text:
Mmm, I'm a bit unsure on how to best describe something that tends to be so far reaching, but I will make the attempt. New Escapologist (NE) magazine, which by the way also has a blog, is best described by its introduction:
New Escapologist is a magazine for white-collar functionaries with escape on the brain.
Drawing from philosophy, literature and original humour; we discuss practical escape routes from the present-day predicaments of demeaning work, status anxiety and urban lethargy.
Each issue is a compendium of funny and practical essays on the subject of escape. We promote freedom, anarchy and the absurd.
So while the Canadian Dream blog tends to fall into the financial independence and early retirement bend, NE takes it a step further of getting out as soon as possible. How? Well in issue #3 which deals with the practical issues it suggests the following format as an option.
- Cut Expenses – Any going for FI would have already done this, but its good to start with stop buying crap that doesn't make you happy and doesn't serve a purpose.
- Save Like Mad – The more you save the sooner you can get out.
- Beef Up Your Skill Set – Use work as a training ground to learn as much as possible on different skills sets.
- After Developing a Escape Fund, Quit – Yes, rather than going for full FI, get a beefy escape fund and get out now.
- Enjoy your Escape – Consider it a sabbatical or long vacation rest and recharge from the horrors of work with up to half your escape fund.
- Seed Money – Use the remainder of your escape fund to start your own cottage industry or small business. The idea is to have a business the covers your basic expenses leaving you time to actual enjoy living your life.
Overall I rather like the concept as it mirrors somewhat my current plan to some degree, but it just gets you out the door sooner. It's more of FI by starting your own business route, but that isn't the only way 'out' and points out what others have done to escape the odd world of work in a cube. I should note that they just released the entire back catalog as a PDF, so I got all six issues for $40 and highly enjoyed reading them.
Since the issues also look at the somewhat absurd things it is a fun read which I often felt was very well written. Some examples include essays on the absurd things they teach us in school in the theory of getting a better job (which of course doesn't get used at all), on the practical side a report card on someone's escape just one year into their plan, and even a essay on the digital bohemian.
If you have any doubts on if you would enjoy it or not I would suggest reading the blog as you can get a good feel for the publication by the posts. I personally enjoyed the entire back catalog and read all six issues in just a few days, but do keep in mind the publication isn't serious all the time. You have to laugh sometimes just to get through the day.


text:
Visualizing Sound With Fire: Pyro Board [VIDEO]
Science is cool... or in this case, flaming hot! For all the kids out there that think science is boring, think again.
Without getting too technical, a couple of Danish (people from Denmark) physicists and chemists developed a new version of the Ruben's Tube, which is a device for measuring sound waves using fire. This new 2D version of the tube has 2500 holes and makes it extremely nifty to watch the effects music makes on the flames.
I can just see your local rave club lit up with these all over.
*If you want to bypass the science and just watch the music and fire show, skip to the 3:38 mark*

subset: commoncrawl


text:
Why are America's sons and daughters dying in Iraq and Afghanistan and why is a new, more terrible front in this war about to be opened up against Iran that will likely be the start of Armageddon?
One word--ISRAEL.
Watch this groundbreaking speech by Dr. Hesham Tillawi of www.currentissues.tv as he lays out the case in lawyerly fashion how all of this is the result of Israel's plans for Middle east hegemony.
Watch the speech by USS Liberty survivor Phil Tourney at the conference.
Friday, November 2, 2007
Keynote speech at No More Wars For Israel Conference by Hesham Tillawi
Posted by mark glenn at 12:08 PM


text:
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This Dodge Durango offers drivers the best of everything. It starts with a well-appointed interior with a generous amount of room for cargo and passengers. Muscular styling sets this Durango apart from the softer crossovers on the market. Impressive confidence comes from a powerful, yet efficient drivetrain. The standard all-wheel drive capability is balanced by plush, upscale interior details making for a well-balanced, family friendly SUV. This SUV has 19301 kms. It's bright white in colour and is completely accident free based on the CARFAX Report. It has a 8 speed auto transmission and is powered by a 360HP 5.7L 8 Cylinder Engine. This vehicle has been upgraded with the following features: Navigation, Leather Seats, Bluetooth, Cooled Seats, Premium Sound Package, Heated Steering Wheel. It has some remaining factory warranty for added peace of mind.
Our Durango's trim level is R/T. Enjoy riding in this 2018 Dodge Durango R/T with its focus on interior comfort and convenience. Relax in the eight-way power driver seat with lumbar support, heating, and ventilation. A heated steering wheel keeps your hands warm in cool weather. Other features that are standard on this trim include Nappa leather seating, dual-zone climate control, remote keyless entry, push button start, cruise control, and Uconnect 8.4 with Bluetooth, navigation, and 9-speaker premium audio.
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Buy this vehicle now for the lowest bi-weekly payment of $314.91 with $0 down
Ad #32457531
PostedOctober 13, 2018
RenewedDecember 14, 2018
ExpiryJanuary 13, 2019
DetailsCommercial Seller
CategoryAutos › Cars › SUVs
EditMy ad
SellerPioneer Chrysler (Mission)
PhoneNot available
|Last hour
|Last 24 hours


text:
Via InstaPundit and Jeff Jarvis, the Pew Internet Trust has tried to measure the reach of the blogosphere. I'm not sure how much credence to give Pew's numbers, but they're probably as good as any.
According to the Pew survey, around 120 million American adults "use the internet." Of that number, 7%, or 8 million people, say they have set up blogs. That seems like a very high number to me.
In Pew's survey, 27% of internet users said they read blogs, and 9% said they read political blogs. That would be nearly 11 million political blog readers, which seems high to me, based on what I know of our traffic and other sites' traffic. Four percent said they read political blogs "regularly." If I understand the numbers correctly, that means there are around 4.8 million self-described "regular" readers of political blogs–again, a higher number than I would expect, based on reported traffic.
On the other hand, 62% of internet users say they still aren't clear on what a blog is. So the bottom line is, there is plenty of room left to grow.
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"Arise and take our stand for freedom as in the olden time." Winston Churchill
"Proclaim Liberty throughout All the land unto All the Inhabitants Thereof." Inscription on the Liberty Bell


text:
The Code of Conduct
Our Sixth form students are ambassadors for our school and we have the highest expectations regarding their conduct and behaviour in school at all times. Our Sixth Form is thriving and we are very supportive of our students. As a result, our Sixth Form students excel in thir studies and play an active role in many areas of extra-curricular life in the school. Underpinning all of this, however, is our insistence on the highest standards in relation to study and behaviour in the school.
