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).     

Observed LATCH use and misuse characteristics of child restraint systems in seven states

INTRODUCTION: Although the LATCH System (Lower Anchors and Tethers for Children) holds the promise of simplifying the installation of a child restraint system (CRS) to the vehicle's seat, many drivers transporting young children have difficulties using this technology. This paper reports on an observation study of LATCH use and misuse. METHOD: Observations of approximately 1,000 children less than 5 years of age in CRSs, in the back seats of vehicles that were equipped with tether and lower anchors, in seven states. RESULTS: Tethers were used for 51% of the children when the forward-facing CRS had tether straps and the vehicle had tether anchors. Lower anchors were used for 58% of the children when the CRS had lower attachments and the vehicle had lower anchors. The most common tether and lower attachment misuses were loose tether straps (18% of cases) and loose lower attachment installation (30% of the cases), respectively. Vehicle safety belts were used in combination with lower attachments in 20% of all lower anchor installations. CONCLUSION: As more caregivers of young children drive vehicles equipped with LATCH, it will be important to promote the proper installation of CRSs using this technology. LATCH education messages must also emphasize that the lower anchors may not always be the safest choice for CRS attachment -- the safest attachment is the one that results in a tight fit and will be used correctly consistently.     

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.     

A new THOR shoulder design: a comparison with volunteers, the Hybrid III, and THOR NT

OBJECTIVE: Since the shoulders are rarely seriously injured in frontal or oblique collisions, they have been given low priority in the development of frontal impact crash test dummies. The shoulder complex geometry and its kinematics are of vital importance for the overall dummy kinematics. The shoulder complex also influences the risk of the safety belt slipping off the shoulder in oblique forward collisions. The first aim of this study was to develop a new 50th percentile male THOR shoulder design, while the second was to compare the new shoulder, mounted on a THOR NT dummy, with volunteer, THOR NT, and Hybrid III range of motion and stiffness data. The third aim was to test the repeatability of the new shoulder during dynamic testing and to see how the design behaves with respect to belt slippage in a 45 degrees far-side collision. METHODS: The new 50th percentile THOR shoulder design was developed with the aid of a shell model of the seated University of Michigan Transportation Research Institute (UMTRI) 50th percentile male with coordinates for joints and bony landmarks (Schneider et al., 1983). The new shoulder design has human-like bony landmarks for the acromion and coracoid processes. The clavicle curvature and length are also made similar to that of a male human, as is the range of motion in the anterior-posterior, superior-inferior, and medial-lateral directions. The new shoulder design was manufactured and tested under the same conditions that T?rnvall et al. (2005b) used to compare the shoulder range of motion for the volunteers, Hybrid III, and THOR Alpha. The new design was also tested in two dynamic test configurations: the first was a 0 degrees full-frontal test and the second was a 45 degrees far-side test. The dummy tests were conducted with an R-16 seat with a three-point belt, the Delta V was 27.0+/-0.5 km/h and the maximum peak acceleration was approximately 14.6+/-0.5 g for each test. RESULTS: A new shoulder design with geometry close to that of humans was developed to be retrofitted to the THOR NT dummy. The results showed that the range of motion for the new shoulder complex during static loading was larger by at least a factor of three, for the maximum load (200 N/arm), than that of either the Hybrid III or the THOR NT; this means it was more similar to the volunteers' range of motion. It was observed that the THOR NT with the new shoulder did not slide out of the shoulder belt during a far-side collision. The performance of the new shoulder was reasonably repeatable and stable during both the static tests and the sled tests. CONCLUSION: A new shoulder for the THOR NT has been designed and developed, and data from static range of motion tests and sled tests indicate that the new shoulder complex has the potential to function in a more human-like manner on the THOR dummy.     

Experience,but not distance,influences the recruitment precision in the stingless bee <Emphasis Type="Italic">Scaptotrigona mexicana</Emphasis>

Recruitment precision, i.e. the proportion of recruits that reach an advertised food source, is a crucial adaptation of social bees to their environment. Studies with honeybees showed that recruitment precision is not a fixed feature, but it may be enhanced by factors like experience and distance. However, little is known regarding the recruitment precision of stingless bees. Hence, in this study, we examined the effects of experience and spatial distance on the precision of the food communication system of the stingless bee Scaptotrigona mexicana. We conducted the experiments by training bees to a three-dimensional artificial patch at several distances from the colony. We recorded the choices of individual recruited foragers, either being newcomers (foragers without experience with the advertised food source) or experienced (foragers that had previously visited the feeder). We found that the average precision of newcomers (95.6 ± 2.61%) was significantly higher than that of experienced bees (80.2 ± 1.12%). While this might seem counter-intuitive on first sight, this “loss” of precision can be explained by the tendency of experienced recruits to explore nearby areas to find new rewarding food sources after they had initially learned the exact location of the food source. Increasing the distance from the colony had no significant effect on the precision of the foraging bees. Thus, our data show that experience, but not the distance of the food source, affected the patch precision of S. mexicana foragers.     

Loss of legs: is it or not a handicap for an orb-weaving spider?

