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Featured MNV Articles:
Syngenta Breaks Above 200-Day Moving Average - Bullish for SYT
Monday, January 9, 11:11 AM ET, by Market News Video Staff
In trading on Monday, shares of Syngenta AG (SYT) crossed above their 200 day moving ...
Materials Select Sector SPDR Fund Experiences Big Outflow
Wednesday, January 11, 11:11 AM ET, by Market News Video Staff
Symbols mentioned in this story: XLB, DOW, DD, MON Exchange traded funds (ETFs) trade just ...
Notable Thursday Option Activity: MNK, CELG, DE
Thursday, January 12, 2:30 PM ET, by Market News Video Staff
Looking at options trading activity among components of the S&P 500 index, there is noteworthy ...
Coty Becomes #65 Most Shorted S&P 500 Component, Replacing Deere
Thursday, January 12, 3:02 PM ET, by Market News Video Staff
The most recent short interest data has been released for the 12/30/2016 settlement date, and ...
Symbols mentioned in this story: MOO, MON, POT, DE
Exchange traded funds (ETFs) trade just like stocks, but instead of ''shares'' investors are actually buying and selling ''units''. These ''units'' can be traded back and forth just like stocks, but can also be created or destroyed to accommodate investor demand. Each week we monitor the week-over-week change in shares outstanding data, to keep a lookout for those ETFs experiencing notable inflows (many new units created) or outflows (many old units destroyed). Creation of new units will mean the underlying holdings of the ETF need to be purchased, while destruction of units involves selling underlying holdings, so large flows can also impact the individual components held within ETFs.
Looking today at week-over-week shares outstanding changes among the universe of ETFs covered at ETF Channel, one standout is the Agribusiness ETF (AMEX:MOO) where we have detected an approximate $22.4 million dollar outflow -- that's a 0.4% decrease week over week (from 111,700,000 to 111,250,000). Among the largest underlying components of MOO, in trading today Monsanto Co. (NYSE:MON) is off about 0.4%, Potash Corp. of Saskatchewan Inc. (NYSE:POT) is up about 0.2%, and Deere & Co. (NYSE:DE) is higher by about 0.1%. For a complete list of holdings, visit the MOO Holdings page �
The chart below shows the one year price performance of MOO, versus its 200 day moving average:
Looking at the chart above, MOO's low point in its 52 week range is $39.86 per share, with $56.22 as the 52 week high point � that compares with a last trade of $49.64. Comparing the most recent share price to the 200 day moving average can also be a useful technical analysis technique -- learn more about the 200 day moving average �.


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Avast Internet Security 5.0.396 + Key |44 MB
Avast Internet Security includes antivirus engine, anti-rootkit protection, anti-spyware protection, script shield, process virtualization, command line scanner, anti-spam filter, and built-in firewall. Avast Internet security includes all the features of the free Avast anti virus and Firewall, and antispam. Once installed, avast runs silently in the background to protect your computer against all known forms of malware with a real time virus protection. Avast Internet Security also fully integrated antispam filter to detect unwanted email.
Información (Cómo descargar - NUEVO: Ver Enlaces de Descarga actualizados al 2017):

subset: commoncrawl


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Vía Laietana, 69, Ciutat Vella, 08003 Barcelona, Spain
On our trip to Spain, we spent four of our 16 nights in Barcelona. There was so much art I had to see there and I didn't even have time to see it all! Two of those nights we stayed at the W Hotel on the beach, and two we stayed at the H10 Cubik.
I researched several hotels in Barcelona, and admittedly booked pretty late so I probably didn't have as many options as I could have. There are so many cool hotels in Barcelona. I had a list of around ten that I really wanted to stay in. I ultimately picked H10 because it was the cheapest of the nice hotels on my list and still looked really cool.
The H10 Cubik is located on one of the main streets and is in a central location that feels more local versus touristy. If you say on La Rambla that is more touristy, but the H10 Cubik is just a few blocks over, so you get a more local feel.
The hotel was really close to what ended up being one of our favorite restaurants in Barcelona — Cafe Lolea. We almost went back to eat there a second time because we couldn't stop thinking about the black truffle risotto. I'm still thinking about it… Continue reading…


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Showcase Schedule Is Up for the Miami Music Festival
With all the Art Basel hype (only one week away, woooo), you can be forgiven if you forgot our previous announcement about the Miami Music Festival. In its inaugural year, it's a three-day shebang styled after bigger industry conferences like SXSW and CMJ, with simultaneous showcases at a bunch of different venues, and panels and workshops with industry folks. You can also be forgiven if that last sentence makes you really skeptical, considering how other large-scale local music festivals have gone off recently.
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TicketsSun., Feb. 26, 2:00pmpowered by goldstar
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iPhone apps
Saddest Zombie
Ghoultide Meets YuletideThey said it couldn't be done, combining zombies and christmas, but novelist Douglas Clegg and artist Glenn Chadbourne have done just that. Follow Tommy Tommy Tomby and his ghoulish brethren as they discover the meaning….. Read more
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Rated 4/5 based on 3 customer reviews.
4/5 stars
Boo hoo
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Littlest zombie
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Contact the Register
Lobby hours: 8 a.m. – 5 p.m. Mon.-Fri
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New Local Customer Service
- The Orange County Register: 714-796-7777
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Central County Newsroom Contact:
(Costa Mesa, Fountain Valley, Huntington Beach, Irvine, Los Alamitos, Newport Beach, Santa Ana, Seal Beach and Tustin)
Susan Vardon, 714-796-7954 email@example.com
625 N. Grand Ave.
Santa Ana, CA 92701
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(Aliso Viejo, Coto de Caza, Dana Point, Ladera Ranch, Laguna Beach, Laguna Hills, Laguna Niguel, Laguna Woods, Lake Forest, Mission Viejo, Rancho Santa Margarita, San Clemente and San Juan Capistrano)
Susan Vardon, 714-796-7954 firstname.lastname@example.com
625 N. Grand Ave.
Santa Ana, CA 92701
North County Newsroom Contact:
(Anaheim, Buena Park, Cypress, Fullerton, Garden Grove, La Habra, La Palma, Orange, Placentia, Stanton, Villa Park, Westminster and Yorba Linda)
Heather McRea, 714-796-7932 firstname.lastname@example.com
625 N. Grand Ave.
Santa Ana, CA 92701
- Brian Calle, VP, Opinion Page Editor firstname.lastname@example.com
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Letters to the Editor
Please provide your name and telephone number (telephone numbers will not be published) to email@example.com. Letters of about 200 words will be given preference. Letters will be edited for length, grammar and clarity. Visit our Letters To The Editor section.
Corporate Communications
Eric Morgan, Dir., Communications firstname.lastname@example.com 714-796-2460
Excélsior / UNIDOS
Southern California's premier Spanish-language weekly newspaper and website. For the newsroom, call 951-368-9330. For advertising, call 714-796-3845.
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Our agents are our #1 priority, and it shows with our average agent growth of 35% every year since we started! We're a team of driven and winning freight broker...
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Safeway Logistics, a United Van Lines Agent,consists of approximate 50 trucks, and 15 owner operators in a flatbed division.....
Michigan City Driver
Dillon Logistics Inc - Michigan City, IN
No more than 2 moving violations in the last 36 months. Including, but not limited to, 1) driving a loaded tractor trailer for up to 11 continuous hours, 2)...
Regional CDL Class A Delivery Driver,Home every week
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KRC Logistics is home to Keller Transfer, RTI Transport, Commerce Distribution and Staley General. Come join our family at KRC Logistics....
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Michigan City Ground Crew
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Up to 12 hours at a time. Ability to squat/ kneel while fully extending arms and supporting 20 lbs. Ensuring product is unloaded into proper silo (as designated...
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Responsibility * collaborate to create solutions to fulfill clients requests within the boundaries of how client's system runs * create web apps * Work with
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Logistics route analyst, multi-task, manage,. Previous experience in logistics planning or dispatching preferred....
