Avoiding chromosome pathology when replication forks collide

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ 2013 Macmillan Publishers Limited.Chromosome duplication normally initiates through the assembly of replication fork complexes at defined origins1, 2. DNA synthesis by any one fork is thought to cease when it meets another travelling in the opposite direction, at which stage the replication machinery may simply dissociate before the nascent strands are finally ligated. But what actually happens is not clear. Here we present evidence consistent with the idea that every fork collision has the potential to threaten genomic integrity. In Escherichia coli this threat is kept at bay by RecG DNA translocase3 and by single-strand DNA exonucleases. Without RecG, replication initiates where forks meet through a replisome assembly mechanism normally associated with fork repair, replication restart and recombination4, 5, establishing new forks with the potential to sustain cell growth and division without an active origin. This potential is realized when roadblocks to fork progression are reduced or eliminated. It relies on the chromosome being circular, reinforcing the idea that replication initiation is triggered repeatedly by fork collision. The results reported raise the question of whether replication fork collisions have pathogenic potential for organisms that exploit several origins to replicate each chromosome.THe MRC, the Leverhulme Trust, and the BBSRC

The impact of modularisation strategies on small modular reactor cost

Small Modular Reactors (SMRs) based on established light-water technology have gained a lot of attention from the nuclear industry; however, the potential that SMRs have to reduce the cost of nuclear construction has been under-studied. Modularisation is a cost reducing mechanism where a SMR power plant is subdivided into smaller units, or modules. These modules can be produced offsite in a controlled environment, potentially offering cost reductions that offset their apparently higher capital costs. This paper will investigate the effects modularisation and standardisation might have on SMR capital costs. Modularisation and standardisation not only reduce direct and indirect costs, respectively, but also enable activation of other cost-reducing mechanisms, such as shifting construction work from site to a factory, transferring learning between tasks, and achieving economies of multiples. It will show that constructing a SMR using the same methods as current large reactors is not economically feasible and will demonstrate how modularisation reduces SMR capital costs. The primary constraints on module size are imposed by weight and height transport limitations, linking reactor size to ease of modularisation. This leads to an analysis of which SMR components and structures should be targeted for modularisation in order to achieve optimal cost benefits

Influences on diet and physical activity choices of 11–13-year-olds in a school setting

This is the author accepted manuscript. The final version is available from SAGE Publications via the DOI in this record.Objective: This paper uses a qualitative approach to explore the factors that influence diet and physical activity choices of 11–13-year-olds with a particular focus on the impact of the school environment. Design: Qualitative focus groups. Setting: Three purposively sampled secondary schools in Devon, UK. Method: A total of 53 students, aged 11–13, took part in six focus groups. Thematic, framework analysis was used to analyse the data. Result: Four overarching themes emerged: (1) health now and in the future; (2) the role of others; (3) provision, temptation and addiction; and (4) boundaries, strategies and support. Participants demonstrated good knowledge of what constitutes a healthy lifestyle and its importance for future health, although it was not necessarily seen as a priority at this stage of life. Key influences on their choices were their peers and family, although participants also identified that the school environment influences the food choices they make while there. Conclusion: In this study, 11–13-year-olds identified that schools could do more to support them to make healthier food choices. However, future research needs to understand the constraints schools face in terms of food provision in order to highlight possible opportunities for intervention.All Saints Educational TrustNational Institute for Health research (NIHR

Variations in soil chemical and physical properties explain basin-wide Amazon forest soil carbon concentrations

We investigate the edaphic, mineralogical and climatic controls of soil organic carbon (SOC) concentration utilising data from 147 primary forest soils (0–30 cm depth) sampled in eight different countries across the Amazon Basin. Sampled across 14 different World Reference Base soil groups, our data suggest that stabilisation mechanism varies with pedogenetic level. Specifically, although SOC concentrations in Ferralsols and Acrisols were best explained by simple variations in clay content – this presumably being due to their relatively uniform kaolinitic mineralogy – this was not the case for less weathered soils such as Alisols, Cambisols and Plinthosols for which interactions between Al species, soil pH and litter quality are argued to be much more important. Although for more strongly weathered soils the majority of SOC is located within the aggregate fraction, for the less weathered soils most of the SOC is located within the silt and clay fractions. It thus seems that for highly weathered soils SOC storage is mostly influenced by surface area variations arising from clay content, with physical protection inside aggregates rendering an additional level of protection against decomposition. On the other hand, most of the SOC in less weathered soils is associated with the precipitation of aluminium–carbon complexes within the fine soil fraction, with this mechanism enhanced by the presence of high levels of aromatic, carboxyl-rich organic matter compounds. Also examined as part of this study were a relatively small number of arenic soils (viz. Arenosols and Podzols) for which there was a small but significant influence of clay and silt content variations on SOM storage, with fractionation studies showing that particulate organic matter may account for up to 0.60 of arenic soil SOC. In contrast to what were in all cases strong influences of soil and/or litter quality properties, after accounting for these effects neither wood productivity, above-ground biomass nor precipitation/temperature variations were found to exert any significant influence on SOC stocks. These results have important implications for our understanding of how Amazon forest soils are likely to respond to ongoing and future climate changes

