The field of conservation physiology strives to achieve conservation goals by revealing physiological mechanisms that drive population declines in the face of human-induced rapid environmental change (HIREC) and has informed many successful conservation actions. However, many studies still struggle to explicitly link individual physiological measures to impacts across the biological hierarchy (to population and ecosystem levels) and instead rely on a ‘black box’ of assumptions to scale up results for conservation implications. Here, we highlight some examples of studies that were successful in scaling beyond the individual level, including two case studies of well-researched species, and using other studies we highlight challenges and future opportunities to increase the impact of research by scaling up the biological hierarchy. We first examine studies that use individual physiological measures to scale up to population-level impacts and discuss several emerging fields that have made significant steps toward addressing the gap between individual-based and demographic studies, such as macrophysiology and landscape physiology. Next, we examine how future studies can scale from population or species-level to community- and ecosystem-level impacts and discuss avenues of research that can lead to conservation implications at the ecosystem level, such as abiotic gradients and interspecific interactions. In the process, we review methods that researchers can use to make links across the biological hierarchy, including crossing disciplinary boundaries, collaboration and data sharing, spatial modelling and incorporating multiple markers  into their research. We recommend future studies incorporating tools that consider the diversity of ‘landscapes’ experienced by animals at higher levels of the biological hierarchy, will make more effective contributions to conservation and management decisions. Through the explicit incorporation of physiology into conservation science, the field of conservation physiology  and population vital rates . Such individual-level data can be inherently problematic for conservation physiology as most conservation action needs to be implemented at the population-level or higher to produce meaningful results . Despitehabitat as the immediate abiotic and biotic environment experienced by an individual, and landscape refers to a collection of habitats used by a population or species . Our knowledge of this species, from genomic architecture, sensory physiology, individual behaviours, population dynamics in natural and disturbed environments, and speciation, could potentially yield a suite of possible links among levels of the biological hierarchy; however, the mechanistic links between these dynamics and applied conservation have yet to be elucidated  scent marks under increased temperatures, thereby reducing the ability of females to find chemosensory cues that play a significant role in sexual selection. This study successfully demonstrates the link between a globally important stressor (elevated temperature) and sensory and behavioural responses. Future studies wishing to scale up to the population level would need to explicitly measure population level consequences. For example, by implementing landscape ecology spatial modelling  are negatively phototaxic and require cold habitat. By linking physiologically determined thermal tolerance and visual sensory requirements, managers can determine critical habitat areas such as the space available in the water column in which walleye can forage , while also characterizing vegetative structure to reveal habitat and nutritional covariates related to potential population declines. While this study does not show that heterophil:lymphocyte ratios predicted local population extirpations, the research design is well-suited to scaling up from individual to population-level effects.Population responses  to environmental stressors are often determined by long-term demographic studies. While there is value in using population demography to make conservation determinations for species, or inferring habitat quality in some cases , such trPusa hispida) population demographics, Landscape physiology offers a new and potentially powerful approach that allows for broader application of certain data by integrating behaviour and physiology that are shaped by environmental and landscape features. However, we argue that without the integration of other disciplines  with landscape physiology, limitations still exist for population-level inference. Population-level responses to changes in habitat quality still elude many studies but can ultimately be reached by increasing spatial scope and integrating other disciplines. For instance, in a longitudinal study on ringed seal (Bufo bufo) moving through a landscape matrix  to assess whether corticosterone is indicative of landscape resistance to animal movement. Using lab- and field-based substrate choice experiments, the authors found that adult toads preferred forest litter over ploughed soil and had higher corticosterone levels when crossing ploughed soil, with no variation in corticosterone observed in juveniles. Overall, the study successfully integrated landscape ecology, behavioural ecology and physiology. However, the authors explained the differences between adult and juvenile corticosterone levels by making assumptions about the cognitive processing and sensory perception of the developmental stages of toads (i.e. the ‘black box’), without any direct data to support these assertions. Despite this ‘black box’, the authors attempt to scale to the population by discussing how corticosterone influences population dynamics, outlining specific effects including changes in amplexus behaviour, fertilization probability and adrenal desensitization. Avoiding such assumptions are key in this case, as elevated glucocorticoids are not universally a signal of stress and may be adaptively elevated to achieve specific life cycle needs . They demonstrated, through investigation of hormone profiles, that poor population growth rate was ultimately linked to poor habitat quality. Despite the obvious strengths of a macrophysiological approach in identifying large-scale stressors and promoting species-level conservation, this approach has primarily been used in assessing thermal tolerances in relation to global temperature change  to impacts on populations or meta-populations ; Table 2e change . We sugge change ; howeverTurdus merula), that have wide distributions overlapping urban areas and diets of ground-dwelling invertebrates, are expected to be at risk of metal toxicity over a large portion of their range, not just from specific areas with significantly elevated metal concentrations. A key next step to scaling up farther is to ground-truth predictions by examining bird tissue samples for heavy metals , or a bird leaving overwintering grounds late because of reduced precipitation (Box 2)) translate to changes in higher levels of the biological hierarchy. In Platymantis vitiana) exhibits significantly reduced body condition, increased urinary corticosterone metabolites, suppressed sex steroid metabolites and reduced reproductive success due to competition with invasive cane toads  and American bullfrogs (Lithobates catesbeianus), invasive in California wetland communities, have reduced native amphibian growth rates and body mass due to a reduction in foraging time activity level  from 1995 to 1996 (Procyon lotor) foraging of intertidal and subtidal prey was reduced following vocalization playbacks of large carnivore predators in the Gulf Islands of BC, Canada (Cancer productus), subsequently leading to a decrease in one of the crab’s competitors, staghorn sculpin (Leptocottus armatus) and one of their prey items, the periwinkle snail (Littorina scutulata). The evaluation