Chikungunya virus (CHIKV) is becoming an increasing global health issue which has spread across the globe and as far north as southern Europe. There is currently no vaccine or anti-viral treatment available. Although there has been a recent increase in CHIKV research, many of these in vitro studies have used a wide range of cell lines which are not physiologically relevant to CHIKV infection in vivo. In this study, we aimed to evaluate a panel of cell lines to identify a subset that would be both representative of the infectious cycle of CHIKV in vivo, and amenable to in vitro applications such as transfection, luciferase assays, immunofluorescence, western blotting and virus infection. Based on these parameters we selected four mammalian and two mosquito cell lines, and further characterised these as potential tools in CHIKV research.
Chikungunya virus (CHIKV), a mosquito-borne alphavirus, causes febrile disease, muscle and joint pain, which can become chronic in some individuals. The non-structural protein 3 (nsP3) plays essential roles during infection, but a complete understanding of its function is lacking. Here we used a microscopy-based approach to image CHIKV nsP3 inside human cells. The SNAP system consists of a self-labelling enzyme tag, which catalyses the covalent linking of exogenously supplemented synthetic ligands. Genetic insertion of this tag resulted in viable replicons and specific labelling while preserving the effect of nsP3 on stress granule responses and co-localisation with GTPase Activating Protein (SH3 domain) Binding Proteins (G3BPs). With sub-diffraction, three-dimensional, optical imaging, we visualised nsP3-positive structures with variable density and morphology, including high-density rod-like structures, large spherical granules, and small, low-density structures. Next, we confirmed the utility of the SNAP-tag for studying protein turnover by pulse-chase labelling. We also revealed an association of nsP3 with cellular lipid droplets and examined the spatial relationships between nsP3 and the non-structural protein 1 (nsP1). Together, our study provides a sensitive, specific, and versatile system for fundamental research into the individual functions of a viral non-structural protein during infection with a medically important arthropod-borne virus (arbovirus).
Friedel-Crafts reactions in the ionic liquid system 1-methyl-3-ethylimidazolium chloride-aluminium(III) chloride can be performed with excellent yields and selectivities, and in the case of anthracene, have been found to be reversible.The possibility of carrying out chemical transformations in low temperature ionic liquids has, to date, received little attention. 1 Ionic liquids such as the [emim]Cl-AlCl 3 ([emim] + = 1-methyl-3-ethylimidazolium cation) system have been shown to demonstrate catalytic activity in reactions such as Friedel-Crafts acylations, 2,3 alkylation reactions, 4 isomerisation of alkanes, 5 and the alkylation of isobutane with butene. 6 Here, we present a series of reactions of AcCl with carbocyclic aromatic compounds in acidic compositions (X = 0.67) ‡ of [emim]Cl-AlCl 3 (Fig. 1) and compare their performance with similar reactions in 'conventional' molecular solvents.
Complex dynamics of language use are in operation within bilingual healthcare settings and organizational as well as individual factors are important in facilitating appropriate language use. Many of the issues highlighted are not peculiar to the Welsh context, but apply to healthcare settings across the world, where other minority languages are in use.
Communication and the interactions that take place between nurse and patient are the foundation of nursing care. However, in practice, cross-cultural communication is often inadequate and therefore the health care needs of many cultural groups are not being met. An examination of language switching within nurse/patient interaction in a bilingual setting provides a further dimension for the study of cross-cultural communication.
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