Refutation of traumatic insemination in the Drosophila bipectinata species complex.

Traumatic insemination (TI) is a rare reproductive behaviour characterized by the transfer of sperm to the female via puncture wounds inflicted across her body wall. Here, we challenge the claim made by Kamimura (Kamimura 2007 Biol. Lett.3, 401-404. (doi:10.1098/rsbl.2007.0192)) that males of species of the Drosophila bipectinata complex use a pair of claw-like processes (claws) to traumatically inseminate females: the claws are purported to puncture the female body wall and genital tract, and to inject sperm through the wounds into the lumen of her genital tract, bypassing the vaginal opening. This supposed case of TI is widely cited and featured in prominent subject reviews. We examined high-resolution scanning electron micrographs of the claws and failed to discover any obvious 'groove' for sperm transport. We demonstrated that sperm occurred in the female reproductive tract as a single-integrated unit, inconsistent with the claim that sperm are injected via paired processes. Laser ablation of the sharp terminal ends of the claws failed to inhibit insemination. We showed that the aedeagus in the complex delivers sperm through the vaginal opening, as in other Drosophila. The results refute the claim of TI in the Drosophila bipectinata species complex.

Direct and indirect effects of male genital elaboration in female seed beetles.

Our understanding of coevolution between male genitalia and female traits remains incomplete. This is perhaps especially true for genital traits that cause internal injuries in females, such as the spiny genitalia of seed beetles where males with relatively long spines enjoy a high relative fertilization success. We report on a new set of experiments, based on extant selection lines, aimed at assessing the effects of long male spines on females in Callosobruchus maculatus. We first draw on an earlier study using microscale laser surgery, and demonstrate that genital spines have a direct negative (sexually antagonistic) effect on female fecundity. We then ask whether artificial selection for long versus short spines resulted in direct or indirect effects on female lifetime offspring production. Reference females mating with males from long-spine lines had higher offspring production, presumably due to an elevated allocation in males to those ejaculate components that are beneficial to females. Remarkably, selection for long male genital spines also resulted in an evolutionary increase in female offspring production as a correlated response. Our findings thus suggest that female traits that affect their response to male spines are both under direct selection to minimize harm but are also under indirect selection (a good genes effect), consistent with the evolution of mating and fertilization biases being affected by several simultaneous processes.

Individual and synergistic effects of male external genital traits in sexual selection.

Male genital traits exhibit extraordinary interspecific phenotypic variation. This remarkable and general evolutionary trend is widely considered to be the result of sexual selection. However, we still do not have a good understanding of whether or how individual genital traits function in different competitive arenas (episodes of sexual selection), or how different genital traits may interact to influence competitive outcomes. Here, we use an experimental approach based on high-precision laser phenotypic engineering to address these outstanding questions, focusing on three distinct sets of micron-scale external (nonintromittent) genital spines in male Drosophila kikkawai Burla (Diptera: Drosophilidae). Elimination of the large pair of spines on the male secondary claspers sharply reduced male ability to copulate, yet elimination of the other sets of spines on the primary and secondary claspers had no significant effects on copulation probability. Intriguingly, both the large spines on the secondary claspers and the cluster of spines on the primary claspers were found to independently promote male competitive fertilization success. Moreover, when large and small secondary clasper spines were simultaneously shortened in individual males, these males suffered greater reductions in fertilization success relative to males whose traits were altered individually, providing evidence for synergistic effects of external genital traits on fertilization success. Overall, the results are significant in demonstrating that a given genital trait (the large spines on the secondary claspers) can function in different episodes of sexual selection, and distinct genital traits may interact in sexual selection. The results offer an important contribution to evolutionary biology by demonstrating an understudied selective mechanism, operating via subtle trait interactions in a post-insemination context, by which genital traits may be co-evolving.

Interactions with ectoparasitic mites induce host metabolic and immune responses in flies at the expense of reproduction-associated factors.

