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1.  3D facial landmarks: Inter-operator variability of manual annotation 
BMC Medical Imaging  2014;14:35.
Background
Manual annotation of landmarks is a known source of variance, which exist in all fields of medical imaging, influencing the accuracy and interpretation of the results. However, the variability of human facial landmarks is only sparsely addressed in the current literature as opposed to e.g. the research fields of orthodontics and cephalometrics. We present a full facial 3D annotation procedure and a sparse set of manually annotated landmarks, in effort to reduce operator time and minimize the variance.
Method
Facial scans from 36 voluntary unrelated blood donors from the Danish Blood Donor Study was randomly chosen. Six operators twice manually annotated 73 anatomical and pseudo-landmarks, using a three-step scheme producing a dense point correspondence map. We analyzed both the intra- and inter-operator variability, using mixed-model ANOVA. We then compared four sparse sets of landmarks in order to construct a dense correspondence map of the 3D scans with a minimum point variance.
Results
The anatomical landmarks of the eye were associated with the lowest variance, particularly the center of the pupils. Whereas points of the jaw and eyebrows have the highest variation. We see marginal variability in regards to intra-operator and portraits. Using a sparse set of landmarks (n=14), that capture the whole face, the dense point mean variance was reduced from 1.92 to 0.54 mm.
Conclusion
The inter-operator variability was primarily associated with particular landmarks, where more leniently landmarks had the highest variability. The variables embedded in the portray and the reliability of a trained operator did only have marginal influence on the variability. Further, using 14 of the annotated landmarks we were able to reduced the variability and create a dense correspondences mesh to capture all facial features.
doi:10.1186/1471-2342-14-35
PMCID: PMC4205300  PMID: 25306436
3D Facial landmarks; Inter-operator annotation variance; Dense point correspondence; Point distribution mode; ANOVA
2.  Integrated phenotypes: understanding trait covariation in plants and animals 
Integration and modularity refer to the patterns and processes of trait interaction and independence. Both terms have complex histories with respect to both conceptualization and quantification, resulting in a plethora of integration indices in use. We review briefly the divergent definitions, uses and measures of integration and modularity and make conceptual links to allometry. We also discuss how integration and modularity might evolve. Although integration is generally thought to be generated and maintained by correlational selection, theoretical considerations suggest the relationship is not straightforward. We caution here against uncontrolled comparisons of indices across studies. In the absence of controls for trait number, dimensionality, homology, development and function, it is difficult, or even impossible, to compare integration indices across organisms or traits. We suggest that care be invested in relating measurement to underlying theory or hypotheses, and that summative, theory-free descriptors of integration generally be avoided. The papers that follow in this Theme Issue illustrate the diversity of approaches to studying integration and modularity, highlighting strengths and pitfalls that await researchers investigating integration in plants and animals.
doi:10.1098/rstb.2013.0245
PMCID: PMC4084533  PMID: 25002693
integration; modularity; variation; phenotype
3.  Genetic constraints predict evolutionary divergence in Dalechampia blossoms 
If genetic constraints are important, then rates and direction of evolution should be related to trait evolvability. Here we use recently developed measures of evolvability to test the genetic constraint hypothesis with quantitative genetic data on floral morphology from the Neotropical vine Dalechampia scandens (Euphorbiaceae). These measures were compared against rates of evolution and patterns of divergence among 24 populations in two species in the D. scandens species complex. We found clear evidence for genetic constraints, particularly among traits that were tightly phenotypically integrated. This relationship between evolvability and evolutionary divergence is puzzling, because the estimated evolvabilities seem too large to constitute real constraints. We suggest that this paradox can be explained by a combination of weak stabilizing selection around moving adaptive optima and small realized evolvabilities relative to the observed additive genetic variance.
doi:10.1098/rstb.2013.0255
PMCID: PMC4084540  PMID: 25002700
evolutionary rate; G-matrix; integration; macroevolution; microevolution; power relationship
4.  Signal honesty and cost of pollinator rewards in Dalechampia scandens (Euphorbiaceae) 
Annals of Botany  2012;109(7):1331-1340.
Background and Aims
Most species of Dalechampia vines (Euphorbiaceae) attract bee pollinators with terpenoid resins secreted by a gland-like structure in the inflorescence. In some species, pollinating bees appear to preferentially visit inflorescences (blossoms) with large resin-producing glands, whereas in other species bees preferentially visit blossoms with large involucral bracts. In this study, the reliability of bract and gland size as signals of the quantity of resin produced in one species, D. scandens, was assessed. Whether resin secretion has a cost with respect to the number or mass of the seeds produced by a blossom was also examined.
Methods
Measurements were made of bract size, gland size and the amount of resin secreted by blossoms of D. scandens maintained in a common environment, and the relationships between these traits were analysed. Resin production was also manipulated, and the effects of the manipulation were tested on seed set and seed mass.
Key Results
The amount of resin produced was better predicted by the size of the gland than by the size of the bract. Furthermore, when the effect of gland size was accounted for, bract size only weakly predicted the amount of resin produced. Neither an increase in resin secretion (by daily removal of the resin) nor a decrease (by removal of the resin gland) affected seed set or seed mass detectably, but resin production correlated positively with mean seed mass at the individual level once the size of the resin gland was accounted for.
Conclusions
Gland size is a better indicator of the amount of reward than bract size, although the latter remained an honest signal of the quantity of resin produced. Resin secretion has no detectable cost in terms of seed production, but may be condition dependent, as suggested by a positive correlation with seed mass at the individual level.
doi:10.1093/aob/mcs091
PMCID: PMC3359932  PMID: 22628366
Dalechampia scandens; blossom; signal honesty; nectar; pollination; reward; seed production; terpenoid resin; condition dependence
5.  Development of microsatellite markers for the neotropical vine Dalechampia scandens (Euphorbiaceae)1 
Applications in Plant Sciences  2013;1(6):apps.1200492.