of the LOF in this system revealed important ecosystem functions that may otherwise have been overlooked, and pursuing LOF studies could be a valuable link in scaling from the individual to community and ecosystem level.The ‘landscape of fear’ (LOF) is another potential indirect predator–prey interaction that could help scale to the community and ecosystem levels. Seminal LOF work has focused on the Yellowstone National Park ecosystem following the reintroduction of grey wolves  . Migratory life history strategies are positively correlated with higher metabolic rates and feeding rates , possiblDirect examples of connections between individual-level stress and community and ecosystem dynamics are key in the implementation of conservation and management schemes; however, such examples are rare and largely comprised of reviews synthesizing existing studies. For example, While examples of successful conservation physiology studies integrated across disciplines are emerging, there are many obstacles that stand in the way of studies successfully scaling up the biological hierarchy. Here, we outline some of these key challenges and suggest potential solutions for researchers and managers trying to increase the scope of their studies. We do not expect that single studies will be able to scale all the way from physiological responses of individuals to ecosystem functioning, as this is an unrealistic expectation. However, collectively, through careful study design and collaboration, we can achieve the goal of scaling up the biological hierarchy. See Lack of funding and personnel: scaling up often requires larger operating budgets and more personnel spread across numerous locations. Working groups or other collaborations  can ofteAvailable time: university research is often carried out by graduate students and thus limited to 2–4 years to allow degree completion. Principal investigators can establish long-term studies, where individual graduate projects take steps to scale up the research, gradually building the dataset. Researchers can also design studies that take advantage of existing longitudinal datasets or previously well-studied systems where some mechanistic understanding already exists but has not been explicitly scaled up.Initial study design (i.e. choosing the ‘right’ metrics): researchers should place an emphasis on collecting metrics appropriate to their study system that can be used to scale up the hierarchy. Often this means collecting metrics from different disciplines that can then be used in conjunction. Currently, there is no consensus on which metrics are best for scaling up; however, focusing on metrics that have previously been successful in similar systems and newly emerging metrics may improve our ability to scale up.Disciplinary expertise: since scaling up requires both depth and breadth of knowledge, it is often necessary to perform research outside of one’s primary area of expertise. Given the extent and depth of literature within fields, we suggest that disciplinary breadth will best be achieved by collaborating with researchers across specializations. However, since communication can be difficult across disciplines (e.g. due to disciplinary jargon and different standards), it is important to communicate frequently with collaborators during every step of the project starting with initial study design.Taxa-specific and logistical challenges: Some research challenges are unique to particular study organisms or systems . Communication with researchers, managers, indigenous communities or local organizations with experience working in that area, or forming working groups to synthesize knowledge and increase group expertise, could help address these logistical challenges.Integration of theoretic approaches: Models used to scale up are often challenging to parameterize and require quantities of data beyond the scope of many studies, though may provide important predictive power in moving toward conservation and restoration goals. This again may be overcome by forming collaborations, such as working groups, or utilizing public access databases.Interdisciplinary research, that combines principles and techniques from multiple physiological  and ecological  disciplines to scale the biological hierarchy, will ultimately be the most successful in addressing large-scale conservation threats currently facing biodiversity. While the need to integrate research across disciplines has been generally acknowledged, there is still much that can be done to more effectively inform conservation practices. Population-level effects are often assumed based on physiological principles, but these ‘black box’ assumptions are often missing mechanisms that would explicitly link levels of the biological hierarchy. Additionally, many population-level studies do not identify the mechanistic, physiological responses exhibited by the individual (though it is often assumed). For some conservation management, simply investigating one level of the biological hierarchy may be sufficient; however, in more complex systems that are experiencing increasing anthropogenic influences and multiple stressors, investigation across multiple levels, achieved through collaboration among disciplines , may be Tachycineta bicolor) to assess their correlation with body condition , habitat quality and fitness. While this study demonstrated that lower quality habitat and body condition can alter baseline glucocorticoids long-term, concomitant effects to fitness (e.g. offspring mass and reproductive output) were not observed. Thus, an understanding of individual responses does not necessarily give a full indication of the magnitude of the population-level response. In order to remove these ‘black boxes’, we must instead determine the intensity at which a response must occur in order to elicit population-level responses  could be considered a poster child for a number of disciplines, including ethology, evolutionary biology, sensory ecology and speciation genomics. For example, the threespine stickleback is a model organism for evolutionary biology, used to understand the process of speciation across its wide geographic range . Previous reviews have highlighted connections made among disciplines where possible  study system.For migratory organisms, successful conservation strategies must address threats occurring in disparate locations across the full annual cycle of the organism to achieve maximum potential for success , is the exception not the rule. It remains unclear how widespread carryover effects and population-level seasonal interactions are, though they have been described in a few other select systems (e.g. black-tailed godwit


The recent editorial by Dr Susan Peters “Although a valuable method in occupational epidemiology, job-exposure matrices are no magic fix” ably describes the strengths and limitations of job-exposure matrix (JEM) approaches in occupational epidemiology research . In addiJEM could assist the compensation process by supporting the assessment of relevant exposures related to specific health conditions . The potSharing JEM across different countries may be useful in providing estimates of exposures across larger populations to calculate global burden of disease related to occupational exposure. The JEMINI  initiative was launched to explore the possibility of developing international JEM that could be used across countries . BeginniAs Peters mentioned, many limitations exist in the use of JEM. Users of JEM must consider the source of exposure data – expert assessments, data collected from individual workers, or environmental sampling. The coding of occupations is time consuming and can introduce error , and morIn conclusion, these JEM developed for research might also be used as a public health tool, provided that their limitations are properly taken into account.