Our Code of Conduct for Sixth Form students sets out our expectations regarding attendance, behaviour, attitudes to learning and many other aspects of Sixth Form life at The Boswells School and can be found at the bottom of this page along with our Home-School Agreeement for Sixth Form students and our Dress Code for Sixth Formers.
|Sixth Form Code of Conduct
|Sixth Form Home School Agreement
|31st Aug 2016
|Sixth Form Dress Code
|17th Sep 2018
|Sixth Form Code of Conduct
|8th Apr 2015


text:
Mission & Vision Statement
About Exceed:
Exceed Extended Services is a partnership of 8 primary schools in BD7 and 8. An extended school is one that provides a range of services and activities often beyond the school day to help meet the needs of its pupils, their families and the wider community. This can range from breakfast clubs, lunchtime, after school and evening clubs, holiday activities, family trips, parent courses, inter-school competitions, joint school projects, working with faith groups, police and health.
Exceed Extended Schools Partnership also has a key role in ensuring school development, joint CPD, staff networking and sharing good practice.
We, the schools of the EXCEED partnership, aim to collaboratively work together and with our local community, to create places of learning where children are happy, healthy and safe. A community of Schools where children can celebrate the successes of their learning and achievements, can maximise their attainment through shared, innovative, inclusive practice, can communicate effectively to a variety of audiences and develop the skills and attributes necessary to be positive future citizens in our changing society.
To provide a World Class education to raise expectations, aspirations and promote achievement for all in our community


text:
Sisi Sinaloa and Lexi Love share a tender kiss before they both starts sucking on his cock and his balls at the same time!
Tag: Sisi Sinaloa
Sisi Sinaloa aka Paris Mayne Goes For A Big Black Cock!by New Sensations
Sisi Sinaloa has her a black cock and she is working it, firts with her hand, then mouth and then her pussy!
Sisi Sinaloa Begs For A Big Cock!
Sisi Sinaloa shows off in her sexy bra and pantie set!
Sisi Sinaloa gets on all fours and lets her tits hang down!
Sisi Sinaloa starts to slip that bra off to show her tits!
Sisi Sinaloa flashes a peek at those rock hard nipples!
Sisi Sinaloa squeezes together those perky tits!
Sisi Sinaloa slips those panties down and shows her ass!
Sisi Sinaloa just can't wait to get her mouth around that cock!
Sisi Sinaloa looks hotter than ever in that bra and pantie set but she gets hotter than ever when she slips out of her bra and shows off those perky titties! Just watch her opening her mouth wide and begging for cock!
Sisi Sinaloa Gets A Mouth Full Of Sticky Cum!
Sisi Sinaloa slowly slides that big fat dick between her warm wet lips!
Sisi Sinaloa licks the tip of that dick with a naughty smile on her face!
Sisi Sinaloa takes more of that throbbing cock shaft between her lips!
Sisi Sinaloa teases her sweet nipples as she begs for more of that dick!
Sisi Sinaloa can't wait to get her mouth filled full of cum!
Sisi Sinaloa sticks her ass in that lucky guys face as she sucks that cock!
Sisi Sinaloa gets a thick load of sticky cum in her mouth!
Sisi Sinaloa sucks cock like a real professional and once she gets her lips wrapped around this throbbing member there is no stopping her! She sucks and tugs on that cock until she gets a mouth full of the sticky sweet stuff.
Sisi Sinaloa Sucks A Fat Dick Beside The Pool!by Throated
Sisi Sinaloa shows off in that cute bikini in the pool!
Sisi Sinaloa flashes those perky titties and sweet hard nipples!
Sisi Sinaloa slips those panties down and flashes her juicy ass!
Sisi Sinaloa slides the tip of that cock between her warm wet lips!
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Sisi Sinaloa loves the feel of that throbbing cock in her mouth!
Sisi Sinaloa can give a blow job like no one you've ever seen and she always drives that cock so deep down her throat that she starts to gag! Watch as Sisi sucks and teases this dick until she just can't take anymore!
Sisi Sinaloa Strips Out Of Her Sexy Bikini!by Throated
Sisi Sinaloa shows off in her sexy new bikini!
Sisi Sinaloa flips around and shows off that sweet ass in her panties!
Sisi Sinaloa stretches out those long legs in her cute high heels!
Sisi Sinaloa teases as she contemplates untying that cute bikini top!
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Sisi Sinaloa gives a nice close look at that delicious naked ass!
Sisi Sinaloa looks gorgeous in her sweet new bikini but once she starts to get horny and unties that bikini top she just can't wait to get it off! Watch Sisi as she slips those bottoms off and spreads her sweet round ass!
See more of Sisi Sinaloa showing off her ass now at Throated
Sisi Sinaloa Gives A Sloppy Hand Job!
Sisi Sinaloa looks delicious as she shows her cleavage for Mano Job!
Sisi Sinaloa flashes her sexy panties underneath her dress!
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Sisi Sinaloa wraps her fingers around that fat cock!
Sisi Sinaloa lubes up her hand to stroke that dick!
Sisi Sinaloa is a kinky babe and when Mano Job asked her to show off for the camera and stroke this fat cock she just couldn't say no! Just watch as she wraps her fingers around that cock and goes to town!


text:
Green light for five new Kyogle bridges
Media Release
01 December 2017
Joint release with:
Danielle Mulholland
Mayor Kyogle Council
Kevin Hogan
Federal Member for Page
- Australian Government approves more than $2 million to replace five timber bridges in Kyogle
- Upgrades to improve safety, flood resilience, travel times and maintenance costs
- Works on the first bridge replacement planned to start before the end of the year
The Australian Government has delivered more than $2 million to replace five timber bridges across Kyogle Shire as part of the Page Bridge Upgrade Package, with funding approved for the upgrades today.
Federal Minister for Infrastructure and Transport Darren Chester said 2016 election commitment would deliver better, safer and more reliable road access for residents in Kyogle and beyond.
"Replacing these five timber bridges is a great example of how investing in regional infrastructure can create longstanding benefits for the region and the state—improving safety, supporting emergency services access and better connecting communities," Mr Chester said.
Federal Member for Page Kevin Hogan said the funding would replace three bridges in urgent need of repairs due to damage from Cyclone Debbie earlier this year.
"Kyogle Shire has one of the highest numbers of ageing wooden bridges in New South Wales, and replacing these five bridges will improve access and connections during natural disasters, improve safety and reduce travel times for motorists," Mr Hogan said.
Kyogle Council Mayor Danielle Mulholland said the upgraded bridges would have higher load limits, which would reduce operating costs for local businesses by opening up more direct routes and accommodating heavier vehicles.
"Future Council budgets will feel the benefit of this investment in the form of lower ongoing maintenance costs, which is a further benefit to the shire's ratepayers," Cr Mulholland said.
The five bridges to be replaced are Sommerville Bridge, Boorabee Park Bridge, Burt Rayner Bridge, J Campbells Number 1 Bridge, and J Campbells Number 2 Bridge.
Work on all five bridges is expected to be completed in late 2019, weather permitting. The $4.04 million Page Bridge Upgrade Package is jointly funded by the Australian Government and Kyogle Council.


