The air we breathe contains microorganisms that can cause infectious respiratory diseases. After two occupants of an apartment were diagnosed with influenza in February of 2013, efforts were made to detect and isolate airborne influenza virus using two different types of active air samplers: a Sioutas Personal Cascade Impactor Sampler (PCIS) and an SKC BioSampler. The PCIS collects size-fractionated particles by impaction on polytetrafluoroethylene filters, whereas the SKC BioSampler collects airborne particles in liquid media. Influenza H3N2 virus was collected by both types of air samplers. The PCIS collected a range of particle sizes containing influenza virus near one of the sick individuals but only ultrafine particles when the samplers were positioned farther away. Viable virus was present in the liquid collection media of the SKC BioSampler and some PCIS filters. These findings suggest that influenza patients produce ultrafine aerosol particles that contain viable virus.

John A. Lednicky

Julia C. Loeb

Detection and Isolation of Airborne Influenza A H3N2 Virus Using a Sioutas Personal Cascade Impactor Sampler

Currently not much is known about the H7N9 strain, and this is the major drawback for a scientific strategy to tackle this virus. Herein, the 3D complex structure of the H7N9 RNA-dependent RNA polymerase has been established using a repertoire of molecular modelling techniques including homology modelling, molecular docking, and molecular dynamics simulations. Strikingly, it was found that the oligonucleotide cleft and tunnel in the H7N9 RNA-dependent RNA polymerase are structurally very similar to the corresponding region on the hepatitis C virus RNA-dependent RNA polymerase crystal structure. A direct comparison and a 3D postdynamics analysis of the 3D complex of the H7N9 RNA-dependent RNA polymerase provide invaluable clues and insight regarding the role and mode of action of a series of interacting residues on the latter enzyme. Our study provides a novel and efficiently intergraded platform with structural insights for the H7N9 RNA-dependent RNA Polymerase. We propose that future use and exploitation of these insights may prove invaluable in the fight against this lethal, ongoing epidemic.

Dimitrios Vlachakis

Argiro Karozou

Sophia Kossida

3D Molecular Modelling Study of the H7N9 RNA-Dependent RNA Polymerase as an Emerging Pharmacological Target

The nature of influenza virus to randomly mutate and evolve into new types is an important challenge in the control of influenza infection. It is necessary to monitor virus evolution for a better understanding of the pandemic risk posed by certain variants as evidenced by the highly pathogenic avian influenza (HPAI) viruses. This has been clearly recognized in Egypt following the notification of the first HPAI H5N1 outbreak. The continuous circulation of the virus and the mass vaccination programme undertaken in poultry have resulted in a progressive genetic evolution and a significant antigenic drift near the major antigenic sites. In order to establish if vaccination is sufficient to provide significant intra- and interclade cross-protection, lentiviral pseudotypes derived from H5N1 HPAI viruses (A/Vietnam/1194/04, A/chicken/Egypt-1709-01/2007) and an antigenic drift variant (A/chicken/Egypt-1709-06-2008) were constructed and used in pseudotype-based neutralization assays (pp-NT). pp-NT data obtained was confirmed and correlated with HI and MN assays. A panel of pseudotypes belonging to influenza Groups 1 and 2, with a combination of reporter systems, was also employed for testing avian sera in order to support further application of pp-NT as an alternative valid assay that can improve avian vaccination efficacy testing, vaccine virus selection, and the reliability of reference sera.

Eleonora Molesti

Adelaide Milani

Calogero Terregino

Giovanni Cattoli

Nigel J. Temperton

Comparative Serological Assays for the Study of H5 and H7 Avian Influenza Viruses

<i>Objectives</i>. The national influenza vaccination rate among healthcare workers (HCWs) remains low despite clear benefits to patients, coworkers, and families. We sought to evaluate formally the effect of a one-hour time off incentive on attitudes towards influenza vaccination during the 2011-2012 influenza season. <i>Methods</i>. All HCWs at the Philadelphia Veterans Affairs (VA) Medical Center were invited to complete an anonymous web-based survey. We described respondents&#x2019; characteristics and attitudes toward influenza vaccination and determined the relationship of specific attitudes with respondents&#x2019; acceptance of influenza vaccination, using a 5-point Likert scale. <i>Results</i>. We analyzed survey responses from 154 HCWs employed at the Philadelphia VA Medical Center, with a response rate of 8&#x25;. Among 121 respondents who reported receiving influenza vaccination, 34 (28&#x25;, 95&#x25; CI 20&#x2013;37&#x25;) reported agreement with the statement that the time off incentive made a difference in their decision to accept influenza vaccination. <i>Conclusions</i>. Our study provides evidence that modest incentives such as one-hour paid time off will be unlikely to promote influenza vaccination rates within medical facilities. More potent interventions that include mandatory vaccination combined with penalties for noncompliance will likely provide the only means to achieve near-universal influenza vaccination among HCWs.