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Holiday Inn Express Marshall
- Tasuta Rootsi lauaga hommikusöök, tasuta wifi ja tasuta parkimine
Peamised teenused/mugavused
- 65 mittesuitsetajatele mõeldud numbrituba
- Sisebassein
- Treeningsaal
- Kontorinurk
- Õhukonditsioneer
- Igapäevane koristusteenus
- Seif vastuvõtulauas
- Pesumajateenus
- Tasuta ajalehed fuajees
- Lastevoodid saadaval
- Lisavoodid saadaval
- Mikrolaineahi
- Külmik
- Privaatne vannituba
- Kvaliteettelekanalid
- Honolulu House (~ 2 km)
- American Museum of Magic (~ 2,5 km)
- U.S. Postal Museum (~ 2,7 km)
- Alwyn Downs Golf Club (~ 3,9 km)
- Oakland Hills Golf Club (~ 12 km)
Out of pretty much everything for breakfast from 8am to 930pm before they got eggs and…
good and convenient - that is it - no more words - if you keep on asking questions you…
Holiday Inn Express Marshall
Teave hotelli kohta: 800 004 4466 TASUTA lauatelefonistalates 86€
- One King Bed With Kids Suite Nonsmoking
- Tuba, suitsetamine keelatud
- Standard tuba
- Üks suur voodi Mittesuitsetajatele Juurdepääs
- Tuba, suitsetamine keelatud
- Tuba, 1 ülilai voodi, suitsetamine keelatud (whirlpool)
- Kaks kaheinimesevoodit
- Tuba, suitsetamine keelatud
- Tuba, suitsetamine keelatud
Rohkem häid valikuid
Hotelli suurus
- Selles hotellis on 65 tuba
- Sellel hotellil on 3 korrust
- Sisseregistreerimine alates 16.00
- Väljaregistreerimine 12.00
Sisseregistreerimisel vajalik
- Nõutav on krediitkaardi- või sularahadeposiit
- Nõutav on ametlik fotoga isikutunnistus
- Lemmikloomad ei ole lubatud
- Kokkupandavad voodid saadaval, võib kehtida lisatasu*
- Hotelli territooriumil saadaval tasuta wifi
- Tubades tasuta wifi
- Tasuta iseteenindusega parkla
Veel teavet
- Hotellis suitsetamine keelatud
Lähedal asuvad 2 tärni hotellid Marshall
- Tasuta Rootsi lauaga hommikusöök
- Sisebassein
- Jõu-/treeningsaal
- Kontorinurk
- Keemiline puhastus ja/või pesumajateenus
- Tasuta ajalehed fuajees
- Lift
- Pangaseif vastuvõtulauas
- Invatualett/vannituba
- Invaabivahendid toas
- Dušinurk ratastoolis liikujale
- Õhukonditsioneer
- Kohvi/teekeetja
- Triikraud/triikimislaud
- Privaatne vannituba
- Föön
- Lameekraantelerid
- Tasulised filmid
- Kvaliteettelekanalid
- Laud
- Tasuta ajaleht
- Tasuta wifi
- Kohalikud kõned tasuta
- Külmik
- Mikrolaineahi
- Igapäevane majapidamis-/koristusteenus
Populaarsemad hotellid Marshall
Lähedal asuvad hotellid
Soovitatud hotellid
Lisaks saadaval
Auhinnad ja kuuluvus ühendustesse
Roheline/keskkonnasäästlik hotell
See majutusasutus on liitunud Green Key Eco-Rating Program hindamissüsteemiga, mis mõõdab majutusasutuse mõju ühele või mitmele järgnevatest: keskkond, kogukond, kultuuripärand, piirkonna majandus.
Viimased arvustused
Suurepärane 4,1 /5 from 71 reviews
the hotel was ok, but upon arrival there were men in a pickup truck and a circle of lawn chairs sitting under the entrance awning (in the driveway intended for front door drop off) drinking cases (yes cases) of beer... management would not tell them to move because they were "guests"..at least 8 grown men drinking that my 16 year old daughter had to walk thru to enter ...and noone could drive a car to door. spoke to manager and she refused to tell them to move to rooms, appropriate area, etc. so gross. management showed zero respect for any guests except this group of drinkers. gross, odd, inappropriate, and possibly dangerous and illegal, too
Hotels.com did not pay for my stay in full and I was told I owed a balance when checking in. This held my check in up by nearly an hour. The hotel was diligent in ensuring I was taken care of, however I would like compensation from Hotels.com as they did not fulfill my request/needs as they have in the past. It was quite disappointing.
Very clean and very comfortable otel with (unfortunately!) terrible night shift front desk person. Everything about this otel was great except the front desk person who has no idea how to welcome and speak to hotel's guests!!
I would have liked to give this hotel a better rating, it was typical, just off the expressway and close enough to the casino to make it work. Our check in was nice, the room was clean and well-appointed. My only complaint regarding the room was a large, ugly cigarette burn on the bathroom counter. The rooms appear to have been re-done recently and I expect you charged whoever did this a large sum for the damage. However, once you collect the damages, you must fix the damage. The other "bad experience" was breakfast. I know the breakfast is advertised as "free", however it is figured into the price of the room so you are really paying about $10 - $12 for this. Breakfast was to be served from 8 am to 10 am. We arrived in the breakfast room about 9:10 am to find all the eggs gone, all the sausage gravy gone, the little egg cooker was broken. The biscuits were gone. The milk was gone, se even cereal wasn't an option. When we asked about refills, we were told "You should have gotten here earlier"! We are reasonable people who travel often, we would have been happy to wait a bit for more food, but this response was unacceptable. Insulting us certainly won't get us back!
The only reason I did not give full points is that the pool was very cold. But everything was great. Clean room, great breakfast and very friendly service!


text: Gym Named In Honor Of Dunn Educator Gym Named In Honor Of Dunn Educator Undercover Investigation Results In Arrest Undercover Investigation Results In Arrest Can You Spell Marathon? Can You Spell Marathon? Averasboro Reenactment This Weekend Averasboro Reenactment This Weekend Campbell, County Connect Campbell, County Connect Attention Print Subscribers: Activate your FREE ONLINE ACCESS now! FEATURED POSTS Gym Named In Honor Of Dunn Educator Feb 240 Undercover Investigation Results In Arrest Feb 240 Can You Spell Marathon? Feb 240 Averasboro Reenactment This Weekend Feb 240 Lillington Chamber Honors Those Who Excel Feb 240 COMMUNITY NEWS Crown Arena To Host Expo March 17-19 Feb 240 Bring The Mediterranean Into Your Kitchen Feb 240 Add Some Greenery To Your Grill Feb 240 Daily Record Photo/Shaun Savarese Feb 240 North Harnett Primary Feb 240 Multiplication Color by Number Feb 240 Enhanced Flu Testing Available at ABC Pediatrics! Feb 240 SPORTS Building Continues For Campbell Feb 240 Raiders Ousted Feb 240 Midway Girls Topped By S. Lenoir Feb 240 Lady Camels Log 20th Win Of Season Feb 240 Overhills' Season EndsFeb 23 at 10:38 PMFeb 230 Poll What is your favorite Girl Scout cookie? Girl Scout S'mores Thin Mints Caramel deLites/Samoas Peanut Butter Patties/Tagalongs Shortbread/Trefoils Do-si-dos/Peanut Butter Sandwich Lemonades Savannah Smiles Thanks-A-Lot Toffee-tastic Trios View Results Loading ... View our E-Edition, a digital replica of our print edition. MOST POPULAR RIGHT NOW Tar Heel Traveler To Be In CoatsFebruary 21Feb 210 Governor Offers Reward In Missing Man CaseFebruary 21Feb 210 New Faces Coming To AngierFebruary 22Feb 220 Overhills Edged In Double OT, Pairings DelayedFeb 19 at 9:04 AMFeb 200 Camels Fall Without Clemons, HamerFeb 19 at 8:43 AMFeb 200 OBITUARIES Doris M. McClam, 65 Feb 240 Joyce P. Lucas, 74 Feb 240 Robin S. Fleming, 40 Feb 240 Velda S. Bass, 63 Feb 240 Ronnie R. Stanley Sr., 60 Feb 240 Daniel R. 'Poppa' Moore, 65 Feb 240 Joyce D. Byrd, 66 Feb 240 Flora L. Clayton, 91 Feb 240 Social Media EDITORIALS Remembering A Giant In American History Feb 240 SOUND OFF Feb 240 Published By RECORD PUBLISHING COMPANY, INC. 99 W. Broad St., P.O. Box 1448 Dunn, N.C. 28335 Feb 240 REMEMBERING WHEN Feb 240 — The Salisbury Post Feb 240 IN THE RECORD St. Matthew's Valentine King And Queen Feb 240 Carla And Redemption In Concert Feb 240 Second Editon To Be At Lee's Chapel Sunday Feb 240 Piney Grove Chapel To Host Jeff & Sheri Easter Feb 240 Janet Paschal To Be At Hickory Grove AC March 4 Feb 240 Archives Older articles are available in the E-Edition. Archives February 2017 January 2017 December 2016 November 2016 October 2016 September 2016 August 2016 July 2016


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No Bass No Fun :) #wojtek_pilichowski #bass #guitar #music
I am new here!
