• To exploit opportunities for short-term profitable deployment,
  • Meet maturing liabilities if expected inflows do not materialize and
  • Ensure uninterrupted completion of day-to-day transactions.

Considering these factors, policies are developed for the amount of cash balance to be maintained at all times.

Short-Term Financing: Supplier credit is a significant source of working capital finance. You can buy raw materials and use it for 60 or so days before paying for it. During this period you might be able to produce and sell products, and generate funds to pay the supplier in part. The factoring operations mentioned earlier can generate additional funds. Any remaining shortfall is typically financed with short-term bank loans or lines of credit (where you draw funds only if and when needed).

Cash Flow Forecasts: Cash flow forecasts are prepared to estimate cash outflows and inflows, and identify the timing and extent of shortfalls in funds availability. Based on these estimates, bank finance or other funding options can be arranged in time.

Working capital is distinct from fixed capital in that it is short-term in nature. There is no long-term commitment of funds. As against this, even a short-term liquidity problem can affect the profitability of the business. If the liquidity problem persists, the very survival of the business can be in danger. This is why working capital management has become a critical function.

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Many American BP Gas Stations Want Amoco Name Back

A bunch of BP gas station owners are trying to revert back to the Amoco name, which was bought by BP, hoping to attract customers as some stations have seen a 40 percent drop in sales since the oil spill.

BP stations across the country have been targets of public anger, protests and sign alterations. And that's bad news for the 11,300 BP gas stations in America. Many owners now want to revert back to Amoco, according to the executive director of BP Amoco Marketers Association, John Kleine. He told The Sunday Telegraph, "They are interested in where the brand is going, and want it to be returned to its premier place."

It's still not known if that is even a possibility, but Klein says the issue will probably come up at the distributor's annual meeting in October, which will be attended by top BP executives. But for now, the owners are pleading with the public. One station owner in Alabama, Savita Kakadia two weeks ago told reporters, "BP doesn't own this station, I do."

Send an email to Jeff Neumann, the author of this post, at jeff@gawker.com.

The Worldwatch Institute sang the praises of BP’s it’s-a-problem, we-can-solve-it approach to climate change. Far Left environmentalist Joe Romm featured John Browne/BP in his book Cool Companies as a leading example of corporations going green for profits and virtue.

Both Worldwatch and Romm were wrong—dead wrong—about BP, just as they were also wrong about climate-alarmist Enron and Ken Lay.

It turns out that a lot of political profit-making and greenwashing was going on at both rogue companies. Remember what Jeff Skilling told one of Enron’s coal executives who complained that the company’s greenwashing was hurting his division:

“We are a green company, but the green stands for money.”  - Jeff Skilling, CEO, Enron Corp., quoted in Capitalism at Work, p. 310.

Corporate altruism? A wise man’s suspicion in 1776 still applies. “I have never known much good done by those who affected to trade for the publick good,” wrote Adam Smith. “It is an affectation, indeed, not very common among merchants, and very few words need be employed in dissuading them from it.” (Wealth of Nations, p. 456)

And Enron, as it turned out, had seven profit centers that stood to benefit from government-priced CO2. Altruism—or naked rent-seeking at the expense of broader society? Wake will Left environmentalists smell the skunk?!

“What Every Business Executive Needs to Know About Global Warming” (by Kimberly O’Neill Packard and Forest Reinhardt)

And in the pages of the esteemed Harvard Business Review, a trendy essay cajoling business executives to buy into climate alarmism as a new corporate ethic put BP and John Browne on a high pedestal.

The article begins by assuming the problem and the solution—two fatal flaws. “The [Kyoto] Protocol underscores the growing consensus among scientists that global climate change is a threat that must be taken seriously” (p. 130). The essay warned of “catastrophes” in-the-making were business to continue as usual. Corporations were urged to get on board or face even more punitive and disruptive regulation (such as the Kyoto Protocol).

