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CrossFit is the principal strength and conditioning program for many police academies and tactical operations teams, military special operations units, champion martial artists, and hundreds of other elite and professional athletes worldwide. Since then it has evolved to be an all inclusive fitness program for people of any athletic capability. Our program delivers a fitness that is, by design, broad, general, and inclusive. Our specialty is not specializing. Combat, survival, sports, and life all reward this kind of fitness and, on average, punish the specialist.
The CrossFit program is designed for universal scalability making it the perfect application for any committed individual regardless of experience. We've used our same routines for elderly individuals with heart disease and cage fighters one month out from televised bouts. We scale load and intensity; we don't change programs. The needs of Olympic athletes and our grandparents differ by degree not kind. Our armed forces, skiers, mountain bike riders and housewives have found their best fitness from the same regimen.
We have lots of memberships for your needs and budget. Get started today with a FREE intro session by clicking here!
Membership Options:
3x per week
Month-to-Month - $175
6 Month - $155
12 Month - $145
Month-to-Month - $199
6 Month - $179
12 Month - $169
Personal Training Sessions
1 - $75
4 - $280
8 - $520
16 - $960
24 - $1200
Fundamentals Course
$159 one time fee for four 1-on-1 sessions
When you first join CrossFit Belmont Heights, you'll start out with a FREE intro session where you'll perform a baseline workout, chat with our coaches, and discuss your goals. Then, you'll attend a fundamentals class or regular group class, where a coach will scale each movement to your fitness level. Afterward, we'll assess your experience and answer any membership questions you may have.
We can't wait to meet you! To get started today, just click here.
In short, yes CrossFit can be dangerous. But not in the ways you might believe. With inexperienced coaches, shoddy equipment, and the wrong attitude, CrossFit can be very dangerous – but so can riding your bike or driving your car. The experienced and proven CrossFit coaches you'll find at our facility will ensure that you're training in the safest most effective and efficient way possible.
A WOD is the workout of the day. We program a different WOD everyday.
We love visitors! To schedule your drop-in session, click here.
Single: $20
3-Class: $55
5-Class: $89
10-Class: $169
Yes! Our coaches would love to meet with you 1-on-1 so you can get the most from your workouts. To get started with personal training, click here.


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I like it by itself, but I also like to make my "breakfast crumble" with it. I slice up some fruit, put some yogurt on top and then sprinkle the granola over it.
I have been sewing. Made myself a top. Wore it, and now it's in the wash, so no picture just yet.

subset: commoncrawl


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Join the movement! Historic structures and homes tell an important story of who we are as a state, as a town, as a community. Your membership is crucial to our continued work of saving Maine's past.
This critical underwriting supports the organization's basic operations and programs, reinforcing our ability to save the historic homes and buildings that give Maine its unique character.
The best way to support our mission is through a donation. Maine Preservation is a federal and state-registered charitable organization. It's only through your support that we are able to promote and preserve Main'e historic places.
It's easy to make a bequest in your will of either property or income to ensure future generations will remember your legacy. Maine Preservation has expertise in selling property and, if it is your wish, can protect buildings for future generations. A planned gift of an annuity can provide you with generated income over the span of your retirement. Please contact us for more information!
You can make a meaningful gift in memory or in honor of a special person in your life or a milestone event with a gift that supports Maine Preservation's valuable work. A letter commemorating your generosity will be sent to loved ones to let them know of your thoughtfulness.
Gifts of securities can be directed to the Maine Preservation brokerage account with Bangor Financial Services. If you have any questions please contact Eric McLeod at 800.585.6145, ext. 1517 or Shelley Witt at ext. 1594.
Please also contact us at when you make a stock gift so it doesn't land in the account silently.
Clearing Firm: LPL Financial Services
Account Registration: Maine Preservation
Account #: 4697-4984
Patricia McGraw Anderson was a noted local author, art historian, teacher and tireless advocate of historic preservation. Maine Preservation's summer fellowship program is dedicated to the professional education of the next generation of historic preservationists.


text:
Macro Man managed to hang on and eke out a small positive return (0.17%) for March, though the drawdown from the intramonth high was substantial. In contrast to prior months, it was the beta portfolio that did the heavy lifting, returning 0.64%. This is actually understated, moreover, as SPY went ex-div during March (taking $0.55 off the price), but payment will not accrue to the portfolio until the end of April.
The alpha portfolio was weighed down by costly equity hedges, the poor performance of commodity proxy plays, and the decoupling of 'risky FX' from equity markets. Equity, commodity, and FX alpha lost 0.59%, 0.14%, and 0.42%, respectively. On the other hand, just about every fixed income bet came off, and the fixed income alpha bets made 0.68%. Macro Man's one regret is that he didn't do more of the Euribor trade, as the view was strong and the risk/reward was solid.
Q1 was clearly a quarter for alpha, rather than beta. All told, Macro Man's beta strategies (ex SPY dividend) broke even on the quarter, so the entirety of the 2% quarterly return was from alpha bets. Fixed income and equities have seen far and away the best alpha returns, with slightly negative performance from commodity and FX bets. If Macro Man's portfolio is anything to judge by, "beta hugging" macro managers will have delivered flat to slightly negative returns for Q1 (assuming 0.50% quarterly management fees.)
It was disappointing but not surprising to see commodity alpha as a singificant drag on portfolio performance. After all, the exisiting positions are both proxy (Goldcorp) or RV (Dec WTI-Brent spread) positions rather delta bets on the underlying. Given that Macro Man recently explored the dangers of proxy trading, some remedial action may be required soon.
In that post, Macro Man offered a legitimate, tangible prize to anyone who could correctly identify the two correlated series in the chart below. The point, of course, was to demonstrate that apparent correlation does not imply causality. Sadly, there were no takers, though it's difficult to know whether that's down to the difficulty of the task or the paucity of interest that Macro Man's new quiz feature is generating.In any event, the two series are about as different as two things can be. Series two is the June 2007 short sterling contract, through which investors can bet on UK cash rates. Series one is an index displaying the long-term average temperature per day in Fukuoka, Japan. The correlation appears strong, but the underlying relationship is actually zero. So the next time somebody shows you a chart that appears to demonstrate a relationship between two series, remember to ask yourself what's the temperature in Fukuoka.


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15 Amp, 14 AWG, 90 Degree Angle Adapter Plug, 24" Inches, BlackHeavy Duty DesignWill not interfere with the backs of furniture or wall mounted displays 24 in. length..
ADP-1520B 15A-20A Adapter Cord
The ADP-1520B Power Adapter Cord is a 12" long adapter cable that features a 20 amp female socket at one end and a 15 amp male plug with an internal breaker at the other. This will allow 20 amp produc..
Panamax GRM2204Rack mounting kit (standard 19" racks) for the MR5100, M5100-PM, M5300-PM, M5400-PM, M7500-PRO...
Options Available: 1
XDB9-TB-H1 Easy Adapters DB-9 Enclosure
Easy Adapters
This plastic shell is designed to enclose any of the Easy Adapters DB9 breakout boards ( FDB9-TB5+, FDB9-TB10+, MDB9-TB5+, MDB9-TB10+ ). Enclosing the breakout board provides additional protection for..
CIS400502 is temporarily out of stock. This product should be available to ship approximately 3/1.Are big, bulky power adapters creating unsightly gaps between your walls and furniture? The Flat..
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Get full use of your power strip outlets... and double the number of available outlets, too, with a Power Strip Liberator Plus II cable! Just plug a bulky AC adapter into a Power Strip ..
Get the full use of your power strips and UPS outlets with our high-quality Power Strip Liberator cables. Just plug those big bulky power adapters into our extension cables and plug the cables into th..
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Temporarily Out of Stock. Expected Ship Date Approximately 1/21/19Connect two AC adapters to one multi outlet extension cord with the Y-Splitter Liberator®. Double the outlets provided by a powe..
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Squirrel's Secrets - Cross Stitch Kit
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Item: NWCS01 - Type: Cross Stitch Kits.
Create a beautiful decoration to display in your home or office! This cross stitch kit contains 14 count Aida, DMC floss, needles, stitch diagrams and instructions. 5.75" x 7".
123Stitch.com ships via the U.S. Postal Service to anywhere in the world. Orders to the U.S. typically arrive 3-5 days after we ship.
Shipping charges are shown in your shopping cart every time you add an item.
If you are not completely satisfied with any of our products, you can return it to us within 30 days for a full refund.
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>> Conference Poster
The conference organizer is Professor Aloyisius Siow, Deparment of Economics, University of Toronto. The on-site organizer at Dalhousie University is Professor Peter Burton, Department of Economics, Dalhousie University. Please direct all inquires and correspondence to the Program Assistant Maureen Church. Postal Address: CEA Program Office, University of Calgary, Department of Economics, 2500 University Drive N.W., Calgary, Alberta, Canada T2N 1N4. Phone/Fax: +1-403-697-9042. E-mail: email@example.com.
The CEA web site is maintained by Werner Antweiler.
valid HTML


