Mud-puddling behavior in tropical butterflies: in search of proteins or minerals?

We experimentally investigated the attraction of adult butterflies to moist soil and dirt places (a behavior termed `mud-puddling') in two species-rich tropical communities on the island of Borneo. At a rain forest site, 227 individuals (46 species) were attracted to the baits, compared to 534 individuals (54 species) at a farmland site. With one single exception, all attracted butterflies were males. Of various salt and amino acid solutions, only sodium was accepted, but overall, albumin solutions turned out to be the most attractive puddling resource. Butterfly families differed consistently in their resource preferences. Representatives of the families Papilionidae and Pieridae more often visited NaCl solutions, but still accepted albumin, whereas representatives of the Nymphalidae, Hesperiidae and, in particular, Lycaenidae preferred the protein resource. In experiments using decoys prepared from pinned butterfly specimens, representatives of the Papilionidae and Pieridae were more strongly attracted to baits provided with decoys made from conspicuous, medium-sized yellow Eurema species (Pieridae), whereas dummies made from small, cryptically colored lycaenids (Prosotas and Caleta species) were ineffective. Decoys did not influence the attraction of lycaenid butterflies towards baits. Hence, visual cues play an important role in locating puddling resources for papilionids and pierids, while for lycaenid butterflies searching for nitrogen sources, olfactory cues emitted by decaying organic matter are more likely to be important. The strong attraction of male butterflies to nitrogen-rich resources suggests that, as in the case of sodium, these nutrients may increase reproductive success. Received: 5 October 1998 / Accepted: 7 December 1998     

A smoking ban in psychiatric units: threat or opportunity?

People with severe mental illness (SMI) experience some of the worst physical health and die younger than almost any section of the population. Mental health professionals have seemed strangely indifferent to this inequality, which in other areas of health would be a national scandal. In this editorial we discuss the recently introduced smoking ban in inpatient mental health service settings, which will offer mental health services an opportunity to implement creative, evidence-based strategies to help people with SMI address smoking and nicotine addiction. In doing this, we refer to National Institute for Health and Clinical Excellence (NICE) guidance. This guidance forms the basis of national smoking policies for the general population and forms a starting point for those with SMI. Such a strategy will necessarily involve close collaboration with primary care, and we specifically examine how this might be achieved.     

Costly courtship or dishonest display? Intensely displaying sand goby males have lower lipid content

Intensely displaying sand goby Pomatoschistus minutus males had lower lipid content, indicating that signalling increased energy expenditure, or that low condition males increased their signalling effort. Display intensity correlated positively with nest‐defence and tended to correlate positively with filial cannibalism.     

Contest behavior in territorial male butterflies: does size matter?

Although intrasexual contests generally favor bigger or stronger individuals, the relevance of body size to war of attrition-typedisputes between weaponless animals such as butterflies isunclear. In this study I aimed to investigate the significanceof size in this context by studying territorial contests inHypolimnas bolina (L.), a species that exhibits consistentseasonal plasticity in body size. In this species adult ageis positively correlated with large size in spring but withsmall size in autumn. This shift allowed independent evaluationof the relevance of each variable (size and age) to intrasexualcontest success. Observation of a population of marked individualsindicated that only age appeared important, with the winnersof pair-wise contests significantly older than losers in both seasons, and with contests lasting longer when the age differencebetween the combatants was small. Age was also linked to residency;residents won 99% of all contests. This study suggests thatsize does not matter in these aerial disputes, but age andresidency do. It is not yet possible to determine whether olderbutterflies are intrinsically better competitors, or whether they simply have greater opportunity to find a vacant territory.     

Does courtship behavior contribute to species-level reproductive isolation in field crickets?

Reproductive behavior influences gene flow within and amongspecies; thus, sexual selection may be a major contributor tothe maintenance of species, and possibly their formation. HereI experimentally manipulate the courtship interactions of thefield crickets Gryllus rubens and G. texensis to examine thepotential of close-range courtship interactions to limit interspecificgene flow. Responses of males to females and of females to malecourtship song and males per se were examined for four pairedsympatric and allopatric populations. Male G. rubens were morelikely to court conspecific females, but male G. texensis courtedfemales of both species equally. If paired with conspecificmales, female G. rubens and G. texensis both preferred conspecificcourtship song. In none of these comparisons were the responsesof males or females from allopatry different from those in sympatry.There was an asymmetry of courtship response across both sexand species: male G. rubens were more discriminating than maleG. texensis, whereas female G. texensis were more discriminatingthan female G. rubens. Despite significant preferences for conspecifics,the net effect of courtship interactions would appear to limitinterspecific gene flow only weakly. These results are consistentwith courtship behavior evolving under the influence of sexualselection and only indirectly promoting species integrity.     