Enter BP and John Browne—the Great Company and Great Man of the new era of social corporate responsibility. The essay states:

“BP Amoco [BP] has been a leader in supporting international efforts to slow climate change….

CEO John Browne and other BP Amoco executives … believe that taking a leadership position on climate change gives the company a distinctive identity in the eyes of government officials, scientists, and environmental groups. Such leadership may give BP Amoco better access to government-controlled oil deposits and more operating flexibility.

Furthermore, the company’s experiments with emissions trading are likely to give it clout at the negotiating table when international regulatory frameworks are being devised; company executives will be able to present hard data on how their system works.”

And still more benefits!

BP Amoco’s leaders also believe that by announcing the 10% cutback they’ll release the creativity of employees and increase their commitment to the company. ‘Do not underestimate the power of preemptive, aspirational target setting,’ says Chris Gibson-Smith, BP Amoco’s executive director for policy and technology. ‘The role of leadership is to invent actions that naturally have the consequence of transforming people’s thinking.’

In other words, confronting the climate challenge will stimulate the company’s employees—line workers and managers alike—to think more imaginatively. And to the extent that the employees see their values reflected in BP Amoco’s goals, they may become more committed to their jobs and to the company.”

- Kimberly O’Neill Packard and Forest Reinhardt, “What Every Executive Needs to Know About Global Warming,” Harvard Business Review, July/August 2000.

Favoritism for drilling on government land? Greater operational flexibility?  Thinking more imaginatively? Company values?

What if carbon dioxide (CO2) was/is the wrong target—the great distraction?

What if the same effort had been expended on clear-and-present environmental and safety dangers? Where would BP be today? And how much better would our environment and our safety be?

Here is my challenge to Kimberly O’Neill Packard (formerly with McKinsey), Forest Reinhardt (the John D. Black Professor of Business Administration at Harvard Business School), and Harvard Business Review: do you care to revisit this article and issue and provide all of us with an update? Start with climate science, and bring us all the way to BP and Enron. And on the physical science, start with Gerald North, a mainstreamer who has privately spilled some beans on the climate alarmists. You will find this to be very interesting—and perhaps even psychologically therapeutic.

 





PSAB ACTIVITIES 

  • ASM Presents Comments to FDA/CDRH at Public Meeting on Oversight of Laboratory Developed Tests
  • ASM Presents Comments at CMS Meeting on Coding for New Clinical Lab Tests
  • ASM Comments on the WMD Prevention and Preparedness Act of 2010
  • ASM Comments for the Trans Atlantic Task Force on Antimicrobial Resistance

ASM NEWS, JOURNAL ARTICLES AND UPDATES

  • ASM ICAAC Meeting
  • ASM 2011 General Meeting Awards
  • ASM Upcoming Audioconferences
  • ASM Journal Articles of Interest

FEDERAL AGENCY UPDATES

  • CDC Health Advisory on Increased Potential for Dengue Infection in Travelers Returning from International and Selected Domestic Areas
  • MMWR Articles of Interest
  • First Smallpox Vaccine for Special Populations Delivered Under Project BioShield
  • Technology Assessment for Quality, Regulation and Clinical Utility of LDTs
  • NIH Scientists Advance Universal Flu Vaccine
  • National HIV/AIDS Strategy

OTHER INFORMATION AND UPDATES

  • New from CLSI
  • NLTN Online Training Course in Packing and Shipping
  • Articles of Interest

 

ASM Presents Comments to FDA/CDRH at Public Meeting on Oversight of Laboratory Developed Tests

On July 20, the ASM presented comments at a public meeting sponsored by the Food and Drug Administration’s (FDA) Center for Devices and Radiological Health (CDRH) on Oversight of Laboratory Developed Tests (LDTs):  Clinical Lab Challenges.  The purpose of the meeting was to provide a forum where interested stakeholders could present comments regarding reasonable and effective regulation of LDTs.  The meeting took place in Hyattsville, MD on July 19-20, 2010 and Laboratory Practices Committee Member, Paula Revell, represented ASM at the meeting and presented ASM’s comments.  To read ASM’s statement, go to http://www.asm.org/index.php/policy/ldtcomments7-20-10.html

