Evolutionary constraints of warning signals: A genetic trade-off between the efficacy of larval and adult warning coloration can maintain variation in signal expression.

To predict evolutionary responses of warning signals under selection, we need to determine the inheritance pattern of the signals, and how they are genetically correlated with other traits contributing to fitness. Furthermore, protective coloration often undergoes remarkable changes within an individual's lifecycle, requiring us to quantify the genetic constraints of adaptive coloration across all the relevant life stages. Based on a 12 generation pedigree with > 11,000 individuals of the wood tiger moth (Arctia plantaginis), we show that high primary defense as a larva (large warning signal) results in weaker defenses as adult (less efficient warning color), due to the negative genetic correlation between the efficacy of larval and adult warning coloration. However, production of effective warning coloration as a larva did not incur any life-history costs and was positively genetically correlated with reproductive output. These results provide novel insights into the evolutionary constraints on protective coloration in animals, and explain the maintenance of variation in the signal expression despite the strong directional selection by predators. By analyzing the genetic and environmental effects on warning signal and life-history traits in all relevant life stages, we can accurately determine the mechanisms shaping the evolutionary responses of phenotypic traits under different selection environments.

All-polymer photonic sensing platform based on whispering-gallery mode microgoblet lasers.

We present an all-polymer photonic sensing platform based on whispering-gallery mode microgoblet lasers integrated into a microfluidic chip. The chip is entirely made from polymers, enabling the use of the devices as low-cost disposables. The microgoblet cavities feature quality factors exceeding 10(5) and are fabricated from poly(methyl methacrylate) (PMMA) using spin-coating, mask-based optical lithography, wet chemical etching, and thermal reflow. In contrast to silica-based microtoroid resonators, this approach replaces technically demanding vacuum-based dry etching and serial laser-based reflow techniques by solution-based processing and parallel thermal reflow. This enables scaling to large-area substrates, and hence significantly reduces device costs. Moreover, the resonators can be fabricated on arbitrary substrate materials, e.g., on transparent and flexible polymer foils. Doping the microgoblets with the organic dye pyrromethene 597 transforms the passive resonators into lasers. Devices have lasing thresholds below 0.6 nJ per pulse and can be efficiently pumped via free-space optics using a compact and low-cost green laser diode. We demonstrate that arrays of microgoblet lasers can be readily integrated into a state-of-the-art microfluidic chip replicated via injection moulding. In a proof-of-principle experiment, we show the viability of the lab-on-a-chip via refractometric sensing, demonstrating a bulk refractive index sensitivity (BRIS) of 10.56 nm per refractive index unit.

Molecular and ecological signs of mitochondrial adaptation: consequences for introgression?

The evolution of the mitochondrial genome and its potential adaptive impact still generates vital debates. Even if mitochondria have a crucial functional role, as they are the main cellular energy suppliers, mitochondrial DNA (mtDNA) introgression is common in nature, introducing variation in populations upon which selection may act. Here we evaluated whether the evolution of mtDNA in a rodent species affected by mtDNA introgression is explained by neutral expectations alone. Variation in one mitochondrial and six nuclear markers in Myodes glareolus voles was examined, including populations that show mtDNA introgression from its close relative, Myodes rutilus. In addition, we modelled protein structures of the mtDNA marker (cytochrome b) and estimated the environmental envelopes of mitotypes. We found that massive mtDNA introgression occurred without any trace of introgression in the analysed nuclear genes. The results show that the native glareolus mtDNA evolved under past positive selection, suggesting that mtDNA in this system has selective relevance. The environmental models indicate that the rutilus mitotype inhabits colder and drier habitats than the glareolus one that can result from local adaptation or from the geographic context of introgression. Finally, homology models of the cytochrome b protein revealed a substitution in rutilus mtDNA in the vicinity of the catalytic fraction, suggesting that differences between mitotypes may result in functional changes. These results suggest that the evolution of mtDNA in Myodes may have functional, ecological and adaptive significance. This work opens perspective onto future experimental tests of the role of natural selection in mtDNA introgression in this system.