text:
"Cutting the cord" sounds like it could be some type of surgical procedure, but really it's slang for getting rid of your subscription TV service. However, like surgery, "cutting the cord" could be painful and not without certain risks. But relax, let BrightReviews guide you through the process so you know exactly what you are in for.
Part One: Why Cut Your Cord?
In a sense, cable is an essential staple of American TV. It has been around in some form or another since at least 1948, when it was used to help over-the-air television stations reach remote areas that couldn't receive the analog signal. But it wasn't until 1975 when HBO became available nationwide by satellite and 1976 when Ted Turner's WTCG came on the air, that there began to be a separate television service that would charge a monthly fee for access.
Of course, since that time there has been an explosion in channels and content. As of 2015, the top players in this game are Comcast (22,375,000 subscribers), DirectTV (20,412,000), DISH (13,844,000), and Time Warner 11,025,000). There were an estimated 900 channels offered in 2014.
That's a lot of channel surfing!
However, all these channels and premiums are not without cost – cable companies are said to have spent almost $307 billion in programming since 1996. This has been passed onto the consumer in the form of ever-rising subscription prices and bundled packages that many complain are overpriced, containing channels they do not want. An average basic cable subscription currently runs $54.92 per month and is expected to rise to $61.76 by 2018.
Rise of the Cord Cutters
But as broadband Internet has become more prevalent allowing the transmission of video, upstart "rebel" services like Netflix, Hulu, and others have attempted to disrupt the traditional cable model and change the way entertainment is consumed. Smart TVs with a WIFI connection and portable devices like smartphones and tablets have also fueled the demand for digital video.
They appear to be winning, or at least gaining a strong foothold: in 2013 the number of cable subscriptions fell by a quarter million the first full-year decline ever reported, noted Bloomberg News. However, they added that the entire pay-TV industry is still huge at approximately 100 million people.
And while these so-called "cord cutters" may still be relatively small, they have banded together and created a very vocal minority that see these cable companies as greedy monsters, robbing them of choice when it comes to what they want to watch, where, and when.
Is Cord Cutting a Good Thing?
If you listen to the cable/satellite/broadband subscription providers, the answer to that question is a resounding "no". The reason the cable bills are so expensive is not their fault, they claim. They must pay expensive fees to carry more popular channels like ESPN and if they didn't bundle so many channels together and offered more a la carte options, some of your favorite channels/shows would become prohibitively expensive to operate and potentially cease to exist.
It could also be argued that given the number of channels provided, including live sporting events and niche programming, it's a relative bargain compared to say, a cellular phone – the average cell phone bill is estimated at $71 and contains essentially NO entertainment content.
Still, something clearly needs to be done to make everyone happy, but the answers are difficult and unclear at the moment. HBO and Showtime have recently broken from the cable mold and have begun offering Internet-based services. Even the pay-TV behemoths themselves are getting into the act. For example Sling TV is a service that proclaims to "liberate" you from an expensive cable package, but it's secretly run by Dish Networks.
Cutting the Cord… and the HEAD?
As we were writing this article, news arrived about the latest blow to the traditional pay-TV model. The Walt Disney Company, considered a stellar money-maker in years past, announced that it would not be as profitable in 2015, because fewer consumers are subscribing to full pay-TV packages. Upon the news, there was a 2-day sell-off of many major media stocks, causing investors to lose $60 billion.
This stock market free fall may be temporary, but it has rattled the industry. These giant companies who once ruled their cable fiefdom are now worth less money, and they may have to cut back on creating more content. This could lead to fewer TV shows, movies, and other events that the Cord Cutters so desperately want (but don't want to pay for).
And who will take their place? Most of these small startups have little experience in content development. Bigger players like Netflix and Amazon have entered the fray, jump-starting their catalogue by spending billions of dollars (they don't have) on programming. Even as their stock prices soar, some feel they are over-valued and could eventually collapse.
This could leave everyone that enjoys movies, sports, concerts, and television shows with less choices overall.
Concluding Thoughts on Part One
We don't want to sound like we're a shill for Comcast, Dish, or for that matter Disney, Viacom, or Netflix. Content is valuable and yet consumers clearly need – and demand – more choices. There are big changes on the horizon, but they could be a mixed blessing for everyone.
In the next installment, we'll look at the key players in the Cord Battle, examine their "weapons" and expose their "weaknesses". Stay tuned!


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Words in English
Published: 14.02.2010, Updated: 02.07.2013
This website is a resource for those who want to learn more about the English language – its history as a language, the origins of its words, and its current modern characteristics. This is an excellent background website to the English language with menus to history, structure, usage, meaning and resources. View the link to The Brief History of English, with chronology http://www.ruf.rice.edu/~kemmer/Words04/history/index.html
Competence aims
Part of
- Tasks and Revision (Link for Engelsk)
- Planning the Year (Link for Internasjonal engelsk)
- Planning the Year (Topic for Internasjonal engelsk)
- Engelsk (Course)


text:
I recently had the opportunity to visit my dear friend, Joe Bowler, in Hilton Head, South Carolina. Before heading to the island, I debated as to whether or not I should take a map with me just in case I got lost or ran into a detour. I realized that there was no need to do so because I had traveled down those familiar roads in the past and knew what lay ahead of me. Even if I were to encounter unexpected detours along the way, I felt confident enough to be able to navigate back to the main roads which would ultimately lead me to my destination.
Upon arriving, I attended the opening of Joe's one man show at Morris & Whiteside Gallery. I spent the next day visiting with him in his studio as we talked about great painters of the past, shared ideas about art and techniques, looked at art books and had my work critiqued as well. We have done this many times before, since the day I first met Joe and his wife, Marilyn, as an art student back in college.
As I sat and listened to Joe's advice, I couldn't help but think of how he has been my artistic map through the years. He has traveled down roads which I have yet to experience and has valuable information about what may lay ahead. There have been times when I have taken detours or even veered off course but, like every great map, Joe has guided me towards the correct path in order to point me in a direction which will help to lead me towards my goals. His work has greatly influenced me over the years and I am constantly learning something new each time that I view his paintings. Not only is he the most talented artist that I have ever met, but he is also one of the greatest men that I have ever known.
I have included some of his work on my Facebook page with the thought of inspiring others as much as Joe has inspired me over the years. I hope that it will encourage you on your own artistic path towards reaching your goals.


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Your donations keep RPGWatch running!
CONSORTIUM is a first-person story and character-based role-playing shooter set in an interactive world defined by your choices.
Rather than endlessly exploring a huge, expansive and ultimately static world, you'll find yourself within a dynamic, bustling, interactive and alive micro-environment: the Consortium Command Vessel, C-3800-D "ZENLIL."
Options: Add / View Comments (0) | Forward this page | Link to this page
Information aboutConsortium
Developer: Interdimensional Games
SP/MP: Single-player
Setting: Sci-Fi
Genre: Shooter-RPG
Combat: Real-time
Play-time: 20-40 hours
Regions & platforms
· Homepage
· Platform: PC
· Released at 2014-01-08
· Publisher: Interdimensional Games


text:
Fetzer Simonson & Booth
50 West Broadway, Ste. 1200 Salt Lake City, Utah 84101 801-328-0266 Office 801-328-0269 Fax
Clark B. Fetzer
Gregory M. Simonsen
Bryan H. Booth
Scott R. Jenkins
John W. Mann
Brooke Ashton
Alan S. Bachman
Collin R. Simonsen
Jaelynn R. Jenkins
Practice Areas
Alternative Dispute Resolution
Business Entity Formation
Charitable Organizations/Non-Profits
Creditor's Rights
Commercial Litigation
Estate Planning/Probate
Family Law
Insurance Defense
Land Use/Zoning
Real Estate
Real Estate Title Dispute
Contact Us
Construction Law
Our attorneys practice in virtually all aspects of construction matters, from project finance and contract negotiations through bid procurement, project performance, and claims such as breach of contract, mechanics liens, construction defects, toxic mold, liquidated damages, delay and acceleration, differing site conditions, and termination. Our clients include general contractors, sureties, owners, developers, subcontractors, and lenders involved in public and private projects, in highway, industrial, school, church, commercial, and residential construction. Construction lawyers at the firm have presented at many seminars on construction law and four of our attorneys serve or have served as leaders in the Utah State Bar's Construction Law Section. We are experienced litigators who also employ alternative dispute resolution and advise construction clients in preventing and avoiding claims and litigation costs.
Copyright © 2019 Fetzer Simonsen Booth Jenkins. All Rights Reserved.
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text:
Ajunta. Verandah of Vihara No 2
Lithographer: Dibdin, Thomas Colman (1810-1893)
Medium: Lithograph, coloured
Date: 1839
This is plate 11 from James Fergusson's 'Illustrations of the Rock Cut Temples of India'.
Cave 2 at Ajanta dates from the 5th century, is one of the most complete examples of the vihara form in the complex. It is known particularly for the complex geometric and floral patterns of the painting on its ceiling, as well as the fine paintings on its walls. Three doorways lead into the hall where columns are arranged around a square space. In the middle of the rear wall an antechamber leads to a shrine, wherein sits a seated figure of the Buddha. The cave is richly decorated with sculptures, and the columns are elaborately carved with scrollwork and jeweled motifs.