text:
FOA Certifications
- Work-to-Cert Certification
Getting started with FOA Certification
Understanding FOA Certification Technical Requirements.
FOA Approved Training Programs
FOA's "Work to Cert" Direct Certification Program
Online Study Guides and References
FOA Online Reference Textbook particularly the section Understanding Fiber Optics, the Basics
FOA Installation Tech Bulletin
The Process of Getting Certified By The FOA
Reference Books (Available
Printed or eBooks)
fiber book is available in Spanish
and French as well as English.
It's now CFOT® The FOA CFOT® (Certified Fiber Optic Technician) is now a registered trademark. With so many fiber optic techs holding CFOTs and the CFOT being recognized worldwide as the foremost certification in fiber optics, the FOA realized the value of the CFOT required trademark protection.
FOA Certification
Note: Techs doing fiber installation for any particular application should have a CFOT to be qualified for doing fiber optic installation work and may also have a specialist certification in that application. The CFOT is the basic certification required for all installation personnel.
FOA also offers specialist certifications in three categories:
* Corporate members are organizations and as such are not certified but these companies/organizations get discounts on certifications and renewals among other benefits.
The FOA is a member of ANSI (the American National Standards Institute). FOA certifications conform to the international standard for certification bodies, ANSI/ISO/IEC 17024 Conformity assessment — General requirements for bodies operating certification of persons.
Is The Value Of Industry-Based Certifications Like The FOA
Quoted from a Georgetown University study on career and technical education:
Industry-based Certifications (IBCs)
As alternative models in higher education—such as do-it-yourself (DIY) education, competency-based education, and e-learning a.k.a. online education—are becoming more prevalent, the need for a reliable credentialing system has risen in tandem. This need arises because of the difficulty in evaluating the reliability of the credentials established by these alternative systems to ensure that workers have the knowledge and skills necessary to perform proficiently on the job.
Industry-based certifications (IBCs) are one model that is being embraced by educators, employers, and policymakers. Market forces, not the educational system, drove the creation of IBCs. They differ greatly from other kinds of postsecondary credentials and awards in that they are exam-based, administered and accredited by third parties, and developed by the private sector with little to no involvement from public institutions. They are not tied to particular programs of study, but can serve as both alternatives and supplements to traditional postsecondary awards, such as college degrees and certificates.
Industry-based certifications address several problems that postsecondary credentials currently face: relevance, accountability, consistency, and portability. Because industry-based certifications are more flexible than traditional degrees, they better reflect current industry standards and competencies, which are in constant flux."
Download the full Georgetown University report here.
Understanding FOA Certifications
How Does the FOA
Program Relate to Other Associations?
FOA Reference Materials For Certification Exams
FOA certifications are all based on FOA published reference books shown below, the FOA Online Reference Guide (available in English, Spanish and French), YouTube Videos, and the FOA's web-based training site Fiber U.
FOA Textbooks
(Available Printed or eBooks - the printed or Kindle fiber book is available in Spanish and French)
Click on any of the books to learn more.
Students in training classes for FOA Certification will be trained in basic hands-on skills appropriate for the certification and the instructor will evaluate the student's abilities to learn these skills. Those applying with field experience will submit documentation of their experience and other training plus references which will be evaluated by the FOA staff.
Studying For FOA Certification Exams
If you are preparing for a FOA exam, either taking a course from an FOA-approved school or directly from the FOA, you can study for the exam using the FOA references. Reference materials for studying for these certification exams are available with the FOA textbooks or online free with the FOA Guide or Fiber U.
The Process of Getting Certified By The FOA
If you want to become FOA-Certified in fiber optics, there are several paths available. First you should understand how FOA Certifications work. Everyone who is FOA-Certified usually begin with a CFOT and can then work toward higher level certifications. To get your CFOT, you must show extensive knowledge of fiber optics and skills in the practices common to the field. How you do this depends on whether you are currently working in the field or are just getting started.
Training - Attending FOA Certification Training At An FOA-Approved School
If you are just getting started in fiber optics and have no practical experience, you should get training and become certified through one of the FOA-Approved Schools. They offer courses that provide classroom training which will provide the knowledge you need for the CFOT exam and the instruction will include labs to develop the basic skills in fiber optic cable preparation, termination, splicing and testing required by the CFOT.
Once you are FOA certified, you can work toward specialist certifications. FOA offers CFOS Specialist Certifications in both skills and applications. To qualify for these certifications, you can apply based on field experience or take a specialist course from one of the FOA-Approved Schools. You should keep a record of your experience in the field to qualify for specialist certifications. When you are ready, you can apply for the CFOS exam in any of the specialties.
Guidelines For Direct FOA Certification
FOA's "Work To Cert" Program
FOA was founded in 1995 by a group of people representing manufacturers, contractors, users, military/government and others, all active in training those who became instrumental in creating the fiber optic industry. As trainers, we were particularly concerned by the quality of training newcomers to the industry were getting, so we developed standards for the knowledge, skills and abilities (KSAs) expected of professionals in the business. Those KSAs became the basis of the FOA CFOT® certification.
Many students have been trained and certified by FOA-approved schools. But in addition, thousands of industry professionals have applied to the FOA directly for certification without the need for classroom training, based on their knowledge and skills developed working the field. Included in that group are most of the current FOA certified instructors teaching in FOA approved schools.
With the increasing acceptance of FOA certifications, many experienced field technicians find themselves being required to have a CFOT® certification for employment, bidding on and/or working on a project. These are generally not people who would need to take a traditional CFOT course.
FOA has always had a policy of allowing experienced people to join the FOA directly and become certified, starting with the founders and including almost all of our current instructors. The requirements for direct certification have been clear, they are simply an extension of our KSAs for students trained at our schools. We require experience in the field and passing our first level exam.
FOA's "Work To Cert" Program makes it easier for experienced fiber techs to qualify for the FOA certification. Qualifying experience in the industry of at least two years minimum is required.
FOA Renewal Policy And Certification Renewal Procedures
Why is it important to renew your certification?
In today's high tech world, certification is considered proof of professional status and is often required for jobs. Employers often contact the FOA to confirm the status of a job applicant. Certification requires actively working in the field. If the person's certification is not current, we confirm when they got their certification, when it expired, and that if expired, we consider them inactive.
Cost to renew: $100US for 3 years
How to renew:
Your expiration date is on your certification ID card. If you have misplaced the card, you can contact the FOA – we can check your status and provide you with a new card.
You can renew online or print a renewal form and send/fax it in:
Be sure to give us your latest address, including e-mail, as that is how we send renewal notices.
How to reestablish expired FOA certifications?
Under 3 years:
If your FOA certification has expired within the last 3 years, you may simply renew. Follow the directions as outlined above.
Over 3 years:
If your FOA certification has expired more than 3 years ago, you have to re-establish your FOA credentials. You start by sending in your resume. We will either arrange for you to re-take the CFOT exam or if you are not working in the fiber optic field, we will work with you to sign up for a CFOT review class.