Securing paternity in spiders? A review on occurrence and effects of mating plugs and male genital mutilation

Low female mating frequencies often appear to be cases of direct male induction that can oppose female interests. Mating plugs are most obvious means leading to low degrees of multiple mating in females. In spiders, mating plugs are formed by a variety of amorphous materials, by the breakage of the male sperm transferring organ, or by the whole male that functions as a mating barrier. Our compilation of the available information on the presence of the various types of mating plugs suggests that plugs predominantly occur in entelegyne spiders. In this group, plugs do not interfere with oviposition since separate openings for insemination and oviposition are present. In contrast, mating plugs seem to be rare in haplogyne spiders that do not possess separate openings. The available experimental studies on the function of the different types of plugs suggest that plugs can be considered as male adaptations to avoid sperm competition. However, females in some cases were shown to have evolved means to prevent or control male manipulation or may selectively favour plug production in specific males, an aspect which has largely been neglected. In order to understand plug evolution and function we need to explore the morphological, behavioural and biochemical aspects involved and extend our approach to interactions between the sexes.     

Grains of truth or transparent blindfolds? A review of current debates in archaeological phytolith analysis

Phytolith analysis has become an increasingly popular archaeobotanical tool in the past few decades. Phytoliths have been used to support key hypotheses relating to the domestication of several food crops and in the study of ancient diet, and they are of particular importance in contexts where other plant remains are poorly preserved. However, the discipline has also been subject to controversy and debate. This paper gives an overview of the technique and three key case studies covering a range of geographical areas. Some of the problems that are common to each are discussed and suggestions are made for how these problems could be resolved in future research. It is suggested that further caution should be taken during interpretation, and a greater consideration given to taphonomy. Despite these criticisms it is concluded that there is still much potential in the technique, particularly when integrated with other lines of microarchaeological evidence.     

Kasha or state selective behavior in the photochemistry of ortho-nitrobenzaldehyde?

The photochemistry of ortho-nitrobenzaldehyde dissolved in tetrahydrofuran was studied by means of femtosecond UV/Vis and IR spectroscopy. Comparison was made of the spectral and temporal signatures for ~400 nm and ~260 nm excitation. The 400 nm excitation promotes NBA to its lowest excited singlet state of nπ* character whereas for 260 nm an upper excited state of ππ* character is addressed. On the picosecond time scale, the molecule undergoes hydrogen transfer, yielding a ketene intermediate, internal conversion recovering the starting material, and intersystem crossing. Time constants and yields of these processes are virtually not affected by the excitation wavelength. For 400 nm excitation a ~100 fs decay component seen in the 260 nm experiment is absent, indicating that this component is due to a ππ* → nπ* internal conversion. In contrast to its formation, the decay of the ketene intermediate is influenced by the excitation wavelength. This can be attributed to different amounts of vibrational excitation.     

Consciousness in dolphins? A review of recent evidence

For millennia, dolphins have intrigued humans. Scientific study has confirmed that bottlenose dolphins are large-brained, highly social mammals with an extended developmental period, flexible cognitive capacities, and powerful acoustic abilities including a sophisticated echolocation system. These findings have led some to ask if dolphins experience aspects of consciousness. Recent investigations targeting self-recognition/self-awareness and metacognition, constructs tied to consciousness on some accounts, have analyzed the dolphin’s ability to recognize itself in a mirror or on a video as well as to monitor its own knowledge in a perceptual categorization task. The current article reviews this work with dolphins and grapples with some of the challenges in designing, conducting, and interpreting these studies as well as with general issues related to studying consciousness in animals. The existing evidence does not provide a convincing case for consciousness in dolphins. For productive scientific work on consciousness in dolphins (and other animals including humans), we need clearer characterizations of consciousness, better methods for studying it, and appropriate paradigms for interpreting outcomes. A current focus on metamemory in animals offers promise for future discovery in this area.     

