Recombinant Subgroup B Human Respiratory Syncytial Virus Expressing Enhanced Green Fluorescent Protein Efficiently Replicates in Primary Human Cells and Is Virulent in Cotton Rats

Human respiratory syncytial virus (HRSV) is the most important viral cause of severe respiratory tract disease in infants. Two subgroups (A and B) have been identified, which cocirculate during, or alternate between, yearly epidemics and cause indistinguishable disease. Existing in vitro and in vivo models of HRSV focus almost exclusively on subgroup A viruses. Here, a recombinant (r) subgroup B virus (rHRSV(B05)) was generated based on a consensus genome sequence obtained directly from an unpassaged clinical specimen from a hospitalized infant. An additional transcription unit containing the gene encoding enhanced green fluorescent protein (EGFP) was introduced between the phosphoprotein and matrix genes (position 5) of the genome to generate rHRSV(B05)EGFP(5). The recombinant viruses replicated efficiently in both HEp-2 cells and in well-differentiated normal human bronchial cells grown at air-liquid interface. Intranasal infection of cotton rats (Sigmodon hispidus) resulted in high numbers of EGFP(+) cells in epithelia of the nasal septum and conchae. When administered in a relatively large inoculum volume, the virus also replicated efficiently in bronchiolar epithelial cells and spread extensively in both the upper and lower respiratory tracts. Virus replication was not observed in ciliated epithelial cells of the trachea. This is the first virulent rHRSV strain with the genetic composition of a currently circulating wild-type virus. In vivo tracking of infected cells by means of EGFP fluorescence in the absence of cytopathic changes increases the sensitivity of virus detection in HRSV pathogenesis studies.

IMPORTANCE

Virology as a discipline has depended on monitoring cytopathic effects following virus culture in vitro. However, wild-type viruses isolated from patients often do not cause significant changes to infected cells, necessitating blind passage. This can lead to genetic and phenotypic changes and the generation of high-titer, laboratory-adapted viruses with diminished virulence in animal models of disease. To address this, we determined the genome sequence of an unpassaged human respiratory syncytial virus from a sample obtained directly from an infected infant, assembled a molecular clone, and recovered a wild-type recombinant virus. Addition of a gene encoding enhanced green fluorescent protein allowed this wild-type virus to be tracked in primary human cells and living animals in the absence of significant cytopathic effects. Imaging of fluorescent cells proved to be a highly valuable tool for monitoring the spread of virus and may help improve assays for evaluating novel intervention strategies.

Ionic liquid gel materials: applications in green and sustainable chemistry

Ionic liquid gel materials offer a way to further utilise ionic liquids in technological applications. Combining the controlled and directed assembly of gels, with the diverse applications of ionic liquids, enables the design of a heady combination of functional tailored materials, leading to the development of task specific / functional ionic liquid gels. This review introduces gels and gel classification, focusing on ionic liquid gels and their potential roles in a more sustainable future. Ionic liquid gels provide the ability to build functionality at every level, the solid component, the ionic liquid, and any incorporated active functional agents. This allows materials to be custom designed for a vast assortment of applications. This diverse class of materials has the potential to yield functional materials for green and sustainable chemistry, energy, electronics, medicine, food, cosmetics, and more. The discussion of the development of ionic liquid gel materials for applications in green and sustainable chemistry centres on uses of ionic liquid gels in catalysis and energy.

Integrating personality research and animal contest theory:Aggressiveness in the green swordtail xiphophorus helleri

Aggression occurs when individuals compete over limiting resources. While theoretical studies have long placed a strong emphasis on context-specificity of aggression, there is increasing recognition that consistent behavioural differences exist among individuals, and that aggressiveness may be an important component of individual personality. Though empirical studies tend to focus on one aspect or the other, we suggest there is merit in modelling both within- and among-individual variation in agonistic behaviour simultaneously. Here, we demonstrate how this can be achieved using multivariate linear mixed effect models. Using data from repeated mirror trials and dyadic interactions of male green swordtails, Xiphophorus helleri, we show repeatable components of (co)variation in a suite of agonistic behaviour that is broadly consistent with a major axis of variation in aggressiveness. We also show that observed focal behaviour is dependent on opponent effects, which can themselves be repeatable but were more generally found to be context specific. In particular, our models show that within-individual variation in agonistic behaviour is explained, at least in part, by the relative size of a live opponent as predicted by contest theory. Finally, we suggest several additional applications of the multivariate models demonstrated here. These include testing the recently queried functional equivalence of alternative experimental approaches, (e.g., mirror trials, dyadic interaction tests) for assaying individual aggressiveness. © 2011 Wilson et al.

Green Space Attachment and Health: A Comparative Study in Two Urban Neighborhoods

The positive relationships between urban green space and health have been well documented. Little is known, however, about the role of residents’ emotional attachment to local green spaces in these relationships, and how attachment to green spaces and health may be promoted by the availability of accessible and usable green spaces. The present research aimed to examine the links between self-reported health, attachment to green space, and the availability of accessible and usable green spaces. Data were collected via paper-mailed surveys in two neighborhoods (n = 223) of a medium-sized Dutch city in the Netherlands. These neighborhoods differ in the perceived and objectively measured accessibility and usability of green spaces, but are matched in the physically available amount of urban green space, as well as in demographic and socio-economic status, and housing conditions. Four dimensions of green space attachment were identified through confirmatory factor analysis: place dependence, affective attachment, place identity and social bonding. The results show greater attachment to local green space and better self-reported mental health in the neighborhood with higher availability of accessible and usable green spaces. The two neighborhoods did not differ, however, in physical and general health. Structural Equation Modelling confirmed the neighborhood differences in green space attachment and mental health, and also revealed a positive path from green space attachment to mental health. These findings convey the message that we should make green places, instead of green spaces.