I'll give props to HVAC Service for being available quickly and having Saturday and Sunday appointments. Tony fixed our air conditioner and was professional. So why only 2 stars? Because we sent the receipt into our homeowner association to see if we could get reimbursed for some costs, and the property manager said that we paid THREE times more than we should have. We got conned. I advise you to call HVAC Service, but also shop around and get other estimates. Learn from our mistake!
The three central functions of heating, ventilation, and air conditioning are interrelated, especially with the need to provide thermal comfort and acceptable indoor air quality within reasonable installation, operation, and maintenance costs. HVAC systems can be used in both domestic and commercial environments. HVAC systems can provide ventilation, and maintain pressure relationships between spaces. The means of air delivery and removal from spaces is known as room air distribution.
For details about apprenticeships or other work opportunities, contact the offices of the state employment service, the state apprenticeship agency, local contractors, or local union–management HVACR apprenticeship committees. Apprenticeship information is available from the U.S. Department of Labor’s Apprenticeship program online or by phone at 877-872-5627.
Alberto said they were busy but could either install on the upcoming Saturday. We were going to be out of town and said unfortunately that day did not work. He then proposed the upcoming Tuesday as a possible solution, but just had to confirm with another appointment. I said great. I will review the estimate and call your office to confirm I want to move forward with the install.
Berkeys is a licensed HVAC contractor and air conditioning company offering AC repair, Air Conditioning tune-up, and air conditioning replacement services in Southlake, TX, Keller, TX, Coppell, TX, Colleyville, TX, Grapevine, TX, Plano, TX, Fort Worth, TX, Arlington, TX, Frisco, TX, Dallas, TX, Addison, TX, Flower Mound, TX, Irving, TX, McKinney, TX, Arlington, TX, Euless, TX, Denton, TX, Highland Village, TX, Grand Prairie, TX, Westlake, TX, Farmers Branch, TX, Lewisville, TX and more. Call to confirm service for your home.
Dichlorodifluoromethane (R-12) was the most common blend used in automobiles in the U.S. until 1994, when most designs changed to R-134A due to the ozone-depleting potential of R-12. R-11 and R-12 are no longer manufactured in the U.S. for this type of application, so the only source for air-conditioning repair purposes is the cleaned and purified gas recovered from other air conditioner systems. Several non-ozone-depleting refrigerants have been developed as alternatives, including R-410A. It was first commercially used by Carrier Corp. under the brand name Puron.
Two weeks go by and no phone call from HVAC Service regarding install date, or any update at all. I call them to check in to see if they have put me on the schedule and when I can expect to have the AC installed. You can tell by the discussion I had with the lady on the phone that they completely forgot about me and have not scheduled anything. She says she needs to get in contact with the owner because she has no idea on the availability of the crane. I was told that she would call me back by the end of the day with an update. Surprise, surprise, no phone call.
When you schedule air conditioning and heating maintenance with The Home Depot, one of our local, licensed, and insured service providers will come to your house and perform a 28-point inspection on your units. Scheduling two seasonal heating and cooling services per year with The Home Depot is a worthwhile investment that can help identify and fix small issues before they lead to bigger, more expensive repairs.
Our A/C was blowing hot air intermittently...it was cold one minute and warm the next. By 8pm I'd have a 90 degree interior temp. After one company just took a cursory glance at our A/C and found nothing wrong, I called HVAC. I called them mostly based on Michael W's post on 8/10/14 where he told us the owner, Alberto, took his feedback and took it SERIOUSLY. That speaks VOLUMES to anyone who has been around long enough to know the characteristics of the serious players are and those who are the wannabes. So HVAC sent Tony Diaz to my house. I was at work so he was subjected to the most excruciatingly exacting woman on the planet (my wife). Tony impressed her with his friendliness, professionalism and the obvious jouneyman-level knowledge of his craft. He found literally --burnt-out-- parts in our system and replaced them. It cost us a bit but I know enough about the parts to see there was no gouging on the parts and a fair rate for the labor. It wasn't cheap but it was fair and worth every dime. And now i have A/C in my house again and a repairman came to my house, actually FIXED something AND managed to do so in a friendly & professional manner. That used to be commonplace...nowadays it deserves a "Wow".. Thank you, Tony!
Before we get into the different AC repairs and replacements that John Moore offers, it’s important to understand the basic operating principles of your AC system. There are two types of systems: electric and natural gas. Electric AC systems use an air handler to blow air through your home while natural gas systems use a furnace. Both furnaces and air handlers are typically located in the attic.
ISO 16813:2006 is one of the ISO building environment standards. It establishes the general principles of building environment design. It takes into account the need to provide a healthy indoor environment for the occupants as well as the need to protect the environment for future generations and promote collaboration among the various parties involved in building environmental design for sustainability. ISO16813 is applicable to new construction and the retrofit of existing buildings.
When seeking emergency heating or cooling services you want experts that actually care about the results and your comfort. You want to choose the HVAC company in Alexandria with extensive skill and top-class tools, but you don’t want them to take advantage of your bad situation! With HVAC Unlimited, you never need to worry. We make it our mission to offer superior services any time you need it, and we guarantee cost-efficient solutions that restore your comfort.
Each spring and fall before you start using your air conditioner, furnace, or other heating and cooling systems, you should get a tune-up to ensure your HVAC system is ready for the season. By spending a little twice a year, you can save a lot in energy bills by keeping your system operating at peak efficiency. You also can avoid costly repairs by identifying and fixing minor problems before they turn into big ones. Plus, many homeowners don't realize that their manufacturer's warranties may be considered invalid if they aren't getting their unit serviced twice a year.
As one of the most trusted commercial HVAC service companies in the Midwest, not only will you receive prompt service, you won’t have to worry about our teams’ knowledge and corrective service capabilities. In the last 18 months, we’ve performed more than 3,790 mechanical preventive maintenance inspections. Whether you're in need of basic air conditioner maintenance or full commercial heating and air conditioning repair, we have yet to find a deficiency we can't fix.
If you need a furnace repaired or an air conditioner serviced, we'll send a local technician out to diagnose the specific problem and recommend solutions for you and your family. There will be a diagnostic fee for the visit, and any specific repairs will be an additional charge. If replacing your system is the best solution, the fee for the diagnosis will be credited toward the purchase and install of a new system.
Before selecting the installation location of air conditioner, several main factors need to be considered. First of all, the direction of air flow from the indoor units should not fall on the place of rest or work area. Secondly, there should not be any obstacles on the way of the airflow that might prevent it from covering the space of the premises as much as possible. The outdoor unit must also be located in an open space, otherwise the heat from the house will not be effectively discharged outside and the productivity of the entire system will drop sharply. It is highly advisable to install the air conditioner units in easily accessible places, for further maintenance during operation.
Call Miller's Air Conditioning & Heating at (925) 583-6387 and always speak with a live and helpful member of our team, who will immediately focus on your requirements. While our competitors punch a strict time clock, we realize heating and cooling challenges often arise outside of regular business hours. Rather than leave a message on a machine, take advantage of personalized service and the prompt response of radio dispatched trucks.
AC air handler squealing or grinding noises. Though most air handlers have direct-drive motors, some older units may be belt-driven. Squealing sounds from a belt-drive air handler generally occur when the belt that connects the motor to the blower slips. In most cases, the belt is improperly aligned or worn and needs replacement. Follow the instructions above under Air Conditioner Not Blowing Air and refer to your owner’s manual.
A heat pump is an air conditioner in which the refrigeration cycle can be reversed, producing heating instead of cooling in the indoor environment. They are also commonly referred to as a "reverse cycle air conditioner". The heat pump is significantly more energy efficient than electric resistance heating. Some homeowners elect to have a heat pump system installed as a feature of a central air conditioner. When the heat pump is in heating mode, the indoor evaporator coil switches roles and becomes the condenser coil, producing heat. The outdoor condenser unit also switches roles to serve as the evaporator, and discharges cold air (colder than the ambient outdoor air).
Fast forward 1 year and 1 week and the circuit board that HVAC Service had installed has failed again. They came out and said that it failed from water condensation dripping onto it, so we needed to replace it and reroute the pipes to prevent future water damage. That would cost double what I paid last year. Why didn't they fix the problem last year at the initial repair instead of putting in a circuit board that was bound to fail again?!? They were strongly recommending that the furnace be completely replace for a huge fee to avoid the same thing happening again. I paid $59 for them to tell me they didn't fix it right last year. This didn't feel right, so I sought a second opinion. I'm very glad that I found another guy to come replace the circuit board and put it away from the water, fix the water leak, and also find and fix another impending failure. All of that was for much less than HVAC Service wanted to charge for repair or replacement. The other guy did a great repair, so I'm confident it will not break down again, but if it does, I will not call HVAC Service. Don't get suckered by their Yelp Deal.
In 1906, Stuart W. Cramer of Charlotte was exploring ways to add moisture to the air in his textile mill. Cramer coined the term "air conditioning", using it in a patent claim he filed that year as an analogue to "water conditioning", then a well-known process for making textiles easier to process. He combined moisture with ventilation to "condition" and change the air in the factories, controlling the humidity so necessary in textile plants. Willis Carrier adopted the term and incorporated it into the name of his company.
With a modern heat pump system installation, you’ll enjoy benefits such as exceptional efficiency and maximum temperature control all year round. These innovative systems combine both heating and cooling into one compact system. The technicians from Sky Heating & Air Conditioning are highly qualified and experienced in heat pump maintenance, repairs, and installations, so you can feel confident in our services. We specialize in WaterFurnace geothermal heat pump systems, which offer unprecedented efficiency and energy savings. Browse our website or give us a call today to learn even more!
Did your furnace suddenly stop working? Call Coolray and relax! Our skilled HVAC technicians can repair all makes and models of furnaces, heat pumps and heating equipment. We serve Cumming, GA with 24-hour heating repair service that you can count on when you need it. Our technicians will arrive on time and will be equipped with the tools necessary to correctly diagnose and repair your heating system. Just need routine maintenance on your furnace? We can do that too. Looking for a new system altogether? We're the HVAC contractor Cumming, Georgia residents can count on for all of their home heating needs!
a/c compressor needed replacing. the A Team Mechanical found a way to return the next day to complete the repairs and made sure the system was running effectively. Even put down another concrete base pad to level the a/c unit. They were professionals and straight forward with the services needed. All the other HVAC companies wanted to charge me three time the amount for the services.
Your air conditioner’s outside unit is home to the compressor, fan, and condenser coils. These components work to release heat from the air, transferring it out of your home to produce a cool indoor climate. When the outdoor fan isn’t working correctly, the heat transfer process is hindered. Not only will this prevent your air conditioner from adequately cooling your home, a broken fan can cause the system’s compressor to overheat. An overheated compressor could trigger the air conditioner’s safety controls, turning your air conditioner off; overheating can also cause damage to the compressor itself, requiring home AC repair or compressor replacement.
Circulating refrigerant vapor enters the compressor, where its pressure and temperature are increased. The hot, compressed refrigerant vapor is now at a temperature and pressure at which it can be condensed and is routed through a condenser. Here it is cooled by air flowing across the condenser coils and condensed into a liquid. Thus, the circulating refrigerant removes heat from the system and the heat is carried away by the air. The removal of this heat can be greatly augmented by pouring water over the condenser coils, making it much cooler when it hits the expansion valve.
ABM is an EOE (Minority / Female / Veteran / Disability / Gender Identity / Sexual Orientation) and is committed to working with and providing reasonable accommodation to individuals with disabilities. If you have a disability and need assistance in completing the employment application, please call 855-350-0226. We will provide you with assistance and make a determination on your request for reasonable accommodation on a case-by-case basis.
A multi-split system is a conventional split system, which is divided into two parts (evaporator and condenser) and allows cooling or heating of several rooms with one external unit. In the outdoor unit of this air conditioner there is a more powerful compressor, ports for connecting several traces and automation with locking valves for regulating the volume of refrigerant supplied to the indoor units located in the room.
When signs of serious trouble arise, it is best to call a professional HVAC technician immediately. Waiting can allow the problem to worsen, which can lead to a higher repair bill. Finding a qualified technician can be as easy as asking family, friends and neighbors for recommendations or consulting online review websites and choosing a service provider that has received a significant number of positive reviews. Consumers must verify that any professional who is hired to do HVAC repair and maintenance has been properly trained and certified. Many service providers list licensing and certification information on their website.Last updated on Jul 11, 2018
For over 65 years, Jacobs Heating & Air Conditioning has been a Portland HVAC leader. Providing comprehensive heating, cooling and ventilation services to the Portland, OR and Vancouver, WA areas, our clients consistently praise our timely and professional work. From air conditioning repair to cooling and safety system installs, trust the professionals at Jacobs Heating & Air Conditioning.
Additionally, new technology and government requirements are pushing manufacturers to build more energy efficient HVAC equipment. January 2006 we observed the end of 10 SEER units with 13 SEER air conditioning units mandated by the government. This only makes sense; however, with the higher efficiency, there comes complexity in the technology engineered into the equipment. A problem can be difficult to troubleshoot and diagnose and may require more time than the lower efficiency, less complex units.
If you ever need a repair or maintenance preformed on your Trane air conditioning and heating equipment, we hope you’ll give R.S. Andrews a chance to earn your business. Literally tens of thousands of homeowners have called our hotline for repairs. While you can’t please everyone, we try and have done so in over 99% of the services calls we have performed. It’s a high standard equaled by the quality of Trane air conditioners, furnaces and heat pumps. Call now for service and repair. And don’t forget that we provide plumbing and electrical service and repair, too.
In the United States, HVAC engineers generally are members of the American Society of Heating, Refrigerating, and Air-Conditioning Engineers (ASHRAE), EPA Universal CFC certified (for installation and service of CFC HVAC devices), or locally engineer certified such as a Special to Chief Boilers License issued by the state or, in some jurisdictions, the city. ASHRAE is an international technical society for all individuals and organizations interested in HVAC. The Society, organized into regions, chapters, and student branches, allows exchange of HVAC knowledge and experiences for the benefit of the field's practitioners and the public. ASHRAE provides many opportunities to participate in the development of new knowledge via, for example, research and its many technical committees. These committees typically meet twice per year at the ASHRAE Annual and Winter Meetings. A popular product show, the AHR Expo, is held in conjunction with each winter meeting. The Society has approximately 50,000 members and has headquarters in Atlanta, Georgia.
With more than 100 years in existence, Lennox has always delivered industry-leading innovation and technology while preserving stringent durability and reliability standards. The professionals at James Heating & A/C, Inc help bring those standards to Lexington so that you can always have total home comfort. If you’re in the market for a new HVAC system or even if you just have a question about what we offer, give us a call at 336-853-6070. We’d love to hear from you.
At Van Haaften Plumbing & Heating, heating and cooling isn’t just our job, it’s our passion. We’re dedicated to providing you with the top HVAC service in Pella. No matter the size of the problem or time of day, you can rely on us to get the job done properly. After all, Van Haaften Plumbing & Heating has been serving the community since 1949, bringing comfort to our customers in every season.
1/18/2014 Hi Jody M- First, as you pointed out, we were the "Only" company to answer your phone call on the… Hi Jody M- First, as you pointed out, we were the "Only" company to answer your phone call on the weekend & provide service for you on the same day. There is a premium for most any service company to come out afterhours or weekends, and being we are one of the very few companies to offer this service, a higher labor rate is understandable, after all, we have to pay our service persons an overtime rate for such calls.
Special chemicals can be used to clean mineral deposits, but one of the safest is vinegar. A 50% mixture of vinegar and water will usually dissolve or help clean away deposits. If the humidifier has a drum with a belt, replacement of the belt at the beginning of the heating season can be the easiest fix if the water has a high mineral content. If the humidifier has screens or is the wick type, some corrosion can occur as well as build up
Strongsville Heating & Air Conditioning simplifies the challenges of keeping a perfectly comfortable home. We provide convenience, cost-savings, and reliability using top of the line products, premium materials, highly skilled technicians and exceptional customer service. Whether you need a new heating/cooling installation, replacement, retrofit, routine maintenance, trustworthy repair or air quality options, count on our team of NATE-certified technicians to eliminate stress and maximize comfort. We protect your busy schedule with flexible appointments, prompt arrival and mess free completion. Strongsville Heating & Air Conditioning is always available to you for Emergency Service, 24 hours a day, 365 days a year across Strongsville & Surrounding Areas.
Serving the entire High Desert and Inland Empire areas - we are about quality service - giving the customer the best options - and delivering what they want. We are a small fully licensed and insured company specializing in residential heating and air conditioning. We are a small fully licensed and insured company specializing in residential and light commercial heating and air conditioning.
It’s a rare occasion for a problem to arise with your heating or cooling equipment at a convenient time. In fact, we’re not even certain such a time exists! That’s why the team at HVAC Unlimited stands ready to provide complete emergency heating and cooling repairs. No matter the time of day or the day itself, we can be on the way and ready to provide prompt, cost-efficient services to our Alexandria clients.
If you’re concerned about dust, allergens or mold, it may be time to have your air ducts cleaned. The national average cost for air duct cleaning ranges from $190 to $250. Air duct cleaning costs will vary based on a number of factors such as your location in the country, the age and condition of your air ducts, the number of linear feet to be cleaned, and any repairs your ducts may need. HVAC cleaning companies should clearly outline what is included in their services and at what rate, so always make sure you understand and have a written agreement before work begins. Here are some examples of average air duct cleaning costs:


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Date: 1909 ca.
Description: The Mazomanie High School.
Date: 1873 ca.
Description: John Theodore Parman and his wife Louise stand second and third from the left behind their home's fence. Their daughters can be seen in the windows and the...
Date: 1908 ca.
Description: View looking north from the depot.
Description: The William Thompson family on the porch roof of their substantial brick bracket style house with shutters and with latticework at its foundation. A girl o...
Description: The Mazomanie School, later the high school. A large group of students and teachers stand in front of the two-story brick building with octagonal cupola a...
Date: date unknown
Description: A Mazomanie school.
Description: Interior view of the dining room in the home of Dr. Isadore Schultz. The turquoise and white wallpaper sets off the blond mahogany furniture.
Description: Scattered young trees, seedlings and saplings, in a field.
Description: View across unpaved road of the Community Church. There are stained glass windows in an arch above the main entrance, and three more along the front. There...
Description: View down street towards the a central business district. There is a T intersection at the end of the main street. Storefronts with awnings line both sides...
Date: 1880 ca.
Description: John Parman's “Blacksmith, Wagon & Carriage” shop, with workmen and others posed with vehicles in front.
Description: Facade detail of a farmhouse with vertical siding, seen through nearby trees. Photographed on the bottomlands of the Wisconsin River north of Mazomanie.
Description: Aerial view of Mazomanie.
Description: A drawing of the private Haskell University that was operating during the 1850's. The drawing was done by a student, Mrs. L. Campbell who spelled the name...
Date: 1907 ca.
Description: Elevated view of church and town.
Date: 1942 ca.
Description: Isabel Bacon La Follette holding a rake on the farm Philip purchased in northwestern Dane County.
Description: Exterior view across unpaved road of the Episcopalian Church, a small brick building with arched windows and doors. Trees in the yard shade the church. A w...
Date: 1874 ca.
Description: A family, perhaps that of D.W. Bronson, is posed by the picket fence in their yard. Two of the women hold small books. A tree stump in the foreground serve...
Description: Probably D.W. Bronson and family. A mounting block cut from a tree stump is in left foreground with carriage and saddle height steps. The family poses in f...
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The Foos to Headline Bamboozle in NJ
The Bamboozle Festival, an annual highlight of summer rock festivals, will be returning to its roots in Asbury Park, NJ in 2012 with a powerhouse trio of headlining acts, headed by Dave Grohl and the Foo Fighters.
Here is a complete look at the Bamboozle Festival, including lineup and tickets.
The festival started in 2003 in the hometown of Bruce Springsteen primarily as a show featuring jam bands but has since grown to be a multi-day event with dozens of bands. In recent years the festival has been held at the Meadowlands Sports Complex, but with the 10th annual festival taking place in 2012, the event’s promoters decided to bring it back to where it all started.
Bamboozle 2012 Headliners
- Foo Fighters
- Bon Jovi
The Bamboozle will take place on May 18th, 19th and 20th right on the waterfront of Asbury Park and will feature the Foo Fighters, Bon Jovi and Blink 182 as headlining acts. The rest of the rumored 100 plus bands and performers will be announced shortly but what is known to date is that the headliners will feature the Foo Fighters, who released one of the most acclaimed rock sets this year with “Wasting Light”; Bon Jovi, a band that is consistently at the top of the list when it comes to touring acts and Blink 182, who has seen recent relevance again since they reformed to tour this past year.
The lineup is sure to be rather diverse. Last year’s version featured LMFAO, Bruno Mars, Motley Crue, Lil’ Wayne, 30 Seconds to Mars and Plain White T’s.
Tickets recently went on sale for three-day packages, which start at $190 per ticket. Be sure to check out SeatGeek for details on tickets and for news on The Bamboozle as the event lineup takes shape. For further information, you can also check out the main site for The Bamboozle. Find more concert tickets right here on SeatGeek.com.
|Friday 3:30 AM||Bamboozle Festival - Three Day Pass Asbury Festival Area – Asbury Park, NJ tickets from $319|
|Friday 12:00 PM||Bamboozle Festival Asbury Festival Area – Asbury Park, NJ|
|Saturday 12:00 PM||Bamboozle Festival Asbury Festival Area – Asbury Park, NJ|
|Sunday 12:00 PM||Bamboozle Festival Asbury Festival Area – Asbury Park, NJ|
Image courtesy of Flickr user SativaBella