Tropical tree branch-leaf nutrient scaling relationships vary with sampling location

Bivariate relationships between plant tissue nutrient concentration have largely been studied across broad environmental scales regardless of their covariation with soil and climate. Comparing leaf and branch wood concentrations of C, Ca, K, Mg, N, Na, and P for trees growing in tropical forests in Amazonia and Australia we found that the concentrations of most elements varied with sampling location, but with foliar and branch woody tissues varying from site to site in different ways. Using a Mixed Effect Model (MEM) approach it was further found that relationships between branch and leaf concentrations within individual plots differed in terms of both slope and/or significance to the ordinary least squares (OLS) estimates for most elements. Specifically, using MEM we found that within plots only K and Mg were correlated across organs, but with the K cross-organ intercept estimates varying significantly between sites. MEM analyses further showed that within-plot wood density variations were also negatively related to wood K and Na, suggesting a potentially important role for these cations in water transport and/or storage in woody tissues. The OLS method could not detect significant correlations in any of the above cases. By contrast, although Ca, N, and P leaf and wood tissue concentrations showed similar patterns when individual elements were compared across sites, MEM analyses suggested no consistent association within sites. Thus, for all these three elements, strong within-tree scaling relationships were inferred when data were analyzed across sites using OLS, even though there was no relationship within individual sites. Thus (as for Ca, N, and P) not only can a pooling of data across sites result in trait (co)variations attributable to the environment potentially being incorrectly attributed solely to the species and/or individual (the so-called “ecological fallacy”), but in some cases (as was found here for K and Na) the opposite can also sometimes occur with significant within-site covariations being obscured by large site-site variations. We refer to the latter phenomenon as “environmental obfuscation.

Securing recruitment and obtaining informed consent in minority ethnic groups in the UK

Background: Previous health research has often explicitly excluded individuals from minority ethnic backgrounds due to perceived cultural and communication difficulties, including studies where there might be language/literacy problems in obtaining informed consent. This study addressed these difficulties by developing audio-recorded methods of obtaining informed consent and recording data. This report outlines 1) our experiences with securing recruitment to a qualitative study investigating alternative methods of data collection, and 2) the development of a standardised process for obtaining informed consent from individuals from minority ethnic backgrounds whose main language does not have an agreed written form. Methods: Two researchers from South Asian backgrounds recruited adults with Type 2 diabetes whose main language was spoken and not written, to attend a series of focus groups. A screening tool was used at recruitment in order to assess literacy skills in potential participants. Informed consent was obtained using audio-recordings of the patient information and recording patients' verbal consent. Participants' perceptions of this method of obtaining consent were recorded. Results: Recruitment rates were improved by using telephone compared to face-to-face methods. The screening tool was found to be acceptable by all potential participants. Audio-recorded methods of obtaining informed consent were easy to implement and accepted by all participants. Attrition rates differed according to ethnic group. Snowballing techniques only partly improved participation rates. Conclusion: Audio-recorded methods of obtaining informed consent are an acceptable alternative to written consent in study populations where literacy skills are variable. Further exploration of issues relating to attrition is required, and a range of methods may be necessary in order to maximise response and participation

Functional near infrared spectroscopy (fNIRS) to assess cognitive function in infants in rural Africa

