mRNA Extraction from Gill Tissue for RNA-sequencing

Adaptation is thought to proceed in part through spatial and temporal changes in gene expression. Fish species such as the threespine stickleback are powerful vertebrate models to study the genetic architecture of adaptive changes in gene expression since divergent adaptation to different environments is common, they are abundant and easy to study in the wild and lab, and have well-established genetic and genomic resources. Fish gills, due to their respiratory and osmoregulatory roles, show many physiological adaptations to local water chemistry, including differences in gene expression. However, obtaining high-quality RNA using popular column-based extraction methods can be challenging from small tissue samples high in cartilage and bone such as fish gills. Here, we describe a bead-based mRNA extraction and transcriptome RNA-seq protocol that does not use purification columns. The protocol can be readily scaled according to sample size for the purposes of diverse gene expression experiments using animal or plant tissue.Peer reviewe

Distribution and Habitats of Mosquito Larvae in the Kingdom of Tonga

Mosquitoes are a significant pest and human health issue in the Kingdom of Tonga. The occurrence of species and habitats used by mosquito larvae were investigated to determine the potential for control through larval habitat management. Forty-two sites, including 22 villages and 20 farm plantations on the six islands of Tongatapu, Pangaimotu, Vava’u, Pangaimotu (Vava’u group), ‘Utungake and Nuku, were surveyed in April 2006. A total of eight mosquito species were collected: Aedes aegypti (Linnaeus), Ae. horrescens (Edwards), Ae. nocturnus (Theobold), Ae. tongae (Edwards), Culex albinervis (Edwards), Cx. annulirostris (Skuse), Cx. quinquefasciatus (Say) and Cx. sitiens (Wiedemann). Several species were widespread, particularly Ae. aegypti and Ae. nocturnus on the main island of Tongatapu, whereas Ae. aegypti dominated sites on islands of the Vava’u group. Comparative sampling of 17 village and 17 rural sites showed that larval habitat was more abundant in towns than in rural areas. Larvae were found in a wide range of habitats but were particularly abundant in artificial water bodies (e.g. disused concrete water tanks, 44-gallon drums and used car tyres). In rural sites, habitats were generally sparse except in rain-filled branch stems of giant taro plants. Mosquito populations in artificial habitats could be markedly reduced by seeding disused water tanks with aquatic predators already present in Tonga, using mesh-net covers over 44-gallon drums, and drilling holes in used car tyres

Distribution and Habitats of Mosquito Larvae in the Kingdom of Tonga

Mosquitoes are a significant pest and human health issue in the Kingdom of Tonga. The occurrence of species and habitats used by mosquito larvae were investigated to determine the potential for control through larval habitat management. Forty-two sites, including 22 villages and 20 farm plantations on the six islands of Tongatapu, Pangaimotu, Vava’u, Pangaimotu (Vava’u group), ‘Utungake and Nuku, were surveyed in April 2006. A total of eight mosquito species were collected: Aedes aegypti (Linnaeus), Ae. horrescens (Edwards), Ae. nocturnus (Theobold), Ae. tongae (Edwards), Culex albinervis (Edwards), Cx. annulirostris (Skuse), Cx. quinquefasciatus (Say) and Cx. sitiens (Wiedemann). Several species were widespread, particularly Ae. aegypti and Ae. nocturnus on the main island of Tongatapu, whereas Ae. aegypti dominated sites on islands of the Vava’u group. Comparative sampling of 17 village and 17 rural sites showed that larval habitat was more abundant in towns than in rural areas. Larvae were found in a wide range of habitats but were particularly abundant in artificial water bodies (e.g. disused concrete water tanks, 44-gallon drums and used car tyres). In rural sites, habitats were generally sparse except in rain-filled branch stems of giant taro plants. Mosquito populations in artificial habitats could be markedly reduced by seeding disused water tanks with aquatic predators already present in Tonga, using mesh-net covers over 44-gallon drums, and drilling holes in used car tyres

How do increasing background concentrations of tropospheric ozone affect peatland plant growth and carbon gas exchange?