No Bass No Fun :) #wojtek_pilichowski #bass #guitar #music
Summer we are waiting for you! Sun, sand, sea, beach and lot of fun. Watch the video and share it with your friends:P
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12 Jul 2012: Tian D, Zhang W, He J, Liu Y, Song Z, et al. (2012) Correction: Novel, Real-Time Cell Analysis for Measuring Viral Cytopathogenesis and the Efficacy of Neutralizing Antibodies to the 2009 Influenza A (H1N1) Virus. PLOS ONE 7(7): 10.1371/annotation/9f718228-cb10-444b-882b-5dc795e9d1e7. doi: 10.1371/annotation/9f718228-cb10-444b-882b-5dc795e9d1e7 View correction
A novel electronic cell sensor array technology, the real-time cell analysis (RTCA) system, was developed to monitor cell events. Unlike the conventional methods labeling the target cells with fluorescence, luminescence, or light absorption, the RTCA system allows for label-free detection of cell processes directly without the incorporation of labels. Here, we used this new format to measure the cytopathic effect (CPE) of the 2009 influenza A (H1N1) virus and the efficacy of neutralizing antibodies in human sera to this virus. The real-time dynamic monitoring of CPE was performed on MDCK cell cultures infected with the H1N1 virus, ranging from 5.50×102 to 5.50×107 copies/mL. The resulting CPE kinetic curves were automatically recorded and were both time and viral load dependent. The CPE kinetics were also distinguishable between different H1N1 stains, as the onset of CPE induced by the A/Shanghai/37T/2009 H1N1 virus was earlier than that of the A/Shanghai/143T/2009 H1N1 virus. Furthermore, inhibition of H1N1 virus-induced CPE in the presence of human specific anti-sera was detected and quantified using the RTCA system. Antibody titers determined using this new neutralization test correlated well with those obtained independently via the standard hemagglutination inhibition test. Taken together, this new CPE assay format provided label-free and high-throughput measurement of viral growth and the effect of neutralizing antibodies, illustrating its potential in influenza vaccine studies.
Citation: Tian D, Zhang W, He J, Liu Y, Song Z, Zhou Z, et al. (2012) Novel, Real-Time Cell Analysis for Measuring Viral Cytopathogenesis and the Efficacy of Neutralizing Antibodies to the 2009 Influenza A (H1N1) Virus. PLoS ONE 7(2): e31965. doi:10.1371/journal.pone.0031965
Editor: Suryaprakash Sambhara, Centers for Disease Control and Prevention, United States of America
Received: September 12, 2011; Accepted: January 19, 2012; Published: February 20, 2012
Copyright: © 2012 Tian et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: The study was supported by the National Major Science and Technology Project–Prevention and Treatment of AIDS, Viral Hepatitis, and Other Major Infectious Diseases (Grant NO. 2009ZX10004-211), China ministry of health, and by the grant from the Shanghai Medical College of Fudan University (Grant NO. 09L-34), Shanghai, China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
With the 2009 global outbreak of pandemic influenza, large-scale vaccination against the new strain of influenza A virus subtype H1N1 was employed to prevent its spread in China, and numerous clinical trials were initiated to estimate the antibody responses elicited with the 2009 influenza A (H1N1) virus vaccine [1], [2], [3]. However, there is still an urgent need for safer and more convenient approaches to monitor influenza stains that may cause pandemic influenza with significant levels of morbidity and mortality. As such, high throughput neutralization assays are required to quantify neutralizing antibodies for serodiagnosis during influenza infection as well as for screening candidate vaccines. The traditional measurement of influenza virus neutralizing antibodies involves hemagglutination, immunological procedures, or various methods for cytopathogenic effect (CPE) identification, such as microscopic, colorimetric, and fluorometric assays [4], [5], [6], [7]. Nevertheless, these methods are somewhat labor-intensive, which may increase the risk of aerosol generation [6], [7], [8].
Since 2004, a real-time cell analysis (RTCA) system based on microelectronic biosensor technology was developed to monitor cell events. The electrode impedance, which is displayed as cell index (CI) values, is measured to characterize cellular status across the electronic cell sensor array integrated on the bottom of the electronic microtiter tissue culture plates (E-Plates) [9], [10]. Due to the the novel electronic cell sensor array technology, the RTCA system can provide dynamic monitoring of nearly all of the cells present in the wells without the label molecules. By this means, a wide range of noninvasive, label-free cell-based assays can be performed, such as monitoring of cytotoxicity and cell death, stem cell proliferation, RNA interference, virus-mediated cytopathogenicity, etc. [11], [12], [13], [14]. Here, we report the use of the RTCA system as a tool for quantifying 2009 H1N1 virus-induced CPE in Madin-Darby canine kidney (MDCK) cells in real time. As a proof of concept, this format was used to measure neutralizing antibodies in pre- and post-immune serum from 21 healthy individuals who had received the 2009 H1N1 vaccination; the data were then compared with those obtained by hemagglutination inhibition tests.
Materials and Methods
MDCK cells (ATCC, Manassas, VA, USA) were maintained in complete DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin (Invitrogen, Carlsbad, CA, USA) at 37°C with 5% CO2. For the virus culture, serum free DMEM (SFM) supplemented with antibiotics and TPCK-trypsin (Sigma, St. Louis, MO, USA) was used.
The following influenza virus strains were used in this study: A/Shanghai/37T/2009 (H1N1) (SH37T) (GenBank Accession No. GQ253489–GQ253496) and A/Shanghai/143T/2009 (H1N1) (SH143T) (GenBank Accession No. GQ340061–GQ340064, GQ411905–GQ411907 and GQ411909). SH37T and SH143T were originally isolated from a 30-year-old and a 24-year-old male patient hospitalized respectively in late-May and early-June 2009 at Shanghai Public Health Clinical Center (SHAPHC), China. The two patients were both imported cases which respectively from Australia and France, presenting with fever, rhinorrhea, sore throat and cough. Viruses were grown and titrated in MDCK cells at 35°C with 5% CO2.
Human sera
The 2009 influenza A (H1N1) vaccine (inactivated split virion) was produced by the Shanghai Institute of Biological Products, contained the WHO recommended vaccine virus A/California/07/2009 NYMC X-179A, and was issued by the U.S. Centre for Disease Control and Prevention [15], [16]. The vaccine was administered intramuscularly in the upper arm. A total of 21 pre- and post-vaccination serum triples were collected from healthy adult subjects. Pre-vaccination sera were obtained before vaccination (S0 samples). Post-vaccination sera were obtained 7 and 21 days later after vaccination (S1 and S2 samples, respectively). Serum samples were stored at −80°C until required and were heat-inactivated at 56°C for 30 minutes before use.
Ethics Statement
Written informed consent was obtained from all subjects. The subjects all confirmed that they understood the study procedures. The overall study was reviewed and approved by the Ethics Committee of SHAPHC.
Real-time PCR
Viral nucleic acid was extracted using a High Pure Viral Nucleic Acid Kit (Roche Applied Sciences, Indianapolis, IN, USA) according to the manufacturer's instructions. Purified viral nucleic acid templates were amplified with real-time PCR using the Diagnostic Kit for 2009 Influenza A/H1N1 Virus RNA (KHB, Shanghai, China) in a Mastercycler ep realplex 2.2 real-time PCR system (Eppendorf, Hamburg, Germany) according to the manufacturer's instructions.
Cell proliferation assays
The RTCA system (xCELLigence; Roche Applied Sciences, Indianapolis, IN, USA) is comprised of four components: an electronic sensor analyzer, a device station, E-Plates (E-Plate 96), and a control unit with applied software. The device station was located inside a tissue culture incubator and could be switched to any one of the wells of the E-Plate for impedance measurement using the electronic sensor analyzer. Under control of the control unit software, the CI values are then derived from the electronic impedances and graphically represented.
For real-time cell proliferation assays, 50 µL of cell culture media was added to each well of a 96-well E-Plate to obtain background readings. Serial two-fold MDCK dilutions from 4.0×104 to 2.5×103 cells/well in 100 µL of media were then seeded into the E-Plate. The E-Plates containing cells were then incubated for 30 minutes at room temperature and placed on the RTCA SP/MP station located in a cell culture incubator. The CI values were measured automatically every 15 minutes.
To determine the optimal TPCK-trypsin concentration, a cell proliferation assay in SFM containing TPCK-trypsin was performed. When the cells had reached confluence, the E-Plate was removed from the RTCA station and washed twice with SFM. A total of 150 µL of SFM containing TPCK-trypsin at a final concentration of 10–0.31 µg/mL was added. The E-Plate was immediately placed back in the station and CI measurements were subsequently resumed.