Several factors may make it difficult, if not impossible, for athletes to train in real sport conditions. These include on the one hand extrinsic factors such as unfavorable weather conditions or costs related to the facilities and materials used, and on the other hand factors intrinsic to the athlete, such as injuries.When confronted to such situations, it becomes necessary for the athletes to be provided with alternative training solutions in order to maintain their level of performance. In this section, we will introduce two of these alternative solutions, namely virtual reality (VR) and motor imagery (MI). The first alternative training solution we will mention here is Virtual Reality (VR). VR is a computer-based simulation that is both interactive and immersive. It provides the opportunity to design fully controlled training procedures that are both adaptable to each athlete and ecological. Coupled with the fact that VR enables athletes to train in environments that may not be readily accessible, these properties make VR a very relevant and promising training tool to improve sport performance. Several studies have already revealed that athletes who had trained in a virtual environment obtained performances similar to those who had trained in real sport conditions , neural signals recorded from the brain are fed into a decoding algorithm that translates these signals into outputs . tDCS applies low-amplitude constant currents via scalp electrodes and modify neuronal transmembrane polarization if applied sufficiently long (10–20 min), thereby inducing cortical excitability changes in humans  phenomenological dimensions of athletes' activity  might be taking into account, (b) portable electroencephalogram systems could record athletes' brain signals during performance, and (c) advanced bioengineering techniques to decode these signals in real time as well as analyzing the experiences with advanced phenomenological techniques should be developed. While neurophenomenological methods have already been used to investigate the neural correlates of several sense-making processes in subjective experience in human activities such as meditation or epileptic seizure  with first-person qualitative analysis of pre-reflective experience emerging during a standing split task in dance. We expected that brain activity signatures underlying the consciousness of various efficient ways of being-and-acting in this task would be identified. Data was collected through (a) video tapes of the athletes' behavior while performing the sporting task, (b) video-recorded and transcribed verbalizations and commentaries elicited post-action during self-confrontation interviews, and (c) 64-electrode EEG recordings made during sport performance. Qualitative analyses were performed on the data from the self-confrontation interviews using the course-of-action method, which consists in identifying elementary units of meaning and their components  (e.g., Hauw and Durand, In conclusion, despite the emergence of some research on the topic, the field of neurotechnologies for sport psychology is still in its infancy and has to face a number of technological difficulties. The conceptual and methodological design might be a relevant direction for future development for sports learning and training.CJ, DH, and JM all took part in the writing of this opinion paper.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Computer-generated characters, so-called avatars, are widely used in advertising, entertainment, human–computer interaction or as research tools to investigate human emotion perception. However, brain responses to avatar and human faces have scarcely been studied to date. As such, it remains unclear whether dynamic facial expressions of avatars evoke different brain responses than dynamic facial expressions of humans. In this study, we designed anthropomorphic avatars animated with motion tracking and tested whether the human brain processes fearful and neutral expressions in human and avatar faces differently. Our fMRI results showed that fearful human expressions evoked stronger responses than fearful avatar expressions in the ventral anterior and posterior cingulate gyrus, the anterior insula, the anterior and posterior superior temporal sulcus, and the inferior frontal gyrus. Fearful expressions in human and avatar faces evoked similar responses in the amygdala. We did not find different responses to neutral human and avatar expressions. Our results highlight differences, but also similarities in the processing of fearful human expressions and fearful avatar expressions even if they are designed to be highly anthropomorphic and animated with motion tracking. This has important consequences for research using dynamic avatars, especially when processes are investigated that involve cortical and subcortical regions. While we are becoming more experienced with computer-generated characters, or avatars, that are used in animated films, social media or as human–computer interfaces, we do not know how we react and adapt to interactions with our virtual counterparts . The useCentral to the processing of facial expressions and facial identity is a distributed network of brain regions that responds more strongly to human faces than to other visual information . In thisBased on the functional characterization of these brain regions for face perception, one may hypothesize that differences in brain responses to dynamic human and avatar expressions depend on the specific functions of these pathways. Thus, the ventral pathway, which is tuned to invariant facial features, may respond equally to anthropomorphic avatar faces and their human counterparts. Especially the FG may show equal responses to human and avatar faces, given its importance in the holistic processing of the facial form, independent of motions and emotions. On the other hand, the dorsal pathway may be activated differently by dynamic human and avatar facial expressions. Computer-generated faces often lack subtle dynamic features, such as expression-related wrinkles. Since dorsal regions are mainly engaged in the processing of facial motion, it is plausible that the STS and the IFG show stronger responses to dynamic human expressions compared to dynamic avatar expressions.In addition to the cortical pathways discussed above, previous research has identified a subcortical route that is particularly involved in the processing of emotional facial expressions . This suAlthough the exact functional role of the subcortical route in face perception remains controversial  who reported no history of psychiatric or neurological disorders. During data collection, we had to exclude four participants from the final analyses for various reasons including excessive movement, vigilance problems and discomfort . Ethical approval for this study was obtained by the local ethics committee and subjects were only tested after given their written informed consent in line with the Declaration of Helsinki.We recruited 30 healthy participants aged between 18 and 62 years  fearful and neutral human facial expressions were recorded from four actors, (ii) for each actor, a customized avatar was created by a graphic artist  to match their appearance see , (iii) bIn addition, we constructed 128 scrambled versions of these videos by using a 40 × 45 grid and randomly scrambling the grid squares with MATLAB 2017a. Thus, we obtained a set of videos containing dynamic information equivalent to the videos with fearful expressions, but without emotional or facial attributes. This condition enabled us to test whether BOLD responses associated with dynamic fearful expressions can be ascribed to the mere processing of low-level dynamic visual information. The set of scrambled videos was divided into two subsets of 64 videos each and one of the two subsets was assigned pseudo-randomly (in the sequence of scanning) to each participant.Before scanning, participants were familiarized with the fMRI task. They were instructed that they would see videos of human and avatar faces showing fearful or neutral expressions and videos with scrambled patterns. They were told to watch the displayed faces or scrambled patterns. Occasionally, a video with a red square centered on the displayed face or scrambled pattern would appear to which they would have to respond with a button press. This task was used to ensure participants’ attention. The simplicity of the task, however, also ensured that the participants could naturally track the videos. Participants completed two functional runs while passively viewing 32 videos of fearful expressions , 32 videos of neutral expressions , 32 scrambled videos and 8 videos with a red square . This resulted in a total of 104 trials per run and 208 trials per session. The total number of button presses and the response times were recorded. Trials were randomly presented in an event-related design with jittered intertrial intervals see . After sStimuli were presented using http://www.fil.ion.ucl.ac.uk/spm/) on MATLAB 2017a. Functional images were realigned to the first image in the series, corrected for slice timing to the middle slice using sinc interpolation, and the mean functional image was co-registered to the individual anatomical image. Next, the anatomical scans were segmented into different tissue types and normalized to the Montreal Neurological Institute template using DARTEL , condition facial expression , as well as condition scramble (non-scrambled > scrambled). Control trials were also modeled as regressors of interest but excluded for group-level analyses. To minimize false-positive findings related to task-correlated motion, realignment parameters were included as regressors of no interest.t-tests. Median rating differences per participant were included as covariates to account for potential BOLD response differences due to dissimilar intensity levels of the videos . As one participant had not completed the rating survey, the median rating difference of the other participants was used as a substitute. Brain activation patterns bigger than a cluster extent of k = 5 and remaining under a voxel-wise FWE corrected P-value of <0.05 will be presented in the results section.To