In this study we have demonstrated that plants originating from upland peat bogs are sensitive to increasing background concentrations of ozone. Peatland mesocosms from an upland peat bog in North Wales, UK were exposed to eight levels of elevated background ozone in solardomes for 4 months from May to August, with 24 h mean ozone concentrations ranging from 16 to 94 ppb and cumulative AOT024hr ranging from 45.98 ppm h to 259.63 ppm h. Our results show that plant senescence increased with increasing exposure to ozone, although there was no significant effect of increasing ozone on plant biomass. Assessments of carbon dioxide and methane fluxes from the mesocosms suggests that there was no change in carbon dioxide fluxes over the 4 month exposure period but that methane fluxes increased as cumulative ozone exposure increased to a maximum AOT 024hr of approximately 120 ppm h and then decreased as cumulative ozone exposure increased further

Workplace interventions to prevent suicide:A scoping review

Objectives: To map organisational interventions for workplace suicide prevention, identifying the effects, mechanisms, moderators, implementation and economic costs, and how interventions are evaluated. Background: Suicide is a devastating event that can have a profound and lasting impact on the individuals and families affected, with the highest rates found among adults of work age. Employers have a legal and ethical responsibility to provide a safe working environment for their employees, which includes addressing the issue of suicide and promoting mental health and well-being. Methods: A realist perspective was taken, to identify within organisational suicide prevention interventions, what works, for whom and in what circumstances. Published and unpublished studies in six databases were searched. To extract and map data on the interventions the Effect, Mechanism, Moderator, Implementation, Economic (EMMIE) framework was used. Mechanisms were deductively analysed against Bronfenbrenner’s socio-ecological model. Results: From 3187 records screened, 46 papers describing 36 interventions within the military, healthcare, the construction industry, emergency services, office workers, veterinary surgeons, the energy sector and higher education. Most mechanisms were aimed at the individual’s immediate environment, with the most common being education or training on recognising signs of stress, suicidality or mental illness in oneself. Studies examined the effectiveness of interventions in terms of suicide rates, suicidality or symptoms of mental illness, and changes in perceptions, attitudes or beliefs, with most reporting positive results. Few studies reported economic costs but those that did suggested that the interventions are cost-effective. Conclusions: It seems likely that organisational suicide prevention programmes can have a positive impact on attitudes and beliefs towards suicide as well reducing the risk of suicide. Education, to support individuals to recognise the signs and symptoms of stress, mental ill health and suicidality in both themselves and others, is likely to be an effective starting point for successful interventions

Surface melt on the Shackleton Ice Shelf, East Antarctica (2003–2021)

Melt on the surface of Antarctic ice shelves can potentially lead to their disintegration, accelerating the flow of grounded ice to the ocean and raising global sea levels. However, the current understanding of the processes driving surface melt is incomplete, increasing uncertainty in predictions of ice shelf stability and thus of Antarctica's contribution to sea-level rise. Previous studies of surface melt in Antarctica have usually focused on either a process-level understanding of melt through energy-balance investigations or used metrics such as the annual number of melt days to quantify spatiotemporal variability in satellite observations of surface melt. Here, we help bridge the gap between work at these two scales. Using daily passive microwave observations from the AMSR-E and AMSR-2 sensors and the machine learning approach of a self-organising map, we identify nine representative spatial distributions (“patterns”) of surface melt on the Shackleton Ice Shelf in East Antarctica from 2002/03–2020/21. Combined with output from the RACMO2.3p3 regional climate model and surface topography from the REMA digital elevation model, our results point to a significant role for surface air temperatures in controlling the interannual variability in summer melt and also reveal the influence of localised controls on melt. In particular, prolonged melt along the grounding line shows the importance of katabatic winds and surface albedo. Our approach highlights the necessity of understanding both local and large-scale controls on surface melt and demonstrates that self-organising maps can be used to investigate the variability in surface melt on Antarctic ice shelves.</p

Predominance of cis-regulatory changes in parallel expression divergence of sticklebacks