Real-time monitoring of virus-induced cytopathogenicity
A total of 1.0×104 MDCK cells/well was seeded into each well of an E-Plate. One of two H1N1 strains, SH37T or SH143T, was 10-fold diluted (10−1 to 10−6) in SFM containing 2.5 µg/mL TPCK-trypsin. Both strains were quantitated by real-time PCR and adjusted to an identical viral load of 5.50×108 copies/mL before dilution. When the CI reached stationary phase after seeding, the cells were washed with SFM and then infected with 150 µL of the virus series. Wells were mock infected with only medium as negative controls. After reloading the E-Plate onto the station, the CI values were measured every 15 minutes for up to 190 hours. Additionally, a colorimetric assay using the cell proliferation reagent WST-1 (Water soluble tetrazolium salts) (Roche Applied Sciences, Indianapolis, IN, USA) was also performed to determine H1N1-induced CPE and titrate the virus as described previously [6], [17]. The assay was based on the cleavage of tetrazolium to formazan salts by mitochondrial dehydrogenases of metabolically active cells, resulting in the color change of the well. Thus the formed formaza dye was quantified spectrophotometrically and correlated directly to the cell viability. The ratio of viral load per 50% tissue culture infectious dose (TCID50) was calculated to determine the number of units required to produce the specific endpoint [18].
Neutralization test (NT) based on the RTCA system
Dilutions of the H1N1 viruses and the test sera were prepared in SFM with TPCK-trypsin to a final concentration of 2.5 µg/mL. The test sera, serially two-fold diluted starting from a dilution of 1∶10, were mixed with an equal volume of medium containing approximately 100 TCID50 of the H1N1 virus. After incubation at 35°C for 60 min, 150 µL of the serum-virus mixture series was transferred to triplicate monolayers of MDCK cells in the E-Plates. Wells containing virus dilutions in the absence of test sera and wells containing only MDCK cells in SFM were included on each assay plate as controls. H1N1-induced CPE was then monitored using the RTCA system (as described above) over 72 hours. The neutralizing titer was defined as the reciprocal of the lowest concentration of serum at which the infectivity of 100 TCID50 of H1N1 was completely neutralized in all the MDCK wells.
Hemagglutination inhibition (HI) test
The HI test was performed according to standard procedures [19]. The test sera were treated with receptor destroying enzyme (RDE) (Sigma, St. Louis, MO, USA) to inactivate nonspecific inhibitors. The sera were then two-fold diluted starting from a dilution of 1∶10. A 1.5% suspension of guinea pig erythrocytes in PBS and four hemagglutinating units of influenza antigen were used.
Statistical analysis
Statistical analysis was performed using SPSS v17.0 (SPSS, Chicago, IL, USA) and GraphPad Prism v5.0 (GraphPad Software, La Jolla, CA, USA) software. Geometric mean titers (GMTs) were calculated by assigning a titer of 5 to the samples HI and NT titers below the lower limit of detection (titer <10). The Wilcoxon signed rank test was used to calculate p values when comparing GMTs between paired data. p values of less than 0.05 were regarded as statistically significant. Spearman's correlation coefficient was used to calculate the correlation between the NT and HI test results. R values of more than 0.5 were regarded as closely related.
Optimization of MDCK cell numbers and trypsin concentration
To identify the most suitable MDCK cell numbers, a series of MDCK cell proliferation curves were performed (Figure 1A). Then the level of 1.0×104 cells/well was selected, for which reached the highest CI value during the grow process. As the cells were in the growth phase or the early stationary phase at 16 to 24 hours after seeding, this period of time was suitable for virus infection.
MDCK cells were seeded in the E-Plates and continuously monitored by measuring the CI to identify the appropriate cell numbers, TPCK-trypsin concentration, and the time point for addition of virus (i.e., during growth or early stationary phase). A) Proliferation curves of MDCK cells in cell culture media. B) Proliferation curves of MDCK cells in SFM with TPCK-trypsin. At the indicated time point the cells were inoculated with different concentrations of virus and TPCK-trypsin.
Since infection of the influenza virus into MDCK cells in vitro is promoted by the addition of trypsin to the medium, the TPCK-trypsin concentration should be optimized. The cell proliferation of 1.0×104 MDCK cells/well under different doses of TPCK-trypsin (Figure 1B) was performed, and the highest TPCK-trypsin concentration that minimally influenced MDCK cell proliferation was determined to be 2.5 µg/mL.
2009 H1N1 virus cytopathogenicity profile
Based on the dynamic monitoring of cell proliferation, 1.0×104 MDCK cells in stationary phase were infected with SH37T or SH143T at the same viral load of 5.50×108 copies/mL after seeding. In the meantime, the presence of CPE was visualized by microscopic examination (Figure 2A). A decline in CI reflected cell death as a consequence of H1N1 virus replication (Figure 2B), while mock-infected cells continued to grow. Parallel WST-1 colorimetric assays were performed to ascertain whether the CIs correlate with CPE. As shown in Figure 2C, the absorbance values representing the viable cell population were decreased due to CPE. These results demonstrated the accurate monitoring of H1N1-induced CPE using the RTCA system. With the virus load decreased an order of magnitude, the initial CPE was delayed by about 8 hours, illustrating the clear correlation between the onset of CPE and the infectious virus load.
A) The CPE on MDCK cells infected with SH37T (1∶10 diluted) at 24, 48, and 72 hours post-infection. B) Real-time monitoring of the CPE on MDCK cells infected with SH37T using the RTCA system. The curve was an average of four independent replicate wells. At the indicated time point the cells were inoculated with different dilutions of virus. C) The colorimetric assays for the CPE on MDCK cells infected with SH37T (1∶10 diluted). OD values at A450 obtained at 24, 48, and 72 hours post-infection are shown. Error bars indicate standard deviation. D) Difference in CPE kinetic patterns between SH37T and SH143T with the same infectious vial load of 5.50×107 copies/mL. The plotted lines indicate the different time points when the CI declined to 0 between stains.
Interestingly, although using the same viral load, cells infected with the different viral strains exhibited different onset times for CPE and complete cell death. For example, for the 10−1 dilutions, complete death occurred at 37 hours post-infection for SH37T-infected cells and at 48 hours for SH143T-infected cells (Figure 2D). As for the colorimetric TCID50 test, the ratio of viral load per TCID50 of SH37T was less than that of SH143T (Table 1), which confirmed the results of the RTCA tests. This demonstrated that the RTCA test could not only quantify H1N1 viruses-induced CPE, but it also reflected the different infectivity of the distinct strains.
Real-time neutralization test
Since H1N1-induced CPE was accurately quantified using the RTCA system, we hypothesized that blocking or delaying CPE by specific neutralizing antibodies from sera could be quantitated using the same method. To explore this concept, the neutralization tests were performed for 21 sera triples using the RTCA System. Neutralization titers were reported as the reciprocal of the highest serum dilution that inhibited virus growth by 100%. MDCK cells on E-Plates were inoculated with serially diluted sera triples that were pre-incubated with SH37T. Initial serum dilutions of 1∶40 for S0, 1∶80 for S1, and 1∶160 for S2 were chosen because at higher serum concentrations virus replication in MDCK cells was clearly inhibited. A total of 100 TCID50 of virus per assay well was achieved to reach maximal cell death. H1N1-induced CPE was monitored in real-time using the RTCA system. The typical results are illustrated in Figure 3. The CI of S0-treated cells declined to 0 entirely, and the resulting curves appeared similar to virus controls (Figure 3A), indicated that the S0 dilution above 20-fold could not inhibit virus growth. However, for the S1 and S2 samples, cell death was not observed until the serum was diluted to 320- and 1280-fold, respectively, and the resulting curves appeared similar to control cells (Figure 3B, C). This indicated that the neutralizing antibody titers in S1 and S2 were 160 and 640, respectively. Interestingly, the average curve of 1280-fold diluted S2-treated cells exhibited difference with virus control (Figure 3C), as the three replicate wells showed distinctly different CI curves (Figure 3D); one started to decline at about 90 hours, whereas the other two had already declined to 0 by this time point. Among the three wells treated with 1280-fold diluted S2, virus in two wells was neutralized completely but the rest one was not. So the finally showed average curve actually resulted from a partial neutralization pattern which was different from those of virus control and complete neutralization wells.