What happens if an individual does not renew after the 3 year deadline?
The FOA follows international standards for certification that require that we verify the relevant professional/work status of everyone we certify. You are either:
1. Active - you have remained in the fiber optic industry and have renewed your certification every one, two or three years. We will confirm your status to anyone who inquires about your credentials.
2. Inactive - If you have not renewed your certification in 3 years , we will inform anyone who asks that you took an FOA exam in the past but your certification is considered inactive - your current professional/work status in the areas in which you were certified is unknown.
To re-establish your FOA certifications, you start by sending in your resume. We will either arrange for you to re-take the CFOT exam online or if you are not working in the fiber optic field, we will work with you to sign up for a CFOT review class.
FOA Certifications Are Recognized By The US Department of Labor
FOA Certifications have been recognized by the US Department of Labor. The DoL reviews certifications and how they relate to job opportunities and training. The DoL Career OneStop website helps individuals explore career opportunities to make informed employment and education choices. The Web site features user-friendly occupation and industry information, salary data, career videos, education resources, self-assessment tools, career exploration assistance, and other resources that support talent development in today's fast-paced global marketplace.
Go to FOA Certifications Listing By US Department of Labor - Career OneStop and search for "Fiber Optic Association" to see all the FOA certifications listed.
See also The DoL Occupational Outlook Handbook which has a page covering fiber optic technicians under "Line Installers and Repairers" and links to FOA certifications.
FOA is listed
in the section "How
to Become a Line Installer or Repairer."
This website lists all sorts of interesting information, including what these workers do in their jobs, how they get trained and certified (where the FOA is referenced) and something everybody is interested in - sections on "Pay" and "Job Outlook" - see the red arrow above.
Where In The US Do Contractors Need Licenses For Fiber Optics?
We often get asked where in the US do contractors doing fiber optic installations need licenses. We found a good website for that information, the NECA -NEIS website. You might remember NECA-EIS, as they are the partner with the FOA in the NECA/FOA 301 Fiber Optic Installation Standard. NECA is the National Electrical Contractors Association and NEIS stands for National Electrical Installation Standards. They have a very easy to use map and table that gives you data on every state in the US, so mark these pages for future reference.
http://www.neca-neis.org (See "State Regulations")
http://www.neca-neis.org/state/index.cfm?fa=state_regs (all electrical licensing)
Low Voltage: http://www.neca-neis.org/state/index.cfm?fa=specialty_licensing
Return to The FOA Home Page


text: James A. Peltier wrote: > > Tronn Wærdahl wrote: > > > > I dont know about apt-cacher, but if its something to do > with apt-get > > you could you --download-only. Like apt-get upgrade --download-only > > > > Tronn > > > > apt-cacher allows you to point all clients to a centralized location, > when one client performs, say and apt-get dist-upgrade, the apt-cache > server caches all of the packages to it's local apt directory. It's > very convienent and easy to setup, whereas the squid solution > proposed > by Dag is a fair bit more conviluted. > > A possible solution would be to possibly extend create-repo to add > functionality to mirror updates and packages for selected > architectures. > Perhaps add the proxy/caching functionality that is required to > perform these operations? I just have a central server mirror the updates repo with wget and have either apache or iis serve them up to clients. Then in the yum repo files, comment out mirrorlist=, uncomment baseurl=, add the url to your update server first for baseurl= and set the failovermethod=priority. -Ross ______________________________________________________________________ This e-mail, and any attachments thereto, is intended only for use by the addressee(s) named herein and may contain legally privileged and/or confidential information. If you are not the intended recipient of this e-mail, you are hereby notified that any dissemination, distribution or copying of this e-mail, and any attachments thereto, is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender and permanently delete the original and any copy or printout thereof.


text:
Whoever ordered this bespoke F12 from Ferrari must have some serious power. The Italian supercar made this F12 TRS specifically for a buyer who requested an "extreme, two-seater, open-top sports barchetta." The TRS is a nod to the 1957 250 Testa Rossa, from which this one-off whip gets its inspiration.
Performance-wise, you're looking at the same engine as the F12berlinetta. The difference is all in the design. A modified aerobridge runs all the way to the back on the 'Rarris sides to create maximum downforce. It also sports a dark band of glass for the wraparound windscreen. Inside the F12's been stripped of everything but the bare essentials, meaning no glovebox, central air vents, stereo system, or even floor mats.
Now that we've seen this F12, we want to make our own requests.
RELATED: 15 Wild Fake Ferraris
RELATED: 25 Wild Custom Ferraris
[via Autoblog]


text:
- Research article
- Open Access
Progressive morphological changes and impaired retinal function associated with temporal regulation of gene expression after retinal ischemia/reperfusion injury in mice
© Kim et al.; licensee BioMed Central Ltd. 2013
- Received: 14 February 2013
- Accepted: 11 June 2013
- Published: 22 June 2013
Retinal ischemia/reperfusion (I/R) injury is an important cause of visual impairment. However, questions remain on the overall I/R mechanisms responsible for progressive damage to the retina. In this study, we used a mouse model of I/R and characterized the pathogenesis by analyzing temporal changes of retinal morphology and function associated with changes in retinal gene expression. Transient ischemia was induced in one eye of C57BL/6 mice by raising intraocular pressure to 120 mmHg for 60 min followed by retinal reperfusion by restoring normal pressure. At various time points post I/R, retinal changes were monitored by histological assessment with H&E staining and by SD-OCT scanning. Retinal function was also measured by scotopic ERG. Temporal changes in retinal gene expression were analyzed using cDNA microarrays and real-time RT-PCR. In addition, retinal ganglion cells and gliosis were observed by immunohistochemistry. H&E staining and SD-OCT scanning showed an initial increase followed by a significant reduction of retinal thickness in I/R eyes accompanied with cell loss compared to contralateral control eyes. The greatest reduction in thickness was in the inner plexiform layer (IPL) and inner nuclear layer (INL). Retinal detachment was observed at days 3 and 7 post- I/R injury. Scotopic ERG a- and b-wave amplitudes and implicit times were significantly impaired in I/R eyes compared to contralateral control eyes. Microarray data showed temporal changes in gene expression involving various gene clusters such as molecular chaperones and inflammation. Furthermore, immunohistochemical staining confirmed Müller cell gliosis in the damaged retinas. The time-dependent changes in retinal morphology were significantly associated with functional impairment and altered retinal gene expression. We demonstrated that I/R-mediated morphological changes the retina closely associated with functional impairment as well as temporal changes in retinal gene expression. Our findings will provide further understanding of molecular pathogenesis associated with ischemic injury to the retina.
