Novel antiviral activity of neuraminidase inhibitors against an avian influenza a virus

<p>Abstract</p> <p>Background</p> <p>Neuraminidase (NA) inhibitors used for influenza therapy are believed to prevent the release of progeny virus from the surface of an infected cell. In this study, we found that NA inhibitors have a novel antiviral function against an avian influenza virus.</p> <p>Results</p> <p>Madin-Darby canine kidney cells, commonly used for the isolation and propagation of the influenza virus, were infected with an avian influenza viral strain A/chicken/German/N/49(H10N7) (H10/chicken) or a human influenza viral strain A/Osaka/981/98(H3N2) (H3/Osaka) virus. Cells were incubated in a medium without or with a NA inhibitor, oseltamivir carboxylate (GS4071), from 1 to 13 h post infection (p.i.). Infected cells were washed 12 h p.i. to remove GS4071, incubated for 1 h without GS4071, and assayed for virus production. Incubation with GS4071 decreased the production of infectious viruses. When H10/chicken virus-infected cells were incubated with GS4071 from 12 to 13 h p.i. (i.e., 1 h before the virus production assay), the inhibitory effect was clearly observed, however, the same was not evident for H3/Osaka virus-infected cells. Furthermore, viral protein synthesis in infected cells was not affected by GS4071. Using a scanning electron microscope, many single spherical buds were observed on the surface of H3/Osaka virus-infected cells incubated without GS4071, whereas many aggregated particles were observed on the surface of cells incubated with GS4071. However, many long tubular virus-like structures, with no aggregated particles, were observed on the surface of H10/chicken virus-infected cells incubated with GS4071. The same results were obtained when another NA inhibitor, zanamivir, was used.</p> <p>Conclusions</p> <p>These results indicate that NA inhibitors interfered with virus particle formation in the H10/chicken virus-infected cells, in which the inhibitor caused the formation of long tubular virus-like structures instead of spherical virus particles.</p

Proteolytic processing of human cytomegalovirus glycoprotein B (gpUL55) is mediatedby the human endoprotease furin

AbstractInhibition of endoproteolytic cleavage of glycoprotein B (gB; gpUL55) of human cytomegalovirus was achieved by treatmentof infected fibroblasts with decanoyl peptidyl chloromethyl ketone (decRVKR-CMK), which inhibits the action of cellular subtilisin-like endoproteases with the amino acid recognition motif R × K/R R. Uncleaved gB precusor molecules of 160 kDa that were accumulated were endoglycosidase H resistant, suggesting that correct cellular transport occurred in the presence of the drug. The inhibitor also prevented endoproteolytic gB processing in CV-1 cells infected with a recombinant vaccinia virus-gB construct (VVgB). Evidence for direct involvement of the ubiquitous subtilisin-like endoprotease furin in gB cleavage was obtained from the observation that coinfection of CV-1 cells with WgB and a recombinant vaccinia-human furin construct reestablished endoproteolytic activity which was normally absent late after infection with WgB alone

Randomized Controlled Trial on Effectiveness of Ultrasonography Screening for Breast Cancer in Women Aged 40–49 (J-START): Research Design

In cancer screening, it is essential to undertake effective screening with appropriate methodology, which should be supported by evidence of a reduced mortality rate. At present, mammography is the only method for breast cancer screening with such evidence. However, mammography does not achieve sufficient accuracy in breasts with high density at ages below 50. Although ultrasonography achieves better accuracy in Breast Cancer detection even in dense breasts, the effectiveness has not been verified. We have planned a randomized controlled trial to assess the effectiveness of ultrasonography in women aged 40–49, with a design to study 50 000 women with mammography and ultrasonography (intervention group), and 50 000 controls with mammography only (control group). The participants are scheduled to take second round screening with the same modality 2 years on. The primary endpoints are sensitivity and specificity, and the secondary endpoint is the rate of advanced breast cancers

Design Study of a Superconducting Insertion Quadrupole Magnet for the Large Hadron Collider

The conceptual design study of a high gradient super conducting insertion quadrupole magnet has been carried out in collaboration between KEK and CERN for the Large Hadron Collider (LHC) to be built at CERN. A model magnet design has been optimized to provide a nominal design field gradient of 240 T/m with a bore aperture of 70 mm and an operational field gradient of 225 T/m at 1.9 K under radiation environment with a beam energy deposit of several watts per meter in the superconducting coils. The design and its optimization process are discussed

Development of an in vitro cell system from zebrafish suitable to study bone cell differentiation and extracellular matrix mineralization

Mechanisms of bone formation and skeletal development have been successfully investigated in zebrafish using a variety of in vivo approaches, but in vitro studies have been hindered due to a lack of homologous cell lines capable of producing an extracellular matrix (ECM) suitable for mineral deposition. Here we describe the development and characterization of a new cell line termed ZFB1, derived from zebrafish calcified tissues. ZFB1 cells have an epithelium-like phenotype, grow at 28 degrees C in a regular L-15 medium supplemented with 15% of fetal bovine serum, and are maintained and manipulated using standard methods (e.g., trypsinization, cryopreservation, and transfection). They can therefore be propagated and maintained easily in most cell culture facilities. ZFB1 cells show aneuploidy with 2n=78 chromosomes, indicative of cell transformation. Furthermore, because DNA can be efficiently delivered into their intracellular space by nucleofection, ZFB1 cells are suitable for gene targeting approaches and for assessing gene promoter activity. ZFB1 cells can also differentiate toward osteoblast or chondroblast lineages, as demonstrated by expression of osteoblast- and chondrocyte-specific markers, they exhibit an alkaline phosphatase activity, a marker of bone formation in vivo, and they can mineralize their ECM. Therefore, they represent a valuable zebrafish-derived in vitro system for investigating bone cell differentiation and extracellular matrix mineralization.FISHCELL project [PTDC/MAR/105313/2008]; Portuguese Science and Technology Foundation (FCT); European Regional Development Fund (ERDF) through the COMPETE Program; National Fund through FCT [PEst-C/MAR/LA0015/2011]; FCT [SFRH/BPD/39189/2007]; Association of European Marine Biological Laboratories through the ASSEMBLE project [FP7/227799]info:eu-repo/semantics/publishedVersio

Test Results of a Variant-Design LHC Twin-Aperture Dipole Magnet

Since 1989, KEK and CERN carried out jointly an experimental program in the frame of the R&D work for the LHC main dipole. The mechanical structure of this design is based on a separate coil/collar and "horizontally split iron" concept. A total of four single aperture and two twin-aperture 1 m long dipole magnets were built. The last twin-aperture magnet was tested at CERN, reaching a maximum field of 9.55 T at 1.9 K. This paper reports the magnet training performance and quench localization at 1.9 K and 4.5 K. The performance as a function of current ramp rate and measurements of the field quality are also reported