CPE kinetic patterns of MDCK cells inoculated with 100 TCID50 virus incubated with a series dilution of sera samples. Antibody titers were calculated as a function of CI with a sequential dilution series of S0 (A), S1 (B), and S2 (C). The curve was an average of three independent replicate wells. At the indicated time point the cells were inoculated with the serial serum-virus mixture. Three replicate wells of 1280-fold diluted S2 treated cells showed different curves (D). Cell control: MDCK cells only; virus control: absence of test sera.
Comparison of the real-time NT and HI test
A total of 21 pre- and post-vaccination sera triples were collected and tested by real-time NT and HI tests (Table S1). Geometric mean pre- and post-vaccination titers (GMT), as determined by NT and HI tests, and the number of serum pairs exhibiting a significant increase in antibody titers (≥4-fold) were calculated (Table 2). The mean NT titer increase was 4-fold (range 1–16) at day 7 and 13-fold (range 2–64) at day 21 after vaccination, while the mean HI titer increase was 4-fold (range 1–16) and 9-fold (range 2–64). The rise in antibody levels between the pre- and post-vaccination sera is illustrated in Figure 4. Almost all of the 21 subjects exhibited significantly increased antibody titers to the H1N1 virus after vaccination, as determined by both the NT and HI test. When the pre- and post-vaccination titers obtained by HI or NT were compared to each other, there was a statistically significant difference (p<0.05, Wilcoxon signed rank test) (Figure 4A, B). The real-time NT and HI antibody titers showed a close correlation (Spearman's correlation coefficient, r = 0.78, p<0.01; Figure 4C). Furthermore, the magnitude of the antibody response to the vaccine was calculated as the maximum difference between the titer at day 21 and the baseline titer (day 0) for both the NT and HI test (Figure 4D). There was no difference in the mean antibody response measured by NT versus the HI test (p>0.05, Wilcoxon signed rank test). As a consequence, the NT assessed using the RTCA system was in good agreement with the HI test. This indicated that the RTCA system could be used to accurately quantify antibody titers.
A, B) There was a statistically significant difference (p<0.01, Wilcoxon signed rank test) when the pre- and post-vaccination titers obtained by real-time NT (A) or HI test (B) were compared to each other. C) The NT and HI antibody titers correlated well (Spearman's correlation r = 0.78, p<0.01). D) Antibody titers on day 21 after vaccination relative to baseline (day 0). There was no significant difference (p>0.05, Wilcoxon signed rank test) in the results among the NT versus HI tests. Low responders: no increase above twofold; high responders: fourfold or more above baseline.
With the emergence of the 2009 influenza pandemic, development of an effective vaccine became critical for the prevention and clearance of influenza virus infections. To evaluate seasonal or pandemic candidate vaccines, a sensitive, specific, and reproducible serological method is necessary to measure the antibody response elicited with a candidate vaccine against wild-type viruses. In this study, we developed a novel real-time method for measuring virus-induced CPE and evaluating the efficacy of neutralizing antibodies using electronic impedance biosensor-based RTCA technology. The RTCA system is based on integrated microelectronic sensor arrays, is non-invasive, and provides kinetic data regarding the dynamic nature of the cellular response to certain challenges, such as drug treatments [20], growth factor stimulation [21], analysis of cytotoxic composite components [22], and virus-induced cytopathogenesis [14].
In this report, this new assay was used to monitor cytolysis induced by H1N1 viral proliferation. Virus-induced CPE kinetic curves could be used to differentiate between different infectious amounts of the same strain as well as to distinguish the infection kinetics of different H1N1 strains. The two strains, SH37T and SH143T, were isolated respectively from two hospitalized patients in SHAPHC received diagnostic of 2009 H1N1 infection at May 2009. While the clinical patterns were essentially similar, these strains revealed different kinetics of viral infection. The time points for both the onset of CPE and complete cell death of SH143T-infected cells were delayed about 10 hours compared with those of SH37T-infected cells. Because an identical viral load was used, SH37T appeared to have a greater infectivity than SH143T. Therefore, the RTCA system can provide an indicator of virus capacity of replication and infectivity on MDCK cell culture. It was deserved to utilize complementary genetic approaches to define the role of specific viral genes and proteins underlying the phenotypes. Using real-time analysis, Ying Fang et al. [14] generated a mathematical model simulating the CI response to different virus species, through which the virus replication doubling time, virus number threshold for host cell death, and average time to cell death was determined. Meanwhile, the RTCA assay could directly distinguish different strains of the same virus on the basis of our results. For quantitation of virulence, it is necessary to compare different viral stains, determine the number of infectious units required to produce the specific endpoint [18]. Taken together, it is indicated that the RTCA system may provide a feasible in vitro format for assessment of virulence by monitoring CPE kinetics in future investigations of the influenza virus. It may further help to elucidate the relationship between genetic alterations and virulent variations for virological surveillance of influenza.
Since virus-induced CPE could be quantitatively monitored using the RTCA system, we also performed a real-time neutralization assay for measuring H1N1-specific antibodies in human sera using the RTCA system. Since the HI test is still used as a standard for epidemiological and immunological studies, as well as for measuring the efficacy of influenza vaccines and potency of neutralizing antibodies, this new assay was evaluated against the standard HI test on a panel of sera collected from adult donors before and after immunization with the H1N1 vaccine. Both assays showed an obvious increase in neutralizing antibodies in the test sera at 3 weeks post-vaccination, and there was a good agreement between the HI and NT antibody titers. However, higher antibody titers were detected using the RTCA-based NT method, indicating the higher sensitivity of this assay. This may have been due to the difference between the CPE-based and HA-based assays, which had been observed in previous reports [23], [24]. Such functional quantitation may provide a valuable platform for serological diagnosis, immunological study, or evaluation of vaccines for influenza.
An HI titer of 1∶40 is commonly recognized as representing protective immunity. Our results revealed 30% of the HI titers in the pre-vaccination sera were equal to 1∶40, and two were even higher (data not shown). The first H1N1 case was identified in May 2009 in Shanghai, and population-wide vaccination followed in October 2009. Hence, widespread exposure to H1N1 in the society may have caused the higher antibody titers in the donors detected here. In addition, other reports have suggested that serum cross-reactive antibody responses to H1N1 occurred after vaccination with seasonal influenza vaccine and that this also existed in the population during the pre-pandemic period [25], [26], [27].
In many settings influenza is recognized as a major cause of disease and death worldwide, which is the first infectious disease with global surveillance. Not the 2009 Influenza A (H1N1) Virus but other concomitant seasonal or highly pathogenic avian influenza viruses have posed considerable threads to public health. Global surveillance and annual vaccination are both of the key strategies and measures for the prevention and control of influenza. Apart from the evaluation of vaccine immunogenicity, the RTCA assay we described here can be used as a fundamental tool in the virological surveillance of influenza.
Assays with 2009 Influenza A (H1N1) Virus and other seasonal influenza viruses are performed biosafety level-2 (BSL-2), whereas assays with replication competent wide-type H5N1 and other potentially pandemic viruses must be performed under biosafety level-3 (BSL-3) containment, requiring convenient and labor-saving assays to minimize the hands-on steps. Conventional virus quantification and neutralization tests for influenza viruses require several manipulations, which are laborious and rather slow. However, the RTCA assay provides data continuously and automatically in a remote manner after the initial virus inoculation, which could save on time and labor for assay development and also decrease the risk of aerosol generation. With the few hands-on manipulations required, this new format allows for label-free, high throughput, and automatic testing, and could easily be adapted to assays under BSL-3 conditions.
Supporting Information
Table S1.
Antibody titers in 21 pre- and post-vaccination serum triples.
The authors wish to thank Prof. Hongzhou Lu at Department of Infectious Disease, Shanghai Public Health Clinical Center Affiliated to Fudan University, Shanghai, China for sharing the serum samples, MS. Tingting Li at Hangzhou High Throughput Drug Screening Center, Hangzhou ACEA Biosciences, Hangzhou, China for providing technical assistance.
Author Contributions
Conceived and designed the experiments: YH. Performed the experiments: DT WZ YL. Analyzed the data: DT WZ JH MZ. Contributed reagents/materials/analysis tools: MZ JH ZS ZZ. Wrote the paper: WZ DT.
- 1. Zhu FC, Wang H, Fang HH, Yang JG, Lin XJ, et al. (2009) A novel influenza A (H1N1) vaccine in various age groups. N Engl J Med 361: 2414–2423.
- 2. Jiang W, Xu F, Lu L, Lu M, Miao L, et al. (2010) Safety and effectiveness of a 2009 H1N1 vaccine in Beijing. N Engl J Med 363: 2416–2423.