 

ASM Presents Comments at CMS Meeting on Coding for New Clinical Lab Tests

Vickie Baselski, Chair of the Professional Affairs Committee presented ASM comments to the Centers for Medicare & Medicaid Services (CMS) on new clinical laboratory tests that will be included on the 2011 Medicare Clinical Laboratory Fee Schedule.  To read ASM’s comments, go to http://www.asm.org/index.php/policy/cmspaymentcomments-2010.html

 

ASM Comments on the WMD Prevention and Preparedness Act of 2010

ASM sent comments to the House Committee on Homeland Security commenting on H.R. 5498, the WMD Prevention and Preparedness Act of 2010.   To read ASM’s comments, go to http://www.asm.org/index.php/policy/wmdactof2010-comments.html

 

ASM Comments for the Trans Atlantic Task Force on Antimicrobial Resistance

On June 7, the ASM presented comments to the Department of Health and Human Services (HHS) at the Stakeholder Listening Session convened to discuss the work of the Trans-Atlantic Task Force on Antimicrobial Resistance (TATFAR).  The ASM commended the establishment of the TATFAR, provided some preliminary comments on addressing antimicrobial resistance and requested more information about how it could assist the Task Force as it pursues its focus on urgent antimicrobial issues world wide. The TATFAR, which is a government to government task force, was formed following the November 3, 2009 US-European Union Summit Declaration which focused on the specific areas of appropriate therapeutic use of antimicrobial drugs in the medical and veterinary communities, prevention of both healthcare and community associated drug resistant infections and strategies for improving the pipeline of new antimicrobial drugs.

The ASM comments are available at http://www.asm.org/index.php?option=com_content&view=article&id=91489. 


ASM NEWS, JOURNAL ARTICLES AND UPDATES

 

ASM ICAAC Meeting

The 2010 Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC) meeting will be celebrating its 50th year in Boston, MA.  The meeting will be held September 12-15.  For more information, visit the ICAAC website at http://www.icaac.org/

 

ASM 2011 General Meeting Awards

Nominations are being accepted for ASM’s 2011 General Meeting Awards.  The awards honor leadership, research and service.  The nomination deadline is October 1.  The 2011 General Meeting will be held in New Orleans, LA May 21- 24, 2011.  To submit a nomination, go to http://www.asm.org/awards

 

ASM Upcoming Audioconferences

Pertussis in the 21st Century: The Laboratory Diagnostic Challenge Continues

Date: August 11, 2010

Time:  1:00 PM, Eastern Time

Speaker:  Mario Marcon

 

Pros and Cons of Screening Assays in the Diagnosis of Tuberculosis

Date:  August 25, 2010

Time:  1:00 PM, Eastern Time

Speaker:  Rebecca Horvat

 

For more information including registration, go to http://www.asmaudio.org/teleconference.asp#sched

 

ASM Journal Articles of Interest

Quitting Smoking May Minimize Harmful Bacteria and Replenish Health Bacteria

Journal of Clinical Microbiology, July 2010

 

Microbicide Containing Engineered Bacteria May Inhibit HIV-1

Antimicrobial Agents and Chemotherapy, July 2010

 

Salmonella Contaminated Pork May Pose Health Risk for Humans

Applied and Environmental Microbiology, July 2010

 

Prior Exposure to Seasonal Influenza May Explain the Mildness of the 2009 H1N1 Pandemic

Journal of Virology, July 2010

 

For as summary of the articles and to download abstracts, go to http://www.asm.org/index.php/news-room/journal-tipsheets.html

 

FEDERAL AGENCY UPDATES 


CDC Health Advisory on Increased Potential for Dengue Infection in Travelers Returning from International and Selected Domestic Areas