domain: artistic


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- Open Access
Bone marrow stromal cell antigen 2 (BST-2) restricts mouse mammary tumor virus (MMTV) replication in vivo
Retrovirology volume 9, Article number: 10 (2012)
Bone marrow stromal cell antigen 2 (BST-2) is a cellular factor that restricts the egress of viruses such as human immunodeficiency virus (HIV-1) from the surface of infected cells, preventing infection of new cells. BST-2 is variably expressed in most cell types, and its expression is enhanced by cytokines such as type I interferon alpha (IFN-α). In this present study, we used the beta-retrovirus, mouse mammary tumor virus (MMTV) as a model to examine the role of mouse BST-2 in host infection in vivo.
By using RNA interference, we show that loss of BST-2 enhances MMTV replication in cultured mammary tumor cells and in vivo. In cultured cells, BST-2 inhibits virus accumulation in the culture medium, and co-localizes at the cell surface with virus structural proteins. Furthermore, both scanning electron micrograph (SEM) and transmission electron micrograph (TEM) show that MMTV accumulates on the surface of IFNα-stimulated cells.
Our data provide evidence that BST-2 restricts MMTV release from naturally infected cells and that BST-2 is an antiviral factor in vivo.
Bone marrow stromal cell antigen 2 (BST-2) protein also known as tetherin/CD317 is a potent restriction factor against a wide range of enveloped viruses such as HIV, FIV, KSHV, MMTV, SIV, Lassa, Marbug, Ebola, and MLV [1–5]. BST-2 achieves its anti-viral effect by connecting both viral and host cell membranes, thus preventing virus egress [6–9]. While BST-2 inhibits virus release, most viruses including HIV-1, HIV-2, and Ebola virus have developed strategies to antagonize BST-2 by degradation, down-regulation of expression, or reduction of its steady-state level [1, 2, 7, 10–14]. In addition to inhibiting virus egress and virus replication in cell culture, there is evidence that, following interferon-induction, BST-2 is up-regulated and incorporated into budding virions [2, 6, 8, 9]. While the antiviral activity of BST-2 has been demonstrated in tissue culture cells, there has been no evidence that BST-2 exerts antiviral activity in vivo. In this context, we evaluated the ability of mouse BST-2 to restrict the replication of the exogenous murine retrovirus mouse mammary tumor virus (MMTV) in cell culture and in mice.
In vivo, MMTV first infects antigen presenting cells (APCs) such as B cells and dendritic cells (DCs) at the site of infection [15–19]. MMTV infected APCs present virus-encoded superantigen (Sag) to T cells expressing Sag-specific T-cell receptor (TCR) Vβ chains. This immunological synapse causes stimulation of Sag-reactive T cells and proliferation of lymphocytes thereby promoting virus replication. Both lymphoid and myeloid cells infected with MMTV are capable of producing infectious virus  and infected lymphoid cells are necessary for virus spread and mammary carcinogenesis [20, 21].
Although MMTV infects and causes mammary cancer in infected mice; nonetheless, infected cells do not produce high virus titer, and time to MMTV-induced cancer is rather long, suggesting that virus replication and spread may be restricted by host factors like BST-2. Indeed, we demonstrate that BST-2 co-localizes with MMTV Gag and Env, and inhibits MMTV particle release in tissue culture cells. Importantly, MMTV infection of mice was significantly inhibited by BST-2.
IFNα induces BST-2 expression and restricts MMTV release
BST-2 expression results in retention of a wide range of virus-like particles (VLPs) assembled in tissue culture [1–4], and IFNα or IFNγ treatment induces the transcription of the gene encoding BST-2 [1, 22]. We hypothesized that IFNα induction of BST-2 in an MMTV producing cell line would enhance BST-2 expression and suppress MMTV release to the culture medium. We treated MMTV-producing GR cells with IFNα or vehicle and then measured BST-2 mRNA and surface protein levels, as well as extra- and intra- cellular viral particles. We observed a significant increase in BST-2 mRNA (Figure 1A) and protein (Figure 1B) levels upon IFNα treatment. This increase in BST-2 levels was accompanied by a reduction in the accumulation of MMTV particles in the culture supernatant, with no change in intracellular viral load as assessed by viral RNA (Figures 1C and 1D) and viral protein (Figure 1E). Additionally, the reduction in particle accumulation in culture supernatant results in lower viral load when such supernatants were used to infect MMTV susceptible cells (Figure 1F). This data demonstrate that IFNα induces endogenous BST-2 in MMTV infected cells and suggest that BST-2 may be the critical factor limiting the accumulation of MMTV particles in the culture supernatant.
Next, we examined whether BST-2 is responsible for IFNα-dependent inhibition of MMTV release. GR cells stably silenced for BST-2 expression with a lentiviral construct carrying BST-2 specific shRNA or control shRNA were stimulated with IFNα or vehicle. Culture supernatants were collected 24 hours later and subjected to Western blot analysis for MMTV capsid protein p27. The corresponding cells were examined for BST-2 protein (Figure 1G). Results show that while shRNA was able to silence BST-2 and enhance accumulation of viral particles in culture supernatant (Figure 1H), IFNα stimulation of shRNA cells results in increased cell surface BST-2 protein expression (Figure 1G) and a reduction in the amount of viral particles accumulated in the culture supernatant (Figure 1H). This result shows that the reduction in the amount of extracellular viral particles observed in IFNα treated cells is linked to BST-2 induction.
IFNα causes accumulation of MMTV structures at the cell surface
In IFNα treated cells, less MMTV particles were detected in the culture supernatant without apparent defect in accumulation of intracellular viral components (Figures 1E, 1H). As BST-2 was shown to block the release of various enveloped viruses/VLPs from the cells [1, 23], we hypothesize that exposure of MMTV-producing GR cells to IFNα will result in accumulation of MMTV virions on the cell surface. To test this, GR cells were stimulated with IFNα or vehicle and 24 hours later, cells were processed for scanning electron microscopy (SEM). In vehicle treated GR cells, 100 nm sized buds were readily observed (Figure 2A). The buds were commonly found on the tip of filamentous structures projecting from the cells surface. IFNα induction drastically increased the number of bud-containing filaments (Figure 2B). To confirm that the buds were indeed virions, thin sections of GR cells prepared under the same treatment conditions were analyzed with transmission electron microscope (TEM). Electron dense virion structures, about 100 nm in diameter (black arrowheads), were detected at the cells surface (Figure 2C and 2D). As expected, in IFNα treated cells, particles that are linked to each other were also observed (Figure 2D). The TEM thus validated that the filamentous structures observed with SEM contain virion(s). Together the electron microscopy data strongly suggest that exposure of MMTV infected cells to IFNα results in retention of MMTV virions on the cell surface.
BST-2 co-localizes with MMTV structural proteins at the cell surface
Consistent with its role in tethering virus particles to the surface of the infected cell, BST-2 has been shown to co-localize with structural proteins of other retroviruses along the plasma membrane of cells [1, 3]. To examine whether BST-2 co-localizes with MMTV structural proteins, BST-2, rat glucocorticoid receptor (RSVGR), and MMTV proviral genome (HP) were co-transfected into 293T cells. Two days later, cells were processed and immuno fluorescently stained to detect BST-2 and MMTV structural proteins. Results show partial co-localization of MMTV Env (Figure 3A) or Gag (Figure 3B) with BST-2 at the cell surface. Our data support previous demonstrations of co-localization of BST-2 with HIV-1 Env in infected cells  and co-localization of SIV Env and BST-2 in activated primary rhesus macaque lymphocytes infected with SIV . In addition, our data are in line with previous report of BST-2 co-localization with the Gag proteins of other retro- and filo-viruses .
Depletion of endogenous BST-2 enhances MMTV particle release and replication in cell culture
To test the hypothesis that BST-2 is responsible for inhibition of MMTV release, we used an RNAi assay to determine whether silencing of endogenous BST-2 expression would lead to higher MMTV particle release and higher levels of infection. GR cells were transfected with either a control siRNA, or a siRNA that targets BST-2 mRNA (Figure 4A). Silencing endogenous BST-2 with siRNA (Figure 4B: upper panel - mRNA, lower panel - protein) enhanced extracellular viral RNA and protein yield compared to cells treated with control siRNA (Figure 4C and 4D respectively), suggesting that endogenous BST-2 affects MMTV particle release. Similar results were obtained when endogenous BST-2 was silenced in GR cells with lentivirus-based shRNA targeting mouse BST-2 (Figures 4E,F,G). Given the ability of endogenous BST-2 to regulate MMTV release, we wondered if BST-2 expression could affect replication and spread of MMTV. Hence, we depleted BST-2 with siRNA in mammary epithelial NMuMG cells (Figure 4H: left panel -mRNA, right panel - protein) and in 3T3 MEFs (Figure 4I: left panel - mRNA, right panel - protein). To evaluate the effect of reduced BST-2 levels on MMTV replication, we infected siRNA treated cells with MMTV, and observed significant differences in viral spread over a course of 96 hours in both NMuMG (Figure 4H: bottom panel) and 3T3 MEFs (Figure 4I: bottom panel). These data suggest that BST-2 impairs MMTV replication and spread in epithelial and fibroblast cells.
Overexpression of BST-2 inhibits MMTV replication
To confirm our finding that BST-2 restricts MMTV replication in cultured cells, we transiently expressed varying amounts of BST-2 or empty vector in TRH3 (293T cells that stably express MMTV entry receptor transferrin receptor 1). Twenty four hours later, a portion of cells were examined for BST-2 expression (Figure 5A), and the remaining cells were infected with MMTV. DNA was extracted from infected cells and used for qPCR examination of virus load 24 hours after infection. Results show that while cells transfected with BST-2 plasmid show lower levels of MMTV DNA across board, transfection of 100 ng of BST-2 plasmid results in about 60% lower infection (Figure 5B), indicating that BST-2 restricts MMTV replication in a dose dependent manner. Whether lower infection observed in BST-2 transfected cells is a result of cells being infected at low multiplicity or lower number of cells being infected is unclear. To address this question, we first generated TRH3 cells stably expressing BST-2 or empty vector (Figure 5C) and infected cells with MMTV.gfp purified from MMTV-producer cell line, CGRES6. These cells produce virions derived from a molecular clone of MMTV (pGR102ES) with a gfp gene inserted into its long terminal repeat (LTR) . Infected cells were collected 24 and 96 hours later and examined for level and intensity of gfp expression. Results show that both at 24 and 96 hours after infection, level of gfp expression was significantly lower in BST-2 expressing cells compared to cells expressing empty vector control (Figure 5D). In addition, gfp mean fluorescence intensity (MFI) was also lower in cells expressing BST-2 at both time points (Figure 5E). These results suggest that BST-2 potently restricts MMTV replication as evidenced by the larger proportion of cells infected in the absence of BST-2 (Figure 5D, solid blue line and solid red line) compared to lower number of cells infected in the presence of BST-2 (Figure 5D, broken blue line and broken red line).
BST-2 specific siRNA depleted endogenous BST-2 in murine lymph nodes
We have demonstrated that loss of BST-2 in tissue culture cells enhances MMTV replication and spread (Figures 4H and 4I lower panels). To examine the role of BST-2 in virus infection in vivo, we first determined whether MMTV target cells and tissues such as dendritic cells (DCs), macrophages (MΦs), lymph nodes and spleens from different mouse strains (C3H/HeN and C57BL/6) that express BST-2 mRNA. We used the mammary epithelial cell line, NMuMG as a reference and show that BST-2 transcript is present in all MMTV targets (Figure 6A). BST-2 mRNA is higher in bone marrow derived DC (BMDC) compared to bone marrow derived macrophages (BMMΦ), popliteal lymph nodes and the spleen, with a modest difference in BST-2 levels between C3H/HeN and C57BL/6 mice. Since lymph nodes express BST-2, we examined whether BST-2 specific siRNA is capable of silencing BST-2 in vivo. Following inoculation of mice with siRNA or siControl subcutaneously in the hind footpad, we evaluated the levels of BST-2 mRNA and protein and show that in 48 hours, siRNA significantly depleted lymph node BST-2 mRNA to about 7 fold (Figure 6B) with a modest reduction in protein level (Figure 6C: upper and lower), and no effect on BST-2 levels of mice inoculated with siControl or no siRNA. We also examined the expression of IFNα, IFNβ, and 2'-5'-oligoadenylate synthetase (OAS) transcripts, and observed that mice treated with siRNA sequences have similar levels of IFNα, IFNβ, and OAS transcripts despite reduced BST-2 mRNA level (Additional file 1). These data suggest that BST-2 specific siRNA reduced BST-2 expression in vivo without eliciting IFN response.
