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.     

Triterpene saponin hemi-biosynthesis of a leaf beetle’s (<Emphasis Type="Italic">Platyphora kollari</Emphasis>) defensive secretion

The adults of the leaf beetle Platyphora kollari (Chrysomelidae) are able to metabolise the oleanane triterpene β-amyrin (1) into the glycoside 3-O-β-d-glucopyranosyl-(1→4)-β-d-glucuronopyranosyl-hederagenin (2) that is stored in their defensive glands. The aim of this study was to test the hypothesis that oleanolic acid (3) is an intermediate in the conversion of 1 into 2 and to check whether the sequestration of pentacyclic triterpenes is selective in favour of β-amyrin (1). To this end, adults of P. kollari were fed with Ipomoea batatas leaf disks painted with a solution of [2,2,3-²H₃]oleanolic acid or [2,2,3-²H₃]α-amyrin and the secretion of their defensive glands analysed by HPLC–ESIMS. The data presented in this work indicated that the first step of the transformation of β-amyrin (1) into the sequestered glycoside 2 is its oxidation into oleanolic acid (3) and that this conversion is selective but not specific in favour of β-amyrin (1).     

Time allocation between work and recreation and the associated injury risks among young people

PROBLEM: The purpose of this study was to examine the relationship between overall risk of injury and time use patterns between work and active recreation among adolescents and young adults. METHODS: Using a representative sample of 9,795 Canadians aged 15-24 years, a multivariate logistic regression on the likelihood of any medically attended injury was conducted, including sociodemographic, individual, and time factors. RESULTS: Young people who engaged in a combination of high work and high activity hours were twice as likely to sustain a medically attended injury compared to those who worked low hours, but did not participate in any recreational activity. Those respondents who were not in school had a 43% increase in injury risk compared to full-time students. SUMMARY: Our findings suggest that injury risk was not a simple function of fatigue and cumulative exposure time. Our findings suggest the importance of considering time use and the associated injury risk tradeoffs. IMPACT ON INDUSTRY: Relevant to state and federal work safety policy makers, our findings suggest the importance of understanding where youth might otherwise spend their time if constraint were placed on their employment opportunities, and the associated injury risk tradeoffs must be taken into consideration.     

A direct observation study of nighttime safety belt use in Indiana

INTRODUCTION: Nearly all direct observation studies of safety belt use are conducted exclusively during daylight hours. Recent work has suggested that safety belt use at night may differ from daytime belt use. METHODS: An observational study of nighttime safety belt use, utilizing specialized night vision equipment, was conducted in Indiana surrounding the Click It or Ticket 2006 safety belt mobilization activities. A pre- and a post-mobilization statewide direct observation survey was conducted at night coinciding with daytime safety belt use data collection conducted by the state of Indiana. Daytime and nighttime belt use rates were compared. RESULTS: The comparisons across the mobilization period revealed a significant increase during the day, but a significant decrease at night. Comparisons between daytime and nighttime belt use revealed no overall difference during the pre wave, but a significant difference during the post wave. Finally, many common daytime trends in belt use were also found at night, with the exception of the typical age and seating position effects. DISCUSSION: The mobilization activities had a positive effect on daytime belt use, but no effect on nighttime belt use, likely resulting in the differences between daytime and nighttime belt use observed during the post wave. IMPACT ON INDUSTRY: The findings of this study suggest that safety belt mobilizations implemented only during the day do not influence nighttime safety belt use. Changes to how these programs are implemented or additional programs specifically targeting belt use at night should be considered, along with continued monitoring of nighttime belt use.     

Male satin bowerbirds (<Emphasis Type="Italic">Ptilonorhynchus violaceus</Emphasis>) compensate for sexual signal loss by enhancing multiple display features

Numerous studies have focussed on the relationship between female choice and the multiple exaggerated sexual traits of males. However, little is known about the ability of males to actively enhance specific components of their display in response to the loss of one component. We investigated the capacity of male satin bowerbirds (Ptilonorhynchus violaceus) to respond to the loss of one of their sexual signals by performing an experiment in which we removed decorations at their bowers. We found that males compensated for decoration loss by increasing bower construction behaviour and decreasing their latency to bower painting. These results are novel because they suggest that males can assess the quality of their own display and make decisions about how to augment their displays. We discuss these results in the context of previous studies of mate choice in satin bowerbirds, as both of the supplementary behaviours we observed are known correlates of male mating success.     