The Centers for Disease Control and Prevention (CDC) sent a Health Advisory to clinicians regarding the increased potential for dengue infection in travelers returning from the many parts of the tropics and subtropics.  Dengue virus transmission has been increasing to epidemic levels in many parts of the tropics and subtropics. Travelers to these areas are at risk of acquiring dengue virus and developing dengue fever (DF) or the severe form of the disease, dengue hemorrhagic fever (DHF).  CDC strongly advises that health care providers in the United States should: 1) consider DF and DHF when evaluating patients returning from dengue-affected areas, both domestic and abroad, who present with an acute febrile illness within two weeks of their return, 2) submit serum specimens for appropriate laboratory testing, and 3) report all presumptive and confirmed cases of DF and DHF to their local or state health department.  To read the Health Advisory, go to http://www2a.cdc.gov/HAN/ArchiveSys/ViewMsgV.asp?AlertNum=00315

 

MMWR Articles of Interest

Emergence of Cryptococcus gattii, Pacific Northwest, 2004 - 2010

July 23, 2010 / 59(28);865-868

 

La Crosse Virus Neuroinvasive Disease, Missouri, 2009

July 23, 2010 / 59(28);869-871

 

Gastrointestinal Anthrax after an Animal-Hide Drumming Event, New Hampshire and Massachusetts, 2009

July 23, 2010 / 59(28);872-877


Notes from the Field: Dengue Epidemic, Puerto Rico, January - July 2010

July 23, 2010 / 59(28);878

 

Use of Anthrax Vaccine in the United States: Recommendations of the Advisory Committee on Immunization Practices (ACIP), 2009

July 23, 2010 / 59(rr06);1-30


West Nile Virus Activity, United States, 2009

July 2, 2010 / 59(25);769-772

 

First Smallpox Vaccine for Special Populations Delivered Under Project BioShield

Delivery to the Strategic National Stockpile of the first 1 million doses of the nation’s first smallpox vaccine for certain immune compromised populations is now complete as the result of a Project BioShield contract. Under this contract, the Danish company Bavarian Nordic is manufacturing and delivering 20 million doses of its next generation smallpox vaccine known as modified vaccinia Ankara (MVA) or Imvamune.  In an emergency, such as the virus being obtained from a secure lab and used in an act of terrorism, the vaccine may be authorized for use to protect people who have weakened immune systems, specifically HIV persons who have not progressed to AIDS. For more information, go to http://www.hhs.gov/news/press/2010pres/07/20100714c.html

 

Technology Assessment for Quality, Regulation and Clinical Utility of LDTs

On May 19, Technology Assessment for Quality, Regulation and Clinical Utility of Laboratory Developed Molecular Tests was published by the ECRI Institute Evidence Based Practice Center under contract for the Agency for Health Research and Quality.  The report, which was requested by the Coverage and Analysis Group at the Centers for Medicare & Medicaid Services, summarizes the available scientific evidence on the quality of laboratory developed tests (LDTs) that are not regulated by the Food and Drug Administration.  The report includes information on the 1) types of LDTs available for conditions relevant to the Medicare population; 2) methodologies and processes that have been developed for the assessment of analytical and clinical performance of molecular tests, 3) summarizes the role of Federal agencies in regulating LDTs, and 4) identifies the quality standards that have been developed for molecular tests by regulatory bodies, industry and the medical community.  The report can be downloaded at http://www.cms.gov/determinationprocess/downloads/id72TA.pdf

 

NIH Scientists Advance Universal Flu Vaccine

A universal influenza vaccine may become a reality because of research led by scientists from the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health.  In experiments with mice, ferrets and monkeys, the investigators used a two-step immunization approach to elicit infection fighting antibodies that attacked a diverse array of influenza virus strains. Current flu vaccines do not generate such broadly neutralizing antibodies, so they must be reformulated annually to match the predominant virus strains circulating each year.  For more information, go to

http://www.nih.gov/news/health/jul2010/niaid-15.htm

 