siRNA mediated silencing of endogenous BST-2 enhances MMTV replication and spread in vivo
Local administration of BST-2 siRNA mediates BST-2 depletion in vivo (Figures 6B and 6C). To determine the role of endogenous BST-2 in virus replication in vivo, we administered either a control siRNA, or siRNA that targets BST-2 mRNA to mice, followed by infection with MMTV. Virus replication was significantly higher in siRNA treated mice compared to their siControl or no siRNA counterparts (Figure 6D). In addition, we observed significantly higher virus load in the spleens of mice that received siRNA (Figure 6E). These data show for the first time that siRNA is capable of mediating BST-2 knock down in vivo (Figures 6B and 6C); and that loss of BST-2 in the draining lymph node for the site of initial infection results in higher rate of virus replication and dissemination in vivo (Figures 6D and 6E).
Here, we describe an important anti-viral function for BST-2 in the mouse. Many in vitro and ex vivo studies have shown that BST-2 inhibits the replication of a number of viruses [1–9]. By using MMTV, we directly demonstrate that BST-2 plays a role in virus replication in vivo. Administration of siRNA sequences into mouse footpad results in knock-down of endogenous lymph node BST-2. This assay allowed us to demonstrate that BST-2 functions as a virus restriction factor in a natural host organism. We chose to silence lymph node BST-2 because lymphocytes play a critical role in the in vivo infection with MMTV and other retroviruses such as HIV-1; and in the absence of a knockout model, siRNA provides the best alternative. The significantly higher virus load observed in BST-2 silenced lymph nodes implies that loss of BST-2 enhances virus replication.
BST-2 expression inhibits the egress of HIV-1, HIV-2, Ebola, Marburg and MLV; and we found that it also inhibits the accumulation of MMTV particles in the culture medium. The inhibitory effect of BST-2 on accumulation of extracellular MMTV particles in culture supernatant is consistent with the effect of BST-2 on virus release from the cell surface as has been observed for other viruses [1–5]. The observation that BST-2 co-localizes with MMTV Env and Gag further suggests that BST-2 prevents the accumulation of MMTV particles in culture supernatants. Our EM data support previously documented studies that MMTV buds off from the plasma membrane and microvilli . The cell surface visualization by SEM provides an opportunity to demonstrate that induction of BST-2 with IFNα facilitates accumulation of MMTV on the cell surface. This finding was backed by our protein analyses showing that stimulation of MMTV producer cells with IFNα results in less accumulation of MMTV particles in the culture supernatant.
We have directly demonstrated in this study that loss of BST-2 enhances replication of MMTV in vivo. However, it remains to be determined why mice become infected with MMTV in the presence of the endogenous BST-2. It is worth noting that MMTV has co-existed with mice for over 20 million years  and perhaps must have evolved to avoid most hosts' anti-viral defense mechanisms. The findings of this study are important as it has shown that BST-2 function as anti-viral in the host. Based on this report, we are now focused on using MMTV as a model to understand the role of BST-2 in viral pathogenesis.
Materials and methods
All experiments involving mice were performed in accordance to NIH guidelines, the Animal Welfare Act, and US federal law. The experiments approved by the University of Iowa Animal Care and Use Committee (IACUC). Mice were housed according to the policies of the Institutional Animal Care and Use Committee of the University of Iowa.
BST-2 plasmids used in the study were previously described  and kindly provided by Dr. Paul Bates, University of Pennsylvania, Philadelphia, PA. The MMTV env (Q61) construct , MMTV molecular clone HYB PRO (HP) , and the rat glucocorticoid receptor (RSVGR)  were kindly provided by Dr. Susan Ross, University of Pennsylvania, Philadelphia, PA.
C57BL/6 and C3H/HeN mice were purchased from the Jackson Laboratory and National Cancer Institute (NCI) respectively.
NMuMG (derived from normal mammary tissue of a Namru mouse strain), GR (a mammary carcinoma cell line), Mm5MT (a mammary carcinoma cell line from C3H/HeN mouse strain), 3T3 MEF and 293T cell lines were purchased from American Type Culture Collection (ATCC). TRH3 cells are 293T-mTfR1 (293T cells that stably express MMTV entry receptor transferrin receptor 1 [mTfr1]) and have been previously described . TRH3-BST-2 and TRH3-Vector cells (TRH3 cells stably expressing BST-2 or empty vector). CGRES6-GFP cells (feline kidney stably transfected with a GFP-tagged MMTV proviral construct  was obtained from Dr. Susan Ross of the university of Pennsylvania. All cells were maintained according to the suppliers' instructions. Both GR and Mm5MT cells are from MMTV-induced mammary tumor and both produce infectious MMTV particles.
Generation of TRH3 cell-line stably expressing mBST-2
To generate a TRH3-BST2 stable cell line, we co-transfected 6 μg of BST-2 and 1 μg of puromycin (selection marker) plasmids into TRH3 cells using polyethylenimine (PEI) transfection reagent. Following overnight incubation, transfection medium was changed, and replaced with medium containing puromycin dihydrochloride (3 μg/ml, Santa Cruz Biotechnology, Inc.). The cells were left in culture with three changes of medium to remove dead cells. By day 10 after transfection, several individual clones of stably transfected cells emerged and clones were picked and expanded. Level of BST-2 expression was examined by FACs analysis. After examination, clones with the highest BST-2 expression were expanded and some cells frozen or used for experiments.
Antibodies, siRNAs, shRNAs, and other reagents
Antibodies used in this study includes goat anti-gp52 (Env) and anti-p27 (capsid) previously described [32, 33]. Others are unconjugated rat anti-mouse BST-2 clone 129c, Alexa flour 647 conjugated anti-mouse BST-2 clone 129c , and FITC conjugated anti-mouse BST-2 clone 927 purchased from eBioscience. Rabbit anti-BST-2 and mouse anti-tubulin were from Abnova and Li-core respectively. The following isotype control antibodies were used: Rat IgG2a and Rat IgG2b, both from eBiosciences. The following secondary antibodies from Li-core were used: RDye 680 donkey anti-mouse, IRDye 680 LT conjugated donkey anti-rabbit, IRDye 800CW donkey anti-goat, and IRDye 800CW conjugated goat anti-rabbit. Rabbit anti-rat Alexa Fluor 594 and chicken anti-goat Alexa Fluor 488 were from Invitrogen. The in vivo ready silencer Select target specific siRNA, control siRNA, and SiPort NeoFX transfection reagent were from Ambion while plasmids for target-specific Mission shRNA, control shRNA, and polybrene were obtained from Sigma. BST-2 shRNA and control shRNA lentiviral particles were produced from plasmids of target-specific Mission shRNA and control shRNA by the University of Iowa Vector Core Facility. Recombinant mouse interferon alpha (IFN-α) was from Miltenyi Biotech. Vector-shield was from Vector Laboratories and Lipofectamine 2000 was from Invitrogen.
Infection of mice
In vivo infection of mice was performed with 50 μl of MMTV.MM5MT inoculated subcutaneously on the hind footpad. At different times after infection, mice were sacrificed and the popliteal lymph node draining the site of infection from each mouse was harvested. Lymphocytes were obtained and used for fluorescence-activated cell-sorting (FACS) analysis or used for DNA and RNA extraction.
BST-2 knockdown by shRNA and siRNA in cell culture
BST-2 shRNA lentiviral particles containing 3 target-specific constructs and shRNA constructs encoding a scrambled sequence (Santa Cruz Biotechnology, Santa Cruz, CA) or target specific Mission shRNA and a corresponding control shRNA (Sigma) were used to knock down BST-2 gene expression. Briefly, mouse mammary epithelial tumor cell line GR were transduced with control or BST-2 shRNA lentiviral particles at a ratio of 5 infectious units of virus per cell in the presence of 8 μg/mL polybrene. The next day, fresh media was added to cells and incubated for another 24 hours. In some experiments, puromycin at 2 μg/ml was added to the culture medium for the selection of cells that had stably incorporated shRNA. BST-2 gene silencing was confirmed at the mRNA and protein levels by qPCR and FACS. Knockdown of BST-2 by siRNA was achieved using Ambion's in vivo ready silencer Select target specific or siRNA in vivo ready scrambled siRNA. Briefly, 250 pico moles of siRNA sequences were mixed with Lipofectamine 2000 following manufacturer's recommendation and used to transfect NMuMG, GR or MEFs. The next day, fresh medium was added to cells and incubated until assayed for BST-2 and MMTV mRNA. NMuMG and MEFs were passaged and used for detection of BST-2 and virus replication experiments. Experiments were repeated at least 3 times with similar results.
Western blots of virus preparations, cell lysates from MMTV infected cells, cells transiently transfected with env and capsid plasmids, or immuno-precipitates of protein complexes were probed with anti-total MMTV, anti-gp52, and anti-p27. The species-appropriate IRDye secondary antibodies were used, followed by detection with the Odyssey Infrared Imaging System (LI-COR Biosciences).
DNA, RNA isolation and real-time quantitative PCR (qPCR)
DNA or total RNA was isolated from cells, tissues, and mice lymphocytes using the QIAGEN-QIAamp DNA Mini Kit or RNeasy Mini Kit (Qiagen, Inc.) respectively according to manufacturer's instructions. For cDNA synthesis, equivalent amounts of RNA treated with DNase I (Qiagen, Inc.) were reverse-transcribed with high capacity cDNA reverse transcription Kit (Applied Biosystems, ABI), and the cDNA was amplified with primers specific to BST-2, MMTV.Mm5MT, IFNα, IFNβ, 2'-5'-OAS, and GAPDH. DNA was used to quantitatively detect integrated proviruses [32, 33], using primers specific to the MMTV long terminal repeats and puromycin. Semi-quantitative PCR was performed using Veriti 96-Well Thermal Cycler, and real time qPCR was carried out using ABI 7500 FAST thermal cycler (ABI). All amplifications were normalized to GAPDH and data are presented as fold change relative WT or naïve sample depending on experiment. Sequence of primer pairs used is available (Table 1).
Immuno-fluorescence and confocal microscopy
293T cells were co-transfected with plasmids expressing BST-2, rat glucocorticoid receptor (RSVGR) and MMTV proviral genome (HP) using lipofectamine 2000. The next day cells were treated with DEX and 24 hours later fixed in 4% paraformaldehyde in PBS for 10 minutes. Cells were then washed once with PBS and incubated for 3 hours in IF buffer (0.5% Triton X-100, 0.5%, sodium deoxycholate, 1% bovine serum albumin, 0.05% sodium azide in PSB). Primary antibodies were diluted in IF buffer as follows: rat anti-BST-2 (1:100), goat anti-p27 (1:100) or goat anti-gp52 (1:100) and detected with rabbit anti-rat Alexa Fluor 594 and chicken anti-goat Alexa Fluor 488. All secondary autobodies were dilated 1:1000 in IF buffer. All confocal microscopy images were taken with Zeiss 510 confocal microscope.
Approximately, 1 × 106 GR, MM5MT, NMuMG, 3T3 MEF, 293T, and lymphocytes were stained in PBS + 1% bovine serum albumin (Sigma-Aldrich) for 30 min on ice with Alexa flour 647 anti-mouse BST-2 (clone 129c) or FITC anti-mouse BST-2 (clone 927) and APC anti-mouse IgG2b. Cells were extensively washed in PBS, fixed with 2% paraformaldehyde and subjected to FACS. At least ten-thousand events were collected for each sample using FACS calibur flow cytometer (BD). Cellular frequency and mean fluorescence intensity (MFI) were determined by Flowjo analysis software (TreeStar).
Virus release assay