obtain results at group level, we analyzed first-level contrast images with one-sample http://www.neuromorphometrics.com/): the FG, the STS, the IFG and the amygdala. Based on the group-level statistical parametric maps, average ROI signals were extracted and compared for the different conditions using MarsBaR  analysis was performed in order to limit the analysis to groups of voxels that have been shown to be functionally coherent in previous studies. The following independent ROIs were a priori defined based on previous literature and using the Neuromorphometrics atlas implemented in SPM12 . First, we analyzed median differences between ratings of fearful and neutral facial expressions separately for human and avatar videos. Second, we investigated median differences between ratings of fearful human and fearful avatar facial expressions as well as median differences between ratings of neutral human and neutral avatar facial expressions. To test for rating differences associated with the gender of the displayed character, we computed median differences between videos with male and female characters separately for human and avatar videos see .To control subject’s task engagement during the scanning session, participants were required to respond with a button press to infrequent presented control videos with a red square centered on the displayed face or scrambled pattern. Average detection rate of control videos was nearly perfect (99%) with a median response time of 754 ± 246 ms.z = −4.47, P < 0.001). The same difference was observable for avatar faces, with fearful avatar expressions rated as more intense (Mdn = 3) in comparison to neutral avatar expressions . Fearful human expressions were judged as more intense (Mdn = 5) than fearful avatar expressions . No significant difference was apparent for neutral human and neutral avatar expressions . Analysis results of the rating of female versus male faces are shown in The statistical analysis of the rating of fearful and neutral videos showed that fearful human expressions were rated as more intense (Mdn = 5) than neutral human expressions  and bilateral IFG . Second, we contrasted neutral human and avatar expressions with one another. They did not evoke significantly different responses in either the whole-brain analysis or the ROI analysis. The interaction contrast face type (human versus avatar) × facial expression  revealed that the right STS showed a larger response difference between human and avatar faces for fearful expressions than for neutral expressions , as well as in the posterior (PCC) and ventral anterior cingulate cortex , right occipital FG , bilateral STS  and bilateral IFG . We then contrasted fearful and neutral avatar expression across the whole brain. Fearful avatar expressions elicited stronger responses in bilateral inferior occipital cortex as well as ventral and dorsal temporal cortex  and right STS .Next, we contrasted fearful and neutral expressions displayed in human faces across the whole brain. Fearful human expressions evoked stronger responses in bilateral inferior occipital cortex, ventral and dorsal temporal cortex, the left aIC and bilateral amygdalae see . The ROIrtex see . The ROIIn favor of a concise presentation of our main results, whole-brain analyses concerning the scrambled videos are summarized in We set out to investigate whether dynamic facial expressions displayed by avatar faces are processed within the same cortical and subcortical brain regions known to be involved in the processing of dynamic human facial expressions. To this aim, we assessed the brain topography of BOLD responses to dynamic human and avatar faces displaying either fearful or neutral expressions. Using videos of actors and their customized avatars derived from the same videos by motion tracking, we were able to demonstrate that the topography and the magnitude of brain response differed between human and avatar faces, yet only when fearful expressions were presented. Our imaging analysis revealed predominantly right-sided brain areas that showed stronger responses to fearful human expressions compared to fearful avatar expressions. This cluster entailed cortical areas such as bilateral posterior and anterior parts of the superior temporal region, the IFG, the right PCC and vACC, as well as the right aIC. For the amygdala, a subcortical core region in face and emotion perception, no response difference was found when comparing fearful expressions between human and avatar faces. Furthermore, our analysis revealed an interaction effect in the right superior temporal region. That is, the response difference between human and avatar faces was larger for fearful than for neutral expressions in the right superior temporal region.When comparing fearful and neutral expressions displayed by human faces, we were able to show stronger BOLD responses to fearful human expressions in dorsal and ventral regions of the face perception network as well as in the left aIC and bilateral amygdala. A qualitatively similar response pattern, but with fewer and smaller activation peaks, was found when comparing fearful and neutral expressions displayed by avatar faces. This cluster entailed posterior dorsal temporal regions, inferior occipital and fusiform regions, as well as the left amygdala.In line with our hypothesis, we found stronger BOLD responses to fearful human expressions than to fearful avatar expressions in posterior and anterior parts of the superior temporal region and the IFG. These differential responses also appeared in comparison to scrambled videos, which indicates that the processing difference between fearful human and avatar expressions is not only associated with dynamic low-level visual features. This is in line with contemporary models of face perception, in which the superior temporal region and the IFG belong to the dorsal face processing pathway specialized for the processing of dynamic features of faces . Since sFurthermore, we observed at a descriptive level that the stronger responses to fearful human in contrast to fearful avatar faces were more apparent in the right brain hemisphere. This is in line with a recognition advantage for facial expressions containing high-spatial frequency information presented to the left visual field and thus processed in the right brain hemisphere (as opposed to the presentation in the right visual field and processing in the left brain hemisphere; In contrast to previous studies applying static avatar stimuli . InvariaTo interpret our results, it is necessary to emphasize that we only found different BOLD responses to human and avatar faces when comparing fearful but not neutral expressions. Thus, emotional expressions exert an important influence on the processing of human and avatar faces. They activate the amygdala via a fast magnocellular channel conveying low-spatial frequency information and thus enabling rapid evaluation of general stimulus features . In lineIn addition, it is plausible that the amygdala has a modulatory effect on subsequent face perception via backward connections to cortical areas, such as the IFG and the STS . CorrespIn contrast to our results, Perceiving emotions in others almost inevitably elicites empathy in us, i.e. the attempt to understand and share the emotional state of another . It is nIt is assumed that mirror neurons, which typically discharge when performing a certain motoric action as well as when the same motoric action is observed, are a potential basis for empathy-related brain responses . This soEquivocally, we should consider that not empathy but rather processes related to emotional intelligence may account for the observed variance in brain responses and rating differences between human and avatar faces. It is possible that our lack of expertise with computer-generated faces is associated with lower measures of emotional intelligence regarding facial expressions of avatars. In line with this, previous studies have shown that measures of emotional intelligence are positively associated with anterior insula activity during face processing (Although our study contributes new findings, it has necessary limitations. First, we only used fearful and neutral facial expressions. Consequently, our results cannot be generalized to other facial expressions, as valence-dependent effects are possible (Future studies may clarify whether the complexity of computer-generated faces influences brain responses and recognition profiles of different emotional expressions. It is plausible that the absence of high-spatial frequency information in computer-generated faces represents a missing cue for emotion recognition. On the other hand, this absence could be advantageous because the facial expression may be less ambiguous. By using computer-generated faces showing only major features of facial expressions, we are able to generate tasks for emotion recognition with a low level of complexity . With inThere are both differences and similarities in the patterns of brain responses to humans and avatars depending on the expressions shown. While amygdalar and cortical responses in ventral temporal regions do not differ in response to fearful human and avatar expressions, dorsal temporal and inferior frontal regions sensitive to dynamic facial information exhibit distinct responses. Together, this means that although avatar expressions evoke amygdala responses associated with the fast processing of emotions similar to human expressions, the artificial facial motion of avatars is likely not to be processed in the same way as human facial motion. Furthermore, empathy-related processes or processes associated with emotional intelligence may interplay with this. These interpretations may have important consequences for the implementation of avatars in various commercial and public applications and research. Nevertheless, we do not believe that the current findings imply that computer-generated faces are of no use considering their methodological benefits. Instead, we consider it necessary that researchers acknowledge these potential limitations and implement avatars according to the overall goal of the study.scan-19-409-File009_nsaa039Click here for additional data file.