Parasites cause harm to their hosts and represent pervasive causal agents of natural selection. Understanding host proximate responses during interactions with parasites can help predict which genes and molecular pathways are targets of this selection. In the current study, we examined transcriptional changes arising from interactions between Drosophila melanogaster and their naturally occurring ectoparasitic mite, Gamasodes queenslandicus. Shifts in host transcript levels associated with behavioural avoidance revealed the involvement of genes underlying nutrient metabolism. These genetic responses were reflected in altered body lipid and glycogen levels in the flies. Mite infestation triggered a striking immune response, while male accessory gland protein transcript levels were simultaneously reduced, suggesting a trade-off between host immune responses to parasite challenge and reproduction. Comparison of transcriptional analyses during mite infestation to those during nematode and parasitoid attack identified host genes similarly expressed in flies during these interactions. Validation of the involvement of specific genes with RNA interference lines revealed candidates that may directly mediate fly-ectoparasite interactions. Our physiological and molecular characterization of the Drosophila-Gamasodes interface reveals new proximate mechanisms underlying host-parasite interactions, specifically host transcriptional shifts associated with behavioural avoidance and infestation. The results identify potential general mechanisms underlying host resistance and evolutionarily relevant trade-offs.

Nutritional geometry of paternal effects on embryo mortality.

Well-established causal links exist between maternal nutritional deficits and embryo health and viability. By contrast, environmental effects operating through the father that could influence embryo mortality have seldom been examined. Yet, ejaculates can require non-trivial resource allocation, and seminal plasma components are increasingly recognized to exert wide-ranging effects on females and offspring, so paternal dietary effects on the embryo should be expected. We test for effects of varying levels of protein (P), carbohydrate (C) and caloric load in adult male diet on embryo mortality in Drosophila melanogaster We demonstrate that macronutrient balance and caloric restriction exert significant effects, and that nutritional effects are more impactful when a prior mating has occurred. Once-mated males produced embryos with marginally elevated mortality under high-caloric densities and a 1 : 8 P : C ratio. In contrast, embryos produced by twice-mated males were significantly more likely to die under male caloric restriction, an outcome that may have resulted from shifts in ejaculate quality and/or epigenetic paternal effects. Body nutrient reserves were strongly and predictably altered by diet, and body condition, in turn, was negatively related to embryo mortality. Thus, sire nutritional history and resultant shifts in metabolic state predict embryo viability and post-fertilization fitness outcomes.

Unitary and dual models of phenomenal consciousness.

There is almost unanimous consensus among the theorists of consciousness that the phenomenal character of a mental state cannot exist without consciousness. We argue for a reappraisal of this consensus. We distinguish two models of phenomenal consciousness: unitary and dual. Unitary model takes the production of a phenomenal quality and it's becoming conscious to be one and the same thing. The dual model, which we advocate in this paper, distinguishes the process in which the phenomenal quality is formed from the process that makes this quality conscious. We put forward a conceptual, methodological, neuropsychological and neural argument for the dual model. These arguments are independent but provide mutual support to each other. Together, they strongly support the dual model of phenomenal consciousness and the concomitant idea of unconscious mental qualities. The dual view is thus, we submit, a hypothesis worthy of further probing and development.

Developmental selection against developmental instability: a direct demonstration.

Developmental selection, the non-random elimination of offspring during development, is hypothesized to alter the opportunity for selection on a given trait at later stages of the life cycle. Here, we provide a direct demonstration of developmental selection against developmental instability, assessed as the incidence of minor, discrete phenotypic abnormalities in the male sex comb, a condition-dependent secondary sexual trait in Drosophila bipectinata. We exposed developing flies from two geographically separate populations to increasing levels of temperature stress, and recovered the males that died during development by teasing them out of their pupal cases. These dead males, the so-called 'invisible fraction' of the population, were more developmentally unstable than their surviving counterparts, and dramatically so under conditions of relatively high temperature stress. We illustrate that had these dead juvenile flies actually survived and entered the pool of sexually mature adult individuals, their mating success would have been significantly reduced, thus intensifying sexual selection in the adult cohort for reducing developmental instability. The data suggest that without accounting for developmental selection, a study focusing exclusively on the adult cohort may unwittingly underestimate the net force of selection operating on a given phenotypic trait.