Regulation of gene expression is thought to play a major role in adaptation but there are conflicting predictions for the relative importance of cis- and trans-regulatory mechanisms in the early stages adaptive divergence. Parallel evolution of marine and freshwater threespine stickleback fish provides an excellent opportunity to dissect whether the same molecular mechanisms underlie repeated adaptive divergence in gene expression. Using RNA-seq of four marine-freshwater ecotype pairs from Scotland and Canada, we first identified genes with parallel divergence in expression and show that these are found near previously reported adaptive loci and show a molecular signature of selection centered around the transcription start site. With allele-specific expression assays in F1 hybrids we next show that expression divergence is predominantly driven by cis-regulatory control in all four river systems, a pattern that is enriched in parallel divergently expressed genes. In particular, for genes whose expression is up-regulated in parallel among freshwater fish the quantitative degree of cis- and trans-regulation is also highly correlated, suggesting a shared genetic basis across populations. This stands in contrast to genes up-regulated in parallel in marine fish, whose degree of cis- and trans-regulation is less correlated and predictable. This observed asymmetry in parallelism in how genes are up-regulated in marine and freshwater fish can be explained by differences in the evolutionary contexts of the diverging ecotypes. Finally, we show that cis-regulation is predominantly additive and shows greater robustness to different in genetic backgrounds and environmental conditions. We argue that these features make cis-regulation well-poised for rapid adaptive divergence of gene expression under conditions of on-going gene flow. Combined our study highlights how natural selection on dispersed cis-regulatory elements can shape the adaptive landscape of the genome.Regulation of gene expression is thought to play a major role in adaptation but the relative importance of cis- and trans- regulatory mechanisms in the early stages of adaptive divergence is unclear. Using RNAseq of threespine stickleback fish gill tissue from four independent marine-freshwater ecotype pairs and their F1 hybrids, we show that cis-acting (allele-specific) regulation consistently predominates gene expression divergence. Genes showing parallel marine-freshwater expression divergence are found near to adaptive genomic regions, show signatures of natural selection around their transcription start sites and are enriched for cis-regulatory control. For genes with parallel increased expression among freshwater fish, the quantitative degree of cis- and trans-regulation is also highly correlated across populations, suggesting a shared genetic basis. Compared to other forms of regulation, cis-regulation tends to show greater additivity and stability across different genetic and environmental contexts, making it a fertile substrate for the early stages of adaptive evolution.Peer reviewe

Building Peace through Sports Projects: A Scoping Review

The term peacebuilding has gained traction in academic works since introduction in the 1960s. In recent decades, sport for development and peace (SDP) has also captured the interest of the academic community, with a growing field of work. This scoping review identifies and considers the academic literature on SDP projects deployed as peacebuilding tools in post-conflict communities, to gain a greater understanding of those projects and draw inferences from them collectively. Using strict inclusion criteria, results of database searches were narrowed down to 30 publications, which the review explored through comparing the publications and their findings, to reveal the range of disciplines this research is emerging from, the countries projects are operating in, the demographics targeted, and other key data. The resulting conclusion is that there is scope for more targeted studies to clarify specific demographics to include, whether there is an ideal age to engage with youth, or an optimal timeframe for involvement. Many of the publications reference the importance of being part of broader initiatives, but the best context in which to utilise sport, and how much of an impact is being made on the wider communities, is yet to be determined

An assessment of a portable cyanoacrylate fuming system (LumiFume™) for the development of latent fingermarks

The effectiveness and suitability of a portable cyanoacrylate fuming system (LumiFume™) with Lumicyano™ at detecting latent fingermarks was assessed. The first phase of the study compared the LumiFume™ system with traditional cabinet fuming and black/white powder suspension for the development of latent fingermarks on a variety of surfaces (glass, plastic bin bag, laminated wood and tile) by means of depletion series’ from 10 donors and four ageing periods (1, 7, 14 and 28 days). The portable fuming system provided superior quality of developed marks on glass and laminated wood whereas powder suspension was better on bin bags and all three techniques were comparable on tile. A decrease in mark quality was recorded from 1-14 days for the fuming techniques before an increase at 28 days. Lumicyano™ fluorescence stability studies over a 28 day period by means of depletion series’ on glass slides and plastic bin bags revealed better quality marks for the portable system LumiFume™; however, storing marks under light conditions expedited deterioration for both systems. All marks developed with Lumicyano™ were subsequently treated with BY40 resulting in further improvement in mark quality for all substrates and ageing periods, with the exception of laminated wood which absorbed the fluorescent stain reducing the contrast in the process. The second phase of the study consisted of a pseudo-operational trial on 300 various substrates (e.g. glass bottles, aluminium cans, plastic bags) recovered from recycling bins. LumiFume™ and Lumicyano™ yielded 1,469 marks whereas Lumicyano™ cabinet fuming and powder suspension yielded 1,026 and 641 marks respectively. Similar to the first phase of the study, further treatment of the Lumicyano™ treated marks with BY40 resulted in further quality improvement as well as additional new marks. The LumiFume™ system produced results at least equivalent to the traditional cabinet fuming with Lumicyano™ highlighting its potential for implementation into casework to process crime scenes