- 3. Liang XF, Wang HQ, Wang JZ, Fang HH, Wu J, et al. (2010) Safety and immunogenicity of 2009 pandemic influenza A H1N1 vaccines in China: a multicentre, double-blind, randomised, placebo-controlled trial. Lancet 375: 56–66.
- 4. Rowe T, Abernathy RA, Hu-Primmer J, Thompson WW, Lu X, et al. (1999) Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays. J Clin Microbiol 37: 937–943.
- 5. Smee DF, Morrison AC, Barnard DL, Sidwell RW (2002) Comparison of colorimetric, fluorometric, and visual methods for determining anti-influenza (H1N1 and H3N2) virus activities and toxicities of compounds. J Virol Methods 106: 71–79.
- 6. Lehtoranta L, Villberg A, Santanen R, Ziegler T (2009) A novel, colorimetric neutralization assay for measuring antibodies to influenza viruses. J Virol Methods 159: 271–276.
- 7. Stephenson I, Das RG, Wood JM, Katz JM (2007) Comparison of neutralising antibody assays for detection of antibody to influenza A/H3N2 viruses: an international collaborative study. Vaccine 25: 4056–4063.
- 8. Tsai C, Caillet C, Hu H, Zhou F, Ding H, et al. (2009) Measurement of neutralizing antibody responses against H5N1 clades in immunized mice and ferrets using pseudotypes expressing influenza hemagglutinin and neuraminidase. Vaccine 27: 6777–6790.
- 9. Atienza JM, Yu N, Kirstein SL, Xi B, Wang X, et al. (2006) Dynamic and Label-Free Cell-Based Assays Using the Real-Time Cell Electronic Sensing System. Assay Drug Dev Technol 4: 597–607.
- 10. Solly K, Wang X, Xu X, Strulovici B, Zheng W (2004) Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. Assay Drug Dev Technol (4): 363–372.
- 11. Huang DY, Mock M, Hagenbuch B, Chan S, Dmitrovic J, et al. (2009) Cytotoxic response to microcystins using microelectronic sensor arrays. Environ Sci Technol 43: 7803–7809.
- 12. Ke Y, Zhang EE, Hagihara K, Wu D, Pang Y, et al. (2007) Deletion of Shp2 in the brain leads to defective proliferation and differentiation in neural stem cells and early postnatal lethality. Mol Cell Biol 27: 6706–6717.
- 13. Arora S, Gonzales IM, Hagelstrom RT, Beaudry C, Choudhary A, et al. (2010) RNAi phenotype profiling of kinases identifies potential therapeutic targets in Ewing's sarcoma. Mol Cancer 2010 18: 218.
- 14. Fang Y, Ye P, Wang X, Xu X, Reisen W (2011) Real-time monitoring of flavivirus induced cytopathogenesis using cell electric impedance technology. J Virol Methods 173: 251–258.
- 15.
Availability of a candidate reassortant vaccine virus for the novel influenza A (H1N1) vaccine development, X-179A. World Health Organization website. Available: http://www.who.int/csr/resources/publications/swineflu/2009_05_27_X179Aa.pdf. Accessed: 2009 May.
- 16.
Pandemic (H1N1) 2009 guidance documents. World Health Organization website. Available: http://www.who.int/csr/resources/publications/swineflu/en/#A.
- 17. Nissen E, Pauli G, Vollenbroich D (1997) WST-1 assay–a simple colorimetric method for virus titration. In Vitro Cell Dev Biol Anim 33: 28–29.
- 18.
Nathanson N (2007) Viral Pathogenesis and Immunity, Second Edition. Publisher: Academic Press. USA.
- 19.
Kendal AP, Pereira MS, Skehel JJ (1982) Concepts and Procedures for Laboratory based Influenza Surveillance. Distributed by the Viral Diseases Unit. Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
- 20. Xing JZ, Zhu L, Jackson JA, Gabos S, Sun XJ, et al. (2005) Dynamic monitoring of cytotoxicity on microelectronic sensors. Chem Res Toxicol 18: 154–161.
- 21. Hahon N, Booth JA, Eckert HI (1972) Further studies on the anti-immunoglobulin G hemagglutination-inhibition test for influenza. Appl Microbiol 23: 485–489.
- 22. Xi B, Yu N, Wang X, Xu X, Abassi Y (2008) The application of cell-based label-free technology in drug discovery. Biotechnol J 3: 484–495.
- 23. Staršíchová A, Kubala L, Lincová E, Pernicová Z, Kozubík A, et al. (2009) Dynamic monitoring of cellular remodeling induced by the transforming growth factor-β1. Biol Proced Online 11: 316–324.
- 24. Hancock K, Veguilla V, Lu X, Zhong W, Butler EN, et al. (2009) Cross-reactive antibody responses to the 2009 pandemic H1N1 influenza virus. N Engl J Med 361: 1945–1952.
- 25. Boon ACM, deBeauchamp J, Krauss S, Rubrum A, Webb AD, et al. (2010) Cross-reactive neutralizing antibodies directed against pandemic H1N1 2009 virus are protective in a highly sensitive DBA/2 mouse influenza model. J Virol 84: 7662–7667.
- 26. Qiu C, Tian D, Wan Y, Zhang W, Qiu C, et al. (2011) Early Adaptive Humoral Immune Responses and Virus Clearance in Humans Recently Infected with Pandemic 2009 H1N1 Influenza Virus. PloS one 6: e22603.
- 27. Qiu C, Huang Y, Wang Q, Tian D, Zhang W, et al. (2012) Boosting Heterosubtypic Neutralization Antibodies in Recipients of 2009 Pandemic H1N1 Influenza Vaccine. Clin Infect Dis 54: 17–24.


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Original image properties:
Camera make: NIKON CORPORATION
Camera model: NIKON D90
Image size: 4288x2848 (12.2 megapixels)
Capture time: 2011-07-15 16:17:57 - 2011-07-15 16:46:31
Aperture: f/29
Exposure time: 0.0125
Focal length (35mm equiv.): 450.0 mm
Digital zoom: off
White balance: Automatic
Exposure mode: Manual
Horizontal overlap: 22.3 to 38.7 percent
Vertical overlap: 23.0 to 55.1 percent
Computer stats: 4094.18 MB RAM, 4 CPUs
Total time 49:45 (6.9 seconds per picture)
Alignment: 19:28, Projection: 7:25, Blending: 22:52
(Preview finished in 31:50)


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4446 Glenwood Drive,Perkiomenville PA, 18074
Sold Price
3 beds , 1 Full/1 Half baths
1,692 sq. ft
Status: Sold
Berkshire Hathaway HomeServices Fox & Roach, REALTORS
(866) 486-5636
Last Updated: 2/25/2017 12:06 AM
Description for 4446 Glenwood Drive Perkiomenville, PA 18074
Welcome to this stunning end unit in Perkiomen Crossing. Back on the market due to buyer remorse. Not a thing wrong with the house. In fact, it's in stellar condition! Looking for something you can "just move into"? Just unpack your bags and enjoy! Prepare to be wowed as you enter through the living room adorned by gorgeous scratch resistant cherry Pergo flooring (2016) that extends into the kitchen. The living room also has a bump out window, crown molding, coat closet and built-in wall shelf. Preparing meals will be a pleasure in the updated oak kitchen offering solid surface counters, smooth top convection cooking, microwave, dishwasher, garbage disposal, double sink, pantry and a new French door slider opening to an expansive deck. A powder room completes the first floor. The second floor offers a master bedroom with walk-in closet with a cherry organizational system. There are two additional nice sized bedrooms with plenty of closet space. For your comfort, all bedrooms have ceiling fans. Enjoy the updated bath with newer everything (sink, toilet, flooring, lighting & a gorgeous tiled shower/bath.) Access to the attic for the potential of added storage space. For added living space, the lower level has a spacious family room, plenty of storage and a laundry area. As an added bonus, this home has new roman shades, windows, flooring and a French door slider. The HVAC was replaced in 2012, as well as the carpeting. The bath was completely updated within the last 5 years, kitchen updated, as well as storm & front doors, and updated lighting and appliances replaced during ownership. Ready to entertain this summer or maybe just want a place for some peace and serenity? This private lot backs up to woods and common space. Tons of mature landscaping and plenty of storage under the deck. Absolutely one of best locations in Perkiomen Crossing! If that's not enough, there's more ? maintenance free living at its best with a low HOA fee of only $75, which includes lawn care, snow & trash removal and common area maintenance. USDA approved. Easy access to Routes 73 and 29, Plymouth Meeting & Lansdale Turnpike. Don't forget to view the virtual tour of this beautiful home.