National HIV/AIDS Strategy

On July 13, President Obama released the new National HIV/AIDS Strategy.  The strategy is an across-the-board approach that calls for a more coordinated national response to achieve three primary, concrete goals:  (1) reduce new HIV infections, (2) increase access to care and optimize health outcomes for people with HIV/AIDS, and (3) reduce HIV-related health disparities.  For more information, go to http://aids.gov/federal-resources/policies/national-hiv-aids-strategy/

 

OTHER INFORMATION AND UPDATES

 

New from CLSI

M100-S20 June 2010 Update, Performance Standards for Antimicrobial Susceptibility Testing; Update (M100-S20-U)

 

To purchase this and other CLSI products, go to http://www.clsi.org

 

NLTN Online Training Course in Packing and Shipping

The National Laboratory Training Network (NLTN) is sponsoring an interactive online training course, “Packaging & Shipping Division 6.2 Materials.”  The intermediate level online course is designed for individual study and is suitable for those seeking recertification.  Participants are provided with information useful for complying with regulations through use of instructional content and the opportunity to apply knowledge using realistic scenarios.  For more information, go to http://www.nltn.org/302-10.htm


Articles of Interest

To fight drug-resistant MRSA, algorithm predicts bacteria’s future mutations

Popular Science, July 20, 2010

Researchers led by Bruce Donald, a professor of computer science and biochemistry at Duke University, are using a computer algorithm to predict MRSA’s next move.

 

Enlist malaria-resistant mosquitoes to stop its spread

New Scientist, July 20, 2010

Michael Riehle at the University of Arizona at Tucson and colleagues put a novel gene into the mosquito species that carries malaria in India. The new gene permanently switched on a set of genes normally affected by insulin and involved in the immune system.

 

Synthetic cell-like microcapsules communicate like biological cells, cooperate like ants

Popular Science, July 20, 2010

Taking cues from slime molds, ants, and living biological cells, a team of University of Pittsburgh researchers has designed a system of artificial cells that can communicate with one another and cooperate to carry out tasks. The computer models they have devised could lead to artificial cellular systems that perform highly specialized jobs at the microscopic level.

 

This won't hurt a bit

Science News, July 18, 2010

Researchers have invented a new vaccine-delivery system that replaces the large single needle with 100 tiny dissolvable ones embedded in a Band-Aid–like patch. The new patch can immunize mice against influenza just as effectively as conventional needle vaccination, its developers report online July 18 in Nature Medicine.

 

Retrovirus replication process different than thought

Science Blog, July 15, 2010

How a retrovirus, like HIV, reproduces and assembles new viruses is different than previously thought, according to Penn State College of Medicine researchers. Understanding the steps a virus takes for assembly could allow development of a way to prevent the spread of retroviral diseases.

 

Few people cover coughs and sneezes

Reuters, July 13, 2010

Lots of people aren't getting the message about covering their coughs and sneezes, at least in New Zealand, according to a study conducted during last year's H1N1 swine flu pandemic.

 

Study suggests link between HPV, skin cancer

U.S. News & World Report, July 8, 2010

The ubiquitous virus linked to cervical, vaginal and throat cancers may also raise the risk of developing squamous cell carcinoma, the second most common form of skin cancer, a new study suggests.

 

Amid the murk of 'gut flora,' vitamin D receptor emerges as a key player

Science Daily, July 8, 2010

Scientists have found that the vitamin D receptor is a key player amid the gut bacteria, what scientists refer to matter-of-factly as the "gut flora, "helping to govern their activity, responding to their cues, and sometimes countering their presence. The findings deliver a new lead to scientists investigating how bacteria might play a role in the development of inflammatory bowel diseases such as Crohn's disease or ulcerative colitis.