GR cells were induced for virus production with 1 μM of dexamethasone (DEX)  and treated with IFNα or vehicle for 24 hours. Culture supernatants from cells were clarified by low speed centrifugation, passed through a 0.45 μm filter, and virions were pelleted through a 20% sucrose layer at 32,000g as previously described (30-31). Pellets of virus particles and corresponding cell extracts were denatured and analyzed by SDS-PAGE and Western blot assays. In some experiments, virus particles were used for qPCR analysis of viral RNA or to infect TRH3 cells.
Statistical analysis of significant differences between experimental groups was tested using paired two-tailed Student's t test, and a p value of 0.05 was considered significant. Error bars represent standard deviations.
*Department of Microbiology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, United States of America.
bone marrow stromal cell antigen 2
human immunodeficiency virus type 1
human immunodeficiency virus type 2
interferon alpha and beta, siRNA: short interfering RNA
short hair pin RNA
normal murine mammary gland
simian immunodeficiency virus
scanning electron micrograph
transmission electron micrograph.
Neil SJ, Zang T, Bieniasz PD: Tetherin inhibits retrovirus release and is antagonized by HIV-1 Vpu. Nature. 2008, 451: 425-30. 10.1038/nature06553.
Goffinet C, Schmidt S, Kern C, Oberbremer L, Keppler OT: Endogenous CD317/Tetherin limits replication of HIV-1 and murine leukemia virus in rodent cells and is resistant to antagonists from primate viruses. J Virol. 2010, 84: 11374-84. 10.1128/JVI.01067-10.
Jouvenet N, Neil SJ, Zhadina M, Zang T, Kratovac Z, Lee Y, McNatt M, Hatziioannou T, Bieniasz PD: Broad-spectrum inhibition of retroviral and filoviral particle release by tetherin. J Virol. 2009, 83: 1837-44. 10.1128/JVI.02211-08.
Kaletsky RL, Francica JR, Agrawal-Gamse C, Bates P: Tetherin-mediated restriction of filovirus budding is antagonized by the Ebola glycoprotein. Proc Natl Acad Sci USA. 2009, 106: 2886-91. 10.1073/pnas.0811014106.
Lopez LA, Yang SJ, Hauser H, Exline CM, Haworth KG, Oldenburg J, Cannon PM: Ebola virus glycoprotein counteracts BST-2/Tetherin restriction in a sequence-independent manner that does not require tetherin surface removal. J Virol. 2010, 84: 7243-55. 10.1128/JVI.02636-09.
Perez-Caballero D, Zang T, Ebrahimi A, McNatt MW, Gregory DA, Johnson MC, Bieniasz PD: Tetherin inhibits HIV-1 release by directly tethering virions to cells. Cell. 2009, 139: 499-511. 10.1016/j.cell.2009.08.039.
Habermann A, Krijnse-Locker J, Oberwinkler H, Eckhardt M, Homann S, Andrew A, Strebel K, Krausslich HG: CD317/tetherin is enriched in the HIV-1 envelope and downregulated from the plasma membrane upon virus infection. J Virol. 2010, 84: 4646-58. 10.1128/JVI.02421-09.
Hammonds J, Wang JJ, Yi H, Spearman P: Immunoelectron microscopic evidence for Tetherin/BST2 as the physical bridge between HIV-1 virions and the plasma membrane. PLoS Pathog. 2010, 6: e1000749-10.1371/journal.ppat.1000749.
Fitzpatrick K, Skasko M, Deerinck TJ, Crum J, Ellisman MH, Guatelli J: Direct restriction of virus release and incorporation of the interferon-induced protein BST-2 into HIV-1 particles. PLoS Pathog. 2010, 6: e1000701-10.1371/journal.ppat.1000701.
Le Tortorec A, Neil SJ: Antagonism to and intracellular sequestration of human tetherin by the human immunodeficiency virus type 2 envelope glycoprotein. J Virol. 2009, 83: 11966-78. 10.1128/JVI.01515-09.
Dube M, Roy BB, Guiot-Guillain P, Mercier J, Binette J, Leung G, Cohen EA: Suppression of Tetherin-restricting activity upon human immunodeficiency virus type 1 particle release correlates with localization of Vpu in the trans-Golgi network. J Virol. 2009, 83: 4574-90. 10.1128/JVI.01800-08.
Goffinet C, Homann S, Ambiel I, Tibroni N, Rupp D, Keppler OT, Fackler OT: Antagonism of CD317 restriction of human immunodeficiency virus type 1 (HIV-1) particle release and depletion of CD317 are separable activities of HIV-1 Vpu. J Virol. 2010, 84: 4089-94. 10.1128/JVI.01549-09.
Hauser H, Lopez LA, Yang SJ, Oldenburg JE, Exline CM, Guatelli JC, Cannon PM: HIV-1 Vpu and HIV-2 Env counteract BST-2/tetherin by sequestration in a perinuclear compartment. Retrovirology. 2010, 7: 51-10.1186/1742-4690-7-51.
Pardieu C, Vigan R, Wilson SJ, Calvi A, Zang T, Bieniasz P, Kellam P, Towers GJ, Neil SJ: The RING-CH ligase K5 antagonizes restriction of KSHV and HIV-1 particle release by mediating ubiquitin-dependent endosomal degradation of tetherin. PLoS Pathog. 2010, 6: e1000843-10.1371/journal.ppat.1000843.
Burzyn D, Rassa JC, Kim D, Nepomnaschy I, Ross SR, Piazzon I: Toll-like receptor 4-dependent activation of dendritic cells by a retrovirus. J Virol. 2004, 78: 576-84. 10.1128/JVI.78.2.576-584.2004.
Beutner U, Kraus E, Kitamura D, Rajewsky K, Huber BT: B cells are essential for murine mammary tumor virus transmission, but not for presentation of endogenous superantigens. J Exp Med. 1994, 179: 1457-66. 10.1084/jem.179.5.1457.
Martin P, Ruiz SR, del Hoyo GM, Anjuere F, Vargas HH, Lopez-Bravo M, Ardavin C: Dramatic increase in lymph node dendritic cell number during infection by the mouse mammary tumor virus occurs by a CD62L-dependent blood-borne DC recruitment. Blood. 2002, 99: 1282-8. 10.1182/blood.V99.4.1282.
Courreges MC, Burzyn D, Nepomnaschy I, Piazzon I, Ross SR: Critical role of dendritic cells in mouse mammary tumor virus in vivo infection. J Virol. 2007, 81: 3769-77. 10.1128/JVI.02728-06.
Dzuris JL, Golovkina TV, Ross SR: Both T and B cells shed infectious mouse mammary tumor virus. J Virol. 1997, 71: 6044-8.
Golovkina TV, Dudley JP, Ross SR: B and T cells are required for mouse mammary tumor virus spread within the mammary gland. J Immunol. 1998, 161: 2375-82.
Okeoma CM, Huegel AL, Lingappa J, Feldman MD, Ross SR: APOBEC3 proteins made in mammary epithelial cells can be packaged into retroviruses and restrict transmission of milk-borne virions. Cell Host Microbe. 2010, 8: 534-43. 10.1016/j.chom.2010.11.003.
Yoo H, Park SH, Ye SK, Kim M: IFN-gamma-induced BST2 mediates monocyte adhesion to human endothelial cells. Cell Immunol. 2011, 267: 23-9. 10.1016/j.cellimm.2010.10.011.
Zhang F, Wilson SJ, Landford WC, Virgen B, Gregory D, Johnson MC, Munch J, Kirchhoff F, Bieniasz PD, Hatziioannou T: Nef proteins from simian immunodeficiency viruses are tetherin antagonists. Cell Host Microbe. 2009, 6: 54-67. 10.1016/j.chom.2009.05.008.
Jolly C, Booth NJ, Neil SJ: Cell-cell spread of human immunodeficiency virus type 1 overcomes tetherin/BST-2-mediated restriction in T cells. J Virol. 2010, 84: 12185-99. 10.1128/JVI.01447-10.
Serra-Moreno R, Jia B, Breed M, Alvarez X, Evans DT: Compensatory changes in the cytoplasmic tail of gp41 confer resistance to tetherin/BST-2 in a pathogenic nef-deleted SIV. Cell Host Microbe. 2011, 9: 46-57. 10.1016/j.chom.2010.12.005.
Indik S, Gunzburg WH, Salmons B, Rouault F: Mouse mammary tumor virus infects human cells. Cancer Res. 2005, 65: 6651-9. 10.1158/0008-5472.CAN-04-2609.
Gonda MA, Arthur LO, Zeve VH, Fine DL, Nagashima K: Surface localization of virus production on a glucocorticoid-stimulated oncornavirus-producing mouse mammary tumor cell line by scanning electron microscopy. Cancer Res. 1976, 36: 1084-93.
Bittner JJ: SOME POSSIBLE EFFECTS OF NURSING ON THE MAMMARY GLAND TUMOR INCIDENCE IN MICE. Science. 1936, 84: 162-
Shackleford GM, Varmus HE: Construction of a clonable, infectious, and tumorigenic mouse mammary tumor virus provirus and a derivative genetic vector. Proc Natl Acad Sci USA. 1988, 85: 9655-9. 10.1073/pnas.85.24.9655.
Miesfeld R, Rusconi S, Godowski PJ, Maler BA, Okret S, Wikstrom AC, Gustafsson JA, Yamamoto KR: Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA. Cell. 1986, 46: 389-99. 10.1016/0092-8674(86)90659-8.
Zhang Y, Rassa JC, deObaldia ME, Albritton LM, Ross SR: Identification of the receptor binding domain of the mouse mammary tumor virus envelope protein. J Virol. 2003, 77: 10468-78. 10.1128/JVI.77.19.10468-10478.2003.
Okeoma CM, Lovsin N, Peterlin BM, Ross SR: APOBEC3 inhibits mouse mammary tumour virus replication in vivo. Nature. 2007, 445: 927-30. 10.1038/nature05540.
Okeoma CM, Shen M, Ross SR: A novel block to mouse mammary tumor virus infection of lymphocytes in B10.BR mice. J Virol. 2008, 82: 1314-22. 10.1128/JVI.01848-07.
Schliemann C, Roesli C, Kamada H, Borgia B, Fugmann T, Klapper W, Neri D: In vivo biotinylation of the vasculature in B-cell lymphoma identifies BST-2 as a target for antibody-based therapy. Blood. 2010, 115: 736-44. 10.1182/blood-2009-08-239004.
Parks WP, Ransom JC, Young HA, Scolnick EM: Mammary tumor virus induction by glucocorticoids Characterization of specific transcriptional regulation. J Biol Chem. 1975, 250: 3330-6.
We are also grateful to Dr. Peter Pfeffer of AgResearch Crown Research Institute, New Zealand for the puromycin primer sequence. This research was supported by University of Iowa Startup funds to CMO. PHJ was partly supported by University of Iowa Startup funds to CMO and Immunology T32 training grant to the University of Iowa. This publication was made possible through core services from the University of Iowa Vector core and Central Microscopy Research Facility.
The authors declare that they have no competing interests.
CMO conceptualized and designed research, PHJ, HVM, MM, CMO performed research, and analyzed data; PHJ, HVM, MM, RJR, CMO wrote and read the paper, RJR, Contributed reagents. All authors reviewed the manuscript and approved the final version.
Electronic supplementary material
Additional file 1: BST-2 siRNA did not elicit interferon response in mice. Age-matched C3H/HeN mice were inoculated with siRNA (n = 3), siControl (n = 3), or PBS (WT, (n = 3) subcutaneously on the hind foot pad. Forty-eight hours after inoculation, mice were sacrificed and cells of the draining popliteal lymph node used for total RNA extraction. Quantitative PCR was used to examine mRNA levels of (A) BST-2 (B) IFNα, (C) IFNβ, and (D) 2'-5'-oligoadenylate synthetase (OAS). Data is presented as fold change relative to WT mice. Error bars are standard deviation, and p is significance level. Experiments were performed with 3 mice per group and repeated at least three times with similar results. (TIFF 261 KB)
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Jones, P.H., Mehta, H.V., Maric, M. et al. Bone marrow stromal cell antigen 2 (BST-2) restricts mouse mammary tumor virus (MMTV) replication in vivo. Retrovirology 9, 10 (2012). https://doi.org/10.1186/1742-4690-9-10
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The 3rd floor of the Nedelkovich House, now their home away from home, was reconditioned and redesigned by Ștefania Țiplea. The building dates back to the 18th century and has a beautiful architecture. Some of the original elements were preserved and introduced into the office’s new design.
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Saying that he and his team still felt “troubled” by his loss to President Obama, Mitt Romney on Wednesday attributed his defeat in part to what he called big policy “gifts” that the president had bestowed on loyal Democratic constituencies, including young voters, African-Americans and Hispanics.
In a conference call with fund-raisers and donors to his campaign, Mr. Romney said Wednesday afternoon that the president had followed the “old playbook” of using targeted initiatives to woo specific interest groups — “especially the African-American community, the Hispanic community and young people.”
“In each case, they were very generous in what they gave to those groups,” Mr. Romney said, contrasting Mr. Obama’s strategy to his own of “talking about big issues for the whole country: military strategy, foreign policy, a strong economy, creating jobs and so forth.”
And he goes on with that blather… making the same case regarding women and contraception.
This is basically doubling down on what he said to his donors behind closed doors in Boca Raton, in the infamous "47%" comments. He thinks certain large segments of society are freeloaders who want free stuff and President Obama gives it to them.
Fuck you, Romney. Here's why you lost: because you can't recognize that the middle and working classes are the backbone of this country, and that people like you wouldn't be rich if you didn't profit off the labor and sweat of the 47%. When you asshole Wall Street types fuck up the economy because of your greed, it's not you who pays through the nose; it's the 47% of people who you so callously condemn. When you start your little wars, it's not you, Mr. Senator's Son, Mr. Vietnam forlough, who fights and dies — it's the working and middle class sons and daughter who do all the dirty work and come back with broken bodies and minds. Do they deserve more from this country than you do? Fuck yeah!
In other words, you elitist douchebag, you lost because you ARE an elitist douchebag. Now it is time to prance off to the Cayman Islands, Little Lord Fauntleroy. Begone with you. And don't ever EVER go near a microphone or camera again.