- Retinal Detachment
- Retinal Ganglion Cell
- Retinal Thickness
- Retinal Layer
- Outer Nuclear Layer
Retinal ischemia, often referred as "stroke of retina", is an important cause of visual impairment in retinal vascular occlusion, diabetic retinopathy, glaucoma, and ocular trauma [1–5]. It is caused by a reduction of the retinal blood supply that decreases the delivery of oxygen and other nutrients to various retinal layers. Reperfusion of blood following ischemia is associated with oxidative stress and inflammatory responses [6]. In particular, resulting retinal ganglion cell (RGC) death is caused by a variety of cell death mechanisms including necrosis, apoptosis, necroptosis and autophagy after ischemia/reperfusion (I/R) injury [7–9]. To better understand the pathophysiological mechanisms associated with retinal I/R injury, several different experimental approaches have been designed in rodent models [8, 10–12].
The "pressure-induced retinal I/R model" involves cannulation of the ocular anterior chamber followed by raising intraocular pressure above systolic blood pressure. After a specified period of time, the cannula is removed allowing restoration of retinal blood flow. This model has been used for the investigation of ischemia-derived ocular pathologies such as glaucoma and diabetic retinopathy [13, 14]. This model currently is the most widely used method to study ocular diseases related to retinal ischemia.
Retinal damage due to I/R injury is associated with the loss of neurons, morphological degeneration of the retina, loss of retinal function, and ultimately vision loss [15–17]. Although degeneration times vary in different experimental conditions, I/R-induced injury and retinal degeneration is initially observed primarily in inner retinal layers [e.g. the inner plexiform layer (IPL) and inner nuclear layer (INL)] that are supplied by the central retinal artery, in contrast to the outer nuclear layer (ONL) that is generally less affected [18–21]. Differently from inner retinal layers, the choroid supplies blood and nutrients to the photoreceptors and ONL [22, 23]. This structural difference may influence the initiation stage of I/R injury. Furthermore, morphological changes are often associated with functional impairment of the retina [24–26]. However, the correlation between morphological and functional changes with molecular mechanisms from different stages of pathogenesis associated with retinal I/R injury in retina is poorly characterized.
Several signaling pathways have been reported as key molecular events related to the degeneration or protection of RGCs and their axons in retinal I/R injury. Increased nuclear factor (NF)-κB p65 immunoreactivity was associated with retinal degeneration following retinal ischemia and reperfusion injury in mice [27], and inactivation of astroglial NF-κB promotes survival of retinal neurons following ischemic injury [28]. In addition to NF-κB, cyclooxygenease-2 appears to play a critical role in RGC death after transient ischemia [29]. Activation of Stat3 protects retinal ganglion cell layer neurons in response to transient retina ischemia in mice [30]. Several reports have indicated that inflammatory responses associated with the innate and adaptive immune system are major pathological processes in retinal I/R injury. These include toll-like receptor 4, complement component C3, tumor necrosis factor receptor, and surface molecule CD40 that are associated with functional impairment, retinal layer morphological changes, and/or loss of RGCs [20, 31–33]. However, further studies are still required to define and better understand pathologic progression including temporal changes in retinal morphology, function, and molecular signaling.
In our study, we used a mouse model of pressure-induced retinal I/R injury and characterized morphological changes in retinal layers associated with retinal ERG functions. Using histological analysis and SD-OCT (spectral domain-optical coherence tomography) scanning, we showed progressive morphological changes of retinal layers at various times after I/R injury. In addition, we used scotopic electroretinography (ERG) to demonstrate functional deficits after I/R injury that corresponded to the morphological changes. We also profiled changes in retinal gene expression associated with I/R injury at various time points using cDNA microarray analysis to correlate the molecular mechanisms with morphological and functional changes.
Material and methods
Female C57BL6/J mice (8-10 weeks of age) were maintained in 12:12 light /dark cycle under optimal temperature and humidity controlled conditions. All studies were approved by University of North Texas Health Science Center's Institutional Animal Care and Use Committee (IACUC) and complied with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.
Pressure induced retinal ischemia
Mice were anesthetized using a ketamine/xylazine cocktail (100/10 mg/kg). While anesthetized, mice were placed on a heating pad to prevent hypothermia. Mydfrin (Alcon, Inc. Fort Worth, TX) was topically administered to the test eye to dilate the iris. A 30-gauge needle connected to saline reservoir was inserted into the anterior chamber through the cornea of left eyes. Intraocular pressure was raised to 120 mmHg for 60 min. Retinal ischemia was confirmed by blanching of the retina using an ophthalmoscope. Contralateral right eyes served as controls. The needle was removed after 60 minutes to allow reperfusion of the retina. Tobrex (Alcon Inc. Fort Worth, TX) was topically administered to prevent ocular infection.
Spectral domain-optical coherence tomography (SD-OCT)
Mouse retinas were scanned with an SD-OCT Ophthalmic Imaging System (Bioptigen Inc. Durham, NC). Briefly, mice were anesthetized using a ketamine/xylazine cocktail (100/10 mg/kg) and placed on the mouse holder to fix the mouse posture for scanning. The retina was scanned with rectangular scanning mode (1.2 mm diameter) consisting of 100 B scans/1000 A scans per B scan using InVivoVue software (Bioptigen Inc, Durham, NC). Superior, center and inferior images were imported and analyzed by ImageJ software (NIH) with four vertical calipers on each retinal layer (n = 4-5 mice per time point).
Flash scotopic electroretinogram (ERG)
Mice were dark adapted for 16 hrs. After anesthesia with Ketamine/Xylazine (100/10 mg/kg), mice were placed in a Gantzfield light chamber on the LKC electroretinogram system (LKC Technologies Inc., Gaithersburg, MD) with temperature control (37°C). Amplitude and implicit times of ERG waveforms were measured at a series of flash intensities (-30, -20, -10, 0, 5, 10, 15 dB) (n = 9 mice per time point).
Histology and cell counting in RGC layer
Eyes were harvested and fixed in neutral buffered 10% or 4% paraformaldehyde. After paraffin embedding, retinal cross sections were prepared (5 μm) followed by Hematoxylin-Eosin (H&E) staining for morphological observation of the retinal layers. Four retinal sections from ora serrata to ora serrata through the optic nerve head were chosen from each eye and the cells in the RGC layer were counted and averaged. Day 0 RGC counts from non-ischemic control eyes were set as 100%, and RGC cell counts in the rest of the eyes were compared to these controls.
Analysis of retinal thickness
H&E stained whole retina or individual retinal layer thicknesses were measured using ImageJ software (NIH). Individual or whole (RGCL to ONL) layer thickness from 4 retinal cross-sections per eye were measured at quarterly points for each retinal cross-section and averaged. For SD-OCT images, layer thickness at 2 different distances from optic nerve head (~0.35 and 0.55 μm) was determined and averaged. Averaged retinal thickness was converted to the percentage of the thicknesses from day 0 non-ischemic contralateral eyes.
