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.     

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.     

Implementation of Washington State's zero tolerance law: patterns of arrests, dispositions, and recidivism

OBJECTIVES: Zero tolerance (ZT) laws have been effective in reducing alcohol-related crashes among underage drivers. However, enforcement in some states has not been rigorous, and ZT offenses may not be viewed as serious offenses. On July 1, 1994, the state of Washington implemented a ZT law that allowed police to request a test for alcohol on suspicion of either a ZT or driving-under-the-influence (DUI) offense. The present study examined effects of the ZT law on arrests and case dispositions among underage offenders as a function of blood alcohol concentration (BAC) and post-law patterns of recidivism. METHODS: Times-series analyses examined the effects of the ZT law on trends in arrests of underage drivers between 1991 and 1999. Based on arrest records matched with driver's license records, the effects of the law on dispositions of alcohol-related offenses among underage drivers were examined, and rates of recidivism among underage offenders were examined for the period following the ZT law. RESULTS: There was a substantial increase in arrests of underage drivers beginning immediately after implementation of the ZT law, especially among drivers with low BACs. The types of court or administrative dispositions received by underage offenders changed markedly after the ZT law was implemented. Underage offenders with lower BACs became far more likely to receive alcohol-related convictions and/or license suspensions. However, the percentage of underage offenders with higher BACs receiving DUI convictions declined as some of these offenders received the lesser ZT disposition. After the ZT law, underage offenders with BACs of 0.10 g/dL or higher were more likely to recidivate than those with lower BACs, but appreciable proportions of drivers were re-arrested for another alcohol offense, whatever the BAC and however they were penalized. CONCLUSIONS: Implementation of Washington's law indicates that a ZT law can increase the likelihood that an underage person will be sanctioned for drinking and driving. However, recidivism remains an issue as more than one in four underage drivers arrested with low BACs subsequently were re-arrested.     

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.     

Preventing alcohol-related convictions: the effect of a novel curriculum for first-time offenders on DUI recidivism

OBJECTIVE: To determine whether DUI offenders can better avoid future drinking and driving by controlling their vehicle usage rather than by controlling their drinking. METHODS: Using a randomized experimental post-test only design, 9,571 first-time DUI offenders were randomly assigned to receive one of two 12-hour educational programs: a traditional DUI curriculum or the PARC (Preventing Alcohol-Related Convictions) curriculum, which uses a novel theoretical approach to preventing DUI recidivism. Whereas traditional programs focus on participants controlling their drinking to avoid future drinking and driving, the PARC curriculum focuses on participants controlling their driving. Instead of trying to control alcohol consumption after driving to a drinking venue (previously found to be a flawed strategy), PARC teaches students to make a decision before leaving home not to drive to a drinking event, thus greatly limiting the possibility of drinking and driving. Driving records were obtained from the Florida Department of Motor Vehicles using driver's license numbers to assess DUI recidivism rates among the students in the PARC and Traditional curricula for the first year following program participation and again at 2 years post-intervention. RESULTS: Binary logistic regression analyses revealed that offenders receiving the PARC curriculum exhibited significantly lower 1-year and 2-year recidivism rates than those receiving the Traditional curriculum. The effect was consistent across two different measures of recidivism, and across gender, race, ethnicity, and location. CONCLUSION: Results suggest that the PARC educational approach may be more effective than the traditional approach in reducing DUI recidivism.     

Nuptial coloration of red shiners (<Emphasis Type="Italic">Cyprinella lutrensis</Emphasis>) is more intense in turbid habitats

Communication is shaped and constrained by the signaling environment. In aquatic habitats, turbidity can reduce both the quantity and quality of ambient light and has been implicated in the breakdown of visual signaling. Here, we examined the relationship between turbidity (quantified with long-term data) and the expression of carotenoid-based nuptial coloration in the red shiner (Cyprinella lutrensis), a small-bodied cyprinid. Males in more turbid habitats displayed redder fins, and an experimental manipulation of adult diet suggested that carotenoid intake alone did not explain among-population color differences. These results run counter to similar studies where signal expression decreased in turbid conditions, and may be explained by the non-territorial red shiner mating system, interactions between the mechanism of coloration and the signaling environment, or reduced cost of color expression in turbid habitats (e.g., reduced predation risk). Our results highlight how the behavioral and ecological contexts in which signals function can shape evolutionary responses to the environment.     

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.     

Anatomical localization and stereoisomeric composition of <Emphasis Type="Italic">Tribolium castaneum</Emphasis> aggregation pheromones

We report that the abdominal epidermis and associated tissues are the predominant sources of male-produced pheromones in the red flour beetle, Tribolium castaneum and, for the first time, describe the stereoisomeric composition of the natural blend of isomers of the aggregation pheromone 4,8-dimethyldecanal (DMD) in this important pest species. Quantitative analyses via gas chromatography–mass spectrometry showed that the average amount of DMD released daily by single feeding males of T. castaneum was 878 ± 72 ng (SE). Analysis of different body parts identified the abdominal epidermis as the major source of aggregation pheromone; the thorax was a minor source, while no DMD was detectable in the head. No internal organs or obvious male-specific glands were associated with pheromone deposition. Complete separation of all four stereoisomers of DMD was achieved following oxidation to the corresponding acid, derivatization with (1R, 2R)- and (1S, 2S)-2-(anthracene-2,3-dicarboximido)cyclohexanol to diastereomeric esters, and their separation on reversed-phase high-performance liquid chromatography at −54°C. Analysis of the hexane eluate from Porapak-Q-collected volatiles from feeding males revealed the presence of all four isomers (4R,8R)/(4R,8S)/(4S,8R)/(4S,8S) at a ratio of approximately 4:4:1:1. A walking orientation bioassay in a wind tunnel with various blends of the four synthetic isomers further indicated that the attractive potency of the reconstituted natural blend of 4:4:1:1 was equivalent to that of the natural pheromone and greater than that of the 1:1 blend of (4R,8R)/(4R,8S) used in commercial lures.     

Heterospecific pairing and hybridization between <Emphasis Type="Italic">Nasutitermes corniger</Emphasis> and <Emphasis Type="Italic">N. ephratae</Emphasis>

The sympatric neotropical termites Nasutitermes corniger and Nasutitermes ephratae are clearly distinguishable based on morphology, nest architecture, defensive secretion composition, and molecular markers. However, given the extensive ecological, geographical, and behavioral overlap of these closely related species, the potential for interbreeding may exist. To explore this possibility, heterospecific pairs were formed experimentally to examine courtship and colony-establishment behaviors, and reproductive potential. Courtship and nest construction behavior occurred in heterospecific pairs in a similar manner to that of conspecific pairs. Survival of pairs depended upon the species of the female partner. N. ephratae females paired with N. corniger males produced as many offspring as conspecific pairs. N. corniger females mated to N. ephratae males, however, produced significantly fewer offspring at 60 days post-establishment than the reciprocal cross or conspecific N. ephratae or N. corniger pairs. This was also the only pairing in which any aggression was observed. Heterospecific pairs and groups formed in mate choice mesocosms, suggesting that species recognition between these two termites is not an important aspect of mate choice. Overall, species mismatch tolerance and hybrid offspring viability are high. The present data, together with previous evidence from defensive secretions and isozyme analysis, suggest that hybridization may periodically occur in nature, and that reproductive barriers between these two species may be incomplete. Hybridization could provide a rare but important source of genetic diversity and may ensure mating opportunities for the more abundant sex of alates in each species.