Agonistic behavior in naïve juvenile lobsters depleted of serotonin by 5,7-dihydroxytryptamine

We have been exploring the role of serotonin in fighting behavior in lobsters using a specific model of agonistic behavior, the establishment of hierarchical relationships between pairs of socially naive juvenile lobsters. We selected this model because the behavior is easily evoked, readily quantifiable, and the effects of experience are eleminated by using socially naive animals. In these studies we injected a specific neurotoxin, 5,7-dihydroxytryptamine, into juvenile lobsters over a 4-week period and then measured the effects on fighting behavior. This treatment reduces the levels of serotonin in the nervous system and immunocytochemical studies show a dramatic reduction in neuropil staining for the amine. Control animals received vehicle injection alone. All injected animals were paired against larger or smaller non-injected opponents, and three successive 30-min fights were carried out and statistically analyzed. The results were surprising: As with elevations of serotonin, reduced levels of serotonin increased the amount of time animals engaged in fighting behavior. No significant effects were seen on who initiated encounters, who retreated first, or who the eventual winner would be. Thus, in this model, elevation or reduction of serotonergic function increases the tendency of animals to engage in agonistic encounters.     

Gene-environment interactions in human disease: nuisance or opportunity?

Many environmental risk factors for common, complex human diseases have been revealed by epidemiologic studies, but how genotypes at specific loci modulate individual responses to environmental risk factors is largely unknown. Gene-environment interactions will be missed in genome-wide association studies and could account for some of the 'missing heritability' for these diseases. In this review, we focus on asthma as a model disease for studying gene-environment interactions because of relatively large numbers of candidate gene-environment interactions with asthma risk in the literature. Identifying these interactions using genome-wide approaches poses formidable methodological problems, and elucidating molecular mechanisms for these interactions has been challenging. We suggest that studying gene-environment interactions in animal models, although more tractable, might not be sufficient to shed light on the genetic architecture of human diseases. Lastly, we propose avenues for future studies to find gene-environment interactions.     

Lonely hearts or sex in the city? Density-dependent effects in mating systems

Two very basic ideas in sexual selection are heavily influenced by numbers of potential mates: the evolution of anisogamy, leading to sex role differentiation, and the frequency dependence of reproductive success that tends to equalize primary sex ratios. However, being explicit about the numbers of potential mates is not typical to most evolutionary theory of sexual selection. Here, we argue that this may prevent us from finding the appropriate ecological equilibria that determine the evolutionary endpoints of selection. We review both theoretical and empirical advances on how population density may influence aspects of mating systems such as intrasexual competition, female choice or resistance, and parental care. Density can have strong effects on selective pressures, whether or not there is phenotypic plasticity in individual strategies with respect to density. Mating skew may either increase or decrease with density, which may be aided or counteracted by changes in female behaviour. Switchpoints between alternative mating strategies can be density dependent, and mate encounter rates may influence mate choice (including mutual mate choice), multiple mating, female resistance to male mating attempts, mate searching, mate guarding, parental care, and the probability of divorce. Considering density-dependent selection may be essential for understanding how populations can persist at all despite sexual conflict, but simple models seem to fail to predict the diversity of observed responses in nature. This highlights the importance of considering the interaction between mating systems and population dynamics, and we strongly encourage further work in this area.     

Agonistic behavior in wild male Magellanic penguins: when and how do they interact?

Game-theory models predict that the frequency and type of agonistic interactions should vary with the value of the resource being contested. We describe bill duels and overt fighting in male Magellanic penguins (Spheniscus magellanicus) at a breeding colony and determine whether these behaviors change with the value of the nests over which they interact. Bill duels represent low levels of aggression while overt fighting high levels of aggression. Consistent with predictions, overt fighting was more common before egg laying when nests have the highest potential value while bill duels were more common at failed nests later in the season when nests are less valuable as they could not be used for reproduction until the next season. Contrary to expectations, overt fights were shorter and resulted in fewer cuts before egg laying than after egg laying. Large size asymmetries between opponents before egg laying may enable losers to quickly assess their opponents and leave before they are hurt. As predicted, the duration and damage occurring during overt fights were positively correlated with nest cover, which is correlated with higher reproductive success. We conclude that male Magellanic penguins have rules of engagement that in the most cases follow game-theory predictions on when and how to interact.     