Listing Information
Perkiomen Crossing
Electric Water Heater, Public Water Service
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Hookups in Basement
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Vinyl, Carpet, Wood
Central Air, Ceiling Fan(s)
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Current market conditions for 4446 Glenwood Drive
Active Market Trends
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Median list price$407,400
Days on market116
Properties for sale20
Sold Market Trends
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Median sold price$310,000
Sales to list price9,600.0%
Properties sold11
Median sq ft price$127
Arrows indicate 90 day trend
Listing Price History for 4446 Glenwood Drive
Perkiomenville Real Estate
Perkiomenville is located in Pennsylvania. Perkiomenville, Pennsylvania 18074 has a population of 6,108.
The median household income in Perkiomenville, Pennsylvania 18074 is $85,943. The median household income for the surrounding county is $78,984 compared to the national median of $53,046. The median age of people living in Perkiomenville 18074 is 41.6 years.
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Dug Begley is the transportation writer for the Houston Chronicle.
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Working against the clock, crews repair giant hole on I-45 in time for evening rush
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Texas lawmakers move to stymie high-speed rail project
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Houston falls in new congestion ranking
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Great Westcott! Sign misspells street name
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Metro drawing up long-term Houston regional transit plan
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TxDOT's long road to centennial winds to Houston
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Town Square
Mustang players kick off season with a third place finish
Original post made on Aug 16, 2010
Read the full story here Web Link posted Tuesday, August 10, 2010, 3:56 PM
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a resident of Danville
on Aug 16, 2010 at 3:26 pm
Congratulations girls!! Way to go!
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Our wallets will feel impact of Oroville dam repairs
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Russian art has a long and storied history. Throughout the years there have been several distinct periods of artistic expression and with each new era came many talented and memorable Russian artists. Russian art has always been a representation of its ancient culture, and there have been many different varieties of Russian artists, including musicians, painters, sculptors, and actors.
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Talk:German Spitz
|WikiProject Dogs / Breeds
|(Rated Start-class, Mid-importance)
|WikiProject Germany
|(Rated Start-class, Low-importance)
Eliminate separate breed pages for sizes[edit]
- The following discussion is closed. Please do not modify it. Subsequent comments should be made in a new section. A summary of the conclusions reached follows.
- The result was do not merge -- AmeIiorate (talk · contribs) 19:56, 18 August 2008 (UTC)
I'm wondering whether, since the breed stds seem to be the same for the 3 sizes in several kennel clubs, this article should just be the one article, like Dachshund is for the various types? Whatchu guys think? Elf | Talk 18:36, 31 January 2006 (UTC)
- I agree with this 2 year old comment. I will place the proposal tags in a moment. ~ Ameliorate U T C @ 00:13, 12 July 2008 (UTC)
- I agree they should be merged. Vortex (talk) 06:06, 12 July 2008 (UTC)
I disagree: I think the dispite being the same breed the differences between the categories results from the categories being quite different both in behaviour and temperament. Take the logic to its limit and all dogs could be put on one page. I belive each of the categories is different enough to warrent their own page. Having a holding page and breaking out to the different sizes makes for a better layout and highlights the differences between the categories.
- My reasoning for the merge is that the German Spitz article itself is sort of a pseudo-disambiguation page, when I feel it would be better to have one "good" article mentioning all the breed variations than 5 shorter articles. ~ Ameliorate U T C @ 21:28, 14 July 2008 (UTC)
- There are actually two article topics here (at least.) According to the article, the German Spitzen is both a breed and a breed grouping in the spitz type. Some of the German Spitz are recognized as separate (from the heading or type of German Spitz) breeds, depending on the registry. The five recognized German Spitz are not the only German Spitz-types; the American Eskimo and the Japanese Spitz are bred from German Spitz, possibly other breeds as well.
"Unter der Rassebezeichnung Deutscher Spitz sind der Wolfsspitz, der Groß-, Mittel-, Klein- und Zwergspitz..."
- If the Zwergspitz and the Wolfsspitz have their own pages separate from "German Spitz" (as well as all the other variants and descendants of the German Spitzen, such as the American Eskimo and Japanese Spitz), why not let the the Großspitz, Mittelspitz, and Kleinspitz have their own pages? Keep separate articles for the separate breeds and use the German Spitz article to explain how they are all related.--Hafwyn (talk) 16:25, 18 August 2008 (UTC)
- The above discussion is closed. Please do not modify it. Subsequent comments should be made in a new section.
Redirections to other languages[edit]
The redirections to other languages are a bit in disarray, I'm afraid. We have:
- Spitz type
- German spitz
- Keeshond
- Pomeranian
The first is a very general dog type represented by many breeds. The last three however represent in FCI terms just 1 breed, the Deutscher Spitz (FCI standard 97). Actually standard 97 recognizes no less than 10 varieties! (see: http://www.fci.be/nomenclatures_detail.asp?lang=en&file=group5#section4).
The Keeshond is the variety called Wolfsspitz in Germany, the Pomeranian is know there as Kleinspitz.
In Holland the German spitz is known as Keeshond. So there is a difference between the English use of the word Keeshond and the Dutch original.
--Cuon 10:05, 29 January 2007 (UTC)
Please, Do not merge the pages of the German Spitz and the American Eskimo Dogs! The American Eskimo Dog may be a deriviate of the German Spitz but they are two distinct breeds and recognized as such within AKC!!!!
Lynn McClure 18:10, 29 September 2007 (UTC)
- This is really confusing. The Pomeranian under the Fédération Cynologique Internationale is breed designation of German Spitz, as the Zwergspitz. Under the Canadian Kennel Club (and others) it is a separate breed in the Toy group. So it would not be right to only have the Pomeranian on the German Spitz page. The same applies to the Keeshond; under the Fédération Cynologique Internationale it is a Geman Spitz breed, the Wolfsspitz, but has been listed by the Kennel Club (UK) as a separate breed since 1926. The Keeshond should not be merged into the German Spitz page either. The "American Eskimo" breed apparently were originally white German Spitz, but are now accepted as a separate breed (also in three sizes) by the major North American breed registries. So where should they be listed? In addition, the Japanese Spitz (Nihon Supittsu) is a white German Spitz that is now accepted as a separate breed by the Fédération Cynologique Internationale. The Volpino Italiano looks almost identical to other white "German Spitz" types. The Indian Spitz may or may not be a variant of the Pomeranian (German Spitz - Zwergspitz.) All of these deserve to have their own breed articles.--Hafwyn (talk) 15:58, 18 August 2008 (UTC)
The etymology of the term "wolfspitz" derives from the conjunction of "wolf" [self explanatory] and "spiti" [greek "home" or "house"]. Many of the endearing characteristics of the breed can be found in their behaviour as a "house wolf".Far fetched. Pomeranian do got it right. Spiz means pointed in german, referring to nose and muzzle.--Tresckow (talk) 01:13, 16 February 2010 (UTC)
- Seems fair enough: I've deleted that claim. Remember, you can edit Wikipedia too! ~dom Kaos~ (talk) 11:10, 6 May 2010 (UTC)
The source for the claim that "German Spitz are descendants of the ancient spitzen found in the Stone Age and they are the oldest breed of dog in Central Europe" is no longer live, so I've had to delete it and add a {citation needed} tag: if anyone has a current link to a reputable source for this, it would really improve the article ~dom Kaos~ (talk)
- I added a citation to a genome sequencing paper that completely contradicts the claim of ancient lineage.Dogface (talk) 15:39, 27 May 2010 (UTC)
- I just linked the word spitzen so that it would take the reader to the article where it's cited. Chrisrus (talk) 19:35, 16 September 2011 (UTC)
Wolfspitz, Disambiguation?[edit]
The link in this article wolfspitz takes the reader to the Keeshound, but if you see where it appears in context within this article, it seems to be referring to the oldest fossil dogs in Eastern Europe/Western Asia which are among the oldest dog skeletons that we know. It is said that these skeletons looked like either a spitz or across between a wolf and a spitz. Please go read the origin section of Spitz, which is where I am getting this from, and also please look at the references for that section. The experts who examined those dogs, especially the ones that were a bit wolfier than spitzish, as wolf/spitz, a transitional animal. I think we have two different referents here and there either needs to be a bit of disambiguation. Or, and this might be a better option, it would be best to use a different word in this article or not to link the word "wolfspitz". Chrisrus (talk) 19:30, 16 September 2011 (UTC)