RNA extraction and DNA microarray analysis
Total retinal RNA was extracted from control and I/R injured eyes. Retinas were collected and homogenized in Iso-RNA Lysis Reagent (5 PRIME Inc. Gaithersburg, MD). Total RNA was further extracted using an RNeasy Micro Kit (QIAGEN Science, Germantown, MD). RNA quality was controlled by determining RNA integrity number (RIN) using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) with nano- or picochip systems. Total RNA with RINs >7 were selected and pooled from either control or experimental eyes at each time point for gene expression microarray analysis. Microarray hybridizations were performed at the University of Iowa DNA Core Facility. Briefly, 50 ng total RNA was converted to SPIA amplified cDNA using the WT-Ovation Pico RNA Amplification System, v2 (NuGEN Technologies, San Carlos, CA, Cat. #3302) according to the manufacturer's recommended protocol. The amplified SPIA cDNA product was purified through a QIAGEN QIAquick PCR Purification column (QIAGEN Cat #28104) according to modifications from NuGEN. Five microgram samples were fragmented (average fragment size = 85 bases) and biotin labeled using the NuGEN FL-Ovation cDNA Biotin Module (NuGEN Technologies, Cat. #4200) per the manufacturer's recommended protocol. The resulting biotin-labeled cDNA was mixed with Affymetrix eukaryotic hybridization buffer (Affymetrix, Inc., Santa Clara, CA), placed onto Affymetrix Mouse Gene 1.0 ST arrays (Part No. 901168) (Affymetrix Inc. Santa Clara, CA), and incubated at 45°C for 18 h with 60 rpm rotation in an Affymetrix Model 640 Genechip Hybridization Oven. Following hybridization, the arrays were washed, stained with streptavidin-phycoerythrin (Molecular Probes, Inc., Eugene, OR), and the signals were amplified with anti-streptavidin antibody (Vector Laboratories, Inc., Burlingame, CA) using the Affymetrix Model 450 Fluidics Station. Arrays were scanned with the Affymetrix Model 3000 scanner with the 7G upgrade and data were collected using the using the GeneChip operating software (GCOS) v1.4.
Bioinformatic analysis
The 16 data CEL files for all 8 time points (0, 6 hr., 1, 3, 7, 14, 21 and 28 days) after I/R injury were imported into the Partek Genomics Suite 6.6 software (Partek Inc., Louis, MO) and normalized based on robust multi-array averaging (RMA). At each time point, the retinal I/R sample was compared to the contralateral control samples to calculate the microarray ratios and log2 fold change values. Using Excel, the selective filter of ≥1.5 fold change was utilized to identify up-regulated genes and ≤ -1.5 for the down-regulated genes per time point for the retina datasets. The genes were further analyzed using the publicly available bioinformatics software DAVID (Database for Annotation, Visualization and Integrated Discovery). Gene ontology (GO) based cluster analysis was performed to identify possible enrichment of genes (GO enrichment score calculated using a χ2 test) using filtered genes from each time point. The Fishers Exact p value is calculated by DAVID to identify GO enrichment based clusters, and p values <0.05 were considered to be significant in the enriched annotation category based on the Benjamini multiple test correction[34, 35]. Clusters of genes were identified at specific time points and changes in gene expression were graphed temporally for each GO category.
Real-time reverse transcription (RT)-polymerase chain reaction (PCR)
Complementary DNA (cDNA) was generated from the individual retinal RNA samples (n = 4-5) that were pooled for the microarray study. 2 μg of total RNA was used for reverse transcription (RT) at 25°C (10 min), 37°C (120 min) and 85°C (5 min) using a Multiscribe reverse transcriptase kit (Applied Biosystems, Life Technologies Corp., Grand Island, NY) and a PTC-100 thermal cycler (MJ Research Inc. Waltham, MA). Gene expression of cryaa, cryba1, ccl12, c3 and gapdh was analyzed using Taqman gene expression assay primer sets (Applied Biosystems, Life Technologies Corp., Grand Island, NY) with Taqman Fast Advanced Master Mix (Applied Biosystems, Life Technologies Corp., Grand Island, NY) on a CFX96 real-time system (Bio-Rad, Life Science Research, Hercules, CA) with recommended thermal cycles by manufacturer (hold 50°C (2 min), hold 95°C (20 sec), 40 cycles of denaturation (3 sec) and annealing /extension (30 sec)). The expression of the housekeeping gene gapdh was used as an internal control to normalize target gene expression between samples. Differences in target gene expression were calculated using the following formula: ΔΔCT = ΔCT (target gene) – ΔCT (gapdh). The ΔΔCT value of cDNA amplification from the I/R eye was normalized to the control right eye. 0 hr was set as 1 and data from other time points were expressed as fold difference compared to 0 hr data.
Sections of paraffin embedded retinas were subjected to antigen retrieval in citrate buffer (pH 6.0), and the slides were washed twice in PBS. Slides were blocked for 1 hr using SuperBlock Blocking Buffer (Thermo Scientific, Rockford, IL), and then incubated with mouse anti-Brn3a (1:50) (Cat # MAB 1585, EMD Millipore Corp. Billerica, MA), mouse anti-NeuN (1:1000) (Cat # MAB 377, EMD Millipore Corp. Billerica, MA), or rabbit anti-GFAP (1:200) (glial fibrillary acidic protein) (Abcam plc. cat.# ab7779, Cambridge, MA) for 16 hr. at 4°C, followed by washing and then incubation with goat anti-mouse antibody (1:500) or goat anti-rabbit antibody (1:500) conjugated with Alexaflour 488 (Cat.# A11001 or A11008, Invitrogen, Life Technologies, Grand Island, NY) for 1 hr. at room temperature. Cover slips were mounted using ProlongGold anti-fade reagent with DAPI (Molecular probes, Life Technologies, Grand Island, NY) for nuclear visualization. Images were acquired using a Nikon Eclipse Ti inverted microscope (Nikon Instruments Inc. Melville, NY) and a CRi Nuance FX multispectral imaging system (Caliper Life Sciences, Hopkinton, MA). Autofluorescence was subtracted using the Nuance 3.0 software.
