Adaptation of High-Growth Influenza H5N1 Vaccine Virus in Vero Cells: Implications for Pandemic Preparedness

Current egg-based influenza vaccine production technology can't promptly meet the global demand during an influenza pandemic as shown in the 2009 H1N1 pandemic. Moreover, its manufacturing capacity would be vulnerable during pandemics caused by highly pathogenic avian influenza viruses. Therefore, vaccine production using mammalian cell technology is becoming attractive. Current influenza H5N1 vaccine strain (NIBRG-14), a reassortant virus between A/Vietnam/1194/2004 (H5N1) virus and egg-adapted high-growth A/PR/8/1934 virus, could grow efficiently in eggs and MDCK cells but not Vero cells which is the most popular cell line for manufacturing human vaccines. After serial passages and plaque purifications of the NIBRG-14 vaccine virus in Vero cells, one high-growth virus strain (Vero-15) was generated and can grow over 108 TCID50/ml. In conclusion, one high-growth H5N1 vaccine virus was generated in Vero cells, which can be used to manufacture influenza H5N1 vaccines and prepare reassortant vaccine viruses for other influenza A subtypes

Predicting Hypnotic Use among Insomnia Patients with the Theory of Planned Behavior and Craving

While long-term hypnotic use is very common in clinical practice, the associated factors have been understudied. This study aims to explore the cognitive factors that might influence the long-term use of hypnotics based on the theory of planned behavior (TPB), and examines the moderating effect of craving between cognitive intention and actual hypnotic-use behavior at follow-up. A total of 139 insomnia patients completed a self-constructed TPB questionnaire to measure their attitude, subjective norm, perceived behavioral control, and behavioral intention of hypnotic use, as well as the Hypnotic-Use Urge Scale (HUS) to measure their craving for hypnotics. They were then contacted through phone approximately three months later to assess their hypnotic use. Hierarchical regression showed that perceived behavioral control was the most significant determinant for behavioral intention of hypnotic use. Behavioral intention, in turn, can predict the frequency of hypnotic use after three months. However, this association was moderated by hypnotic craving. The association was lower among the participants with higher cravings for hypnotic use. The findings suggest that the patients’ beliefs about their control over sleep and daily life situations, and their craving for hypnotics should be taken into consideration in the management of hypnotic use

Clinical observations on enzyme replacement therapy in patients with Fabry disease and the switch from agalsidase beta to agalsidase alfa

Background: Fabry disease is an X-linked inherited lysosomal storage disease that can be treated with the enzymes of agalsidase beta (Fabrazyme) and agalsidase alfa (Replagal). Since June 2009, viral contamination of Genzyme's production facility has resulted in a worldwide shortage of agalsidase beta, leading to the switch to agalsidase alfa for patients with Fabry disease in Taiwan. Methods: The medical records were retrospectively reviewed for nine male patients with Fabry disease from the start of agalsidase beta treatment until the switch to agalsidase alfa for at least 1 year. Results: After 12–112 months of enzyme replacement therapy (ERT), decreased plasma globotriaosylsphingosine (lyso-Gb3) was found in five out of seven patients, indicating improvement in disease severity. Among the six patients with available echocardiographic data at baseline and after ERT, all six experienced reductions of left ventricular mass index. Renal function, including microalbuminuria and estimated glomerular filtration rate, showed stability after ERT. Mainz Severity Score Index scores revealed that all nine patients remained stable at 12 months after switching to agalsidase alfa. ERT improved or stabilized cardiac status and stabilized renal function, while reducing plasma lyso-Gb3. ERT was well tolerated, even among the three patients who had hypersensitivity reactions. Conclusion: The switch of ERT from agalsidase beta to agalsidase alfa appears to be safe after 1 year of follow-up for Taiwanese patients with Fabry disease

Plaque morphology of NIBRG-14 and Vero cell-adapted (Vero-15) H5N1 viruses grown in Vero cells in different days post infection (DPI).

<p>Plaque morphology of NIBRG-14 and Vero cell-adapted (Vero-15) H5N1 viruses grown in Vero cells in different days post infection (DPI).</p

Growth efficiency of the NIBRG-14 and Vero-adapted influenza H5N1 (Vero-15) viruses with different multiplicity of infection (MOI) in T-flasks.

<p>TCID₅₀ (50% tissue culture infectious dose) was measured in triplicate and shown as geometric mean ± standard error of mean.</p><p>Boldfaces indicate peak infectious virus titers.</p

Growth efficiency of Vero-adapted influenza H5N1 virus (Vero-15) in microcarrier-based Vero cell cultures in 100-ml spinner flasks.

<p>HA: hemagglutination.</p><p>TCID₅₀ (50% tissue culture infectious dose) was measured in triplicate and shown as geometric mean ± standard error of mean.</p

Genetic differences between the NIBRG-14 and Vero-adapted H5N1 (Vero-15) viruses.

<p>*Based on numbering of A/PR/8/34 (accession no. CAA23855).</p