However, a lack of integrated studies deconstructing complex adaptations and linking candidate alleles with fitness consequences leaves room for much deeper exploration. Thus, we still seek the crucial direct link between the phenotypic effect of candidate alleles and their measured adaptive value – a prize that is exceedingly rare to achieve in any study of adaptation. We expect that closing this gap is not far off using the promising model systems described here.Serpentine barrens are among the most challenging settings for plant life. Representing a perfect storm of hazards, serpentines consist of broadly skewed elemental profiles, including abundant toxic metals and low nutrient contents on drought-prone, patchily distributed substrates. Accordingly, plants that can tolerate the challenges of serpentine have fascinated biologists for decades, yielding important insights into adaptation to novel ecologies through physiological change. Here we highlight recent progress from studies which demonstrate the power of serpentine as a model for the genomics of adaptation. Given the moderate – but still tractable – complexity presented by the mix of hazards on serpentine, these venues are well-suited for the experimental inquiry of adaptation both in natural and manipulated conditions. Moreover, the island-like distribution of serpentines across landscapes provides abundant natural replicates, offering power to evolutionary genomic inference. Exciting recent insights into the genomic basis of serpentine adaptation point to a partly shared basis that involves sampling from common allele pools available from retained ancestral polymorphism or  Further, the variation in elemental soil composition between serpentine barrens suggests no single “basis” of serpentine adaptation. On the other hand, as compared to other multi-hazard “extreme” environments , serpentines are fairly well-defined, dominated by the effect of few major elements. This, together with strong selective pressures, makes discovery of major-effect candidates seem likely, similar to what has been found for other soils such as metal contaminated and saline sites  studies applied to serpentine ecotypes so far have provided evidence for a simple genetic architecture of single traits  and parameters screened (the entire serpentine syndrome in nature: both known and unknown factors). In other words, genome scans can be both genetically and phenotypically agnostic. Such scans have been performed in two wild, outcrossing  arenosa  and A. l. lyrata , 2010. BM. guttatus innovatively combined field and genomic approaches by assessing survival differences of F2 mapping populations grown on serpentines by bulk segregant analysis  and between them (convergence). Given the many independent evolutionary paths that species (and indeed populations within species) have taken to serpentine-adaptive phenotypes, it is reasonable to expect reuse of common pathways, or even proximal molecular actors to the common serpentine challenges.M. guttatus, where two serpentine populations share a major QTL important for serpentine adaptation  the species . Yet, thArabidopsis and Mimulus genera have provided proof-of-concept that the genetic basis of serpentine adaptation is accessible, and the time is now ripe for synthetic studies (Given the perfect storm of challenges that serpentine presents, a comprehensive understanding of serpentine adaptation is a challenging task that requires multidisciplinary investigation . However studies . Here, wWe suggest that ongoing ecological and population genomic studies of serpentine adaptation hold strong potential to contribute to our understanding of fundamental evolutionary principles, similar to the contributions of early eco-evolutionary studies achieved in the 1950s and 1960s , 1967. AAll authors listed have made a substantial, direct and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The improvement of exercise performance encountered in sports not only represents the enhancement of physical strength but also includes the development of psychological and cognitive functions. Accumulating evidence has showed that physical exercise is a powerful way to improve a number of aspects of cognition and brain functions at the system and behavioral levels. Yet, several questions remain: what type of exercise program is the most optimal for improving cognitive functions? What are the real effects of some innovative exercise protocols on the relationship between behavior and the brain? To what extent do ergogenic aids boost cognitive functions? What about the efficacy of neuromodulation techniques on behavioral performance? Answers likely require combined insights not only from physiologists and sports scientists, but also from neuroscientists and psychologists. Published manuscripts (sixteen research papers and one perspective article from various academic fields) within this Special Issue “Studying Brain Activity in Sports Performance” did bring novel knowledge and new directions in human exercise-cognition research dealing with performance. Here, we summarize the main insights provided by the contributions and showcase the multiple relationships between cognitive functions, brain activity, and behavioral performance with applications in sports and exercise science.First, the cognitive benefits of acute and chronic (training) physical exercise need to move beyond simple aerobic activities and resistance training. From a multidisciplinary perspective, this Special Issue proposes some encouraging evidence for other types of physical exercise  or interventions (ergogenic help), improving executive functions for various populations  that have received far less attention. Following acute endurance exercise of 30 min at constant moderate intensity on a cycle ergometer, Wang et al.  observedBesides healthy adults, a few original articles in this Special Issue were related to other target clinical populations and aimed to test the beneficial effects of novel exercise protocols on cognition. Wen and Tsai  investigExecutive functioning is important for athletic performance and can be affected by ergogenic aids. The ergogenic effect of caffeine might not only enhance physical strength but also the development of cognitive functions during exercise. Wang et al.  in a croSecond, after establishing a link between exercise and the outcomes of some specific cognitive function tests, using neuroimaging techniques is required to reveal the electrophysiological (EEG) processes that are associated with some behavioral changes, or evaluate the physiological changes  over the course of the intervention or training program, that have the potential to improve executive functions. In their perspective article, Herold et al.  overviewThird, enabling enhanced physical exercise may benefit from muscle strength and whole-body movement improvements. Within this context, non-invasive brain stimulation  approaches have been employed to uncover strength-related brain–muscle (movement) associations. By applying tDCS over the primary motor cortex and the cerebellum with traditional montages, a randomized counter-balanced sham-controlled double-blinded cross-over design study from Kenville et al.  providedTo conclude, this Special Issue showed that the acute or chronic exercise effects upon cognitive functions are varied and dependent upon exercise duration, intensity, and mode, as well as the type of cognitive tasks assessed. The considerable advancement in wearable neuroimaging methods coupled with neural recording and non-invasive brain stimulation for studying human brain physiology are now enabling the gaining of new insight into the complexities of the behavior–exercise relationship, where the prefrontal cortex as a whole plays a cardinal role in the temporal organization of behavior and cognitive activities. This Special Issue makes the point that exercise and sport sciences are a joint research initiative of exercise scientists, psychologists, neuroscientists, neurophysiologists, and other related disciplines such as neuroimaging.