I/R induced retinal detachment after 3 and 7 days
I/R injury induced progressive degeneration of inner retinal layers (IRLs) and decreased cell numbers in the retinal ganglion cell layer
I/R induced retinal functional impairment beginning 3 days after injury
I/R injury differentially altered expression of retinal genes at different time points
Gene ontology clusters in the mouse retina upregulated at various time points after I/R injury
Enrichment score
P value (P < 0.05)
0 day
No significant changes
6 hr
eye development
inflammatory response
blood Bessel morphogenesis
structural constituent of eye lens
cytokine activity
extracellular space
1 day
eye development
inflammatory response
cell death
structural constituent of eye lens
transcriptional factor activity
extracellular region part
3 day
inflammatory response
eye development
cell death
structural molecule activity
extracellular region
7 day
defense response
eye development
peptide antigen binding
extracellular region part
antigen processing and presentation
14 day
cell-cell signaling
inflammatory response
cell death
tissue regeneration
neuron differentiation
glutamate receptor activity
extracellular region
neuron projection
postsynaptic membrane
gap junction
21 day
defense response
inflammatory response
no significant changes
extracellular region part
extracellular region
proteinaceous extracellular matrix
28 day
immune response
protein maturation
complement activation
glycosaminoglycan binding
extracellular region
proteinaceous extracellular matrix
Gene ontology clusters downregulated in the mouse retina at various time points after I/R injury
Enrichment score
P value (P < 0.05)
0 day
No significant changes
extracellular region
1 day
No significant changes
3 day
protein-DNA complex assembly
DNA binding
protein-DNA complex
7 day
No significant changes
14 day
eye development
structural constituent of eye lens
21 day
No significant changes
28 day
G-protein coupled receptor protein signaling pathway
integral to membrane
I/R injury time-dependently increased retinal GFAP
Ischemic damage to different tissues such as brain and kidney share many similar pathologies [41, 42]. In particular, retinal I/R results in neuronal degeneration associated with visual impairment and irreversible destruction of many layers of the structurally complex retina. Common morphological features in rodent models of retinal I/R include morphological degeneration of retinal layers, RGC death, and impairment of retinal function [18–21, 24–26]. In order to discover overall pathological mechanisms, we observed pathological progression of I/R injury over 35 days, studying impairments in retinal morphology, function, and temporal changes in gene expression. There was a significant increase in thickness of inner retinal layers 3 days after I/R injury, most likely due to initial retinal edema. This histological finding was confirmed using non-invasive SD-OCT scanning. SD-OCT is now frequently used clinically and experimentally to detect morphological features of retina [43–47]. This technique allows live monitoring of the retina without mechanical invasion or damage. In contrast, traditional histological assessment is terminal and can introduce artifacts due to tissue processing. SD-OCT scanning allowed us to detect retinal detachment 3 days post injury in all mice, which was confirmed by histology (Figure 1). Both techniques confirmed time-dependent changes in retinal morphology after I/R injury.
Previous studies have suggested diverse molecular events promoting or attenuating I/R-induced retinal damage. A number of neuroprotective approaches have been tested in retinal I/R models. One major effort was to define protective mechanisms using pharmacological approaches. Ueda et al. reported that retinal neuronal injury occurred by both necrosis and apoptosis, which were inhibited by treatment with the cognition-enhancer, Nefiracetam [48]. Sun et al. also demonstrated that Cobalt protophorphyrin-induced Heme Oxygenase-1 attenuated I/R induced injury in retina [49]. The hypolopodemic drug Simvastatin also improved retinal ganglion cell survival in mouse model of retinal I/R injury [50]. More recently, Fujita et al. showed that pharmacological blocking of Angiotensin II type 1 receptor signaling produced neuroprotection via suppression of ROS production [51]. Genetic disruption of specific genes in mice has also been used to identify pathogenic and neuroprotective pathways. Aquaporin-4 null mice were protected from ischemia-induced retinal functional impairment and cell death [52]. Deletion of complement component C3 also induced retinal protection against I/R injury [32].
It is also important to examine molecular mechanisms associated with progression of retinal injury. cDNA microarray analysis is often used to provide extensive mRNA expression data [53–56]. Gene arrays provide profiles of functional gene clusters using a variety of bioinformatic approaches [34, 35]. Youshimura et al. previously reported the temporal and spatial expression of immediate early genes in retinal neurons after retinal I/R injury [57]. Hollborn et al., also specifically identified inflammatory and immune-response-related genes activated in the early stage of experimental retinal detachment [58]. In addition, Kamphuis et al. evaluated changes in retinal gene expression following ischemic preconditioning [59]. In our study, we performed cDNA microarray analysis at 8 different time points after retinal I/R injury and verified their differential expression with real-time RT-PCR (Figures 8 and 9). Based on our gene clustering data, we observed temporal changes of several genes related with signaling pathways, structure/cellular stress and inflammation, based on their relation with ischemic diseases in the retina or other tissues. For example, Stat3 is protective in various ischemic diseases including retinal I/R injury [30]. Modulation of glutathione peroxidase (gpx) expression has been reported under ischemic environments in various tissues [60–63]. bcl6 was originally known as a modulator of STAT-dependent interleukin-4 (IL-4) response in B cells [64]. bcl6 is induced in circulated leukocytes after ischemic stroke, but its precise role in this condition is unknown [65]. Caspase8 is a major molecule in the apoptotic cascade involved in ischemia-induced cell death [66, 67]. Although crystallins were originally known as structural proteins in lens [68–70] crystallins also are molecular chaperones structurally similar to small heat shock protein (hsp) with the ability to prevent protein aggregation [70–73]. Altered expression of the crystallins has been observed in various ocular diseases such as diabetic retinopathy, uveitis and glaucoma [74–78]. However, no clear changes in crystallin expression has been shown previously in retinal I/R injury.
One of the most interesting changes was the up-regulation of various inflammatory genes including c3, c4b, ccl12, and gfap (Figures 8, 9, 10). In particular, several researchers discovered that complement components play important roles in eye development and ocular pathology such as glaucoma [79]. In addition, genetic disruption of c3 protected mice against retinal I/R injury [32]. Both c3 and c4b genes encode C3 and C4b proteins, essential for the classical complement cascade [80, 81]. Expression of these proteins is temporally regulated and may play differential roles at different times during I/R injury. Retinal GFAP expression is mainly observed in Müller cells during retinal injury [39, 82, 83]. Like other glial cells in neuronal system, Müller cells play a pivotal role to maintain retinal neuron homeostasis, such as scavenging neurotransmitter/waste products, supplying energy for retinal neurons, and other protective and maintenance roles for neurons [84, 85]. Under pathological conditions, Müller cells are activated, undergoing functional and morphological changes associated with gliosis [39, 82]. Hirrlinger et al. demonstrated that transient retinal ischemia in mice induced Müller cell gliosis accompanied by altered protein expression and changes in membrane properties [86]. Our data provide further support for Müller cell-dependent retinal gliosis, involving changes in gene and protein expression as well as Müller cell morphological changes that correlate with progression of I/R injury.
As previously mentioned, retinal thickness, especially in the inner retina, significantly increased within 3 days of retinal I/R injury. It is likely that this increased retinal thickness was due to retinal edema. Others have shown that the increased retinal thickness in retinal ischemia is due to retinal edema, which may be mediated by ET-1 in endothelial cells [12]. Retinal edema plays a major role in the pathogenesis of other types of retinal injury, including retinal vessel occlusion and diabetic retinopathy. In addition, progressive retinal degeneration following edema in our model was also strongly correlated with significant cell loss in the RGCL, including RGC and displaced amacrine cells. Immunohistological assessment using RGC specific antibodies to Brn3a and NeuN showed RGC loss induced by I/R injury. Interestingly, a similar cell loss ratio from histology data (~30%) was also shown in our immunohistochemistry data (~30%) at 28 days after I/R injury. These data suggest that I/R injury caused cell loss of both RGC and displaced amacrine cells in the RGCL. In support, Kim et al., previously showed that retinal I/R injury induced apoptotic cell death to both RGCs and displaced amacrine cells [87].