Heritability and preadult survivorship costs of ectoparasite resistance in the naturally occurring Drosophila-Gamasodes mite system.

Our understanding of the evolutionary significance of ectoparasites in natural communities is limited by a paucity of information concerning the mechanisms and heritability of resistance to this ubiquitous group of organisms. Here, we report the results of artificial selection for increasing ectoparasite resistance in replicate lines of Drosophila melanogaster derived from a field-fresh population. Resistance, as ability to avoid infestation by naturally co-occurring Gamasodes queenslandicus mites, increased significantly in response to selection and realized heritability (SE) was estimated to be 0.11 (0.0090). Deployment of energetically expensive bursts of flight from the substrate was a main mechanism of host resistance that responded to selection, aligning with previously documented metabolic costs of fly behavioral defenses. Host body size, which affects parasitism rate in some fly-mite systems, was not shifted by selection. In contrast, resistant lines expressed significant reductions in larva-to-adult survivorship with increasing toxic (ammonia) stress, identifying an environmentally modulated preadult cost of resistance. Flies selected for resistance to G. queenslandicus were also more resistant to a different mite, Macrocheles subbadius, suggesting that we documented genetic variation and a pleiotropic cost of broad-spectrum behavioral immunity against ectoparasites. The results demonstrate significant evolutionary potential of resistance to an ecologically important class of parasites.

Efficacy of Lidocaine Infusion in High-Risk Vascular Surgery-A Randomized, Double-Blind, Placebo-Controlled Single-Center Clinical Trial.

BACKGROUND: In perioperative pain control, adjuvants such as lidocaine can reduce opioid consumption in a specific type of surgery. The aim of this single-center prospective double-blinded randomized controlled trial was to determine opioid consumption in the perioperative period in patients receiving continuous lidocaine infusion. METHODS: Patients undergoing elective abdominal aorta and/or iliac arteries open surgery were randomized into two groups to receive 1% lidocaine or placebo at the same infusion rate based on ideal body weight (bolus of 0.15 mL/kg during the induction of anesthesia followed by continuous infusion of 0.2 mL/kg/h during surgery; postoperatively 0.1 mL/kg/h for 24 h) additionally to standard opioid analgesia. RESULTS: Total opioid consumption within 24 h after surgery was 89.2 mg (95%CI 80.9-97.4) in the lidocaine and 113.1 mg (95%CI 102.5-123.6) in the placebo group (p = 0.0007). Similar findings were observed in opioid consumption intraoperatively (26.7 mg (95%CI 22.2-31.3) vs. 35.1 mg (95%CI 29.1-41.2), respectively, p = 0.029) and six hours postoperatively (47.5 mg (IQR 37.5-59.5) vs. 60 mg (IQR 44-83), respectively, p = 0.01). CONCLUSIONS: In high-risk vascular surgery, lidocaine infusion as an adjunct to standard perioperative analgesia is effective. It may decrease opioid consumption by more than 20% during the first 24 h after surgery, with no serious adverse effects noted during the study period.

Shifted levels of sleep and activity under darkness as mechanisms underlying ectoparasite resistance.

Parasites harm host fitness and are pervasive agents of natural selection. Host defensive traits in natural populations typically show genetic variation, which may be maintained when parasite resistance imposes fitness costs on the host coupled with variability in parasite prevalence in space and/or time. Previously we demonstrated significant evolutionary responses to artificial selection for increasing behavioral immunity to Gamasodes queenslandicus mites in replicate lines of Drosophila melanogaster . Here, we report transcriptional shifts in metabolic processes between selected and control fly lines based on RNA-seq analyses. We also show decreased starvation resistance and increased use of nutrient reserves in flies from mite-resistant lines. Additionally, mite-resistant lines exhibited increased behavioral activity, reduced sleep and elevated oxygen consumption under conditions of darkness. The link between resistance and sleep was confirmed in an independent panel of D. melanogaster genetic lines exhibiting variable sleep durations, showing a positive correlation between mite resistance and reduced sleep. Experimentally restraining the activity of artificially selected mite-resistant flies during exposure to parasites under dark conditions reduced their resistance advantage relative to control flies. The results suggest that ectoparasite resistance in this system involves increased dark-condition activity and metabolic gene expression at the expense of nutrient reserves and starvation resistance.

