationship in which two or more events or variables are not causally related to each other, yet it may be wrongly inferred that they are, due to either coincidence or the presence of a certain third, unseen factor (referred to as a "common response variable", "confounding factor", or "lurking variable")."
There is a web site devoted to such correlations (www.tylervigen.com) where you can choose the variables to see how they may be statistically related. I always knew that there was a strong, positive correlation between the price of Jamaican rum and the average salary of Methodist ministers. Did you know, however, that there is a strong positive correlation between:
The divorce rate in Maine and per capita consumption of margarine
The age of Miss America and number of murders by steam, hot vapours, and hot objects.
Per capita cheese consumption and the number of people who died by being tangled in their bed sheets.
Total revenue generated by arcades and the number of computer science doctorates awarded in the US.
What alarms me is that some people will believe anything and will try to find relationships between things that can't possibly be related, just to support their beliefs or prejudices. I, personally, try to remain rational and sceptical. I think I will go now and eat some bread crusts. I am trying to make my hair curly.
Come up with your own spurious correlations.


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News & Blog
Congrats David Farmer!
Congrats on the record breaking year.
AP Radi-CAL equipped!
Ferodo DSUNO for the win.
Shaving time & weight
A peek behind the curtain
Another assist for the Radi-CAL
AP Racing brakes help secure the million dollar purse!
Kudos on another win for Team Penske
If you ain't first, you're last!
Supporting all levels of motorsport since 1981!
Ever wonder what a Swedish AutoX looks like?
Why the biggest names in motorsport trust Essex for brake solutions and advice.
Another week, another record assist for AP Racing.
The Corvette Daytona Prototypes add another chapter to the AP Racing Radi-CAL's legendary performance.
Only the best for the best.
Ferodo DSUNO pads get the job done under hours of intense pressure.
Going behind the scenes of NASCAR's important players


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On September 16th, the world of boxing was paralyzed. Gennady Golovkin and Saul "Canelo" Alvarez starred in the most anticipated fight of the year and did not disappoint anyone after 12 rounds of battle in which they finished in a draw.
The debt of what was promised by both was paid in full. The T-Mobile Arena brought together 22,358 people, many of them from Mexico to support both fighters. Thanks to the amount of fans that "GGG" has on Mexican soil, it can be said that the support of the public was relatively divided between both fighters.
The fastest combinations were on the side of Alvarez, who took the beginning of the assaults and surprised Golovkin who went from less activity to more in each round.
That was the strength of the Kazakh, to hold steady early in the rounds and let his rival make the fight and then attack without mercy. The blows of power were present, but both proved to have the necessary resistance to support what their rival offered.
Around a million and a half people bought the "Pay Per View" service for a fight that was seen almost all over the world and with which the critics were more than happy.
Although the negotiations for the rematch suffered many problems, in the end it was possible to hold the fight for a second time, something that seemed obligatory if you consider how the first match was carried out.
Golovkin threw 707 shots and landed 205, compared to 505 thrown and 169 landed by Alvarez. However, the Mexican counted for a 33% land percentage, which was greater than the 31% from the Kazakh native.
In the end, the opinions of the specialists and the fans were shared, which shows the parity of the combat. The cards were 118-110, 113-115 and 114-114, which determined the tie between both and left everyone wanting a rematch, which will be fulfilled this Saturday, September 15th, on the same stage.