One of our novel observations is I/R-induced retinal detachment (Figure 1). Retinal detachment is a major cause of vision loss in various ocular pathologies, including age-related macular degeneration (AMD) [88–90]. We first observed the retinal detachment in all ischemic eyes by SD-OCT scanning at days 3 and 7 after I/R injury, but this detachment disappeared at 14 days. We took advantage of SD-OCT scanning to monitor the real-time morphological status of the retina without sacrificing mice. Interestingly, Zeng et al. developed a novel mouse model of retinal detachment using a similar cannulation method [91]. In contrast to our data, retinal detachment completely recovered within 24 hrs in their model. In support, Uckermann et al. also suggested that transient retinal ischemia in rabbits can cause exudative detachment of the retina through days 3 and 8, which was accompanied by changes in Müller cell K+ conductance [92]. They suggested fluid-mediated retinal detachment as a novel ischemia-mediated damage to the ONL. Our findings also support retinal detachment as another potential pathologic mechanism for temporal retinal dysfunction and degeneration. Our ERG a-wave data, which is associated with outer retinal photoreceptor function, also support this finding (Figure 6). Retinal detachment was associated with ERG a-wave amplitude deficits from days 21-28 after I/R injury. Interestingly, a-wave amplitudes recovered at 35 days, correlating with recovery of retinal detachment. These results suggest that early retinal detachment causes delayed outer retina ERG deficits that are reversible after retinal reattachment. Therefore, retinal detachment in the mouse model of retinal I/R may contribute to overall ischemia-mediated retinal damage.
Interestingly, all of our data showed strong temporal correlations. We detected thickening of the whole retina, especially the inner retina, 3-7 days after I/R injury. Fourteen days post I/R injury, four significant changes simultaneously occurred including: (1) decreased inner retinal layer thickness, (2) significant loss of cells in RGC layer, (3) significant changes in gene expression profiles, and (4) increased GFAP immunostaining (gliosis) throughout entire retina. Functional impairment (i.e. decreased ERG responses) began 7 days after I/R injury, suggesting decreased retinal function was due to early retinal edema and/or damaged retinal cells prior to morphological degeneration. Our findings are the first to demonstrate temporal morphological changes accompanied with functional and molecular changes associated with progression of retinal I/R injury.
In conclusion, transient I/R induced morphological changes mainly in the inner retina that were strongly associated with functional impairment as well as temporal changes in retinal gene expression. Our data also indicated that retinal detachment was induced by retinal ischemia in the early stages of injury. Our characterization of temporal retinal changes produced by retinal ischemia will lead to a better understanding of molecular pathogenesis associated with this injury as well as suggest novel therapeutic approaches to mitigate this retinal damage. Future studies will identify cellular and molecular mechanisms associated with I/R damage to the optic nerve and visual axis in the brain. This will lead to the discovery of new neuroprotective strategies and agents for the treatment of the retina, optic nerve, and visual axis in the brain associated with retinal I/R injury.
The authors would like to thank Tasneem P. Sharma for technical help and advice on gene array analysis with Partek and DAVID. This work was supported by a grant (W81XWH-10-20-0003) from the Department of Defense (DOD).
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Reasons To Visit An Art Musem
Art museums are one of the most well-known types of museums throughout the world. Regardless of if you are an art enthusiast or not there are actually many reasons to go to an art museum. It may sound quite strange to the average person who may not know much about art but it will sound become apparent why art museums are an excellent place to explore.
Some of the many reasons to visit an art museum are as follow:
Art museums make people happy
Studies have shown that art museums actually can increase people's happiness. Staring at bright colors and beautiful images is known to release dopamine in the brain. This is the same hormone which is responsible for people falling in love.
They help develop empathy
Art museums have actually shown to increase a person's level of empathy. All art can be analyzed in a different way. Analyzing pieces of art has been shown to increase feelings of understanding and empathy in unique ways.
In many cases art exhibitions are a great way to meet new people and to socialize. Discussing different interpretations and views on art can be a wonderful way to meet all kinds of new and exciting people.
Art makes a statement
Many art pieces make a very clear statement. It teaches people the different ways to say things and to express ourselves. Visiting art museums can be beneficial in teaching people how to communicate and spread messages in a more effective manner.
Art pieces in history
Attending art museums is a great way to learn more about the famous historical pieces of art. The well-known artists are recognized again and again throughout history. Throughout your life it will be very beneficial for you to understand, know and be able to recognize the famous artists and their work.


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Cloud Computing is Good, But Hybrid Cloud Computing is Best
Hybrid Cloud Computing is the only way to backup your business data.
Shopping through the aisles of the internet for help with your company's data recovery options and cloud computing needs, can feel a little like being an ant in a Super Walmart. However, that's no longer necessary as hybrid cloud computing has proven to be the way to go. Remember, running a small business shouldn't have to mean running an inadequate one. Just because a company is operating on a smaller scale with limited resources, doesn't mean it can't enjoy the privileges and luxuries like the big guns in the game.
Small and medium-sized businesses (SMBs) run the same risks and challenges with their data recovery and backup as larger companies, especially in the case of an emergency. Too many companies have learned the hard way after a disaster that one backup plan just isn't enough. The benefits of a hybrid cloud computing backup firm can give a young business, small start-up, or industry innovator, the opportunity to thrive in competitive markets, build a better businesses, and continue to grow.
Without proper business continuity and data backup strategy, businesses can suffer severely from the effects of natural disasters, human error, or hacker attacks. Businesses can't afford downtime or unhappy customers and lost revenue in a time when people have little patience for the compromised security of their data and personal records. When a company's infrastructure is compromised, business owners need professionals to formulate and execute a plan that will enable them to remain functional and operational.
Imagine your company's servers sitting under five feet of water after a hurricane and the infrastructure of your entire business being compromised. Regardless of size, you need a plan that affords you with the preparedness, that until recently, only big companies could afford to deploy. A plan that ensures your networks can be virtualized, your data will be backed-up in multiple locations, and one that gives you a way to run your business remotely from the cloud.
With the cloud, businesses can expect to enjoy improved communication, higher customer satisfaction, and heightened security as well. According to Forbes, 25 percent of executives say they unexpectedly experienced improved communications within their organizations. 22 percent reported increased revenues they did not anticipate and 22 percent say they experienced greater customer satisfaction. It's also notable that 21 percent say cloud has actually delivered improved security."
As the challenges of protecting systems and data have become more complex as company's IT infrastructures have expanded, the risks of not having adequate business continuity and data backup strategies have increased dramatically as well.
Please contact us for any additional information on hybrid cloud computing and be prepared for whatever mother nature, or the next hacker attack throws your way.
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