Diversification of host use in two polyphagous butterflies: differences in oviposition specificity or host rank hierarchy?

Novel host usage may represent an initial step towards diversification or radiation onto novel hosts within an evolutionary lineage, particularly if a shift in host plant preference ranking takes place. Polyphagous stages of evolutionary lineages may represent transitional states in which novel host associations are more likely to develop, but may be more difficult to detect experimentally. The polyphagous sister species Papilio glaucus L. and Papilio canadensis (Lepidoptera: Papilionidae; these Papilio = Pterourus) are known to exhibit differences in host‐plant use, despite significant overlap in host‐use abilities, providing an opportunity to examine how host shifts in polyphagous species may occur and what the implications for future divergence may be. In particular, we were interested in (i) determining whether differences in oviposition behavior of these species were due to changes in specificity or shifts in host‐plant hierarchy, (ii) whether the varying preference for primary hosts also affected the preference for secondary hosts, and (iii) what the oviposition preferences of a new hybrid swarm population are. We examined more than 40 000 oviposition bouts from more than 400 P. glaucus, P. canadensis, and hybrid females placed in seven‐, three‐, or two‐choice assays. In each of the choice assays, leaves from plants in different plant families of varying suitability for P. glaucus and P. canadensis larvae were used. We found the primary difference between P. glaucus and P. canadensis to be limited to a Z‐linked shift in host rank hierarchy due to an acceptance of Populus tremuloides Michx. (Salicaceae) and reduced specificity for Liriodendron tulipifera L. (Magnoliaceae) in P. canadensis. In addition, we found the absence of the Z‐linked oviposition acceptance of P. tremuloides in a recently formed allochronically separated hybrid swarm population found in P. canadensis territory at the northern border of the P. glaucus and P. canadensis hybrid zone.     

Sexual conflict over mating in Gnatocerus cornutus? Females prefer lovers not fighters

Female mate choice and male–male competition are the typical mechanisms of sexual selection. However, these two mechanisms do not always favour the same males. Furthermore, it has recently become clear that female choice can sometimes benefit males that reduce female fitness. So whether male–male competition and female choice favour the same or different males, and whether or not females benefit from mate choice, remain open questions. In the horned beetle, Gnatocerus cornutus, males have enlarged mandibles used to fight rivals, and larger mandibles provide a mating advantage when there is direct male–male competition for mates. However, it is not clear whether females prefer these highly competitive males. Here, we show that female choice targets male courtship rather than mandible size, and these two characters are not phenotypically or genetically correlated. Mating with attractive, highly courting males provided indirect benefits to females but only via the heritability of male attractiveness. However, mating with attractive males avoids the indirect costs to daughters that are generated by mating with competitive males. Our results suggest that male–male competition may constrain female mate choice, possibly reducing female fitness and generating sexual conflict over mating.     

Nitrite in nitric oxide biology: cause or consequence? A systems-based review

All life requires nitrogen compounds. Nitrite is such a compound that is naturally occurring in nature and biology. Over the years, the pharmacological stance on nitrite has undergone a surprising metamorphosis, from a vilified substance that generates carcinogenic nitrosamines in the stomach to a life-saving drug that liberates a protective agent (nitric oxide or NO) during hypoxic events. Nitrite has been investigated as a vasodilator in mammals for over 125 years and is a known by-product of organic nitrate metabolism. There has been a recent rediscovery of some of the vasodilator actions of nitrite in physiology along with novel discoveries which render nitrite a fundamental molecule in biology. Until recently nitrite was thought to be an inert oxidative breakdown product of endogenous NO synthesis but the past few years have focused on the reduction of nitrite back to NO in the circulation as a possible mechanism for hypoxic vasodilatation. Nitrite has evolved into an endogenous signaling molecule and regulator of gene expression that may not only serve as a diagnostic marker but also find its role as a potential therapeutic agent of cardiovascular disease. These data therefore warrant a reevaluation on the fate and metabolism of nitrite in biological systems. This review serves to encompass the history and recent evolution of nitrite, the compartment-specific metabolism of nitrite and its role in plasma as a biomarker for disease, the role of nitrite as a potential regulator of NO homeostasis, and the future of nitrite-based research.     