Cortical mapping of cognitive function during infancy is poorly understood in low-income countries due to the lack of transportable neuroimaging methods. We have successfully piloted functional near infrared spectroscopy (fNIRS) as a neuroimaging tool in rural Gambia. Four-to-eight month old infants watched videos of Gambian adults perform social movements, while haemodynamic responses were recorded using fNIRS. We found distinct regions of the posterior superior temporal and inferior frontal cortex that evidenced either visual-social activation or vocally selective activation (vocal > non-vocal). The patterns of selective cortical activation in Gambian infants replicated those observed within similar aged infants in the UK. These are the first reported data on the measurement of localized functional brain activity in young infants in Africa and demonstrate the potential that fNIRS offers for field-based neuroimaging research of cognitive function in resource-poor rural communities

Implementing a set of health promoting processes in English secondary schools: A comparative case study

This is the final version. Available on open access from Elsevier via the DOI in this recordObjective To understand the enablers and barriers to implementing a set of adaptive processes aimed at supporting secondary schools to reflect on and subsequently address how they could adjust school practices, culture and the environment to create a whole school approach to promoting healthy lifestyles. Study design A qualitative, comparative case study. Methods Two in depth case studies were created of two purposefully selected schools in low socio-economic areas of South West England. Data were collected via meetings, observations, field notes, interviews and audit. Interviews were transcribed verbatim. Individual thematic analyses were conducted for each school and a comparative analysis approach was used to understand the barriers and enablers across both cases. Results Schools were supported to use a health-promoting lens and identify feasible improvements through an adaptive and context specific process. The school environment and ethos were identified as the areas where schools could conceive the most adjustments to enhance the promotion of healthy lifestyle choices. With the lack of government policy for health promotion in schools (HPS), the Head teacher's approach to health was key to making meaningful changes. Conclusions Health promoting school approaches need to be adaptive to local context, actively involve community partners and link to local initiatives where possible, with support from Head teachers and business managers. Starting with what teachers, pupils and parents see as the barriers to health can create a whole school ethos for broad reaching and sustainable HPS programmes.All Saints Education Trust (ASET)National Institute for Health Research (NIHR

Functional near infrared spectroscopy (fNIRS) to assess cognitive function in infants in rural Africa

Cortical mapping of cognitive function during infancy is poorly understood in low-income countries due to the lack of transportable neuroimaging methods. We have successfully piloted functional near infrared spectroscopy (fNIRS) as a neuroimaging tool in rural Gambia. Four-to-eight month old infants watched videos of Gambian adults perform social movements, while haemodynamic responses were recorded using fNIRS. We found distinct regions of the posterior superior temporal and inferior frontal cortex that evidenced either visual-social activation or vocally selective activation (vocal > non-vocal). The patterns of selective cortical activation in Gambian infants replicated those observed within similar aged infants in the UK. These are the first reported data on the measurement of localized functional brain activity in young infants in Africa and demonstrate the potential that fNIRS offers for field-based neuroimaging research of cognitive function in resource-poor rural communities

The pestivirus N terminal protease N(pro) redistributes to mitochondria and peroxisomes suggesting new sites for regulation of IRF3 by N(pro.)

The N-terminal protease of pestiviruses, N(pro) is a unique viral protein, both because it is a distinct autoprotease that cleaves itself from the following polyprotein chain, and also because it binds and inactivates IRF3, a central regulator of interferon production. An important question remains the role of N(pro) in the inhibition of apoptosis. In this study, apoptotic signals induced by staurosporine, interferon, double stranded RNA, sodium arsenate and hydrogen peroxide were inhibited by expression of wild type N(pro), but not by mutant protein N(pro) C112R, which we show is less efficient at promoting degradation of IRF3, and led to the conclusion that N(pro) inhibits the stress-induced intrinsic mitochondrial pathway through inhibition of IRF3-dependent Bax activation. Both expression of N(pro) and infection with Bovine Viral Diarrhea Virus (BVDV) prevented Bax redistribution and mitochondrial fragmentation. Given the role played by signaling platforms during IRF3 activation, we have studied the subcellular distribution of N(pro) and we show that, in common with many other viral proteins, N(pro) targets mitochondria to inhibit apoptosis in response to cell stress. N(pro) itself not only relocated to mitochondria but in addition, both N(pro) and IRF3 associated with peroxisomes, with over 85% of N(pro) puncta co-distributing with PMP70, a marker for peroxisomes. In addition, peroxisomes containing N(pro) and IRF3 associated with ubiquitin. IRF3 was degraded, whereas N(pro) accumulated in response to cell stress. These results implicate mitochondria and peroxisomes as new sites for IRF3 regulation by N(pro), and highlight the role of these organelles in the anti-viral pathway