Sex-specific selection on energy metabolism--selection coefficients for winter survival.

Selection for different fitness optima between sexes is supposed to operate on several traits. As fitness-related traits are often energetically costly, selection should also act directly on the energetics of individuals. However, efforts to examine the relationship between fitness and components of the energy budget are surprisingly scarce. We investigated the effects of basal metabolic rate (BMR, the minimum energy required for basic life functions) and body condition on long-term survival (8 winter months) with manipulated densities in enclosed populations of bank voles (Myodes glareolus). Here, we show that survival selection on BMR was clearly sex-specific but density-independent. Both the linear selection gradient and selection differential for BMR were positive in females, whereas survival did not correlate with male characteristics. Our findings emphasize the relative importance of individual physiology over ecological factors (e.g. intra-specific competition). Most current models of the origin of endothermy underline the importance of metabolic optima in females, whose physiology evolved to fulfil demands of parental provisioning in mammals. Our novel findings of sex-specific selection could be related to these life history differences between sexes.

Flexible fluidic microchips based on thermoformed and locally modified thin polymer films.

This paper presents a fundamentally new approach for the manufacturing and the possible applications of lab on a chip devices, mainly in the form of disposable fluidic microchips for life sciences applications. The new technology approach is based on a novel microscale thermoforming of thin polymer films as core process. The flexibility not only of the semi-finished but partly also of the finished products in the form of film chips could enable future reel to reel processes in production but also in application. The central so-called 'microthermoforming' process can be surrounded by pairs of associated pre- and postprocesses for micro- and nanopatterned surface and bulk modification or functionalisation of the formed films. This new approach of microscale thermoforming of thin polymer film substrates overlaid with a split local modification of the films is called 'SMART', which stands for 'substrate modification and replication by thermoforming'. In the process, still on the unformed, plane film, the material modifications of the preprocess define the locations where later, then on the spatially formed film, the postprocess generates the final local modifications. So, one can obtain highly resolved modification patterns also on hardly accessible side walls and even behind undercuts. As a first application of the new technology, we present a flexible chip-sized scaffold for three dimensional cell cultivation in the form of a microcontainer array. The spatially warped container walls have been provided with micropores, cell adhesion micropatterns and thin film microelectrodes.

A large panel of novel microsatellite markers for the bank vole (Myodes glareolus).

We describe a set of 66 highly polymorphic microsatellite loci isolated from the bank vole, Myodes (Clethrionomys) glareolus. These microsatellites were characterized for a long-term study on periodically fluctuating density of the bank vole population in Central Finland. We detected six to 38 alleles per locus in the population sampled at two different density phases, and the levels of observed and expected heterozygosities varied between 0.17 and 1.00, and between 0.72 and 0.95, respectively. This microsatellite panel serves as an informative tool for population and molecular genetic studies.

Eimeria-parasites are associated with a lowered mother's and offspring's body condition in island and mainland populations of the bank vole.

This study, based on correlative data, tests the hypothesis that infections with Eimeria spp. parasites exert a significant loss of fitness of bank voles (Clethrionomys glareolus) reflected in lower reproductive success and survival, declining host population densities and are associated positively with population size. The study was conducted in 20 mainland and 27 island populations in central Finland during May-September in 1999. Faecal samples showed that 28% of 767 individuals were infected with Eimeria spp. The presence of Eimeria parasites was higher in dense mainland populations than in sparsely populated islands. Eimerian infections increased during the course of the breeding season, probably as a result of the high infection rate of young individuals. Accordingly, the body masses of bank voles were negatively related to the presence of Eimeria spp. Reproductive output, as measured by the breeding probability of females and litter size, was not associated with the presence of eimerian infection. Interestingly, the body condition of the infected mothers appeared to be low. Moreover, mother's body condition was the single most important variable studied that showed a positive correlation to pup's body condition at birth. On small islands (< or =3.2 ha) that were comprehensively trapped, the mean number of Eimeria spp. in the bank vole population was negatively related to density changes of the bank vole population during the study. Our data are consistent with the idea that infection with coccidian parasites may be one of the factors responsible for declining host populations in small, isolated populations.