Why do males emerge before females? protandry as a mating strategy in male and female butterflies

Summary The reproductive strategy of butterfly males can be defined as being to maximize the number of females mated. We have earlier shown that, if the eclosion period of females is regarded as given, males should emerge before females to achieve maximal reproductive success. However, females may also be considered to have a reproductive strategy with respect to the issue when to emerge. In this paper we assume that females are selected to minimize the time spent unmated (to minimize prereproductive death), and analyze when females should optimally emerge in relation to males to achieve this end. We show that there is no conflict between the sexes with respect to the timing of eclosion when the length of the eclosion period is approximately equal for males and females. Thus, protandry should be considered a reproductive strategy of both males and females.     

Towards precision medicine in diabetes? A critical review of glucotypes

In response to a study previously published in PLOS Biology, this Formal Comment thoroughly examines the concept of ’glucotypes’ with regard to its generalisability, interpretability and relationship to more traditional measures used to describe data from continuous glucose monitoring.

Although the promise of precision medicine has led to advances in the recognition and treatment of rare monogenic forms of diabetes, its impact on prevention and treatment of more common forms of diabetes has been underwhelming [1]. Several approaches to the subclassification of individuals with, or at high risk of, type 2 diabetes have been published recently [2–4]. Hall and colleagues introduced the concept of “glucotypes” in a research article [3] that has received enormous attention in the highest impact scientific journals [5–8], mostly in relation to precision medicine. The authors developed an algorithm to identify patterns of glucose fluctuations based on continuous glucose monitoring (CGM). They named the 3 identified patterns: “low variability,” “moderate variability,” and “severe variability” glucotypes. Each individual was characterised by the proportion of time spent in the 3 glucotypes and was assigned to an overall glucotype based on the highest proportion. They argued that glucotypes provide the advantage of taking into account a more detailed picture of glucose dynamics, in contrast to commonly used single time point or average-based measures, revealing subphenotypes within traditional diagnostic categories of glucose regulation. Even though the study was based on data from only 57 individuals without a prior diabetes diagnosis, others have interpreted the results as indicating that glucotypes might identify individuals at an early stage of glucose dysregulation, suggesting a potential role in diabetes risk stratification and prevention [5]. However, before glucotypes can become “an important tool in early identification of those at risk for type 2 diabetes” [3], the concept requires thorough validation. Therefore, we explore the generalisability and interpretability of glucotypes and their relationship to traditional CGM-based measures.We used data from The Maastricht Study [9] and the PRE-D Trial [10] comprising a total number of 770 diabetes-free individuals with a 7-day CGM registration. We observed that the average proportion of time spent in the low variability glucotype was low both in The Maastricht Study (6%) and the PRE-D Trial (4%), compared to 20% in the original study. A reason for the difference may be that our study populations were on average 11 to 12 years older and that the PRE-D Trial (n = 116) included only overweight and obese individuals with prediabetes. In The Maastricht Study, the median (Q1 to Q3) body mass index was 25.9 kg/m² (23.4 to 28.7), and 72% had normal glucose tolerance. As a logical consequence, the severe glucotype was most common in the PRE-D Trial (55%). Regardless, our data show that the initial estimates of the different glucotype prevalences do not necessarily generalise to other populations, especially in age groups at increased risk of type 2 diabetes.Hall and colleagues described glucotypes as a new measure of glucose variability, a clinically relevant metric of glycaemic patterns [3]. In the figures accompanying the original publication, the low variability pattern was characterised by both the lowest mean glucose level and variation, while the severe pattern had both the highest mean glucose level and variation. As such, these examples did not give an intuition whether glucotypes were predominantly driven by glucose variability or by mean glucose levels. We therefore present 3 examples from the PRE-D Trial (Fig 1). The first 2 profiles are very similar with regard to glucose variability. Thus, the driver of the most severe glucotype of the second participant is clearly the slightly higher mean glycaemic level. Also, even though the third participant has a much larger variation than the first two, the proportion of time in the severe glucotype is not higher than for the second participant as one would expect from a classical measure of glucose variability. To investigate this further, we assessed the association between glucotypes and classical CGM measures, i.e., the mean CGM glucose level (Fig 2A) and the coefficient of variation (Fig 2B) in The Maastricht Study. The