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Middletons Steakhouse & Grill - Norwich
Vegan steak, tomato, mushroom, house salad
Lamb Rump Steak
Lamb rump, tomato & mushroom
Available 12-4pm. Our ever popular roast dinners feature on a separate menu offering two or three courses. The perfect Sunday treat! Please ask your server for more details. (Vegetarian alternative available)
First time attending the restraunt and they didn't dissapointed, we recieved excellent service and the quality of food was outstanding! Will definitely be returning again!
The steaks we had were tremendous and cooked to perfection. They even prepared a small steak for our daughter as she didn’t want anything from the kids menu !! Top service as usual !!
Lovely meal as always . Good good . Good staff . Always meets our expectations.
Never had a bad meal here. Staff always friendly and helpful.
We have been to Middletons a few times now, and it never disappoints. We visited yesterday for our daughter's 12th birthday, and it was beautiful. The staff are so incredibly friendly and the food is sublime. Three of us tried the roast dinner for the first time, and it was delicious! To be able to enjoy a two course meal for four people, with drinks for under £60, is fantastic value for money, especially when you know the meat is top quality. We can't wait to come back again.
Great food and lovely staff looking after us. The mixed grill was a huge plate of delicious meats.
Had a lovely evening, we were told there was a wait for food but it didn’t take long at all and it was all delicious!
Lovely food as always. My favourite place to eat. Second time for breakfast and booked for lunch next week.
Excellent meal and service Extremely good value.
Great service excellent food great place very reasonably priced
The only fault, which can’t be helped. It was a bit too hot in there for my liking
It was the most awful food we ever had. My partner ordered chilli burger, received without spring onion and sauces inside burger, so was funny when staff with afro style bringed to use plate with onion ring and sauce and told us - it goes on the burger by the way. Was so funny and rude. Meat was cold.
Love Middletons, we go at least once a fortnight, staff are friendly, great with children and always make you feel special.
The lunch menu 2 courses 9.95 it’s great if you can’t or don’t eat much but the portions are very small for big eaters.
I was slightly disappointed with the fish grill, not as good as I was expecting.
We all enjoyed our meals and thought the choice was very good indeed.
Lovely place to eat, food was fabulous and service was prompt, will definitely go again!
Nice buzzy place . Nice atmosphere. Good service . We waited a bit of time for food . My chicken was tasty but a bit dry without veg . Otherwise recommend.
Food arrived quickly,all correct,both members of staff encountered were polite and efficient,food was delicious,price is very good for set menu,will be returning,
Everything was nice! The food, the service, the cocktails. No doubt, will come back again next time.
Weekend breakfast, great choice for all tastes and beautifully presented. Really relaxing Sunday morning, cannot fault.
The restaurant was a not that busy, only around 6 tables occupied. The table service was very good, waitress carried out a quality check to ensure we were happy with the food. We did have to wait 20-25 minutes for our food. We could see the chefs preparing our breakfast with the beans being first thing to go on the plate. The next items to be put on the plate was at 10 minutes later so when the order eventually arrived to our table the beans were Luke warm. The bacon was under cooked and the fat not rendered down. Not sure how the bacon was cooked but looked as if it had been steamed, not grilled. Finally I asked for the bill and found that there was a 10% discretionary service charge added. I didn’t have a problem with this and decided to pay with cash, only to be asked if I wanted any change. The change would have been a third of the bill, a little cheeky I thought.
Finally, in hindsight we should have pointed out these failures to the manager and for 5hat we apologise.
Nice Breakfast, quality of the produce was excellent, only thing I would like next time for it to be a bit hotter, it was all rather tepid.
Food was lovely steak cooked how we asked it to be. Waiters quick to get drinks and clear tables.
Great food, service and atmosphere we thoroughly enjoyed our evening
A bit pricey but you don’t mind as the food is always so lovely, really enjoyable evening great service too will definitely be back thanks and we’ll done
We went for a pre theatre meal and chose the 3 course lunch menu. Found it excellent value for money and more than enough choice.
A frequent visitor here, great table service and super food as always. This time we had the lunchtime set menu. Will be back again.
Been here. Few times and we always enjoy it. Staff very nice and the food is always fresh and lovely. We alway have the set menu and it's great value for money.
We had been recommended and we were not disappointed. It was excellent on all levels,food ,service and atmosphere.
Their new breakfast is lovely! Staff were amazing and service was very good
We have always enjoyed the food and ambience at Middletons and any reservations we may have had about a safe eating environment melted away on arrival. We were warmly welcomed, adhering to the usual Covid measures, and ushered upstairs to where the layout has been well-spaced and spotlessly prepared. We were able to enjoy dinner with both privacy and the Middletons atmosphere that makes the restaurant a popular choice for our families. Well done to Middletons for safely retaining the old familiarity, yet a step nearer "new normality'.
Our 44th anniversary and we had a lovely few hours with delicious food, good cocktails and excellent service, the staff were attentive and very friendly , but not in your face. We have booked to take visiting friends in a few weeks.
My husband had the Middleton mixed grill and was very happy with everything he had. I had lobster for the first time and it was delicious. Desserts were lovely, the staff did a special strawberries and cream anniversary plate which was very cute.
A very good evening
We wanted a value for money 2/3 course meal that was in stylish surroundings & included alcohol in the city centre. Somewhere that felt like dining out rather than takeaway & wasn't all "modern" food. Middletons fitted the bill perfectly & we were not disappointed. I choose the 3 course meal at £10.99 whilst my partner choose the same at £18.99. I thought my cheaper meal was just as good as his & excellent value for money. Food was lovely & portions adequate without leaving us feeling overfull. Service & (Covid) safety couldn't be faulted & it was nice to have water included without having to request it. It is nice to find a restaurant that caters for all occasions & pockets under one roof & we are very much looking forward to a return visit.
As usual lovely food and great service. However starters are very small for what you pay for.
Shame about the surroundings as people kept walking in from outside to use the toilet without any masks. Some of these weren’t even customers.
Always good apart from always getting cold black and unappetising mushrooms with the steaks Don’t like the added on staff tips!
Very attentive staff, great food, that was great value - fixed price lunch.
Have been several times and it never disappoints.
very expensive not worth the money expensive expensive
Very nice evening.
Staff were excellent and deserve a pay rise!!