Effective treatment interventions for childhood obesity involve parents, are multicomponent and use behavior change strategies, but more information is needed on the mechanisms influencing behavioral outcomes and the type of parental involvement that is efficacious in behavioral treatment interventions with school-age children. This review aimed to understand key characteristics of programs that contribute to dietary and physical activity behavioral outcomes, and through which key mechanisms. This was a systematic review with narrative synthesis following PRISMA guidelines and realist analysis using RAMESES guidelines to explain outcome patterns and influence of parental involvement. Overall, the findings contribute to understanding the complex relationship between family barriers to behavior change, strategies employed in treatment interventions and behavioral outcomes. Implications for enhancing future policy and practice include involving parents in goal setting, motivational counselling, role modeling, and restructuring the physical environment to promote mutual empowerment of both parents and children, shared value and whole-family ownership in which intrinsic motivation and self-efficacy are implicit. These characteristics were associated with positive dietary and physical activity behavior change in children and may be useful considerations for the design and implementation of future theory-based treatment interventions to encourage habitual healthy diet and physical activity to reduce childhood obesity. Reducing the prevalence of childhood obesity is a global public health focus. In 2016, over 340 million school-aged children and adolescents aged 5–19 were overweight or obese  even the most authoritarian parents began to relinquish unnecessary levels of control by week 8 as they witnessed their children making reasonable choices”.Siwik et al.’s strategy was congruent with the premise in the literature that a key parenting practice is to create a protective environment in the home applicable to the whole family to avoid stigmatizing obese children . This su Epstein  that chiAn emphasis on being non-judgmental, reducing the stigma of the “obese child”, and being supportive and sensitive to socioeconomic determinants of obesity ,50 was aThree studies included in the realist review used strategies based on the assumption that positive parent health-related outcomes plus involvement in the intervention are key tools to achieving sustainable change for their children ,45,51 F. SCT posTwo key barriers to behavior change these studies sought to address were (i) individual differences in rates of learning and parenting behaviors , and   employedIn GOALS , parentsIn terms of whether significant changes by parents accounted for children’s results, the findings were mixed. Epstein et al. (1994)  demonstrThis review aimed to determine the key mechanisms for the ways in which family-based behavioral treatment interventions for overweight and obesity in community settings contribute to diet and physical activity behavioral outcomes. Overall, more than half of the 36 identified family-based interventions promoting diet and physical activity were effective in positively changing behaviors. In terms of key mechanisms by which family-based behavioral treatment strategies resulted in their outcomes, there was considerable heterogeneity and the final program theory was not able to establish clear links between the most prominent BCTs, psychological factors and outcomes. This highlighted the complexity and multifaceted nature of treatment interventions for childhood obesity and the multitude of barriers to behavior change as starting premises for the intervention components.This review found no clear set of ‘ingredients’ for an ideal program but importantly certain program features were common across successful dietary and/or physical activity behavior outcomes. These features were promoting intrinsic motivation and self-efficacy through empowering children and their parents, fostering shared value between children and parents, and whole-family ownership. Social Cognitive Theory was a prominent theoretical foundation in this review. The use of such theories of motivation has been common in child obesity interventions aiming to influence engagement in positive health behaviors by promoting self-efficacy and intrinsic motivation ,98. TherThis review reinforces the impact and importance of parental involvement on mechanisms associated with behavioral outcomes. Although there was no standout types of parental involvement, promoting parent support of the child’s behavior demonstrated a slightly higher proportion of diet and/or physical activity behavior change. Cohesion and agreement within families was the key to successful behavior change, which echoes the wider literature . This reThis review and realist analysis offers important opportunities for future research. For example, the mechanisms and clinical implementation of the novel approach to parental involvement in child obesity treatments referred to as ‘Conjoint Behavioral Consultation’ offers potential . FurtherThis review has several strengths including the use of a standardized reporting format  helping to minimize variations and aid information synthesis in future reviews. A number of weaknesses must be acknowledged as well. Firstly, the cause of childhood obesity is not at all simple and we did not consider factors such as genetic predisposition, sociodemographic factors, and psychological and environmental factors. A social ecological approach would be ideal for the focus of further reviews to acknowledge social and environmental influences on behavior change. However, the realist analysis did not draw upon stakeholder expertise as modelled by a recent realist analysis  which inImplications of this research include involving parents in goal setting, motivational counselling, role modeling, and restructuring the physical environment to promote mutual empowerment of both parents and children, shared value and whole-family ownership in which intrinsic motivation and self-efficacy are implicit. These characteristics were associated with positive dietary and physical activity behavior change in children and may be useful considerations for the design and implementation of future theory-based treatment interventions to encourage habitual healthy diet and physical activity to reduce childhood obesity.


Enhanced spreading of semantic activation has been hypothesized to underlie some of the most significant symptoms of schizotypal personality, like thought disorder, odd speech, delusion, or magical thinking. We applied the Deese/Roediger-McDermott false memory task to the study of semantic activation in a group of 123 non-clinical individuals varying in the three dimensions of schizotypal personality: positive, negative and disorganized schizotypy. In the study phase, we presented them with lists composed of words semantically associated to unpresented critical words. Then, they responded to a recognition questionnaire including previously presented words and critical unpresented lures, as well as weakly related and unrelated unpresented lures. Participants rated their confidence in recognizing each word. They also filled in a standardized schizotypal personality questionnaire. Confirming the false memory effect, recognition ratings provided in response to critical words were higher than those produced for both weakly related and unrelated items. Crucially, scores in the positive dimension increased recognition percentages and confidence ratings for weakly related and unrelated lures. This study indicates that high levels of positive schizotypy might influence the tendency to accept false memories of unrelated unpresented material. Schizotypal personality has been considered to be an indicator of vulnerability to psychosis , based oAtypical patterns of semantic activation in individuals with schizotypal personality have been hypothesized to underlie some of the most significant schizotypal symptoms, like thought disorder , odd speThe Deese-Roediger-McDermott (DRM) paradigm  has alsoDespite several previous attempts to clarify the relation between schizotypal personality and semantic processing, it remains uncertain whether schizotypy affects false memory rates or, if it does, whether one specific schizotypal dimension is responsible for that effect. Factor analyses-based studies have suggested that three dimensions can be distinguished within the schizotypy construct: positive, negative and disorganized . The pos1 ].Although schizotypal personality has proven to be a useful construct to capture individual variability, it also presents problems due to a vague operationalization of its constituent dimensions based on various measurement instruments, often inconsistent with each other . This prIn sum, previous studies have failed to provide a consistent pattern of results regarding the possible