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text: taxes Kansas House Preparing To Debate Proposals To Raise Taxes By Associated Press • May 29, 2015 Kansas House members are preparing to debate proposals for raising sales, tobacco and business taxes to close a projected budget shortfall. Republican leaders in the GOP-dominated chamber put three tax bills on Friday's agenda. Top Republicans hope the House passes at least one. Lawmakers must close a $406 million budget shortfall for the fiscal year beginning July 1. The gap arose after they cut income taxes in 2012 and 2013 at Republican Gov. Sam Brownback's urging to stimulate the economy. One key issue is increasing the state's 6.15 percent sales tax. Kansas House Members Block Tax Debate By Stephen Koranda • May 28, 2015 The Kansas House had been planning to debate a tax bill Thursday, but dissatisfied members of the chamber were able to stop the debate before it even started. The bill failed to get the two-thirds vote needed to be brought up for debate. It appears conservative Republicans and a small number of Democrats were able to block the discussion. House Republican Majority Leader Jene Vickrey says the bill will be back on the agenda Friday. Tax Plan Fails In Kansas Senate After Hours Of Debate By Stephen Koranda • May 28, 2015 Stephen Koranda / KPR/File photo The Kansas Senate spent hours yesterday debating a tax plan and eventually there was agreement: Virtually everyone in the chamber agreed that they did not like the bill. The plan failed with 30 votes against it and only one in favor. KPR's Stephen Koranda reports on the proposal, which would have raised a variety of taxes to fill a budget hole. Kansas Senate Begins Debate On Tax Bill By Stephen Koranda • May 27, 2015 The Kansas Senate has spent today debating a bill that would raise taxes to close a projected budget shortfall. The work started with Republican Sen. Dennis Pyle moving to kill the tax bill, saying increasing taxes is not the way to fix the state's budget. He wants lawmakers to look for more budget cuts. "By not raising taxes, you're going to empower the private sector," Pyle says. "Or are you going to empower big government, are you going to empower more government consumption?" Kansas Lawmakers Back In Topeka To Wrap Up Tax And Budget Issues By Stephen Koranda • May 26, 2015 Stephen Koranda / KPR/File photo Kansas lawmakers return to Topeka Tuesday after a four-day weekend. As Stephen Koranda reports, they'll have to wrap up tax and budget issues before they can end the session. Lawmakers can't leave until they balance the budget. They seem to be leaning towards a tax increase following tax cuts in recent years, but there's still division among lawmakers over what kinds of taxes should be increased. The Senate will take up their first tax proposal this week, after abruptly canceling a debate on a bill last week and then leaving town for the holiday. Gov. Brownback Says He's Not Drawing Clear Lines On Taxes By Associated Press • May 21, 2015 Governor Sam Brownback says he will not draw clear lines on what he'll accept from the Kansas Legislature, which is considering rolling back one of his major economic initiatives. Lawmakers are working to close a projected budget deficit of $406 million dollars for the next fiscal year, which begins July 1st. The House Taxation Committee was working on a plan to raise the state's sales tax to 6.5 percent from 6.15 percent. Kansas Governor, Leaders Meet Amid Work On Tax Proposals By Associated Press • May 19, 2015 Stephen Koranda / KPR/File Photo Republican Gov. Sam Brownback has met with top GOP leaders as the Kansas Legislature continues its discussions about raising taxes to erase a state budget shortfall. Brownback had a private session Tuesday morning with Senate President Susan Wagle, House Speaker Ray Merrick and the majority leaders in both chambers. Their gathering came before the Senate tax committee convened to discuss revenue-raising proposals. Kansas House Lawmakers Back To Work On Taxes By Stephen Koranda • May 18, 2015 Stephen Koranda / KPR/File photo House lawmakers are going back to the drawing board on Monday after the chamber roundly rejected a tax proposal last week. The bill they rejected would have mostly relied on a sales tax increase to fill a budget hole of more than $400 million. Statehouse reporter Stephen Koranda explains... Kansas House Panel To Debate Proposals For Raising Taxes By Associated Press • May 11, 2015 Stephen Koranda A Kansas House committee is preparing to debate proposals for raising taxes to erase the budget shortfall. The Taxation Committee plans to discuss various revenue-raising proposals today. Legislative researchers say lawmakers must eliminate a $422 million deficit in the state budget for the fiscal year that begins July 1. Budget problems arose after the Legislature aggressively cut personal income taxes at Gov. Sam Brownback's urging in 2012 and 2013 in an effort to stimulate the economy. Urban/Rural Divide Mucking Up The Works In Kansas Senate Tax Committee By Stephen Koranda • May 11, 2015 Stephen Koranda / Kansas Public Radio/File photo A divide between rural and urban lawmakers appears to be holding up work in the Senate's tax committee. The panel has had two meetings where there was little agreement on tax plans. As Stephen Koranda reports, lawmakers are looking for ways to fill a budget gap of more than $400 million. Senator Jeff Melcher is a Republican from Leawood in Johnson County. He says the Johnson County lawmakers on the tax committee will not support any major tax plans until another issue is settled. Pages« first ‹ previous … 4 5 6 7 8 9 10 11 12 … next › last »


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NEW YORK (AP) - Braves outfielder Jason Heyward sustained two jaw fractures when he was struck by a pitch and will undergo surgery on Thursday.
Heyward was beaned by a 90 mph fastball from New York Mets left-hander Jonathon Niese in the sixth inning of the Braves' 4-1, 10-inning win on Wednesday. The impact made a loud noise, and the crowd groaned as Heyward crumpled to the ground.
He stayed down on his back for about three minutes before getting helped to his feet. Heyward walked slowly off the field under his own power, escorted by a trainer, and was replaced by Jordan Schafer.
Visibly concerned, Niese tapped himself with his glove as Heyward walked off, and Heyward appeared to acknowledge him.
An inning later, the Mets announced that Heyward was examined by their doctors before he was taken to the hospital for X-rays.
On a tear at the plate since moving into the leadoff spot last month, Heyward began the day batting .254 with 13 homers and 37 RBIs. Atlanta entered with a 15-game lead in the NL East.
Copyright 2017 by STATS LLC and Associated Press.
Any commercial use or distribution without the express written
consent of STATS LLC and Associated Press is strictly prohibited.


text:
Endlessly Adorable - Scene 4
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DVD Title Endlessly Adorable
Category Twinks
Added November 02, 2010
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Duration 25m, 48s
Tags Anal Licking, Anal Sex, Blowjobs, Cum on Hands, Cum on Stomach, Fingering, Handjobs, Hardcore
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The gay stud in the dorm has got his sights set on a geeky guy with a curly hair. At first, the guys just relax on the couch together, and then the stud leans over to give his friend's cock a little rub. The geek looks around, worried that someone will find them in the middle of a homosexual cocksucking experience, but the stud pushes ahead, overcoming his buddy's objections with his hot lips and tongue. These two queers end up sucking each other off on the couch, and then the geeky guy bends over, willingly pulling his ass cheeks apart in order to get sodomised. He moans while his virgin anus gets torn apart by his friend's thrusting.
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Endlessly Adorable
November 02 2010
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- Scene 4 - Geeky Guy Has His First Anal Experience
- Scene 1 - Man Falls In Love With Sweet Brown Lover
- Scene 2 - Horny Gay Hunks in Romantic Fuck
- Scene 3 - Big Dick Euro Guys Getting Gay
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text:
Catalogue Number: VA4-VB35
- Author: Edward Little,Anthony Batchelor,Gareth Davies
- Format:Paperback
- Publisher:Pearson Education Limited
- Published:14 July 2008
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Product description
- Total Pages: 160
- Genre: Interdisciplinary studies
Edexcel's AS Student Book is an ideal companion to help students succeed in this qualification. * Provides complete coverage of the revised AS specification. * Helps students understand the requirements of the exam. *.Includes an examzone section, featuring worked exam questions with examiners' comments that help students understand the standards they will need to meet and really help to raise attainment.


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Scientists dismiss Paltrow's shampoo cancer claims
"A couple of years ago I was asked to give a quote for a book concerning environmental toxins and their effects on our children. While reading up on the subject, I was seized with fear about what the research said," the Sun quoted Paltrow, as saying.
"Foetuses, infants and toddlers are basically unable to metabolize toxins the way that adults are, and we are constantly filling our environments with chemicals that may or may not be safe. The research is troubling; the incidence of diseases in children such as asthma, cancer and autism have shot up exponentially and many children we all know and love have been diagnosed with developmental issues like ADHD," she added.
However, scientists at Cancer Research UK insist that there is absolutely no evidence to back up the actress" findings. And medical chiefs have been quick to wade into the debate, with bacteriologist Professor Hugh Pennington calling her claims 'rubbish" and 'loopy". "It"s a load of nonsense. Shampoo is perfectly safe, unless you drink it in large quantities," he said.