Quantitation of viral RNA levels of H5N1 RNP complexes containing PB2 mutations.

<p>293T cells were co-transfected with expression plasmids of NP, PA, PB1 and either wild type (WT) of indicated PB2 mutants with amino acid substitution of E158G, T271A or E627K, together with pPolI-NA plasmid. Total cellular RNA was isolated after 48 hours post-transfection and subjected to quantitative RT-PCR for segment 6 (NA genes) transcripts. Cells were incubated at (A) 33°C, (B) 37°C. RNA levels were expressed as relative activity to wild-type. Results shown are means with standard deviations from three independent assays. *indicates <i>P</i><0.05 when compared to wild-type.</p

Comparison of <i>in vitro</i> polymerase activity of reconstituted RNP complexes.

<p>Activities of polymerase complexes with a single gene replacement following expression in human 293T cells. (A) H5N1 polymerase complexes substituted with one H3N2 gene, (B) H3N2 polymerase complexes substituted with one H5N1 gene, (C) H5N1 polymerase complexes substituted with one H1N1pdm09 gene, and (D) H1N1pdm09 polymerase complexes substituted with one H5N1 gene, were analyzed in 293T cells transfected with the indicated plasmids of NP, PA, PB1 and PB2 together with pPolI-vNP-Luc and a reporter plasmid pGL4.73. Cells were incubated at 33°C and 37°C. Polymerase activity was normalized with the expression of a reporter plasmid. Relative polymerase activity (%) was expressed as relative activity to the corresponding parental vRNPs. Results shown are means with standard deviations from three independent assays.</p

Comparison of <i>in vitro</i> polymerase activity of H1N1pdm09, seasonal H3N2, avian H5N1, and WSN H1N1.

<p>293T cells were co-transfected with expression plasmids of NP, PA, PB1 and PB2 together with pPolI-vNP-Luc and a reporter plasmid pGL4.73 [hRluc/SV40], encoding a <i>Renilla</i> luciferase gene. Cells were incubated at (A) 33^o and (B) 37°C. Polymerase activity was normalized with the expression of a reporter plasmid. Relative polymerase activity (%) was expressed as relative activity to WSN H1N1 in percentage. Results shown are means with standard deviations from three independent assays. *indicates <i>P</i><0.05 when compared to WSN H1N1, and ***indicates <i>P</i><0.001 when compared to H5N1.</p

Polymerase activity of H5N1 RNP complexes containing mutations E158G, T271A and E627K in PB2.

<p>293T cells were co-transfected with expression plasmids of NP, PA, PB1 and either wild type (WT) or PB2 mutants with the indicated amino acid substitution of E158G, T271A or E627K, together with pPolI-vNP-Luc and a reporter plasmid pGL4.73. Cells were incubated at (A) 33°C, (B) 37°C. Polymerase activity was normalized with the expression of a reporter plasmid. Relative polymerase activity (%) was expressed as relative activity to WT. Results shown are means with standard deviations from three independent assays. *indicates <i>P</i><0.05 when compared to WT.</p

Cervical intraepithelial neoplasia grade 2 across continents.

<p>Cervical intraepithelial neoplasia grade 2 across continents.</p

Cervical adenocarcinoma and adenosquamous cell carcinoma across continents.

<p>Cervical adenocarcinoma and adenosquamous cell carcinoma across continents.</p

Study flow chart.

<p>Study flow chart.</p

Prevalence and attribution of HPV52 and HPV58 across different lesion grades in Eastern Asia and other parts of the world.

1<p>Relative prevalence, no. of HPV52-positive cases regardless of single- or multiple-type infection/total no. of HPV-positive cases.</p>2<p>% of cases with HPV52 single-type infection +% of cases with HPV52 multiple-type infection × attribution factor. Attribution factor  =  no. of cases with HPV52 single-type infection/no. of cases with single-type infection of any HPV type.</p>3<p>Relative prevalence, no. of HPV58-positive cases regardless of single- or multiple-type infection/total no. of HPV-positive cases.</p>4<p>% of cases with HPV58 single-type infection +% of cases with HPV58 multiple-type infection × attribution factor. Attribution factor  =  no. of cases with HPV58 single-type infection/no. of cases with single-type infection of any HPV type.</p><p>CIN, cervical intraepithelial neoplasia; SCC/UNSPEC, squamous cell carcinomas and invasive cervical cancers of unspecified histology; adenocarcinoma includes cervical adenocarcinoma and adenosquamous cell carcinoma.</p><p>Prevalence and attribution of HPV52 and HPV58 across different lesion grades in Eastern Asia and other parts of the world.</p

Prevalence and attribution of HPV52 and HPV58 among cervical lesions in Eastern Asia.

<p>Prevalence and attribution of HPV52 and HPV58 among cervical lesions in Eastern Asia.</p

Levels of anti-hepatitis B surface antibody (anti-HBs) after booster vaccination for young adults.

<p>Each data point represents one student aged 17–23 years, born after the adoption of universal neonatal immunization for hepatitis B. All these 69 students were negative for HBsAg and anti-HBs before booster vaccination. Anti-HBs levels were determined one month after the first dose and 2–4 months after the third dose of booster vaccination with 20 mcg of ENERGIX-B (GlaxoSmithKline Biologicals, Belgium).</p