association between false memories and schizotypal traits. Those assessing the relation between false memories and positive symptoms have reported positive , negativOne possible explanation for the incongruences observed in previous studies addressing the association between schizotypal personality and false memories is that the former is not related with false memory rates, but with overconfidence in them . According to Thus, it could be the case that schizotypal traits are not directly related to specific semantic-related memory processes, but to metacognitive awareness. The degree of confidence in one’s memories is of utmost importance because memories considered unreliable will probably be dismissed and will differentially affect our behavior compared to those we consider reliable see . In thisPrevious studies from the pattern recognition domain have observed that individuals showing greater belief in paranormal phenomena, which is considered to be a positive trait, tend to require a lower amount of objective evidence for the perception of meaningful patterns . The resFinally, the origin of the discrepancies between previous studies could also be related to a lack of control of certain characteristics known to influence false memories. In their review of emotion effects on false memory, The aim of our study was to clarify the association between the different dimensions of schizotypal personality and semantic processing by means of a DRM task. The recognition questionnaire included target or presented words, critical or related unpresented words, weakly related unpresented words and unrelated unpresented words. We hypothesized that the volunteers should accurately recognize presented words based on both item-specific and semantic memory traces. Following spreading-of-activation interpretations of DRM effects , we expeMoreover, our recognition questionnaire consisted of a scale in which the volunteers had to state how confident they were that each word had or had not been presented. We could expect that if schizotypal personality is associated with differential metacognitive processes, then variability in the measures of schizotypal traits should be related with confidence in the recognition responses. Following previous studies , we coulSD = 3.25) took part in the experiment in exchange for course credits. They were all native speakers of Spanish or Spanish-Catalan native bilinguals. The university’s ethics committee  approved the study protocols (IRBOOOO3O99) and written informed consent was obtained from all the volunteers before their participation in the study. We treated all data anonymously.A group of 123 Psychology students from the University of Barcelona  for 18 critical lures selected from Spanish free association norms . The norSD = 0.05) and 5th  positions in each list to be presented as target (presented) words in the recognition test . For each list, two not presented words from positions 13th and above of each list not appearing in any of the other lists were selected as weakly related lures . Items in this category correspond to words for which, when presented in free association tasks, only a low percentage of individuals produce the critical word . Finally, two unrelated words per list were also selected for the recognition test . For each of the 126 words used in the recognition questionnaire, we gathered valence and arousal data using values from Emofinder , we selected the words in the 1st  and recognition confidence ratings. We started by analyzing the recall and recognition percentages corresponding to each word category in relation to the schizotypy measures by means of correlation analyses. Regarding recall, we analyzed the percentages of words produced by each participant which corresponded to target words (presented) and critical lures  as well as other unpresented words different from the critical lures. As for recognition, we started analyzing percentages of responses indicating that the word had been presented (aggregating responses 3 and 4 in the recognition questionnaire) for each word category.d’true values comparing hits to presented items with false alarms to unrelated unpresented items, as well as d’critical comparing the false recognition of critical items to false alarms to unrelated unpresented items, and d’weak, comparing the false recognition of weakly related items to false alarms to unrelated unpresented items. In order to correct our data when any of our participants presented zero hits or zero false alarms, which prevents the calculation of d’ scores, we followed the procedure recommended by d’-values close to zero, whereas those who tend to accept target words  and reject unrelated unpresented lures obtain higher d’-values.Second, given that recognition percentages can be misleading because recognition responses depend on how liberal or conservative the participants are, we calculated three sensitivity scores for each participant following the signal detection theory approach . We calcThird, we analyzed the confidence in the ratings separating between responses indicating that a given word had not been presented (responses 1 and 2) and those indicating that the item had been previously presented (responses 3 and 4). For this analysis, we recoded the responses so that those showing high confidence  were assigned 2 points, whereas those responses showing low confidence  were assigned 1 point. This way, we obtained separate measures for confidence in rejection and acceptance responses.k = 1.99, indicating acceptable levels of interrelation between the predictor variables  and participant-level  variables. Standardizing continuous numeric predictors eliminates non-essential collinearity due to scaling  and alloGiven the ordinal nature of our dependent variable, we analyzed the recognition ratings through cumulative link mixed models using the clmm2 function furnished by the ordinal package . CumulatWe present the results of the model including the maximal random effects structure justified by our design , and between recognition percentages for unrelated stimuli and scores in the disorganized dimension (p = 0.045).All data are available from the OSF databaseyses see  conducteyses see  showed nyses see , barely d’ and schizotypy measures indicated decreased discriminability for true items for participants with higher disorganized scores. Interestingly, we observed a significant negative correlation between discriminability of critical lures and scores in the positive dimension , indicating that the volunteers were, on average, prone to mistakenly consider them as presented. New correlation analyses without these two items yielded the same results as those conducted with the full item set , ruling out possible influence of specific characteristics of these items in the results.The presence of an association between schizotypal traits and confidence ratings for weakly related stimuli led us to conduct a by-items examination of our results in order to ascertain whether the observed effects could be influenced by the inclusion of specific weakly related words in the analyses. Two of the weakly related items elicited mean confidence ratings higher than three  = 507.96, p < 0.001, and Akaike information criterion .The datasets generated for this study can be found in the OSF (The studies involving human participants were reviewed and approved by the Comissió de Bioètica de la Universitat de Barcelona (CBUB – IRBOOOO3O99). The participants provided their written informed consent to participate in this study.JR-F designed the study, supervised the data collection, and wrote the first draft. MA collaborated in data collection. JR-F and RD analyzed the data. JR-F, MA, and RD contributed to the manuscript revision, read, and approved it.