domain: food


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3-2: Virada no 'el Clásico' e 11 pontos de vantagem na liderança
Ancelotti: "É um passo muito importante para vencer a LaLiga"
Lucas Vázquez: "We've taken a big step towards the league title"
Madrid's equaliser to make it 2-2: Vini Jr. crossed for Lucas Vázquez who applies the perfect finish
Bellingham: “It's always great to play a Clásico and even better to win”
Bellingham and Linda Caicedo in the running for the Laureus Awards, which are being presented today in Madrid.
Djokovic, Tom Brady and Andy Garcia at the Clásico
Chus Mateo: "We're going into the playoffs full of energy, enthusiasm and optimism"
Real Madrid starts the Euroleague playoffs tomorrow

domain: news


text:
Once considered to fulfill no other purpose than that of a physical barrier between blood and tissue, the multifunctional nature of the endothelium was discovered in the later half of the 20th century. In cardiology, the dysfunctional nature of the endothelium has received even more attention, initially mainly within the research community but later also in the clinical community, serving as a prime example for the translation of bench research to patient care. In this review, the entity of endothelial dysfunction, its modes of diagnosis in clinical practice, its prognostic implications, and its treatment options will be defined. From past conceptual ideas to current practical applications to the road ahead, the endothelium is to be viewed as the cardiovascular health barometer.
|Original language||English (US)|
|Number of pages||11|
|State||Published - Jul 1 2008|
- Coronary artery disease
- Endothelial dysfunction
ASJC Scopus subject areas
- Cardiology and Cardiovascular Medicine

domain: medical


text:
ATG NEWS RELEASE: Department of the Attorney General releases report on sexual assault evidence collection kitsPosted on Dec 2, 2016 in Latest Department News
HONOLULU –Attorney General Doug Chin announced that the Department of the Attorney General provided a report to the State Legislature today that includes a statewide inventory of sexual assault evidence collection kits, as well as a state plan for how to move forward with untested kits. Act 207 (H.B. 1907) required the Department of the Attorney General to involve community stakeholders in drafting the report, including representatives from each county. The Honolulu, Hawaii, Maui and Kauai Police Departments that are charged with the maintenance, storage and preservation of the kits and other community stakeholders worked with the Department of the Attorney General to produce the report.
Attorney General Chin said, “This report is the product of a great deal of hard work and coordination between the county police departments, county prosecutors, sexual assault treatment providers, and the Department of the Attorney General, among others. I want to thank everyone involved and I also want to thank the Legislature for working with us to make sure this information is available to the public.”
The report provides updates on untested kits from each county, information relating to crime lab DNA testing capacity and related issues, plans for outreach to the public, explanations as to when and why sexual assault kits are tested, and recommendations for legislative action.
In addition to the information provided in the report, the Act 207 working group also developed Malāma Kākou Project, a state plan to:
- Test untested sexual assault kits and new sexual assault kits;
- Identify the criteria for testing and not testing sexual assault kits and the order of testing;
- Provide active outreach and public notification to ensure that information and services are provided to impacted survivors; and
- Establish a tracking system for sexual assault kits.
The report is attached to this news release, but a summary of the inventory of sexual assault kits is as follows:
Honolulu Police Department
Total No. of Kits = 1512
Total No. of Kits Tested = 137
Total No. of Untested Kits = 1375
Kauai Police Department
Total No. of Kits = 209
Total No. of Kits Tested = 74
Total No. of Untested Kits = 135
Hawaii Police Department
Total No. of Kits = 353
Total No. of Kits Tested = 63
Total No. of Untested Kits = 290
Maui Police Department
Total No. of Kits = 166
Total No. of Kits Tested =15
Total No. of Untested Kits = 151
General Chin added, “We will be using this information to better understand the issues that resulted in some kits being untested, and to ensure that all kits that should be tested are tested.”
The Department of the Attorney General previously announced that it was awarded a competitive grant, which should further assist with the efforts to track sexual assault kits. The $2 million National Sexual Assault Kit Initiative (SAKI) grant was awarded to the Department of the Attorney General by the United States Department of Justice, Bureau of Justice Assistance.
The report is available online at https://ag.hawaii.gov/wp-content/uploads/2016/12/Act-207-FINAL-Rpt-12-1-16.pdf
# # #
For more information, contact:
Joshua A. Wisch
Special Assistant to the Attorney General

domain: law


text:
Chronological Guide to the Bible
Published by Thomas Nelson
(Note: This book was provided for review by Thomas Nelson. This did not affect my review in any way.)
Let’s be honest, here – I am not a history buff in any way. It was one of my worst subjects in school. I almost failed both semesters in college! There has been almost no other subject that I have approached with as much dread as history; World, U.S., Social, Heritage – you name it, I hated it. And yet…
At the same time I have always wished that I knew what was going on in the world at large during the Biblical stories I’ve heard since I was a child. Questions like, “Which Pharaoh didn’t know the great Joseph (Exodus 1:8) and why?” have haunted me.
The Guide divides the Bible into nine epochs, beginning with undated creation and ending with the Revelation. It is designed to
“provide you with a guide through the history and culture of the Bible. It will help you follow the flow of events in the Scriptures and see where sacred history and secular history converge into one story of salvation.” (from the “Introduction”)
Each epoch is introduced with an overview of the time period followed by archaeological insights, information about the peoples and groups living then, and the Biblical books that fit within that time.
While it’s not necessarily meant to be read from front to back, it reads like the story that it is – the story of our world. And the pages (which feel like thick magazine pages) are full-color bursting with charts, timelines, pictures, articles, and outlines on every page.
My only criticism of this excellent reference guide comes in two forms:
- There are a few dates that I do know (and they are my litmus test when it comes to books like this) that it gets wrong. But they are not many and certainly not enough to ruin my praise.
- This book is designed to be a wide reference. العاب بوكر اون لاين That means that it provides more information than is necessary for someone who just wants to study the Bible. لعبة بوكر حقيقية For example, instead of just sticking with the Biblical Job, this Guide tells us about other “Job” stories from Mesopotamian legend.
I cannot say that I have read every page, because I haven’t. I have, however, looked at every page and read several of the articles, searched the timelines, and verified many things that do line up with the way I teach the Scriptures. لعب بوكر At this point I would recommend this Guide to anyone who wants to see how the Biblical story fits within the story of the world.
Anyone want to study some history with me?