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Tourism, Accommodation and Historical Attractions in Natal, South Africa
Mabibi: isiMangaliso Wetlands Park
Clear water, offshore reefs and tropical fish make Mabibi a paradise for snorkellers and scuba divers and underwater photography.
Zululand: Mabibi
|Advertise Now -it's Free!
|Change text size:
It is also well known for its offshore angling and the profusion of life in the intertidal pools. The beach is reached through the coastal dune forest and then by means of a ladder.
Accommodation: Ten camping sites, a communal braai and ablution block.


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Radical changes including a sugar tax and regulation of junk food promotions have been recommended by government advisers as a new report revealed sugar consumption in Scotland has failed to improve in five years.
Food Standards Scotland (FSS) made a raft of recommendations yesterday on how to tackle Scotland's "health time-bomb" of obesity and poor diet, including reductions in promotion, advertising and portion control.
Experts have predicted Scotland could face adult obesity levels of 40 per cent by 2030 if current trends continue.
The independent food body will advise ministers to consider how a sugar tax could be brought in, allowing the industry a 12-month grace period to come up with an alternative to reduce sugar consumption levels.
Despite reductions a 21 per cent drop in sugary drink purchases since 2010, the amount of sugar purchasing has not changed. Industry representatives said punishing the industry which employs 34,000 people in Scotland would be a "retrograde step" and reformulation of products would be a better move.
Ross Finnie, FSS chair, said: "There can be few in any doubt now as to the gravity of the health timebomb related to poor diet and obesity facing our nation.
"Individual responsibility around food choices, exercise and activity levels remain important, but this cannot be left to individuals alone."
A recent study revealed that a sugary drinks tax in Mexico reduced consumption rates by 12 per cent in its first year.
The report also scrutinised Scots buying habits which confirms large quantities of discretionary foods such as biscuits, cakes and chocolate are being consumed, particularly in the lead-up to Christmas where an additional 9,000 calories is consumed by every adult in Scotland.
Obesity campaigners hailed the measures as a turning point in addressing Scotland's battle with poor diet but urged FSS to go further through reformulation of products and better labelling.
Lorraine Tulloch, programme lead for Obesity Action Scotland, said: "If we are to turn the tide on the crisis we face with obesity and overweight in Scotland we need such bold actions."
David Thomson, chief executive at the Scottish Food and Drink Federation, said: "Punishing and legislating against an industry that employs 34,000 people in Scotland would be a retrograde step."


text: I like the dual boot solution too. There are a lot of very hairy issues involved in cycle stealing...especially if the machines crash more frequently with your software than without. Desktop support people notice things like that (as well they should!). The TCO argument seems silly to me when based on hardware costs. Seems to me that you need to buy the disk anyway, it's whether or not you have to buy a computer to go with it. :) The real savings from moving to a dedicated cluster is on the admin / support end. In terms of OS coherency, there are folks on this list with great experience in that area. The short form is that you're going to need to address the issue of pushing out updates to compute nodes whether they're dedicated to a cluster or not. The answer is to do it at boot time, from a boot image server. > > If, on the other hand, it's concern over trying something new, the > > Engine system recently implemented at Novartis is a decent example of > > a corporation gaining a great deal of horsepower this way. > > Can you provide more details on this or point me in the right direction > to get more info please? United Devices (http://www.ud.com) has a commercial implementation of cycle stealing software a la Seti@home. Their page has a number of good links. Included there is a PDF describing a major install at Novartis. Basically, Novartis has already plugged 2,700 workstations into their UD grid, and they plan to include 62,000 more as they finish standardizing every single workstation across all their sites. It's a cool project by any measure, but (of course) the devil is in the details. -Chris Dwan


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In the event of an Emergency, please dial 000 to reach Police, Fire and Ambulance services.
Carers provide care and support to family members and friends who have a disability, mental illness, chronic condition, terminal illness or who are frail aged. We work closely with government and other support organisations to improve the lives of caring families throughout Victoria.
Case Management Services provide a single point of contact for people with a moderate level acquired brain injury to remain living at home, and their families. Case managers work one on one with clients, families and carers to identify needed services and supports for the person; offering support; advocacy; and assisting to organise and coordinate community care services. Funding may be used to provide respite, and fund social support groups.
159 Langtree Ave, Mildura. Central location point offering information and referral, drop in centre, advocacy, training and resource library. Support groups include; Parents and Carers Support group; Sunraysia Autism Spectrum Support Group; Sunraysia ADHD Support Group; Sunraysia Disability Group; and Sunraysia Downs Syndrome Support Group.
67 Pine Ave, Mildura.
Northern Mallee CAMHS. ASSET (Autism Assessment & Treatment Service) is a service offered by CAMHS. This assessment service is designed for children whose parents, teachers or doctors are concerned that they may have autism or a related problem in development.
115 Twelth St, Mildura. Adult Training & Support Service (ATSS) and Supported Employment Services (SSES) which provides programs for people with disabilities.
253 Eleventh Street, Mildura. Working with children identified as having a disability & their families; offering support, information and referrals. Loddon Mallee Regional Office Tel: (03) 5434 5888 or 1800 783 783 or TTY 1800 008 149
Dareton Primary Health Centre 44-46 Tapio Street. Working with children identified as having a disability & their families.
148 Lime Avenue, Mildura Host and volunteer program offering respite for carers, including; weekend activities; camps and holidays.
58 Sturt Place,Dareton NSW. Provides services assisting those with disabilities, the aged and the frail to continue living independently.
181 Walnut Ave, Mildura. Provides private Fee charging services to children and adults. Professionals at the facility include a Speech therapist; Occupational therapist; Psychologist and Dietician. Issues may involve Autism Spectrum Disorder, Dyslexia, speech difficulties, and assistance with physical disabilities.
108-116 Madden Ave, Mildura. Aged & Disability Services. Provides services to support and promote the safety and well being of older people, the frail, those with disabilities and their carers in Mildura who are in need of support to retain their independence and quality of life.
66 Ninth Street, Mildura. A transportation service offered to the frail aged who are not able to use public transport to travel to appointments and services.
341 Deakin Avenue, Mildura. Provides education to students with disabilities, aged between 5-18 who are eligible for funding.
148 Lime Ave, Mildura. This is an advocacy service for people with a disability, their families and carers, to be provided with representation, information and support.
Ramsay Crt, Mildura. The Adult Day Activity Centre for the aged can assist in preventing inappropriate/premature admission into residential care. The centre also provides respite for carers.
877/Fifteenth Street, Mildura. Services include respite for carers, supported accommodation for people with disabilities and related services. Supported Accommodation-Tel: (03) 5023 2268 Respite-Tel: (03) 5023 6703.
123 Langtree Avenue, Mildura. Offers support services for the visually impaired.
24 Drummond St, Carlton VIC 3053. Autism Victoria is the first place to call for information and advice for anyone interested in finding out more about autism spectrum disorders.
Level 30, 570 Bourke Street, Melbourne 3000. The Commissioner has been created to work with people with a disability and disability service providers to resolve complaints.
53 Eighth Street, Mildura Vic 3500. Working with children identified as having a disability & their families