Improving graduated driver licensing systems: a conceptual approach and its implications

Graduated driver licensing (GDL) is a concept for how to transform non-drivers into reasonably safe drivers while minimizing the risks as they learn. Several state GDL programs can be improved by moving their structures closer to an adequate implementation of that concept. The learner stage of a GDL system needs to be long enough for beginners to obtain a thorough introduction to the vagaries of driving. The second or intermediate stage needs to effectively limit exposure to known high risk conditions as novices adapt to being fully in charge of the vehicle. The benefits of GDL to date are due almost entirely to the risk-reducing conditions it implements. To improve the functioning of GDL will probably require a better understanding of teen driving than we presently have. The likelihood of further gains will be enhanced by efforts to learn more about the actual causes of teen crashes, the nature and type of teen driver exposures, and what parents do with their teens during the supervised driving stage of GDL. Without a better understanding of these, and other, phenomena it will be difficult to further reduce crashes among young beginning drivers, whether through GDL enhancements or with other approaches.     

Ultrastructure of spermatozoa of <Emphasis Type="Italic">Onthophagus taurus</Emphasis> (Coleoptera,Scarabaeidae) exhibits heritable variation

Sperm competition is thought to be an important selective pressure shaping sperm form and function. However, few studies have moved beyond gross examinations of sperm morphology. Sperm length is subject to sexual selection via sperm competition in the scarab beetle Onthophagus taurus. Here, the structure and ultrastructure of spermatozoa in this species were investigated using light and electron microscopy. Spermatozoa were found to be filiform, measuring about 1,200 mm in length. The sperm head consists of a three-layered acrosome and a nuclear region bearing the anterior extension of the centriole adjunct. Acrosome and nuclear regions are bilaterally symmetric, with their axes of symmetry being orthogonal to each other. Head and flagellar structures are connected by a well-developed centriole adjunct. The sperm heads are asymmetrically surrounded by accessory material and embedded into the cytoplasm of the spermatocyst cell. The accessory material is produced inside the spermatids and then transferred to the outside due to a new membrane formed around the sperm’s organelles. The old spermatid membrane separates the accessory material from the cyst cell. The flagellum contains a 9+9+2 axoneme, two accessory bodies, and two mitochondrial derivatives of unequal size. The major mitochondrial derivative is significantly larger than the minor one. The axoneme is arranged in a sinusoidal manner parallel along the major mitochondrial derivative. The spermatozoa show no progressive motility when released in buffer solution which is likely to be the result of the flagellar arrangement and the structure of the major mitochondrial derivative. The cross-sectional area of the minor and the major mitochondrial derivatives show different patterns of genetic variation. The data provide the first estimates of genetic variation in sperm ultrastructure for any species, and give evidence for the persistence of genetic variation in ultrastructure required for the rapid and divergent evolution that characterizes spermatozoa generally.     

Impact on sward composition and stock performance of grazing Molinia-dominant grassland

A comparative experiment was conducted to test the extent to which grazing of rank Molinia caerulea-dominated grassland increases botanical diversity and improves animal performance. The study was carried out on an area of under-grazed Molinia-dominant pasture that had not been stocked for 20 years. Three treatments were compared from 2001 until 2008: (1) no grazing, (2) grazing by Welsh Mountain sheep and (3) grazing by Welsh Black heifers. Patterns of change in sward surface composition across the grazing season were consistent with greater consumption of Molinia by the cattle, but there was little evidence of management regime influencing changes in sward composition year-on-year. There was a significant effect of year on animal performance (P < 0.001). Although the growth rates of the heifers improved over the course of the first three years of the experiment, this performance was not sustained. In contrast, any weight changes for the sheep were positive. The results highlight the slow pace of change within upland swards, and the need to develop site-specific grazing guidelines.     

Application of winery and distillery waste composts to a Jumilla (SE Spain) vineyard: Effects on the characteristics of a calcareous sandy-loam soil

The effects on different physico-chemical, chemical and biological soil properties of the incorporation of different organic materials into a calcareous vineyard soil were studied in an experiment during three growing seasons. The organic materials used were sheep manure and four composts derived from the treatment of winery and distillery wastes. A multivariate technique, factor analysis (FA), was also used for interpreting the data obtained from the samplings carried out throughout the three vine growing seasons. The application of the organic materials induced an increase in the activity of the soil microorganisms and in the soil macro and micronutrient contents, as well as a slow release of inorganic N. Through FA of the soil properties studied, three factors that explained 74.7% of the variability were established and the effects of the treatments after the whole experimental period were assessed.