Evolution: Natural selection, sexual selection, and the jaws of death.

Traits that increase reproductive success in males can have negative fitness consequences in females. A new study shows that natural selection by a predator that targets males with larger secondary sexual traits drives an evolutionary increase in female fitness.

Analysis of correlated responses in key ejaculatory traits to artificial selection on a diversifying secondary sexual trait.

Positive genetic covariance between male sexual display traits and fertilizing capacity can arise through different mechanisms and has important implications for sexual trait evolution. Evidence for such genetic covariance is rare, and when it has been found, specific physiological traits underlying variation in fertilization success linked to trait expression have not been identified. A previous study of correlated responses to bidirectional artificial selection on the male sex comb, a secondary sexual trait, in Drosophila bipectinata Duda documented a positive genetic correlation between sexual trait size and competitive fertilization success, and found that transcript levels of multiple seminal fluid proteins (SFPs) were significantly increased in the large sex comb (high) genetic lines. These results suggest that changes in SFP activity may be a causal factor underlying the increased fertilizing capacity of high line males. Here, we tested for correlated responses to this selection in a suite of additional reproductive traits, measured in the context of variation in male age and exposure to rivals. Whereas several traits including sperm length, number and viability, and accessory gland size, increased with age, only sperm viability was influenced by selection treatment, but in complex fashion. Sperm viability of high line males surpassed that of their smaller-combed counterparts when they had been housed with rivals and were 5-6 days old or older. Interestingly, this interaction effect was evident for sperm sampled from the female seminal receptacle, but not from the male seminal vesicles (where sperm have yet to be combined with accessory gland products), consistent with the differential SFP activity between the lines previously found. Our results suggest that differences in sperm quality (as viability) may be a contributing factor to the positive genetic correlation between sexual trait size and competitive fertilization capacity in D. bipectinata.

Apical amplification-a cellular mechanism of conscious perception?

We present a theoretical view of the cellular foundations for network-level processes involved in producing our conscious experience. Inputs to apical synapses in layer 1 of a large subset of neocortical cells are summed at an integration zone near the top of their apical trunk. These inputs come from diverse sources and provide a context within which the transmission of information abstracted from sensory input to their basal and perisomatic synapses can be amplified when relevant. We argue that apical amplification enables conscious perceptual experience and makes it more flexible, and thus more adaptive, by being sensitive to context. Apical amplification provides a possible mechanism for recurrent processing theory that avoids strong loops. It makes the broadcasting hypothesized by global neuronal workspace theories feasible while preserving the distinct contributions of the individual cells receiving the broadcast. It also provides mechanisms that contribute to the holistic aspects of integrated information theory. As apical amplification is highly dependent on cholinergic, aminergic, and other neuromodulators, it relates the specific contents of conscious experience to global mental states and to fluctuations in arousal when awake. We conclude that apical dendrites provide a cellular mechanism for the context-sensitive selective amplification that is a cardinal prerequisite of conscious perception.

Positive genetic covariance between male sexual ornamentation and fertilizing capacity.