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The Royal Mint
The history of minting in Great Britain is longer than you can imagine. The early coins, minted towards the end of 2nd century BC, were cast in moulds. Later coins were hand-struck, and were produced in this manner for the next fifteen centuries.
200 years after the end Roman conquest, and therefore Roman coins being in circulation in all of the Western Empire, the London mint started operating again in the 7th century. The same mint, coincidentally, produced the Roman coins sometime in the third century. The mint's operating history during that time was short-lived, and it was only open for a mere 40 years before closure.
Fast forward to two centuries ago, and the "modern" Royal Mint opened in 1810 on Tower Hill. Large steam-powered heavy machinery was used in production, which signified the fact that the Royal Mint had definitely entered the industrial age. Time flew, and progress went with it, and at the beginning of the 20th century the steam-powered equipment was fully replaced by electricity.
The British Sovereign coin is has been in production in the UK on and off since 1817. Several commonwealth countries, including Australia, South Africa, and Canada have also minted the British Sovereign in the past. Currently they are minted at the Royal Mint location in Wales, and also in India under full control of the Royal Mint.
Other products minted at the government-owned Royal Mint include items intended for export, and it is known as the world's leading exporting mint. The items include, but are not limited to commemorative and military medals, and similar items intended for schools, businesses, and governments.
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News and information for schools and their communities.
A regular newsletter to keep you up to date with latest news and information about Wellington's parks, sport and recreation sector.
A quarterly update for people with an interest in the city's parks, coast, hills, reserves and streams. Includes reports from environmental community groups about their volunteer work.
A monthly update on what's happening in the arts at the Council. Includes news about workshops, exhibitions and other activities at Toi Pōneke.
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Updates about and from the Strathmore community.
Your weekly guide to life in the capital.
Update about Johnsonville Community Hub.
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Oh, that disturbing police video out of Rohnert Park last week . . .
In case you missed it (and how could you have missed it?): It's July 29, and Don McComas is out in front of his house on what appears to be a neat little cul-de-sac in the so-called Friendly City, when a police officer in a cruiser sort of slow-rolls in front of his driveway and stops.
McComas, as is increasingly the case in a citizenry that's had it with police violence against the very citizens they are sworn to protect and serve, filmed the officer as he sat in his car.
The officer opens his car window and films McComas right back. Then, a few minutes later, he gets out of his car and tells McComas to get his hand out of his pocket. The officer unholsters his weapon, at which point the internet let out a collective "WTF, dude?"
McComas appears to put a phone and set of keys on the roof of his car. He points at the officer with one hand and films with the other. (Does McComas perhaps have three hands?)
As McComas films him, the officer approaches McComas with his weapon, at which point McComas freaks out. He retreats in apparent fear from the officer and makes some comments about past interactions, presumably of the negative variety, at the hands of Rohnert Park police.
McComas has not responded to a request for an interview with this paper.
The unpleasant and tense interaction divided people who commented on the video into two main groups. The online narrative about the episode syncs up with others we've seen, though as police-citizen interactions go, this one was pretty mild.
Nobody got shot in the back while running from a cop, for example. Nobody died screaming that they couldn't breath while being subdued by the police. But it's still a disturbing video and the points of view we've seen on it basically come down to, "Don't get sassy with the police and this won't happen" and "There's no justification for this officer unholstering his weapon."
This paper sent a list of questions to the Rohnert Park police department, but they didn't respond. Elected officials took to the internet a day or two later, once the video had gone viral, to announce that there was an investigation. The gist: We take these things very seriously. The money quote from Assistant City Manager Don Schwartz: "The incident portrayed on the video is not a typical interaction between our Public Safety Officers and the public." Well, good on you!
The interaction is a poignant example of how far off the rails citizen-police interactions have gone in this country. The officer seems to take a page from the "open carry" movement: He deploys a gun to force respect from an unarmed citizen, which is otherwise known as intimidation.
McComas' posture here—freaked out and filming everything—appears to have been influenced by a growing catalogue of highly questionable police activities in and around the presence of, mostly, black Americans. Critics of McComas have called him an "agitator" for filming the police officer, who at one point wonders, mockingly, if McComas is a "Constitutionalist crazy guy." No, that would be Ted Cruz.
The police officer has been placed on paid administrative leave pending the investigation. There's still this thing called "due process" in the criminal justice system, so there's that.
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Search results for author:"Rudolf Kammerl"
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Will an Online Based Development Centre Optimize Teacher Graduation in Germany
Susanne Günther; Rudolf Kammerl
Society for Information Technology & Teacher Education International Conference 2007 (Mar 26, 2007) pp. 71–76
In Germany, for nearly 40 years the first stage of the teacher training program has basically been offered at universities and colleges. Students' graduation for the teaching profession is a public examination conducted by the state examination...


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Herded, saddled, loaded by traders; cared for, sung about and even sung to—one-humped dromedaries prove extraordinary in deserts where the sun is hot; two-humped Bactrians rank no less in others where it is not. Let the rhythmic gaits of our calendar camels carry you along for the year.
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