Power matters in closing the phenotyping gap

Much of our understanding of physiology and metabolism is derived from investigating mouse mutants and transgenic mice, and open-access platforms for standardized mouse phenotyping such as the German Mouse Clinic (GMC) are currently viewed as one powerful tool for identifying novel gene-function relationships. Phenotyping or phenotypic screening involves the comparison of wild-type control mice with their mutant or transgenic littermates. In our study, we explored the extent to which standardized phenotyping will succeed in detecting biologically relevant phenotypic differences in mice generated and provided by different collaborators. We analyzed quantitative metabolic data (body mass, energy intake, and energy metabolized) collected at the GMC under the current workflow, and used them for statistical power considerations. Our results demonstrate that there is substantial variability in these parameters among lines of wild-type C57BL/6 (B6) mice from different sources. Given this variable background noise in mice that serve as controls, subtle phenotypes in mutant or transgenic littermates may be overlooked. Furthermore, a phenotype observed in one cohort of a mutant line may not be reproducible (to the same extent) in mice coming from a different environment or supplier. In the light of these constraints, we encourage researchers to incorporate information on intrastrain variability into future study planning, or to perform advanced hierarchical analyses. Both will ultimately improve the detectability of novel phenotypes by phenotypic screening. Carola W. Meyer and Ralf Elvert contributed equally to this work.     

Impaired visibility: the air pollution people see

Almost every home and office contains a portrayal of a scenic landscape whether on a calendar, postcard, photograph, or painting. The most sought after locations boast a scenic landscape right outside their window. No matter what the scene – mountains, skyscrapers, clouds, or pastureland – clarity and vividness are essential to the image. Air pollution can degrade scenic vistas, and in extreme cases, completely obscure them. Particulate matter suspended in the air is the main cause of visibility degradation. Particulate matter affects visibility in multiple ways: obscures distant objects, drains the contrast from a scene, and discolors the sky. Visibility is an environmental quality that is valued for aesthetic reasons that are difficult to express or quantify. Human psychology and physiology are sensitive to visual input. Visibility has been monitored throughout the world but there are few places where it is a protected resource. Existing health-based regulations are weak in terms of visibility protection. Various techniques, including human observation, light transmission measurements, digital photography, and satellite imaging, are used to monitor visibility. As with air pollution, trends in visibility vary spatially and temporally. Emissions from the developing world and large scale events such as dust storms and wildfires affect visibility around much of the globe.     

Impacts after four years of experimental trampling on alpine/sub-alpine environments in western Tasmania

Experimental trials were undertaken over four years to assess the impact of recreational trampling in undisturbed alpine and sub-alpine vegetation communities in the Western Arthur Range, western Tasmania. Data on 'pad' formation due to human trampling were collected using vegetation cover assessments, biomass estimates and detailed cross-sectional surface profiles. In sub-alpine buttongrass and alpine herbfield, prolonged and sustained damage may occur after 100 passes by walkers. The environmental threshold of the flat alpine herbfield site was breached after 200 passes. Plant morphology was one determinant of resistance and resilience, with upright woody shrubs and tall tussock graminoids most vulnerable to sustained trampling damage. Cushions are susceptible to trampling impacts at 500 passes. Loss of vegetation cover peaks 6-12 months after trampling. Our results show that pads formed with as few as 30-100 passes per annum and tracks form at between 100 and 500 passes per annum. Two years after the cessation of trampling, there is some small recovery in vegetation cover after 30 and 100 passes per annum applied for three years, but no evidence of recovery at the 500 pass treatments. The low trampling threshold and slow recovery rates in western Tasmania suggest that concentrating walkers on a minimal number of sites may be the best management option for these untracked alpine and sub-alpine environments.     

N-nitrosodimethylamine and trihalomethane formation and minimisation in Southeast Queensland drinking water

This study assesses the prevalence of disinfection by-product (DBP) precursors in some Southeast Queensland drinking water sources by conducting formation potential experiments for the four regulated trihalomethanes (THMs), and the potent carcinogen, N-nitrosodimethylamine (NDMA). NDMA formation potentials were consistently low (<5-21 ng/L), and total THM (tTHM) formation potentials were consistently below the Australian Drinking Water Guideline (250 μg/L). NDMA concentration of finished drinking waters was also monitored and found to be <5 ng/L in all cases. The effect of coagulation and advanced oxidation on the formation of NDMA and THMs is also reported. UV/H(2)O(2) pre-treatment was effective in producing water with very low THMs concentrations, and UV irradiation was an effective method for NDMA degradation. H(2)O(2) was not required for the observed NDMA degradation to occur. Coagulation using alum, ferric chloride or poly(diallyldimethylammonium chloride) (polyDADMAC) was ineffective in removing DBPs precursors from the source water studied, irrespective of the low dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) attained. Rather, coagulation with polyDADMAC caused an increase in NDMA formation potential upon chloramination, and all coagulants led to an increased tTHM formation potential upon chlorination due to the high bromide concentration of the source water studied.     

Anti-sense expression of putrescine N-methyltransferase confirms defensive role of nicotine in Nicotiana sylvestris against Manduca sexta

Summary. Several lines of evidence support the defensive function of nicotine production in the Nicotiana genus against a range of herbivores, but the evidence is largely correlative. To suppress nicotine production in planta and to test its defensive function, we expressed DNA of putrescine N-methyl transferase in an anti-sense orientation (AS-PMT) in N. sylvestris and fed leaf material from two lines of transformed and wild type plants to Manduca sexta larvae. Larvae consumed more leaf area and gained more mass on the foliage of plants with low PMT expression and low nicotine levels as compared to plants with high PMT expression and high nicotine levels and wild type plants. Overall, larval consumption and performance were negatively correlated with constitutive nicotine levels. We conclude that nicotine decreases the palatability of N. sylvestris leaves to the nicotine-resistant M. sexta larvae.