Postcopulatory sexual selection results from variation in competitive fertilization success among males and comprises powerful evolutionary forces that operate after the onset of mating.1,2 Theoretical advances in the field of sexual selection addressing the buildup and coevolutionary consequences of genetic coupling3-5 motivate the hypothesis that indirect postcopulatory sexual selection may promote evolution of male secondary sexual traits-those traits traditionally ascribed to mate choice and male fighting.6,7 A crucial prediction of this hypothesis is genetic covariance between trait expression and competitive fertilization success, which has been predicted to arise, for example, when traits subject to pre- and postcopulatory sexual selection are under positive correlational selection.8 We imposed bidirectional artificial selection on male ornament (sex comb) size in Drosophila bipectinata and demonstrated increased competitive fertilization success as a correlated evolutionary response to increasing ornament size. Transcriptional analyses revealed that levels of specific seminal fluid proteins repeatedly shifted in response to this selection, suggesting that properties of the ejaculate, rather than the enlarged sex comb itself, contributed fertilizing capacity. We used ultraprecise laser surgery to reduce ornament size of high-line males and found that their fertilizing superiority persisted despite the size reduction, reinforcing the transcriptional results. The data support the existence of positive genetic covariance between a male secondary sexual trait and competitive fertilization success, and suggest the possibility that indirect postcopulatory sexual selection may, under certain conditions, magnify net selection on ornamental trait expression.

Microscale laser surgery reveals adaptive function of male intromittent genitalia.

The leading hypothesis for the evolution of male genital complexity proposes that genital traits evolve in response to post-insemination sexual selection; that is, via cryptic female choice or sperm competition. Here, we describe a laser ablation technique for high-precision manipulation of microscale body parts of insects, and employ it to discern the adaptive function of a rapidly evolving and taxonomically important genital trait: the intromittent claw-like genital spines of male Drosophila bipectinata Duda. We demonstrate experimentally and unambiguously that the genital spines of this species function to mechanically couple the genitalia together. The excision of the spines by laser ablation sharply reduced the ability of males both to copulate and to compete against rival males for mates. When spineless males did succeed to copulate, their insemination success and fertilization rate were not statistically different from controls, at odds with the post-insemination sexual selection hypothesis of genital function and evolution. The results provide direct experimental support for the hypothesis that genital traits evolve in response to sexual selection occurring prior to insemination.

How to Mitigate the Hard Problem by Adopting the Dual Theory of Phenomenal Consciousness.

In this paper, we propose the following hypothesis: the hard problem of consciousness is in part an artifact of what we call the unitary approach to phenomenal consciousness. The defining mark of the unitary approach is that it views consciousness and phenomenality as inseparable. Giving up this conceptual commitment redefines, in a productive way, the explanatory tasks of the theory of consciousness. Adopting a non-unitary conception of experience does not make the hard problem of consciousness go away completely but it shifts the locus of where the explanation of experience gets difficult, and cuts down the mystery of consciousness to size. Other advantages of the non-unitary account of consciousness are sketched as well.

A Novel Image-Based Screening Method to Study Water-Deficit Response and Recovery of Barley Populations Using Canopy Dynamics Phenotyping and Simple Metabolite Profiling.

Plant phenotyping platforms offer automated, fast scoring of traits that simplify the selection of varieties that are more competitive under stress conditions. However, indoor phenotyping methods are frequently based on the analysis of plant growth in individual pots. We present a reproducible indoor phenotyping method for screening young barley populations under water stress conditions and after subsequent rewatering. The method is based on a simple read-out of data using RGB imaging, projected canopy height, as a useful feature for indirectly following the kinetics of growth and water loss in a population of barley. A total of 47 variables including 15 traits and 32 biochemical metabolites measured (morphometric parameters, chlorophyll fluorescence imaging, quantification of stress-related metabolites; amino acids and polyamines, and enzymatic activities) were used to validate the method. The study allowed the identification of metabolites related to water stress response and recovery. Specifically, we found that cadaverine (Cad), 1,3-aminopropane (DAP), tryptamine (Tryp), and tyramine (Tyra) were the major contributors to the water stress response, whereas Cad, DAP, and Tyra, but not Tryp, remained at higher levels in the stressed plants even after rewatering. In this work, we designed, optimized and validated a non-invasive image-based method for automated screening of potential water stress tolerance genotypes in barley populations. We demonstrated the applicability of the method using transgenic barley lines with different sensitivity to drought stress showing that combining canopy height and the metabolite profile we can discriminate tolerant from sensitive genotypes. We showed that the projected canopy height a sensitive trait that truly reflects other invasively studied morphological, physiological, and metabolic traits and that our presented methodological setup can be easily applicable for large-scale screenings in low-cost systems equipped with a simple RGB camera. We believe that our approach will contribute to accelerate the study and understanding of the plant water stress response and recovery capacity in crops, such as barley.