Saima Cheema

Christopher Vinnard

Sarah Foster-Chang

Darren R. Linkin

A Time Off Incentive Was Not Associated with Influenza Vaccination Acceptance among Healthcare Workers

The impact of the seasonal influenza and 2009 AH1N1 pandemic influenza on mortality is not yet completely understood, particularly in tropical and subtropical countries. The trends of influenza related mortality rate in different age groups and different outcomes on a area in tropical and subtropical climate with more than 41 million people (State of S&#xe3;o Paulo, Brazil), were studied from 2002 to 2011 were studied.  Serfling-type regression analysis was performed using weekly mortality registries and virological data obtained from sentinel surveillance. The prepandemic years presented a well-defined seasonality during winter and a clear relationship between activity of AH3N2 and increase of mortality in all ages, especially in individuals older than 60 years. The mortality due to pneumonia and influenza and respiratory causes associated with 2009 pandemic influenza in the age groups 0&#x2013;4 years and older than 60 was lower than the previous years. Among people aged 5&#x2013;19 and 20&#x2013;59 years the mortality was 2.6 and 4.4 times higher than that in previous periods, respectively. The mortality in all ages was higher than the average of the previous years but was equal mortality in epidemics of AH3N2. The 2009 pandemic influenza mortality showed significant differences compared to other years, especially considering the age groups most affected.

Andr&#xe9; Ricardo Ribas Freitas

Priscila M. S. Bergamo Francisco

Maria Rita Donalisio

Mortality Associated with Influenza in Tropics, State of S&#xe3;o Paulo, Brazil, from 2002 to 2011: The Pre-Pandemic, Pandemic, and Post-Pandemic Periods

<i  >Objectives</i>. Disease surveillance combines data collection and analysis with dissemination of findings to decision makers. The timeliness of these activities affects the ability to implement preventive measures. Influenza surveillance has traditionally been hampered by delays in both data collection and dissemination. <i  >Methods</i>. We used statistical process control (SPC) to evaluate the daily percentage of outpatient visits with a positive point-of-care (POC) influenza test in the University of Utah Primary Care Research Network. <i  >Results</i>. Retrospectively, POC testing generated an alert in each of 4 seasons (2004&#x2013;2008, median 16 days before epidemic onset), suggesting that email notification of clinicians would be 9 days earlier than surveillance alerts posted to the Utah Department of Health website. In the 2008-09 season, the algorithm generated a real-time alert 19 days before epidemic onset. Clinicians in 4 intervention clinics received email notification of the alert within 4 days. Compared with clinicians in 6 control clinics, intervention clinicians were 40&#x25; more likely to perform rapid testing (<svg     style="vertical-align:-0.1638pt;width:66.875px;" id="M1" height="11.375" version="1.1" viewBox="0 0 66.875 11.375" width="66.875"  xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg">
	
		
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</svg>) and twice as likely to vaccinate for seasonal influenza (<svg     style="vertical-align:-0.1638pt;width:66.875px;" id="M2" height="11.375" version="1.1" viewBox="0 0 66.875 11.375" width="66.875"  xmlns:xlink="http://www.w3.org/1999/xlink" xmlns="http://www.w3.org/2000/svg">
	
		
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</svg>) after notification. <i  >Conclusions</i>. Email notification of SPC-generated alerts provided significantly earlier notification of the epidemic onset than traditional surveillance. Clinician preventive behavior was not significantly different in intervention clinics.

Lisa H. Gren

Christina A. Porucznik

Elizabeth A. Joy

Joseph L. Lyon


							Catherine J. Staes

Stephen C. Alder

Point-of-Care Testing as an Influenza Surveillance Tool: Methodology and Lessons Learned from Implementation

H5N1 influenza viruses cause high mortality in avian and mammalian species, including humans. Antigenic drift in H5 sequence poses challenges in the development of vaccine and therapeutic antibody. In this study, a monoclonal antibody 11G12 was produced from inactivated H5N1 immunized mice. Results from IFA, ELISA, HI, and virus neutralization indicated that Mab 11G12 can specifically recognize and neutralize H5 type hemagglutinin from clade 1 and 0 without any cross-reaction to any other clades of H5N1 viruses. Mab 11G12 was used to differentiate and quantify the expression of H5N1 strain A/VietNam/1203/04 from a trivalent vaccine mix in ELISA. Sequencing of escape mutants identified that Mab 11G12 targets a major neutralizing epitope of influenza H5 hemagglutinin. The study indicated that some major neutralizing epitopes in H5s of early strains were mutated due to antigenic drift.

Fang He

Jimmy Kwang

Monoclonal Antibody Targeting Neutralizing Epitope on H5N1 Influenza Virus of Clade 1 and 0 for Specific H5 Quantification

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Influenza Research and Treatment