 

Surgery linked to Creutzfeldt-Jakob disease

Science Blog, July 8, 2010

The finding reveals that, with a few exceptions, the risk of having contracted the sporadic form of CJD manifests itself at least 20 years after having undergone an operation.

 

Souped-up antibody fends off HIV

Nature, July 8, 2010

An antibody that can block more than 90% of strains of HIV-1, the most common form of the disease, has been discovered using a method for fishing specific proteins out of the blood.

 

Map of herpes virus protein suggests a new drug therapy

Newswise, July 6, 2010

New research reveals the unusual structure of the protein complex that allows a herpes virus to invade cells. This detailed map of a key piece of the herpes virus “cell-entry machinery” gives scientists a new target for antiviral drugs.

 

Revolutionary medical dressing uses nanotechnology to fight infection

Science Daily, July 6, 2010

Researchers are using nanotechnology to develop a medical dressing which will detect and treat infection in wounds.

 

Bacterial communication encourages chronic, resistant ear infections

Science Blog, July 6, 2010

Ear infections caused by more than one species of bacteria could be more persistent and antibiotic-resistant because one pathogen may be communicating with the other, encouraging it to bolster its defenses. Researchers from Wake Forest University Baptist MedicalCenter publish their findings today in mBio, the online open-access journal of the American Society for Microbiology

 

Vaccine for Marburg virus passes monkey test

Science News, July 5, 2010

A devastating tropical virus that has no cure can be ambushed by vaccination a day or two after exposure, tests in monkeys show.

 

Discovery of Controlled Swarm in Bacteria: Could Help Design New Strategies to Increase Sensitivity to Antibiotics

ScienceDaily, June 30, 2010

A study led by researchers from Universitat Autònoma de Barcelona (UAB) describes one of the mechanisms in which pathogenic bacteria populations control the way they spread over the surface of the organs they infect and stop when they detect the presence of an antibiotic, only to resume again when the effect wears off. The star of this process is the RecA protein, which significantly increases its concentration at the start of the bacteria DNA repair mechanism induced by antibiotics.

 

Deaths in the family cause bacteria to flee

PhysOrg, June 29, 2010

The deaths of nearby relatives has a curious effect on the bacterium Caulobacter crescentus, surviving cells lose their stickiness. Indiana University Bloomington biologists report in an upcoming issue of Molecular Microbiology that exposure to the extracellular DNA (eDNA) released by dying neighbors stops the sticky holdfasts of living Caulobacter from adhering to surfaces, preventing cells from joining bacterial biofilms. Less sticky cells are more likely to escape established colonies, out to where conditions may be better.

 

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Conclusions

In conclusion, this study demonstrates that h-SH3 can inhibit the replication of influenza A viruses (H1N1, H3N2, and H9N2) at different degrees, and targeting NS1-p85β interaction (PI3K/Akt pathway) or other virus-host interaction may be an attractive strategy against the infection of various influenza A virus subtypes/strains.

Methods

Cell lines, viruses, plasmids, and reagents

Madin-Darby canine kidney (MDCK) cell lines were routinely cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum and antibiotics (100 U penicillin and 100 ug/ml streptomycin) at 37℃ in 5% CO2.

Influenza A virus strains A/PR/8/34(H1N1), A/ST/1233/2006(H1N1), A/ST/364/2005(H3N2), A/Ph/ST/2246/2006(H9N2), and A/Qa/ST/199/2006(H9N2), abbreviated herein to PR8, ST1233, ST364, Ph2246, and Qa199, respectively, were used in this study.

SH3 domain (9-79aa) of p85β subunit was cloned into PNF vector (a modified pcDNA3 plasmid with an N-terminal Flag tag) at the BamHI and EcoRI sites to generate pSH3. Full-length NS1 sequence of H1N1, H3N2, or H9N2 viruses was cloned into pEGFP-c1 at the BamHI site to generate pNS11, pNS32, or pNS92 plasmids, respectively. The constructs were verified by DNA sequencing.