text:
Design a High Power LED Light Fixture
We are seeking to develop a light fixture designer to aid in developing a high power LED light fixtures. The most immediate need is to first produce a conceptual design for a high powered (1000W) industrial light fixture. Our in house electrical engineer will collaborate with you so electrical skills are not necessary.
The project will have three phases:
1.) Conceptual design and refinement.
2.) Detailed mechanical design.
3.) DFM refinement.
The light we will be designing is meant to be an economical fixture so we will be looking for opportunities to strip out cost and simplify the design. The most challenging aspect of the project will likely be thermal management and then the DFM to make an easy to produce and assemble fixture.
Please share an relevant experience working in designing high powered LED fixtures.
I look forward to your application!
Proposal: Work Bid: Rate: $1000 Time Delivery: X days (Negotiable Based On Your Deadline) Your project will be developed with speed and quality, as well as all my previous work. I have experience in the もっと
Dear sir I am TOP RATED Product/Industrial designer with over 8 years of experience. I usually use SolidWork or Autodesk Inventor which is a very powerful tools and here is what I will do for your project: I can もっと
Dear Client. I am an Architect and have over 10 years experience in designing buildings and Interiors as well as any form of Graphic Designing Please note that I have all the knowledge and skill to take up and complet もっと
Hello. I'm a designer with an extensive experience in consumer products design. I already designed some LED light products. I also have a 3D printer to make some prototypes. I'd be glad to speak with you further abo もっと
Hi, I am Zzee, an experienced freelance industrial / product designer with many proven achievements. I hope everything is fine for you. I can assist you in designing products from your idea and rough sketches or もっと
Hi Where there is a Challenge, We Create the Solution. We have tackled challenges like yours and we can help you on your project and make it a reality!!! We are a Team of quality and solution oriented engineerin もっと
Hi, thanks for consider my proposal. I am extremely interested and fully understand the job requirement of LED fixture design I will provide complete conceptual and detail design services as per specifics. Here i もっと
well i would like to do this project for you . we have professional group of engineers and are interested to do this project .let me know the details .regards
Hi, I have read and understood the project outline and will gladly offer an outstanding service. Please give me a chance. A trial will convince you. Looking forward to work with you. Thanks
Dear Sir/Madam I trust you are well; I have design a 100 watt LED fixture and I will send you technical drawings on request.I will offer the following services 1.) Conceptual design and refinement using Creo Paramet もっと
Hi, I would love to know more about this Project. I am an Industrial Design Engineer, with around 4 years of experience. I mostly work on CATIA, AutoCAD, SolidWorks. I have in past designed die cast Aluminium light もっと


text:
One for the Spring Carnival? Kangaroo intrudes on race
Intruders on the racecourse have been a hot topic in recent weeks but it would be a shock to see anything like this on a British or Irish track as a crafty kangaroo almost caused chaos at Cessnock this week.
The runners and riders in the final race of the day at the New South Wales venue got more than they bargained for as the kangaroo joined in midway through the ten-and-a-half-furlong contest.
Perhaps a potential Spring Carnival candidate, the kangaroo showed good speed, but jockey Serg Lisnyy was more concerned about the creature's impact on his mount The Blues Brother.
A shocked Lisnyy pulled his horse out away from the rail to take evasive action, which saw The Blues Brother swerve across the path of Sebring Magic, before the kangaroo veered sharply left through a gap behind those two runners and the rest of the field and off the track.
Sebring Magic's jockey, Luke Cumberland, could hardly believe his eyes, turning his head to make sure Skippy had departed the scene before returning to a position closer to the rail.
There was no kangaroo involved in the finish but the race did go the way of The Cunning Tiger – who is definitely a horse.


text:
Working on some clown knitting. I do not have pictures.Still there is cat News of various types. So it is time to drag out the Hitler Kitty. By the way the Hitler Kitty can speak properly. So if you have too much time on your hands visit: www.catsthatlooklikehitler.com. Yes That is a real site. It is sites like that that keep me going on this blog. That way I do not feel like I am destroying western civilization by writting this foolishness.
These are kittens. Namely the Thre kittens we have at the moment. The striped one is Max. That is Walt's new cat. He is energetic and likes to climb in my shoes. So in the next couple weeks Walt should be taking about foot injuries from cats on his blog.
This varmit is Lydia. She is going to be my next cat. Actually I suppose she is already my cat. She is supper cute as well and not as fascinated with my foot wear.
We are still looking for a home for the third kitten. But we have them for at least another week. Be safe out there and keep you strick on the ice.


text:
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text:
Here's a little offering from my short story The Great Rite of the Golden Coven. This is a tale about an ancient grimoire written by a very old Celtic god, and it's affect on a coven of Romany Chohawnis (witches) who obtain its possession for the Long Night (Yule) sabbat.
* * * * *
"Come, Gueri Chohawni!" the voice called me again and, reluctantly, I stepped towards the fire. The heat grew, and I almost stepped back, but, gathering my courage, and feeding off the growing passion in my womb, I stepped closer and closer to the flames. My legs trembled as I approached the edge, and I stopped and looked up into the centre of the fire. There, at the heart of the flames, glistening and radiant, right in the middle of the stone slab, a large golden phallus, identical in design to those that capped the rods of the scroll, stood beckoning to my desire. The vision filled my eyes and inflamed my heart and yoni with a wanton, aching lust!
I swallowed hard, and stepped forward again.
* * * * *Looks like someone is heading for a LOT of fun!