Optimal allocation of reproductive effort: manipulation of offspring number and size in the bank vole.

The number of offspring attaining reproductive age is an important measure of an individual's fitness. However, reproductive success is generally constrained by a trade-off between offspring number and quality. We conducted a factorial experiment in order to study the effects of an artificial enlargement of offspring number and size on the reproductive success of female bank voles (Clethrionomys glareolus). We also studied the effects of the manipulations on growth, survival and reproductive success of the offspring. Potentially confounding effects of varying maternal quality were avoided by cross-fostering. Our results showed that the number of offspring alive in the next breeding season was higher in offspring number manipulation groups, despite their smaller body size at weaning. Offspring size manipulation had no effect on offspring growth or survival. Further, the first litter size of female offspring did not differ between treatments. In conclusion, females may be able to increase the number of offspring reaching reproductive age by producing larger litters, whereas increasing offspring size benefits neither the mother nor the offspring.

Does risk of predation by mammalian predators affect the spacing behaviour of rodents? Two large-scale experiments.

Predator-prey interactions between small mammals and their avian and mammalian predators have attracted much attention. However, large-scale field experiments examining small-mammal antipredatory responses under the risk of predation by mammals are rare. As recently pointed out, the scale of experiments may cause misleading results in studies of decision-making under predation risk. We studied the effect of small mustelid predators on the spacing behaviour of the gray-tailed vole (Microtus canicaudus) and the bank vole (Clethrionomys glareolus) in two separate field enclosure experiments. The experiments were conducted during the breeding season in North America and northern Europe, where small mustelids have been suggested to be important mammalian predators of voles. As in most of the earlier laboratory studies, predation risk was simulated using fresh mustelid faeces and urine. This made it possible to compare the results from experiments at different spatial scales. We did not find any effect of increased predation risk on spacing behaviour (mean and/or maximum distance moved and home range size) or trappability in either vole species. Simulated predation risk did not affect the breeding of females in gray-tailed voles, as has previously been shown in bank voles. The results disagree with most of the studies conducted in laboratory conditions with small mammals. We discuss whether this discrepancy could be caused by differences in the scale of the experiments.

Maternal effort and male quality in the bank vole, Clethrionomys glareolus.

Parental investment in reproduction is adjusted according to potential benefits in terms of offspring survival and/or mating success. If male quality affects the reproductive success of a female, then females mating with high-quality males should invest more in reproduction. Although the subject has been of general interest, further experimental verification of the hypothesis is needed. We studied whether female bank voles (Clethrionomys glareolus) adjusted their maternal effort according to male quality, measured as mating success. To enable the measurement of maternal effort during nursing separately from male genetic effects the litters were cross-fostered. Further, the genetic background of male quality was examined. Male quality did not correlate with litter size or offspring size at birth. Offspring growth was positively related to food consumption and milk production of mothers. However, these direct measurements of maternal effort were independent of male quality. Male mating success appeared to be significantly heritable indicating that there are genetic benefits. Still, females did not adjust maternal effort according to the genetic quality of their offspring. We suggest that female bank voles gain significant genetic benefits from mating with high-quality males whereas they cannot improve their reproductive success by increasing maternal effort.

High population density in bank voles stimulates food hoarding after breeding.

The effects of conspecific density (i.e. risk of intraspecific competition) on hoarding behaviour were studied in breeding pairs of bank voles, Clethrionomys glareolus. I simulated high population density by using odours of conspecifics, to exclude the direct effects of exploitation or interference competition for food. The pairs of bank voles hoarded only at the end of their breeding season. Hoarding was not correlated with whether the voles had a litter during the experiment or their litter size, but was more likely at the high population density and for pairs with small females. This may indicate that smaller females especially make caches in the autumn, possibly to ensure better winter survival under high population density. I discuss the hoarding behaviour of bank voles in relation to their territorial behaviour and food resources during the breeding season and predictable population changes during winter. Copyright 1998 The Association for the Study of Animal Behaviour. Copyright 1998 The Association for the Study of Animal Behaviour.