Leg loss is a common phenomenon in spiders, and according to the species 5% to 40% of the adults can present at least one missing leg. There is no possibility of regeneration after adult moult and the animal must manage with its missing appendages until its death. With the loss of one or more legs, female orb-weaving spiders can be penalized twice: firstly, because the legs are necessary for web construction and secondly, the legs are essential for the control of the prey after its interception by the web. During development, spiders may be also penalized because regeneration has energetic costs that take away resources for survival, growth and reproduction. All these consequences should influence negatively the development of the spider and thus its fitness. We investigated the impact of leg loss in the orb-weaving spider, Zygiella x-notata by studying its frequency in a natural population and web building and prey capture behaviours in laboratory. In field populations, 9.5% to 13%, of the adult females presented the loss of one or more legs; the majority of individuals had lost only one leg (in 48% of cases, a first one). Leg loss seems to affect all the adult spiders, as there is no difference of mass between intact spiders and those with missing leg. Data obtained with laboratory-reared spiders, showed that the loss of legs due to the moult is rare (less than 1%). Considering changes in web design, spiders with missing legs decreased their silk investment, increased the distance between spiral turns but did not change the capture surface of the web. Under our laboratory experimental conditions, spiders with one or two lost legs did not present any difference in prey capture efficiency. In laboratory conditions, spiders with lost leg(s) did not show any difference in egg sac production or in longevity (adult lifespan) compared to intact spiders.     

An individual and a sex odor signature in kittiwakes? Study of the semiochemical composition of preen secretion and preen down feathers

The importance of olfaction in birds’ social behavior has long been denied. Avian chemical signaling has thus been relatively unexplored. The black-legged kittiwake provides a particularly appropriate model for investigating this topic. Kittiwakes preferentially mate with genetically dissimilar individuals, but the cues used to assess genetic characteristics remain unknown. As in other vertebrates, their body odors may carry individual and sexual signatures thus potentially reliably signaling individual genetic makeup. Here, we test whether body odors in preen gland secretion and preen down feathers in kittiwakes may provide a sex and an individual signature. Using gas chromatography and mass spectrometry, we found that male and female odors differ quantitatively, suggesting that scent may be one of the multiple cues used by birds to discriminate between sexes. We further detected an individual signature in the volatile and nonvolatile fractions of preen secretion and preen down feathers. These results suggest that kittiwake body odor may function as a signal associated with mate recognition. It further suggests that preen odor might broadcast the genetic makeup of individuals, and could be used in mate choice to assess the genetic compatibility of potential mates.     

Improvement of soil aggregate stability by repeated applications of organic amendments to a cultivated silty loam soil

The objective of this study was to compare the effects of repeated field applications of three urban compost amendments and one farmyard manure amendment over a 9-year period on aggregate stability in a silty loam soil initially characterized by low clay and initial organic matter contents and poor aggregate stability. Three different aggregate stability tests with increasing disruptive intensities (fast wetting > mechanical breakdown > slow wetting tests) and different disaggregation mechanisms, were used. All of the amendments, which were applied at approximately 4 Mg C ha⁻¹ every other year, increased the organic carbon content and improved the stability of the aggregates against the disruptive action of water, as determined by each of the stability tests. However, the year-to-year variations in the aggregate stability that related to factors other than the organic inputs were greater than the cumulative increase in aggregate stability relative to the control. The positive effects of the tested amendments on aggregate stability were linked to their contribution to soil organic C contents (r = 0.54 for the fast wetting test and r = 0.41-0.42 for the mechanical breakdown and slow wetting tests; p < 0.05). The addition of urban composts had a larger positive effect on aggregate stability than farmyard manure at the majority of sampling dates. The addition of biodegradable immature compost, such as municipal solid waste (MSW), improved the aggregate stability through an enhanced resistance to slaking. The addition of mature composts, such as the co-compost of sewage sludge and green wastes (GWS) or biowaste compost (BW), improved the aggregate stability by increasing interparticular cohesion. The MSW compost was the most efficient in improving aggregate stability during the first 6 years of the experiment (average improvements of +22%, +5% and +28% in the fast wetting, mechanical breakdown and slow wetting tests, respectively, compared to the control treatment); this result was likely due to the larger labile organic pool of the MSW compost that was highly effective at stimulating soil microbial activity. After the first 6 years, the two other composts, GWS and BW, became more efficient (average improvements of +25%, +61% and +33% in the fast wetting, mechanical breakdown and slow wetting tests, respectively, compared to the control treatment), which was probably linked to the greater increase in soil organic C contents. Therefore, the application of urban compost to silty soil that is susceptible to water erosion was effective at improving aggregate stability and thus could be used to enhance the resistance of soil to water erosion.     

Associated skeletons of a new middle Triassic "Rauisuchia" from Brazil

For more than 30 million years, in early Mesozoic Pangea, ??rauisuchian?? archosaurs were the apex predators in most terrestrial ecosystems, but their biology and evolutionary history remain poorly understood. We describe a new ??rauisuchian?? based on ten individuals found in a single locality from the Middle Triassic (Ladinian) Santa Maria Formation of southern Brazil. Nine articulated and associated skeletons were discovered, three of which have nearly complete skulls. Along with sedimentological and taphonomic data, this suggests that those highly successful predators exhibited some kind of intraspecific interaction. Other monotaxic assemblages of Triassic archosaurs are Late Triassic (Norian-Rhaetian) in age, approximately 10 million years younger than the material described here. Indeed, the studied assemblage may represent the earliest evidence of gregariousness among archosaurs, adding to our knowledge on the origin of a behavior pattern typical of extant taxa.