Ectoparasite resistance is correlated with reduced host egg hatch rate in the Drosophila-Macrocheles system.

We test for fitness costs of resistance in a natural host-parasite system, involving Drosophila nigrospiracula and ectoparasitic mites, Macrocheles subbadius. We contrasted rates of mortality at embryonic and pupal stages of host ontogeny between replicate-resistant and -susceptible (control) lines at different temperatures (24, 28, and 34 degrees C). Evidence for a cost of resistance was shown as a 17% overall reduction in egg hatch rate in replicate-resistant lines, although this effect was heterogeneous across replicate selection experiments. This cost of resistance was not magnified under thermal stress. Pupa survivorship was statistically invariant between resistant and control lines, at either temperature. Embryo and pupa mortalities were significantly elevated at the high temperature, confirming that the thermal treatment was physiologically stressful. The results suggest differential sensitivity of life history traits to the pleiotropic effects of genetic resistance against ectoparasitic mites.

Neural Correlates of Consciousness Meet the Theory of Identity.

One of the greatest challenges of consciousness research is to understand the relationship between consciousness and its implementing substrate. Current research into the neural correlates of consciousness regards the biological brain as being this substrate, but largely fails to clarify the nature of the brain-consciousness connection. A popular approach within this research is to construe brain-consciousness correlations in causal terms: the neural correlates of consciousness are the causes of states of consciousness. After introducing the notion of the neural correlate of consciousness, we argue (see Against Causal Accounts of NCCs) that this causal strategy is misguided. It implicitly involves an undesirable dualism of matter and mind and should thus be avoided. A non-causal account of the brain-mind correlations is to be preferred. We favor the theory of the identity of mind and brain, according to which states of phenomenal consciousness are identical with their neural correlates. Research into the neural correlates of consciousness and the theory of identity (in the philosophy of mind) are two major research paradigms that hitherto have had very little mutual contact. We aim to demonstrate that they can enrich each other. This is the task of the third part of the paper in which we show that the identity theory must work with a suitably defined concept of type. Surprisingly, neither philosophers nor neuroscientists have taken much care in defining this central concept; more often than not, the term is used only implicitly and vaguely. We attempt to open a debate on this subject and remedy this unhappy state of affairs, proposing a tentative hierarchical classification of phenomenal and neurophysiological types, spanning multiple levels of varying degrees of generality. The fourth part of the paper compares the theory of identity with other prominent conceptions of the mind-body connection. We conclude by stressing that scientists working on consciousness should engage more with metaphysical issues concerning the relation of brain processes and states of consciousness. Without this, the ultimate goals of consciousness research can hardly be fulfilled.

Fruit flies may face a nutrient-dependent life-history trade-off between secondary sexual trait quality, survival and developmental rate.

Optimal life-history strategies are those that best allocate finite environmental resources to competing traits. We used the geometric framework for nutrition to evaluate life-history strategies followed by Drosophila melanogaster by measuring the condition-dependent performance of life-history traits, including the morphology of male secondary sexual characters, sex combs. We found that depending on their rearing environment flies faced different forms of trait trade-offs and accordingly followed different life-history strategies. High-energy, high-carbohydrate, low-protein diets supported development of the largest and most symmetrical sex combs, however, consistent with handicap models of sexual selection these foods were associated with reduced fly survival and developmental rate. Expressing the highest quality sex combs may have required secondary sexual trait quality to be traded-off with developmental rate, and our results indicated that flies unable to slow development died. As larval nutritional environments are predominantly determined by female oviposition substrate choice, we tested where mated female flies laid the most eggs. Mothers chose high-energy, high-protein foods associated with rapid larval development. Mothers avoided high-carbohydrate foods associated with maximal sex comb expression, showing they may avoid producing fewer 'sexy' sons in favour of producing offspring that develop rapidly.