text:
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- Research article
- Open Access
Differential stepwise evolution of SARS coronavirus functional proteins in different host species
© Tang et al; licensee BioMed Central Ltd. 2009
- Received: 23 September 2008
- Accepted: 05 March 2009
- Published: 05 March 2009
SARS coronavirus (SARS-CoV) was identified as the etiological agent of SARS, and extensive investigations indicated that it originated from an animal source (probably bats) and was recently introduced into the human population via wildlife animals from wet markets in southern China. Previous studies revealed that the spike (S) protein of SARS had experienced adaptive evolution, but whether other functional proteins of SARS have undergone adaptive evolution is not known.
We employed several methods to investigate selective pressure among different SARS-CoV groups representing different epidemic periods and hosts. Our results suggest that most functional proteins of SARS-CoV have experienced a stepwise adaptive evolutionary pathway. Similar to previous studies, the spike protein underwent strong positive selection in the early and middle phases, and became stabilized in the late phase. In addition, the replicase experienced positive selection only in human patients, whereas assembly proteins experienced positive selection mainly in the middle and late phases. No positive selection was found in any proteins of bat SARS-like-CoV. Furthermore, specific amino acid sites that may be the targets of positive selection in each group are identified.
This extensive evolutionary analysis revealed the stepwise evolution of different functional proteins of SARS-CoVs at different epidemic stages and different hosts. These results support the hypothesis that SARS-CoV originated from bats and that the spill over into civets and humans were more recent events.
- Severe Acute Respiratory Syndrome
- Severe Acute Respiratory Syndrome
- Receptor Binding Domain
- Strong Positive Selection
- Severe Acute Respiratory Syndrome Coronavirus
Severe acute respiratory syndrome (SARS) emerged in Guangdong province of China in November 2002 and subsequently spread rapidly to 25 countries across five continents within 3–4 months [1]. Soon after its first outbreak, the etiological agent of SARS was identified as a novel coronavirus [2–4], and its complete genome sequence was determined [3, 5, 6]. The identification of SARS-CoV in Himalayan palm civets and raccoon dogs in live animal markets in Guangdong, China, provided the first clue of an animal-to-human transmission [7, 8]. Further studies indicated that civets were unlikely to be the natural reservoir [9]. Instead the detection of different SARS-like-CoVs in horseshoe bats (Rhinolophus spp.) seemed to suggest that bats might be the natural reservoir of SARS-CoV and many other closely related coronaviruses [10–13].
Similar to all RNA viruses, SARS-CoV replication is associated with genomic and antigenic variation. The ω ratio (dN/dS ratio of non-synonymous to synonymous substitutions) can measure the selective pressure at protein level, with ω = 1, < 1, > 1 indicating neutral selection, negative selection and positive selection, respectively [18, 19]. Previous studies have suggested that the S protein of SARS-CoV experienced positive selection during SARS epidemic [20–22]. However, these studies did not find or did not analyze for positive selection among the replicase or accessory proteins, which may be equally important for SARS-CoV's adaptation to a new host. In order to systematically investigate the adaptive evolutionary process of SARS-CoVs, we employed the branch-site model to analyze the selective pressures that may act upon some key SARS-CoV functional proteins involved in virus entry, replication and assembly. Our results suggest that diversified selective forces act upon different proteins and during different epidemic phases.
Sequence data
A total of 156 sequences of SARS-CoVs or bat SARS-like-CoVs were retrieved from GenBank (129 complete genomes and 27 partial genomes) (see additional file 1). Based on these sequences, three datasets were constructed. Dataset 1 contains all Spike genes. Dataset 2 is a merged dataset that includes sequences of 4 main replicase domains of SARS-CoV: papain-like protease (PLpro), 3C-like protease (3CLpro), RNA dependent RNA polymerase (RdRp) and Helicase (Hel). Dataset 3 is a merged dataset that includes sequences of 7 ORFs: ORF3a, E, M, ORF6, ORF7a, ORF7b and N genes.
These protein-coding sequences are aligned based on translated protein sequences using Clustal W program implemented in BioEdit [23, 24]. Prior to analysis all sequences that were identical to another within the dataset were removed, since previous studies have shown to have little effect on the detection of positive selection and contribute little evolutionary information [25]. Alignment gaps were manually removed based on the reference sequence of 31-HP03L_Tor2 (NC_004718).
The final composition of each dataset is as follows: dataset 1 contains 3765 bp of 59 S gene sequences; dataset 2 includes 35 sequences of replicase domains, 6435 bp in total (945 bp for PLpro, 918 bp for 3CLpro, 2769 bp for RdRp, 1803 bp for Hel) [17, 26–28]; and dataset 3 contains 56 combined sequences, 3666 bp in total (822 bp for ORF3a, 228 bp for E, 663 bp for M, 189 bp for ORF6, 366 bp for ORF7a, 132 bp for ORF7b and 1266 bp for N).
Phylogenetic analysis and reclassification of SARS-CoVs
Detection of recombination and positive selection
Since recombination can influence the detection of positive selection, we first tested for recombination in our data sets by using a genetic algorithm for recombination detection (GARD) [30]. Identified breakpoints by GARD were then assessed for significance by using Kishino-Hasegawa (KH) test in HYPHY package [31]. Since most sequences in SARS group have high similarity and increasing the number of sequences may dilute the signal, for each dataset, we choose 10 sequences for GARD analysis (four from BSL group: 124-Bat_SARS-273, 125-Bat_SARS-279, 126-Bat_SARS-HKU3, 127-Bat_SARS-Rp3; six from SARS group: 3-HP03E_GZ02, 15-HP03M_BJ02, 31-HP03L_Tor2, 106-HP04_GZ0402, 110-PC04_PC4-136, 130-PC03_SZ13).
To test for diversifying selection and to infer codon sites under positive selection, we mainly used CODEML program in the PAML 4.1 software package, which is based on the maximum likelihood algorithm of Yang and coworkers [32]. Three kinds of models (branch-specific, site-specific and branch-site) were employed to detect selective pressure among different branches and at different sites. The likelihood ratio test (LRT) was used to investigate whether the null hypothesis, where no positive selection is allowed, can be rejected against the alternative hypothesis, where positive selection is allowed [32]. The one ratio model (M0) assumes the same ω ratio for all branches and sites in the phylogeny. The free-ratio (FR) model assumes an independent ω ratio for each branch in the phylogeny. M0 and FR can be compared using LRT to examine whether ω ratios are different among lineages. The discrete model (M3) estimates ω for three classes of codon. Comparing M0 and M3 can test the variability of selective pressure among sites. When evidence for positive selection (ω > 1) was detected, the naïve empirical Bayes (NEB) method was used to calculate posterior probabilities for site classes. A higher posterior probability suggests strong support for a site to be under positive selection. In brief, the branch-specific model assumes variation among branches, but not among sites; the site-specific model assumes variation of selective pressure among sites, but not among branches. Both models are widely used to investigate selective pressure. However, if adaptive evolution occurs at a few time points and affects a few amino acids, these two models might lack power in detecting positive selection. To overcome this limitation, we also used branch-site model, which assumes that the ω ratio varies both among sites and among lineages [33, 34]. In the branch-site model A (model A), the lineages of interest are set to be foreground, and the other lineages to be background. Selective constrains are assumed to vary across sites both along foreground and background, and a small fraction of sites only vary along foreground lineages. There are 3 ω ratios for foreground (0 < ω0 < 1, ω1 = 1, ω2 > 1) and 2 ω ratios for background (0 < ω0 < 1, ω1 = 1) in branch-site model A. When evidence for positive selection (ω > 1) was detected, both naïve empirical Bayes and Bayes empirical Bayes (BEB) were used to calculate posterior probabilities for site classes. Since NEB does not account for sampling errors, we used the BEB outputs as suggested by Yang [35]. The null model (model A') is same as model A, but ω2 = 1 is fixed. Branch-site model tends to be the most powerful of the three tests. In order to investigate the variation of selective pressure in different epidemic periods and hosts, we set each group of SARS-CoVs as foreground in turn to implement branch-site model. However, in such multiple tests, the probability of false rejection of at least one null hypothesis can be high. So we used Bonferroni correction to control the false positive rate, as it has been shown to be powerful when applied to the branch-site test [36]. As to dataset 1 and 3, we applied branch-site model to 6 groups on the tree, and for dataset 2, we applied branch-site model to 5 groups. So we used 0.0083 as the significance level for each branch-site tests in dataset 1 and 3, used 0.01 as the significance level for dataset 2. As indicated previously by Yang [33], these models sometimes fail to converge to maximum likelihood estimates. We thus performed each analysis at least twice using different starting values. Only identical data produced from both runs were considered reliable. All data are available upon request.
In order to examine the robustness of those positive selections identified by PAML, we also analyzed our datasets using HYPHY package accessed through the Datamonkey facility http://www.datamonkey.org[37]. Datamonkey includes three methods for detecting sites under selection: single likelihood ancestor counting (SLAC), fixed effects likelihood (FEL) and random effects likelihood (REL). REL method is often the only method that can infer selection from small (5–15 sequences) or low divergence alignments and tends to be the most powerful of the three tests. So this method was run using the GTR substitution model on a neighbor-joining phylogenetic tree by the Datamonkey web server. In order to investigate selective pressure among different hosts and epidemic phases, we split each dataset (S protein, replicase domains, 3'-end ORFs) into appropriate groups for analysis.
Phylogenetic analysis
For all genes analyzed, represented by S, replicase and 3'-end ORFs gene trees, at least four groups are apparent: BSL, PC03, HP03, PCHP04. As to the HP03 group, it can be subgrouped into HPEM and HPL in S gene tree, and HPEM and HPML in 3'-end ORF tree. It should be noted that the posterior probabilities for several nodes are low and there are some polytomies. These uncertainties could be due to some sequences in SARS group have high amino acid similarity, especially for replicase and 3'-end ORFs which are more conservative. However, previous studies suggested that the LRTs and qualitative results of ML parameter estimation were rather insensitive to tree topology [35, 38–42]. For branch-site model, Bayesian site identification might be affected by tree topology [40]. Remarkably, one isolate (38-HP03L_GD01), which was isolated in the later epidemic phase in 2003, always clustered with the early phase isolates. A possible explanation could be that this patient was infected in the early epidemic phase, which is supported by sequence analysis; this isolate has 29 extra nucleotides between ORF8a and ORF8b, a feature shared among isolates from civets and early phase patients. Another isolate (5-HP03E_HGZ8L1-B), which was isolated in the early epidemic phase, tends to cluster with the middle phase isolates (Fig 2 and 4). This virus may be a transitional virus because it does not have the extra 29 nucleotides like the middle phase isolates.
Detection of recombination
KH tests verify the significance of breakpoints estimated by GARD analysis
Number of breakpoints
AICc improvement
Breakpoint location
3'-end ORFs
Positive selection on the S protein of SARS-CoV
Maximum likelihood (ML) estimates for 59 S genes of SARS-CoV
Branch Model