text:
Lybrate.com has an excellent community of Physiotherapists in India. You will find Physiotherapists with more than 41 years of experience on Lybrate.com. You can find Physiotherapists online in Delhi and from across India. View the profile of medical specialists and their reviews from other patients to make an informed decision.
Book Clinic Appointment with Dr. Saroj Bawaria
Spinal Surgery Disorders
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Treatment of Nerve And Muscle Disorders
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Neuro Physiotherapy Treatment
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Treatment of Sports Injuries
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Treatment of Spondylosis
Arthritis And Pain Management Treatment
Heat Therapy Treatment
Post Pregnancy Classes
Orthopedic Physical Therapy
Treatment of Shin Splints
Submit a review for Dr. Saroj BawariaYour feedback matters!
I am suffering from low back pain I can't bend when I am playing Shuttle I started Shuttle playing since 1 week how could I reduce.
When I go down stairs my knee pains. But when I go up no. I am involved I alot o physical activities at work and so I need my legs strong what should I do?
I feel pain on my both the wrists when I break the sleep in the morning while in the bed But it is getting disappeared when I start my day. This is there for the last one month.
Mere Ek pair me kamar se lekar aedi tak bahut pain hota h uthne bethne me chalne firne me dikkat hoti h.
A broken bone is medically termed as a fracture.Fractures are linked with age as bones tend to become weak as one ages, so, the chances of a fracture occurring also increases. Fractures are usually classified into four types-
1. Closed fracture - In this type of fracture, the breaking of the bone does not result in an open wound
2. Open fracture- The opposite of closed fracture, the bone breaks through the skin. This may result in infections
3. Non-displaced fracture - in this type of fracture, the bone cracks but is not shifted out of its regular position.
4. Displaced fracture - A displaced fracture occurs when the affected bone is shifted out of its natural position.
Fractures can be caused by a number of factors, they are
1. Road accidents
2. When someone falls from a height
3. When the bone is directly exposed to a strong blow
4. When the bone is exposed to a repetitive force from activities such as sprinting
5. Osteoporosis, which is a condition where the bones become weak
6. The bones in the body tends to weaken with age, so at this time they become highly susceptible to fractures
1. Swelling in the area accompanied by pain
2. Limited movement of the affected joint
3. Muscle spasms
4. The affected area may also become numb
The first step to diagnose a fracture is the knowing about the medical history of the patient and conducting a physical exam. The diagnosis is further confirmed by using imaging tests such as -
1. Magnetic resonance imaging (MRI) - In this procedure, images of internal body parts are taken with the help of a magnetic field and pulses of radio energy. MRIs can pick up signs of fractures very quickly, that is within the first week of injury. It is also easier to distinguish fractures from other related injuries in case of MRI.
2. X-rays -X-rays are a procedure that involves sending x-ray particles through an area to detect a fracture. Signs of a fracture may take several weeks to show up in x-rays.
3. Bone scan - In bone scans, some radioactive material is put inside the body through intravenous methods. These material tend to accumulate in the fractured area which then shows up in the images as a bright spot. If you wish to discuss about any specific problem, you can consult an Orthopedist.

domain: health


text:
In this fine cookery course, legendary chef Rowley Leigh traces the history of cooking from hunter-gatherer times and gives simple recipes from each period in humankind’s culinary evolution. Rowley Leigh’s ‘History of Cooking’ is divided into six chapters. Each chapter is 20 to 40 minutes long, and is accompanied by a printable recipe sheet. You can view each film as many times as you like.
You will find Rowley to be delightful company, and warm and witty. You would expect no less from the man who led the Idler team to victory against the Financial Times on University Challenge. And the recipes are divine as well.
Our films were made in Rowley’s kitchen.
Chapter One: The Hunter-gatherer
In this first chapter on wild food, Rowley teaches you how to cook rabbit with ceps.
Chapter Two: The Conquest of Fire
How to cook the perfect steak with béarnaise sauce.
Chapter Three: The Triumph of Farming
How to cook khichiri, the ancestor of kedgeree, plus garam masala and courgette risotto.
Chapter Four: The Egg
How to make an omelette, plus oeufs à la neige
Chapter Five: The Chicken
Roast chicken with tarragon, and chicken à la king
Chapter Six: Salt
The advent of preservatives is celebrated with recipes for Parmesan custard and anchovy toast, and spaghetti Amatriciana


For many mums it's the tipple of choice so the news that Aldi is launching a THREE LITRE bottle of prosecco is causing quite a stir.
The Prosecco Jeroboam DOCG will be available in stores and online from November 14 and will be priced £39.99.
Previous releases of magnum size bottles of champagne and wines have proved extremely popular with shoppers, and with this one being the equivalent size of four bottles or 24 glasses, it's set to be a huge hit.
It’s filled with the supermarket’s DOCG Prosecco from the Italian region of Veneto, which is a light bubbly with an 11 per cent ABV.
Last year around the Christmas period, the supermarket reported it sold more than two million bottles of prosecco - that's 69,000 bottles a day.
No wonder they decided to supersize their options.


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The real world and the fairy world intersect in Tanya, a teenager who can see fairies. These fairies are often darkly malevolent creatures, but she keeps her strange ability secret, unaware that anyone else shares it. When, however, she goes to stay in her grandmother’s decrepit manor house near sinister Hangman’s Wood, the legacy of old tragedies intrude into her life and she finds her very strange family history unravelling and absorbing her in the process.
Two mysterious girls are encountered by Tanya: the very material Red and the unearthly Morwenna, both of them bound up with mysteries involving changelings and events in the grim woodland 50 years earlier; events that had such an impact on Florence, Tanya’s grandmother, and on old Amos, the father of Warwick, the caretaker, that Florence will not speak about them and Amos cannot. The nerdy Fabian, Warwick’s son, becomes Tanya’s ally in her attempts to understand a complex and sinister tangle, often made more dangerous by the intervention of the fairies.
Harrison keeps her plot moving along nicely; the antics and utterances of many varieties of fairy provide some humour, leavening the darker aspects of the plot and ensuring it does not spill over into melodrama. An ambitiously conceived and executed first novel, The 13 Treasures, leaves readers hoping for more from a promising new author.


text:
It has familiar faces at the top — our top nine teams played in the New Year's Day Six in Preseason college football rankings: Top 25 for 1. 1 pick in the NFL Draft, but injuries along an offensive line that already. College football polls and rankings at carpetcleaninginlondon.info include the AP Top 25 College Updated 09/06/ Rank. Team. Move. Schedules/Results. Points. carpetcleaninginlondon.info ranks college football's Top 25, offers reliable predictions, provides expert analysis, reacts to Funny Fantasy Football Team Names.
Ncaa team college football top 25 picks - sportsbookIf Bryant, Johnson or Zerrick Cooper settles into the starting job, the Nov. Ranking the Best College Football Uniforms in Who else made the list? College Football The Purdue Boilermakers nearly pulled off a stunning upset over the Louisville Cardinals last Saturday, and now the North Carolina Tar Heels are going to be the next ones to take a shot at Lamar Expert Predictions for Every Game in Week 2 College Football's Top 20 Quarterback Debuts of USC Seeks to End Losing Streak to Stanford with Statement Win Wake Forest Demon Deacons vs. NFL Spread Picks NBA Spread Picks MLB Spread Picks College Football Spread Picks College Basketball Spread Picks NFL Rankings NBA Rankings MLB Rankings College Football Rankings College Basketball Rankings NFL Stats NBA Stats MLB Stats College Football Stats College Basketball Stats NFL Odds NBA Odds MLB Odds College Football Odds College Basketball Odds.
Nfl: Ncaa team college football top 25 picks
|Ncaa bcs football standings college football boards||962|
|FOODBAL GAMES ESPN NCAA FOOTBALL LINES||Coolege football scores cfb championship game|
|Ncaa team college football top 25 picks||College football scores update yahoo football scores|
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|Ncaa team college football top 25 picks||College football all scores ncaa football top 25|


text:
One always has to know when a stage comes to an end. If we insist on staying longer than the necessary time, we lose the happiness and the meaning of the other stages we have to go through.
Closing cycles, shutting doors, ending chapters – whatever name we give it, what matters is to leave in the past the moments of life that have finished.
Did you lose your job? Has a loving relationship come to an end? Did you leave your parents’ house? Gone to live abroad? Has a long-lasting friendship ended all of a sudden? You can spend a long time wondering why this has happened.
You can tell yourself you won’t take another step until you find out why certain things that were so important and so solid in your life have turned into dust, just like that. But such an attitude will be awfully stressing for everyone involved: your parents, your husband or wife, your friends, your children, your sister.
Everyone is finishing chapters, turning over new leaves, getting on with life, and they will all feel bad seeing you at a standstill.
Things pass, and the best we can do is to let them really go away.
That is why it is so important (however painful it may be!) to destroy souvenirs, move, give lots of things away to orphanages, sell or donate the books you have at home.
Everything in this visible world is a manifestation of the invisible world, of what is going on in our hearts – and getting rid of certain memories also means making some room for other memories to take their place.
Let things go. Release them. Detach yourself from them.
Nobody plays this life with marked cards, so sometimes we win and sometimes we lose.
Do not expect anything in return, do not expect your efforts to be appreciated, your genius to be discovered, your love to be understood.
Stop turning on your emotional television to watch the same program over and over again, the one that shows how much you suffered from a certain loss: that is only poisoni
ng you, nothing else.
Nothing is more dangerous than not accepting love relationships that are broken off, work that is promised but there is no starting date, decisions that are always put off waiting for the “ideal moment.”
Before a new chapter begins, the old one has to be finished: Tell yourself that what has passed will never come back.
Remember that there was a time when you could live without that thing or that person – nothing is irreplaceable, a habit is not a need.
This may sound so obvious, it may even be difficult, but it is very important.
Closing cycles. Not because of pride, incapacity or arrogance, but simply because that no longer fits your life.
Shut the door, change the record, clean the house, shake off the dust.
Stop being who you were, and change into who you are.
~ Paulo Coelho.
I agree with my friend, Paulo. His words are truly exquisite. He’s a doctor to our losses, and of our souls.
Life is sometimes difficult to understand. Sometimes, we can’t close a cycle because we don’t know if we should. Sometimes, we don’t get answers. Sometimes, our wounds don’t heal, even though time is supposed to do that for us. I don’t agree. We might not burst into tears at the memory five years from now because the pain has lessened. But the wounds are never gone. Paulo is right, you never go back to the way you were. This is a problem for most of us. We want to go back to what we had, what we felt, to our unfinished business, just go back to the old days where everything seemed easier. In other words, we want to go back to the past.
But we can’t. So we hold on. We Hope. We Believe. Because It’s hard to accept that things will never be the same.
In a world that is constantly moving, no one should stand still. How do you keep your feet planted on a ground that is always shifting? How is one to keep up? How can you rely on anybody if everyone changes? What does it take to make it in this rat race?
Oh, what a rat race.
Copyright © 2014 Elisa Marie Hopkins. All rights reserved.

domain: literature