Reproductive costs and litter size in the bank vole.

The potential reproductive costs for free-ranging bank vole (Clethrionomys glareolus) offspring and mothers were assessed by manipulating litter size and by determining the effects of nursing varied numbers of offspring. Litter enlargement did not increase the number of weanlings per mother. The mass of juveniles was significantly lower in the enlarged litters and higher in the reduced litters, compared to the control group. However, the survival of juveniles from weaning aged three months did not depend on their mass at weaning. Data from a previous study (Mappes et al. 1995) indicated that a higher mass at weaning may increase juveniles' abilities to maturate and breed during their summer of birth. Manipulation of litter size did not significantly affect the mass or survival of mothers or the success of subsequent breeding. The size of home ranges did correlate positively with the initial litter size. However, space use by females did not change with the degree of manipulation. Our results indicate that females nursing enlarged litters produce smaller offspring at weaning with no residual effects on future maternal survival or reproduction. Mothers did not seem to compensate the nursing costs with increased parental effort (which should be reflected in the condition of mothers or in the use of resources), for example, in an enlarged size of home range. Probably the possibility of obtaining a larger home range is constrained by the other breeding females, in a saturated breeding population. These results may support the optimal investment hypothesis that a female will produce a particular litter size which gives the best reproductive success in the particular environment where offspring are nursed.

Patient choices, family interests, and physician obligations.

Recent articles in biomedical ethics have begun to explore both the relevance of family interests in treatment decisions and the resultant ramifications for physicians' obligations to patients. This article addresses two important questions regarding physicians' obligations vis-a-vis family interests: (1) What should a physician do when the exercise of patient autonomy threatens to negate the patient's moral obligations to other family members? (2) Does respect for patient autonomy typically require efforts on the part of physicians to keep patients' treatment decisions from being influenced by family considerations? A series of clarifications about the concept of autonomy is also presented.

Ectoparasites, nest site choice and breeding success in the pied flycatcher.

It has recently been suggested that nest box studies might bias the measurement of behavioural and life-history traits, because the removal of old nests may reduce the load of ectoparasites. This experimental artefact may have notable effects on nest site choice and breeding success in cavity-breeding birds. We tested (i) if pied flycatchers Ficedula hypoleuca prefer clean nest boxes and (ii) if old nest material affects the number of parasites and the breeding success of pied flycatchers. In the first experiment we offered birds one cleaned nest box and one nest box with old nest material from the previous year. The two nest boxes were placed in very similar sites near each other. In this experiment all pied flycatchers clearly preferred "dirty" nest boxes. In the second part of the study we distributed clean nest boxes and dirty ones on the same study area. After breeding was over we counted the number of fleas Ceratophyllus gallinae in the nest material. This flea species was the most abundant and probably the most hazardous parasite in the nests. Surprisingly, we found that there were significantly more fleas in the nest boxes with nests of the current year only than in the boxes with nests of both current and previous year. This might explain the preference for the dirty boxes. However, our results do not indicate that the number of fleas affects breeding success in the pied flycatcher.

Different demography of friends and strangers: an experiment on the impact of kinship and familiarity in Clethrionomys glareolus.

We examined demographic effects of familiarity and relatedness in the bank vole Clethrionomys glareolus (Schreber) in four 0.5-ha enclosures in Central Finland. In two enclosures were mature voles which had overwintered together and some of their mature off-spring (hereafter referred to as "Friends"), and in the other two individuals of the same species captured from different localities near the study area ("Strangers"). The experiment lasted from June to September. The populations of Friends reached densities twice as high as those of Strangers with a significantly higher rate of recruitment and survival of the young. This may have been due to mutual familiarity decreasing antagonism towards the juveniles. The conflicting results obtained from studies of Clethrionomys and Microtus are discussed. We believe that these genera represent behavioural adaptations to different habitats and ways of life. Most behavioural population regulation hypotheses are based on studies of Microtus. We conclude that these results should be applied with great caution to other rodent genera.