M0 vs. FR
M0 (one ratio)
ω = 0.081
Free Ratio
ω = 0~∞
< 0.001
Not allowed
Site Model
M0 vs. M3
M0 (one ratio)
ω = 0.081
M3 (discrete, K = 3)
p0 = 0.732, ω0 = 0.015
p1 = 0.255, ω1 = 0.285
p2 = 0.013, ω 2= 3.214
< 0.001
142,311, 430,
462, 479, 540
609, 626, 665
Branch-site model A
BSL group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.912, ω0 = 0.047
p1 = 0.088
(p2a+p2b = 0)
p0 = 0.912, ω0 = 0.047
p1 = 0.088, ω2 = 1.000
(p2a+p2b = 0)
PC03 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.707, ω0 = 0.046
p1 = 0.069
(p2a+p2b = 0.224)
p0 = 0.782, ω0 = 0.046
p1 = 0.076, ω 2= 1.592
(p2a+p2b = 0.142)
HPEM group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.587, ω0 = 0.044
p1 = 0.055
(p2a+p2b = 0.358)
p0 = 0.885, ω0 = 0.045
p1 = 0.083, ω 2= 28.756
(p2a+p2b = 0.032)
< 0.001
49, 75, 344,
360, 501, 778
794, 860, 861
1001, 1148, 1163
1179, 124
HPL group
as foreground
MA' vs. MA
MA' (fix ω2 = 1)
p0= 0.400, ω0 = 0.045
p1= 0.038
(p2a+p2b = 0.562)
p0 = 0.400, ω0 = 0.045
p1= 0.038, ω2 = 1.000
(p2a+p2b = 0.562)
PCHP04 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.718, ω0 = 0.045
p1= 0.057
(p2a+p2b = 0.225)
p0= 0.901, ω0= 0.045
p1= 0.072, ω 2= 57.205
(p2a+p2b = 0.027)
< 0.001
78, 91, 108,
113, 147, 227,
243, 425, 440,
462, 479, 609,
613, 632, 743,
765, 839, 844,
856, 900, 1052
SARS group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.753, ω0 = 0.024
p1= 0.035
(p2a+p2b = 0.212)
p0= 0.792, ω0= 0.027
p1= 0.034, ω 2= 1.989
(p2a+p2b = 0.174)
< 0.001
2, 7, 9, 12, 14, 20,
27, 33, 37, 43, 58, 68,
70, 75, 84, 107, 108,
131, 134, 137, 139
147, 151, 154, 163, 165,
167, 169, 174, 199, 201,
214, 227, 230, 237, 239
242, 243, 244, 248, 249,
294, 333, 336, 344, 353,
391, 392, 426, 431,
440, 442, 457, 459, 462,
479, 480, 487, 488,494, 607,
613, 644, 729, 732, 743,
754, 758, 765, 778,
1052, 1080, 1148, 1163
The results of branch-site model revealed that no evidence of positive selection was found in the BSL, PC03 and HPL groups. For the HPEM group, the results indicated that 3.2% sites of S gene are subjected to strong positive selection with ω = 28.756. At p > 90% level, 14 specific sites were identified as potentially under positive selection (Table 2). For the PCHP04 group, 2.7% codon sites of the S gene are driven by strong positive selection with ω = 57.205. Twenty two positively selected sites were identified in this group (p > 90%). Fourteen of them are in S1 and eight in S2 domain. For the selection of entire SARS-CoVs from the two epidemics, the branch-site model A analyses indicated that 17.4% sites are under positive selection with ω = 1.989. A total of 74 sites were identified as potentially under positive selection along these lineages at 90% cutoff. In order to intuitively represent the distribution of these positively selected sites, we constructed the additional file 3, from which we can find that most of these sites distribute in S1 domain.
REL analysis results for three datasets
No. of sequencesd
Mean d N -d S e
Positively selected site(s)
49, 75, 77, 144, 239, 244, 311, 344, 778, 860, 861, 1001, 1148, 1163, 1179, 1247
75, 147, 227, 239, 243, 244, 311, 462, 479, 609, 613, 743, 765, 778, 1080, 1163
3'-end ORFs
Positive selection on replicase domains of SARS-CoV
PLpro, 3CLpro, RdRp and Hel are the major domains for coronavirus replication [43, 44]. We merged these four domains into one supergene for analysis because: 1) Yang et al. reported that gene concatenating analysis produced same outcomes as those obtained from analysis of separate genes [42]; 2) separate analysis results in mechanical repeats; 3) concatenating analysis can provide additional information because of additional number of sequences for the merged dataset, compared to separate dataset. Therefore, dataset 2 consists of 35 concatenated sequences from 129 complete genomes.
Maximum likelihood (ML) estimates for 35 merged replicase genes of SARS-CoV
Parameters under
Branch Model
M0 vs. FR
M0 (one ratio)
ω = 0.024
Free Ratio
ω = 0~∞
< 0.001
Not allowed
Site Model
M0 vs. M3
M0 (one ratio)
ω = 0.024
M3 (discrete, K = 3)
p0= 0, ω0= 0
p1= 0.972, ω1= 0.016
(p2= 0.028), ω2 = 0.360
< 0.001
Branch-site model A
BSL group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.993, ω0 = 0.020
p1 = 0.007
(p2a+p2b = 0)
p0 = 0.993, ω0 = 0.020
p1 = 0.007, ω2 = 1.000
(p2a+p2b = 0)
(-14449. 205)
PC03 group
as foreground
MA' vs. MA
MA' (fix ω2 = 1)
p0 = 0.993, ω0 = 0.020
p1 = 0.007
(p2a+p2b = 0)
p0 = 0.993, ω0 = 0.020
p1 = 0.007, ω2 = 1.000
(p2a+p2b = 0)
-14449. 205
(-14449. 205)
HP03 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.322, ω0 = 0.015
p1 = 0.002
(p2a+p2b = 0.676)
p0= 0.913, ω0= 0.015
p1= 0.006, ω 2= 11.093
(p2a+p2b = 0.081)
23, 123, 222, 236, 237, 250,
266, 375, 377, 409, 504, 563,
646, 654, 884, 1234, 1259, 1482,
1491, 1786, 1866, 1869, 1878,
1963, 1995, 2010, 2032, 2034
PCHP04 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.761, ω0= 0.018
p1= 0.006
(p2a+p2b = 0.234)
p0= 0.760, ω0= 0.018
p1= 0.006, ω2 = 1.000
(p2a+p2b = 0.234)
SARS group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0 = 0.850, ω0 = 0.012
p1 = 0.005
(p2a+p2b = 0.145)
p0 = 0.857, ω0 = 0.012
p1 = 0.005, ω2= 1.061
(p2a+p2b = 0.138)
Utilizing the branch-site model A analysis indicated that there is no positive selection in the BSL, PC03 and PCHP04 groups. However, the model A analysis revealed that among HP03 group about 8.1% codon sites of these 4 domains are potentially under strong positive selection with ω = 11.093 and 28 sites were identified (7 in PLpro, 5 in 3CLpro, 7 in RdRp, 9 in HEL). Weak positive selection (d N -d S = 0.001) was also detected from HP03 group by using HYPHY package but not other groups (Table 3).
Positive selection on 3'-end ORFs of SARS-CoV
The 3'-end of SARS-CoV genome encodes 11 ORFs: ORF3a, ORF3b, ORF4 (E), ORF5 (M), ORF6, ORF7a, ORF7b, ORF8a, ORF8b, ORF9a (N), and ORF9b. The E, M, N proteins are structural proteins of SARS-CoV and the other proteins are accessory proteins. Because the coding regions of ORF3b and ORF9b overlap partially or completely with those of ORF3a and N, we excluded the ORF3b and the ORF9b from this analysis. The ORF8a and ORF8b are present as two separate ORFs in most human isolates but as a single ORF (ORF8ab) in isolates from animals and early phase human due to the presence of extra 29 nt in this region, thus resulting in the fusion of ORF8a and ORF8b. Because of the difficulty in obtaining a reliable alignment in this region, ORF8 (a, b or ab) was excluded from our analysis as well. For similar reasons as mentioned above, we merged the 7 remaining ORFs into a supergene for analysis.
Maximum likelihood (ML) estimates for 56 merged 3'-end ORFs of SARS-CoV
Branch Model
M0 vs. FR
M0 (one ratio)
ω = 0.170
Free Ratio
ω = 0~∞
< 0.001
Not allowed
Site Model
M0 vs. M3
M0 (one ratio)
ω = 0.170
M3 (discrete, K = 3)
p0= 0, ω0= 0
p1= 0.866, ω1= 0.058
(p2= 0.134),ω2= 0.986
< 0.001
Branch-site model A
BSL group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.868, ω0= 0.059
p1= 0.132
(p2a+p2b = 0)
p0= 0.868, ω0= 0.059
p1= 0.132, ω2= 1.000
(p2a+p2b = 0)
PC03 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.868, ω0= 0.059
p1= 0.132
(p2a+p2b = 0)
p0= 0.868, ω0= 0.059
p1= 0.132, ω2 = 1.000
(p2a+p2b = 0)
HPEM group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.855, ω0= 0.059
p1= 0.130
(p2a+p2b = 0.015)
p0= 0.861, ω0= 0.059
p1= 0.131, ω 2= 4.300
(p2a+p2b = 0.008)
HPML group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.125, ω0= 0.047
p1= 0.017
(p2a+p2b = 0.858)
p0= 0.772, ω0= 0.046
p1= 0.106, ω 2= 9.863
(p2a+p2b = 0.122)
11, 29, 85, 129, 136,
222, 279, 280, 304, 319,
377, 388, 418, 423, 436,
449, 463, 469, 504, 584,
696, 850, 932, 954, 993
PCHP04 group
as foreground
MA' vs. MA
MA' (fix ω2= 1)
p0= 0.690, ω0= 0.055
p1= 0.097
(p2a+p2b = 0.213)
p0= 0.862, ω0 = 0.057
p1= 0.119, ω 2= 22.447
(p2a+p2b = 0.019)
< 0.001
25, 259, 609, 1184
SARS group
as foreground
MA' vs. MA
MA' (fix ω2 = 1)
p0 = 0.664, ω0= 0.033
p1= 0.066
(p2a+p2b = 0.270)
p0= 0.804, ω0= 0.037
p1= 0.074, ω 2= 3.138
(p2a+p2b = 0.122)
< 0.001
11, 15, 81,117, 120, 121,
171, 193, 259, 355, 361, 560,
609, 628, 830, 850, 1184
Natural selection generally leads to a reduction in deleterious mutations while promoting advantageous mutations. If a gene is highly divergent, there are two main explanations: first, it may be due to high mutation rate or relaxed selective constraint, in which case the gene may be free to mutate mainly because it has no fitness or function; or second, due to positive selection which is promoted by natural selection and the gene usually has highly important functions [45]. Virus entry, replication, assembly and release are the main steps of viral life cycle. Proteins involved in each of these steps may undergo adaptive evolution after a virus invades a new host.
Recombination and mutation are two important evolutionary mechanisms driving gene diversity and adaptation. Since recombination can affect the detection of positive selection, we first tested for recombination in our datasets [46]. GARD detected no evidence of recombination within the replicase and 3'-end ORFs, while one putative breakpoint in spike protein was detected. Whether there is any recombination among SARS-CoV is still debatable [13, 22, 47–49]. Previous studies suggested putative recombination only when analysis of SARS-CoV sequences were put together with other coronaviruses [47–49]; however, when analyses were focused solely on SARS-CoV sequences, recombination could not be detected [13, 22]. We also tested recombination in SARS group alone for each dataset. No evidence of recombination was detected by GARD. These results might imply that there could be some ancient recombination events occurred between bat SARS-like-CoV and the ancestor of SARS-CoV, which drove the bat SARS-like-CoV adaption to civet and human. Nevertheless, previous studies had revealed that detection of positive selection by LRT method was robust to low levels of recombination (with fewer than three recombination events), and identification of sites under positive selection by the empirical Bayes method appeared to be less affected than the LRT by recombination [46]. Overall, the issue of recombination among RNA viruses is highly controversial because the putative recombination events described were detected only by utilizing computationally-demanding phylogenetic analyses (split decomposition and/or maximum likelihood methods). Therefore, caution should be used when inferring conclusions about putative recombination events that are based only on such analyses. Because viable clonal recombinant viruses have been rarely observed in nature, for natural recombination leading to the transmission of a recombinant strain to be conclusively confirmed, the following prerequisites should be met: (i) the recombinant crossover should be demonstrated in a single PCR amplicon following cloning to ensure it occurs in a single DNA molecule; (ii) the recombination should be demonstrated repeatedly in clonal populations of viable virus (e.g. a plaque harvest or limited endpoint dilution; and (iii) the recombinant should maintain adequate sequence conservation during post-recombination evolution [50].
The S protein is a structural protein of coronavirus. It has a crucial role in the binding of virus to host receptor and subsequent fusion between the viral and host membranes, both processes being important for virus entry into host cell. In the case of several mammalian and avian coronaviruses, the S protein is cleaved into S1 and S2. The former contains receptor attachment sites and the later is involved in the fusion of CoV onto host cell. The S1 subunit, which usually has high divergence, contains a receptor binding domain (RBD); the S2 subunit, which is comparatively more conserved, contains two heptad repeat (HR) domains [51]. Several studies revealed that the S gene of SARS experienced noticeable positive selection during the SARS epidemics, especially in the early and middle phases [20–22]. However, our analyses indicated that the S protein of SARS-CoV underwent a stepwise adaptive process subsequent to its spillover into the civet and human populations. In the BSL group, our analyses suggest that the bat SARS-like-CoVs experienced purifying selection, indicating that the S gene is relatively stable in bats. In palm civet, SARS-CoV experienced strong positive selection as indicated by the results of PCHP04 group. The failure to identify significant positive selection in PC03 group was most likely due to the limited number of sequences available for analyses (only four sequences for PC03 group and two of them have identical S gene sequences). During the early and middle phases of the 2002–2003 SARS epidemic in human population, a small fraction of sites among the S protein were under strong positive selection. In contrast, isolates from the late phase showed no sign of positive selection, implying the S protein became stable again after two stages of adaptive evolution. Our analysis using the HYPHY package accessed through Datamonkey facility also revealed similar evolutionary patterns for the S protein (Table 3). Taken together, these results support the hypothesis that SARS-CoVs originated in bats, that the spill over into civets and humans were recent events and that the two SARS epidemics that took place one year apart, were results of independent animal-to-human transmissions [7, 20]. The major sequence difference of the S genes between bat SARS-like-CoVs and civet/human SARS-CoVs suggests that there may be other more closely related SARS-CoVs in bats or there may be other unknown intermediate animal host(s) in the transmission of the virus(es) from bats to civets [49].