• Premise of the study: Microsatellite markers were developed to assess polymorphism and level of genetic diversity in four Mexican populations of the neotropical vine Dalechampia scandens (Euphorbiaceae).
• Methods and Results: Thirty-seven microsatellite markers representing bi-, tri-, tetra-, and pentanucleotide microsatellite repeats were developed. In total, 166 alleles were identified across 54 individuals. The number of alleles varied from one to 11 with an average of 4.49 alleles per locus. All loci except one were highly polymorphic between populations, whereas considerably less variation was detected within populations for most loci. The average observed and expected heterozygosities across study populations ranged from 0 to 0.63 and 0 to 0.59, respectively, for individual loci, and a deviation from Hardy–Weinberg equilibrium was observed for most loci.
• Conclusions: The developed markers may be useful for studying genetic structure, parentage analysis, mapping, phylogeography, and cross-amplification in other closely related species of Dalechampia.
doi:10.3732/apps.1200492
PMCID: PMC4105026  PMID: 25202553
Dalechampia scandens; Euphorbiaceae; genetic diversity; microsatellite loci
6.  Prevalence of infective endocarditis in patients with Staphylococcus aureus bacteraemia: the value of screening with echocardiography 
Aims
Staphylococcus aureus infective endocarditis (IE) is a critical medical condition associated with a high morbidity and mortality. In the present study, we prospectively evaluated the importance of screening with echocardiography in an unselected S. aureus bacteraemia (SAB) population.
Methods and results
From 1 January 2009 to 31 August 2010, a total of 244 patients with SAB at six Danish hospitals underwent screening echocardiography. The inclusion rate was 73% of all eligible patients (n= 336), and 53 of the 244 included patients (22%; 95% CI: 17–27%) were diagnosed with definite IE. In patients with native heart valves the prevalence was 19% (95% CI: 14–25%) compared with 38% (95% CI: 20–55%) in patients with prosthetic heart valves and/or cardiac rhythm management devices (P= 0.02). No difference was found between Main Regional Hospitals and Tertiary Cardiac Hospitals, 20 vs. 23%, respectively (NS). The prevalence of IE in high-risk patients with one or more predisposing condition or clinical evidence of IE were significantly higher compared with low-risk patients with no additional risk factors (38 vs. 5%; P < 0.001). IE was associated with a higher 6 months mortality, 14(26%) vs. 28(15%) in SAB patients without IE, respectively (P < 0.05).
Conclusion
SAB patients carry a high risk for development of IE, which is associated with a worse prognosis compared with uncomplicated SAB. The presenting symptoms and clinical findings associated with IE are often non-specific and echocardiography should always be considered as part of the initial evaluation of SAB patients.
doi:10.1093/ejechocard/jer023
PMCID: PMC3117467  PMID: 21685200
Infective endocarditis; Echocardiography; Staphylococcus aureus; Screening
7.  The adaptive accuracy of flowers: measurement and microevolutionary patterns 
Annals of Botany  2009;103(9):1529-1545.
Background and Aims
From Darwin's time onward, biologists have thought about adaptation as evolution toward optimal trait values, but they have not usually assessed the relative importance of the distinct causes of deviations from optima. This problem is investigated here by measuring adaptive inaccuracy (phenotypic deviation from the optimum), using flower pollination as an adaptive system.
Methods
Adaptive accuracy is shown to have at least three distinct components, two of which are optimality (deviation of the mean from the optimum) and precision (trait variance). We then describe adaptive accuracy of both individuals and populations. Individual inaccuracy comprises the deviation of the genotypic target (the mean phenotype of a genotype grown in a range of environments) from the optimum and the phenotypic variation around that genotypic target (phenotypic imprecision). Population inaccuracy has three basic components: deviation of the population mean from the optimum, variance in the genotypic targets and phenotypic imprecision. In addition, a fourth component is proposed, namely within-population variation in the optimum. These components are directly estimable, have additive relationships, and allow exploration of the causes of adaptive inaccuracy of both individuals and populations. Adaptive accuracy of a sample of flowers is estimated, relating floral phenotypes controlling pollen deposition on pollinators to adaptive optima defined as the site most likely to get pollen onto stigmas (male inaccuracy). Female inaccuracy is defined as the deviation of the position of stigma contact from the expected location of pollen on pollinators.
Key Results
A surprising amount of variation in estimated accuracy within and among similar species is found. Some of this variation is generated by developmental changes in positions of stigmas or anthers during anthesis (the floral receptive period), which can cause dramatic change in accuracy estimates. There seem to be trends for higher precision and accuracy in flowers with higher levels of integration and dichogamy (temporal separation of sexual functions), and in those that have pollinators that are immobile (or immobilized) during pollen transfer. Large deviations from putative adaptive optima were observed, and these may be related to the effects of conflicting selective pressures on flowers, such as selection against self-pollination promoting herkogamy (spatial separation of pollen and stigmas).
Conclusions
Adaptive accuracy is a useful concept for understanding the adaptive significance of phenotypic means and variances of floral morphology within and among populations and species. Estimating and comparing the various components of adaptive accuracy can be particularly helpful for identifying the causes of inaccuracy, such as conflicting selective pressures, low environmental canalization and developmental instability.
doi:10.1093/aob/mcp095
PMCID: PMC2701747  PMID: 19429671
Adaptive accuracy; Collinsia; Dalechampia; fitness; floral precision; Linum; optimality; pollination; Stylidium

Results 1-7 (7)