scatterplots show a clear association between the mean CGM glucose and glucotypes. They also suggest that participants with a high proportion of time in the moderate glucotype do not have high variation in glucose. Rather than a biological feature, this may well be a methodological consequence of being assigned to the middle cluster. If large fluctuations were present, glucose levels would reach either low or high values, resulting in a higher proportion of time spent in the low or severe glucotypes, respectively (assuming a strong association between glucotypes and mean CGM glucose). Therefore, we decided to quantify this association using regression analysis where glucotype proportions were the outcomes, and the mean CGM glucose concentration was the independent variable modelled with natural cubic splines (more details on the specification of the models are given in Supporting information S1–S3 Codes). Then, we used the equation estimated in The Maastricht Study to predict glucotypes in the external validation sample (PRE-D Trial, Fig 2C). First, similarly to Hall and colleagues, we assigned individuals to the pattern with the highest proportion of time and then compared the predicted and the observed glucotypes. We found that in 107 out of 116 individuals, the glucotype was predicted correctly when using only the mean CGM glucose value. When considering the glucotypes as continuous proportions of time, the root mean squared errors (RMSEs) were 0.05, 0.09, and 0.07 for the low, moderate, and severe variability glucotypes, respectively, indicating good predictive ability. These results demonstrate that glucotypes either mainly reflect the mean CGM glucose level or do not translate to external datasets (e.g., due to overfitting). To investigate this further, we conducted the same analyses as described for the PRE-D Trial in the original data from Hall and colleagues and found a slightly weaker, but still strong association between mean CGM glucose levels and glucotypes. Using the regression model from The Maastricht Study, we could correctly predict 79% of the glucotypes, while the RMSEs were 0.11, 0.15, and 0.13.Open in a separate windowFig 1Example CGM profiles of participants in the PRE-D Trial with corresponding proportion of time spent in different glucotypes and conventional measures (mean and CV).CGM, continuous glucose monitoring; CV, coefficient of variation.Open in a separate windowFig 2Observed proportion of time spent in the 3 glucotypes by mean CGM glucose (A) and coefficient of variation (B) in The Maastricht Study, and by mean CGM glucose in the PRE-D Trial (C) alongside predicted proportions based on the regression analysis in The Maastricht Study. CGM, continuous glucose monitoring.Although the transformation of continuous measures into categorical ones is a common procedure in clinical research, assigning individuals to the glucotype with the highest proportion of time runs very much against the “precision” tenet of precision medicine. In line with this, a recent study has demonstrated how simple clinical features outperformed clusters in predicting relevant clinical outcomes [11]. This is especially problematic when a method does not provide clear separation between clusters, which can be quantified by calculating relative entropy [12]. A relative entropy of zero would mean that all individuals spend one-third of the time in each of the 3 glucotypes, while a value of one would indicate that each individual spends the entire time period in only one of the 3 glucotypes. In the original cohort of Hall and colleagues [3], we calculated a relative entropy of 0.24 indicating that cluster separation is far from optimal and together with the previous results question the claim that the glucotype is really a “more comprehensive measure of the pattern of glucose excursions than the standard laboratory tests in current use” [3].In conclusion, we demonstrate in 2 large, external datasets, that the assessment of glucotypes does not offer more novel insights than the mean CGM glucose, highlighting the importance of large development datasets and external validation for data-driven algorithms. As CGM is becoming more widely used in large clinical studies also among individuals without diabetes, glucose patterns derived from CGMs will be an important focus area in future diabetes research. However, it is important that scientific scrutiny precedes the introduction of emerging tools with a promise of identifying individuals at high risk of type 2 diabetes and its late complications at an earlier stage of disease progression, especially in an observational setting. Furthermore, future efforts towards precision medicine for diabetes prevention and treatment should go beyond the glucocentric approach we have seen so far. We know that hyperglycaemia is a late feature of diabetes development and that patients benefit most from a multifactorial treatment approach [13]. A multifactorial approach, with relevance to the aetiology of micro- and macrovascular complications, may also yield a more clinically useful risk stratification of nondiabetic individuals [14]. Even so, if we aim for precision medicine, we should aim to retain as much precision as possible at every step of the process, by treating determinants and outcomes as continuous measures if possible and by retaining information on the uncertainty of any hard classification such as cluster membership.