Among the SARS-CoVs from human and palm civets, numerous sites are identified to be potentially under positive selection (Table 2 and see additional file 3A). Those sites inferred from different SARS epidemic phases reflect the adaptation process of SARS-CoVs. Those sites identified from the PCHP04 group may be important for SARS-CoV adaptation to palm civets. Those sites identified from the HPEM group may be important for SARS-CoV adaptation to human. Most of these sites, especially from the entire SARS group, are located in the S1 domain that contains the receptor binding domain. Zhang et al. [22] previously identified 12 positively selected sites in the SARS-CoV group, all of which were confirmed in our current study. The greater number of sites identified in our study is likely due to the fact that the branch-site model is more powerful than the site-specific model. Some of these sites have been confirmed by experimental data to be crucial for the adaption of SARS-CoVs to human. For example, it has been found that adaptation of S protein to human angiotensin converting enzyme 2 (ACE2) is facilitated by alteration of residue 479 to asparagine and of 487 to threonine [52, 53]. Also, using site-directed mutagenesis, Zhu and Chakraborti identified that residues 344, 392, 426, 431, 479, 480, 487, 488 and 494 are important for the binding of RBD with ACE2 and SARS-CoV antibody [54, 55]. In RBD, there are eight newly identified sites (333, 336, 353, 391, 440, 442, 457, 459, and 462) which have not been proved to be critical for RBD and ACE2 interaction. Furthermore, there are ~60 sites to be under positive selection beside RBD. Although evaluation of every observed site under positive selection by reverse genetics would not be realistic or feasible, generation and evaluation of mutant viruses based on sites located within or adjacent to functional domains could provide clues for the genetic aetiology of SARS adaptation to new hosts and emergence.
The first two thirds of coronavirus genome encode two large polyproteins: P1a and P1ab, which are cleaved by virus-encoded proteinases (PLpro and 3CLpro) into 16 non-structure proteins (nsp1–nsp16) playing important role during coronavirus replication. Because the P1ab is too big (~21 kb), we analyzed four most important domains related to viral replication: PLpro, 3CLpro, RdRp and Hel, which correspond to nsp3, nsp5, nsp12 and nsp13, respectively [17, 43, 44]. Our results revealed that, unlike the adaptive evolutionary pattern of S protein, these replicase domains did not experience positive selection in bat and palm civet, but underwent strong positive selection in human patient. Moreover the evidence of positive selection is stronger in the later phases than that observed in the early and middle phases (data no shown). Furthermore, our analysis using HYPHY package observed very weak positive selection in the HP03 group but not among the other groups (Table 3). These results to some extent differ from the observations derived from several previous studies. Using pairwise analysis of the dN/dS, Song et al. found that the average ω ratio of S protein for the early phase was larger than that for the middle phase, which in turn was larger than the ratio for the late phase. A similar pattern was found in ORF1a and ORF3a but not in ORF1b and nsp3, which were suggested under purifying selection during the whole course of the epidemic. They identified over 200 single-nucleotide variations (SNVs) and inferred the importance of some SNVs on SARS-CoV adaptive evolution [21]. Zhang et al. also investigated the adaptive evolution of the S protein employing the site specific model, but they did not observe any positive selection in RdRp, Hel or nsp3 [21, 22]. The best explanation for this apparent discrepancy is that the methods used in their studies were more conservative than the branch-site model used in our study, and hence were not able to detect positively selected changes among the highly conserved genes. Alternatively, it may be due to the use of concatenating analysis in our study, which can provide additional information due to the compiling of more dissimilar sequences for datasets. For the replicase genes of bat and civet isolates, there was no sign of positive selection. This is probably due to the following reasons: 1) the civet isolates were collected within a very short time period and thus there was not enough time for adaptive evolution; 2) civet cells are very suitable for SARS-CoV replication which may imply that the civet is a perfect intermediate host for SARS-CoV; and 3) the bat isolates might have completely adapted to their hosts and hence were under no further selective pressure for evolution.
As to the 3'-end ORFs, the most diversifying selection happened in the middle and late phases of the SARS epidemic in 2003–2004. No positive selection was found in the BSL, PC03 and HPEM groups. When the isolates from two epidemics of 2003 and 2004 were investigated together, 12.2% sites in these 7 ORFs were shown to be under positive selection (Table 5). In addition to a few sites identified at p > 90% level, many sites are inferred to be under positive selection at 70–80% posterior probability (see additional file 3B). Most of these sites are located in ORF3a, E, M and ORF6, implying these four genes may play a more important role for SARS-CoV adaptation in a new host. Our results based on the REL method showed weak positive selection in HPML group, but not in other groups (Table 3). The failure to identify specific sites under positive selection could be due to weak and thus undetected positive selection. These results suggested that the 3'end-ORFs underwent positive selection after SARS-CoV spilled out into civet and human populations, and adaptive evolution mainly happened in the middle and later phases in 2003. Song et al. previously suggested that the 3a protein evolved adaptively as S protein [21]. By estimating mutation rates, Zhao et al. suggested that the non-synonymous substitution rates were comparatively high in E, M and N [56]. ORF3a encodes a protein of 274 amino acids. A recent study indicated that the 3a protein forms a potassium sensitive channel that may promote virus release and may be important for modulating expression of S on the cell surface [16, 57]. 3a protein also interacts with the structural proteins S, E, M [57, 58]. Therefore, amino acid changes in 3a protein might be necessary to maintain the interaction between 3a and other proteins. E and M protein play a pivotal role in viral morphogenesis, assembly and budding. Co-expression of E and M was shown to produce virus-like particles, roughly the same size and shape as virions [59]. N protein is important for viral packaging which is the first step in the assembly of infectious SARS viruses [60]. Thus the amino acid changes in these three structural proteins may be critical for virus assembly in the new host. SARS-CoV ORF 6 protein can enhance the virulence of attenuated murine coronavirus (MHV) [61], as well as stimulate cellular DNA synthesis [62]. A recent study showed that ORF 6 protein inhibited both interferon synthesis and signaling [63]. These findings suggested that ORF 6 may have a role in enhancing virus replication or assembly. ORF 7a protein inhibits cellular protein synthesis and blocks cell cycle progression at G0/G1 phase, suggesting that 7a may play important roles in the life cycle of SARS-CoV and the pathogenesis induced by SARS-CoV [64, 65]. The function of other accessory proteins remains to be determined. Overall, these findings suggest that the 3'-end ORFs play important roles for SARS-CoV replication, assembly and release. Collectively, amino acid changes in these proteins could play a role in modulating the host switch of SARS-CoV.
We systematically analyzed the individual SARS-CoV proteins important for virus entry, replication and assembly. The results suggested that SARS-CoVs experienced a stepwise adaptation to humans. In palm civets and humans during the early and middle epidemic phases, virus entry-mediating protein S experienced strong positive selection. In contrast, the replicase proteins experienced positive selection only in human patients but not in palm civets, implying that the palm civet is a suitable intermediate host for SARS-CoV replication. The proteins involved in virus assembly and release mainly underwent positive selection during the middle and later epidemic phases. These results highlight the spectacular dynamics of SARS-CoV evolution in a narrow time window, period of epidemic, support the zoonotic origin of SARS and suggest that some amino acid sites may be critical for viral adaptation in different hosts. Collectively, these results suggest that the development of SARS-CoV reverse genetics system will facilitate further molecular and/or epidemiological investigations to elucidate role of adaptive virus evolution in future emergence events.
We are very grateful to Drs. Yi Guan, Ronald C. Montelaro, and Honglin Chen for critical review of the manuscript. This work was supported by the State Key Program for Basic Research grant (2005CB523004), the Sixth Framework Program EPISARS from the European Commission (no. 51163) to S.Z and a grant (07DZ22940) from the Shanghai government to S.Z. NV was supported by a grant by the Fine Foundation.
We thank Sergei L. Kosakovsky Pond for his help with the HYPHY software package and two anonymous reviewers for valuable suggestions.
Authors' Affiliations
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Apple release Xcode 1.2 and removes eMac model from holdApple releases Xcode 1.2
After seeding developers with build 7K249 of its Xcode 1.2 development environment this week, Apple quickly reviewed last minute feedback from developers and declared the build gold master, sources said.
"Xcode 1.2 is the latest version of Apple's new Integrated Development Environment," the company reportedly told developers in the developer tools release notes. Th new version provides several enhancements over Xcode 1.0 and Xcode 1.1, including improved stability and user experience, an enhanced build system, better dependency analysis, improved cross-project reference support, and improved native target support for yacc and assembler.
Apple also noted a handful of known issues with the release of Xcode 1.2 that could cause the application to crash, but provided a list of workarounds to help developers avoid problems.
The release build is current available to 'AppleSeed' and Apple Developer Connection (ADC) account holders in the form of several 30 MB segments.
eMac off short-term quality hold
Earlier this week, Apple removed a short-term quality hold it had placed on early inventory shipments of the new SuperDrive equipped eMac G4.
Apparently, engineers were able to vindicate the quality of the model, once suspected of containing issues with its SuperDrive implementation.
Apple had previously sent a memo to its retail stores, asking that the models be put on hold, and not be sold to customers until further notice. The company officially removed the SuperDrive eMac G4 from hold with a followup memo days later, though no formal explanation of the suspected issue was given.
The new eMac G4 SuperDrive model is the first Mac to ship with an 8X SuperDrive. 8X SuperDrives are also expected to ship in new Power Mac G5 models when the company gathers enough 90-nanometer chips to announce and begin shipping the models.