Rabbit monoclonal anti-phospho-Akt (Ser473) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA), Rabbit monoclonal anti-Flag antibody from Sigma (St. Louis, MO, USA), Rabbit monoclonal anti-Akt antibody, Cy3-labeled goat anti-rabbit antibody, peroxidase-conjugated goat anti-rabbit antibody, and LY294002 (PI3K inhibitor) from Beyotime Biotechnology (Jiangsu, China), and Lipofectamine 2000 from Invitrogen (Carlsbad, CA, USA).

Co-localization analysis by confocal microscopy

MDCK cell cultures on the glass coverslips were transfected with indicated plasmids. 24 h post-transfection, the cells were washed twice with phosphate-buffered saline (PBS), fixed for 10 min with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100 for 7 min, and incubated for 30 min in PBS containing 3% Bovine Serum Albumin (BSA). Cells were subsequently incubated at room temperature with anti-Flag antibody (1/500) for 2 h and Cy3-labeled secondary antibody (1/300) for 1 h. Images were captured with the Olympus confocal microscopy.

Transient transfection, viral infection, and determination of antiviral activity

MDCK cells were seeded onto six-well plates and transfected with pSH3 or PNF empty vector at 60% confluence as previously described [26]. After 48 h of transfection, MDCK cells were infected with influenza A viruses at a multiplicity of infection (MOI) of 0.001 in serum-free DMEM containing 1 ug/ml TPCK-trypsin (Worthington, Freehold, NJ, USA) and incubated at 37°C. An aliquot of the supernatant was harvested every 12 h and virus yield was titrated by plaque assay in MDCK cells.

Western blot-based phosphorylation assay

MDCK cells transfected with indicated plasmids for 24 h were infected with different influenza A virus strains at an MOI of 2. At 8 h post infection the cells were lysed with Laemmli sample buffer containing 5 mM of NaF for 5 min in boiling water followed by brief sonication. Protein concentration was determined using an RC-DC kit (Bio-Rad, Hercules, CA, USA). Proteins (20 ug/lane) were separated in 10% SDS-PAGE and transferred onto a nitrocellulose membrane. The membrane was blocked with Tris-buffered saline containing 0.1% Tween 20 (TBST) with 10% non-fat milk for 2 h and incubated overnight at 4°C with anti-phospho-Akt (Ser473) (1/500) or anti-Akt (1/700) antibody. The membrane was rinsed extensively in TBST and incubated for 2 h with peroxidase-conjugated secondary antibody (1/1000). Immunoblots were developed using the Enhanced chemiluminescence (ECL) reagents (Pierce, Rockford, IL, USA). Quantity One (Bio-Rad) software was used to analyze images.

Determination of PI3K effect on the replication of ST364 and PR8 viruses

MDCK cells with 100% confluence were pretreated with 10 uM or 50 uM LY294002 (a specific PI3K inhibitor). After 2 h, MDCK cells were infected with ST364 virus or PR8 virus, and plaque assay was performed.

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

Conceived and designed the experiments: KSL, DGZ, WZL, GFW. Performed the experiments: DGZ, WZL, GFW, CZ, XXC, YXX. Analyzed the data: DGZ, WZL, KSL, GFW, YS, XXZ. Wrote the paper: DGZ, WZL, KSL. All authors read and approved the final manuscript.

Acknowledgements

We thank Dr. William Ba-Thein for critical discussion and manuscript editing. This study was supported by National Natural Science Foundation of China (30771988, 30972766), Guangdong Natural Science Foundation(8151503102000022, 9451503102003499), Specialized Research Fund for the Doctoral Program of Higher Education (20094402110004), Outstanding Young Scientists Foundation of Guangdong Province Education Department (LYM08056), State Key Lab of Agriculture Microbiology Open Foundation (AML200910), Shantou University Medical College Research Foundation, and 211 Project of Guangdong Province (Mechanism and Prevention of Emerging Infectious Diseases).

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