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27th November 2016
This is Nala my friends' cat which loves me so much whenever I visit them. Nala lives on the 10th floor and never leave her flat.
|HELLO from wardrobe
|She loves sleeping in the wardrobe..
Enjoy Sunday!!!
I like travelling, family history and reading books. I am ESL teacher in High Junior school and at University.
|HELLO from wardrobe
|She loves sleeping in the wardrobe..
Second last photo, Nala is ignoring the nearby mouse. My sister's cat sleeps in a wardrobe.ReplyDelete
Ouch. Nala is a pretty cat. Jewel loved the wardrobe too.ReplyDelete
A very clever cat!ReplyDelete
She seems happy in her flat!!ReplyDelete
Cats do sleep in some weird and wonderful places and positions.ReplyDelete
She has a good life.ReplyDelete
So sweet.
Enjoy your Sunday too!
All the best Jan
Happy Sunday to you, Gosia. Nala is certainly a very pretty cat. She must see the warmth and love in you.ReplyDelete
She is beautiful and looks lovely and soft. Our cat likes to sleep in my daughter's wardrobe.ReplyDelete
Beautiful cat. She poses for your camera!ReplyDelete
Pretty kitty, but I think I'm allergic to them. :(ReplyDelete


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Sorry, only registred users can create playlists.
During Mill Creek's Spring Remodeling Sale, Shaw Floors offers two very popular flooring options Installed at an exceptional value.
Wood look Tile, just $6.89 a sq ft; or Beautiful 5" Hand scraped Hickory for $6.99 SF.
Leave the details to us shop Mill Creek
This video is for demonstration purposes only. Sale has expired.
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Chocolate souffle or soufflé (French: [su.fle]) is a light baked cake made with egg yolks and beaten egg whites combined with various other ingredients and served as a savory main dish or sweetened as a dessert.
The word soufflé is the past participle of the French verb "souffler" which means "to blow up" or more loosely "puff up"—an apt description of what happens to this combination of custard and egg whites.
For this recipe you need dark chocolate coating (or "Couverture" in French). This is not a brand or type of chocolate but rather a term used to describe professional-quality coating chocolate with a high percentage of cocoa butter -- at least 32 percent and often as high as 39 percent for good-quality couverture. The extra cocoa butter allows the chocolate to form a thinner coating shell than non-couverture chocolate. Ghirardelli, Callebaut, Lindt, Droste or Valrhona are offering this coating. Happy Cooking!
Ingredients Chocolate Souffle
(serves 4)
25 g soft butter
25 g flour
5 cardamom pods or ground cardamom
(If you cannot get cardamom use cinnamon or vanilla instead)
125 ml milk
30 g dark chocolate coating or couverture
20 g cocoa powder
2 egg yolks
3 egg white
35 g sugar
50 g powdered sugar
Cooking Instructions Chocolate Souffle
- Grease 4 souffle forms or cups (glass) and add some sugar which will stick on the butter; remove sugar which is not sticking.
- Knead butter and flour.
- If you use the pods remove cardamom from pods and crush it in a mortar.
- Chop the dark chocolate coarsely; in a pan heat milk, add the chocolate; add cardamom and cocoa powder as well, while stirring bring it to a boil - add the kneaded flour-butter but not at once, bit by bit. Let it cool off.
- With a mixer add the egg yolks one after the other until you have a smooth cream.
- Beat egg white until firm, trickle the sugar into the egg white. Fold firm egg white into the cream.
- Fill the cups or forms until 1 cm underneath the edge. Place them on a dripping pan and add as much hot water so the forms are 2/3 in the water (using the double-boiler method here).
- Bake them for 40 minutes on 400 degrees F (200 C) on the 2. level.
- Take the souffles out and sprinkle them right away with powdered sugar.
Serve them with whipped cream that can be spiced with cardamom.
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2 Responses to "Chocolate Souffle Dessert – After a German Recipe"
Love the temp converter, now how about one for measurements? I
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