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Fetal membranes or amniochorionic membranes are one of the most intriguing tissues in the intrauterine cavity that are essential for the protection of the fetus, maintenance of pregnancy, and as a signaler to initiate parturition  a sin utero, mechanisms underlying its longevity, and cellular biological properties are essential to determine the pathological complications contributing to its dysfunctions and adverse pregnancy outcomes.pregnancy as well as promote parturition  was formed in 2019. This Research Topic has been assembled in response to encouragement by the FMS to highlight the latest data and chart future prospects in fetal membrane research.The challenges to understand fetal membrane functions and dysfunctions are substantial. A major issue is the impracticality of obtaining fetal membrane tissues during pregnancy unless delivered preterm. In addition, pregnancy complications associated with preterm deliveries can confound membrane studies. Animal models often do not mimic human fetal membranes. These circumstances impede research and limit the translational impact of work in the field. To help addressing the needs of interested scientists, the Fetal Membrane Society  cells by 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1). A review on this topic is provided by Wang et al. which discusses the physiologic and pathologic contributions of 11β-HSD1 in fetal membranes and its contributions to preterm and term parturitions. Another key mediator that maintains fetal membrane homeostasis during pregnancy is progesterone. Marinello et al. describes how amnion membrane matrix mesenchymal cells use cell membrane-associated progesterone receptors in providing a progesterone dependent anti-inflammatory environment during pregnancy.The collection includes 16 articles covering the basic, functional, and translational aspects of fetal membrane biology to show how these tissues contribute to pregnancy and pregnancy associated pathologies. Three manuscripts discuss the in utero and aging progresses during gestation. Premature senescence of the fetal membranes leading to dysfunction is one of the reasons underlying pPROM and PTB. The review by Polettini and Guimarães da Silva focuses on the telomere-dependent aging of fetal membranes and its association with telomerase dynamics.Like all other tissues, fetal membranes start aging inflammation. One of the classic signs that describe the severity of adverse pregnancy outcomes and a factor often used to design management strategies of preterm neonates is based on the histologic assessment of fetal membrane inflammation. Infiltration by neutrophils, termed histologic chorioamnionitis, is an indicator of the severity of inflammation in the intrauterine cavity. Compromising immune defense is one of the mechanisms causing fetal membrane inflammation, immune cell infiltration, and membrane dysfunctions leading to chorioamnionitis-associated preterm birth or pPROM. Exogenous risk exposures and their interactions with endogenous factors like the microbiome often amplify pregnancy risk. An article about maternal environmental pollutant exposures that can weaken innate immune defense is discussed by Harris et al. In this review, authors report risk modification by an interaction between the environment and pathogenic vaginal colonizers such as Group B streptococcus. Wheeler and Oyen review data related to the interesting hypothesis that fetal membranes may actually behave as an inflated balloon, and pressure changes during severe weather events can promote pPROM. Inflammation and inflammatory cytokines are well-studied in the fetal membranes. A report by Pardon et al. in this Research Topic and multiple reports by Vincent Sapin and Loïc Blancho's group have provided novel ideas to this area by discussing the roles and contributions of damage associated molecular patterns (DAMPs) and their receptors. By analyzing DAMPs and their receptors on maternal biological samples and exosomes, Sapin's group report a lack of association between these markers and pPROM. As reported, high abundance of DAMPs in fetal membranes and compartments is suggestive of fetal contributions to preterm birth and pPROM.Fetal membrane pathologies are often associated with growth, remodeling, and repair. A better understanding of these mechanisms can improve outcomes of fetoscopy and fetal surgeries as these procedures often contribute to membrane rupture and preterm delivery. Based on their own reported findings, Mogami and Word summarize how epithelial mesenchymal transition (EMT) of amnion epithelial cells can be a mechanism in fetal membranes that can heal membrane wounds. Approaches to enhance this process are also discussed. Multiple factors have been described to promote EMT in various cells. One of them is IL-6, a cytokine produced in high abundance during fetal membrane dysfunctions. IL-6 has been considered one of the cytokines contributing to preterm birth and pPROM as its levels are very high in all pregnancy associated biological fluids and tissues. A basic science report is included in this Research Topic by Omere et al. where they studied IL-6's effect on amnion epithelial cells. IL-6 lacks cell fate determining properties even at levels seen during pPROM or PTB and it does not appear to promote EMT in fetal membrane cells.An understudied area of fetal membrane research is its Wang et al. to determine membrane pathologies. The potential of this approach as an imaging-based biomarker to test membrane function is reviewed by Qi et al. In their report, Omere et al. also describe the role of IL-6 as a potential biomarker associated with adverse events, although IL-6 does not seem to have any specific impact on fetal membrane cells.Lack of biomarkers indicative of membrane dysfunction hampers clinical management of membrane-associated complications such as pPROM and spontaneous preterm birth. An innovative approach using MRI has been introduced recently by Richardson et al. points out how innovative approaches like fetal membrane organ on a chip (OOC) can be used to replace traditional 2D cell cultures, transwells, and organ explant models to study fetal membrane biology and function. In their review, three phases of fetal membrane OOCs development have been outlined and its advantages and disadvantages are discussed. Without a doubt, the in vitro reconstruction of fetal membrane tissues is an exciting futuristic approach with great potential.Fetal membrane research is notoriously obstructed by a lack of specialized techniques and tools. In summary, the Research Topic “The Role of the Fetal Membranes in Pregnancy and Birth” provides a comprehensive overview of fetal membrane biology, endocrine, mechanical, immune functions, normal and risk-associated changes to the membranes, development of biomarkers to diagnose membrane functions or dysfunctions, and tools to study fetal membranes. The collection of papers, under the auspices of the Fetal Membrane Society, will certainly promote fetal membrane biology to take its deserved place of prominence in perinatal science.RM, ML, and TZ contributed to this editorial. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Defining neuronal cell types and their associated biophysical and synaptic diversity has become an important goal in neuroscience as a mechanism to create comprehensive brain cell atlases in the post-genomic age. Beyond broad classification such as neurotransmitter expression, interneuron vs. pyramidal, sensory or motor, the field is still in the early stages of understanding closely related cell types. In both vertebrate and invertebrate nervous systems, one well-described distinction related to firing characteristics and synaptic release properties are tonic and phasic neuronal subtypes. In vertebrates, these classes were defined based on sustained firing responses during stimulation (tonic) vs. transient responses that rapidly adapt (phasic). In crustaceans, the distinction expanded to include synaptic release properties, with tonic motoneurons displaying sustained firing and weaker synapses that undergo short-term facilitation to maintain muscle contraction and posture. In contrast, phasic motoneurons with stronger synapses showed rapid depression and were recruited for short bursts during fast locomotion. Tonic and phasic motoneurons with similarities to those in crustaceans have been characterized in Drosophila, allowing the genetic toolkit associated with this model to be used for dissecting the unique properties and plasticity mechanisms for these neuronal subtypes. This review outlines general properties of invertebrate tonic and phasic motoneurons and highlights recent advances that characterize distinct synaptic and plasticity pathways associated with two closely related glutamatergic neuronal cell types that drive invertebrate locomotion. With few exceptions, every cell in an animal has the same gene set encoded in their chromosomal DNA. Yet different cells navigate unique paths to differentiation and express only a subset of the individual genes that define what that cell becomes and how it works within the organism as a whole. There is no place where that diversity is on display more than in the nervous system. Thousands of individual cell types are found in the brain, each forming connections with many other neurons. This developmental feat gives rise to a biological machine that processes external stimuli and combines it with internal motivation states and prior experiences to guide ongoing behavior. Beyond the remarkable diversity of cell types, neurons can also rapidly alter the genes they express to guide modifications in their activity and structure that contribute to behavioral plasticity . How neuA goal for many in the field has been to unravel the genomic complexity of neurons and create comprehensive brain cell atlases . Can oneSynapses are key sites where information is transferred between neurons. Changes to their structure or function can alter local information flow or circuit activity as a general mechanism for behavioral plasticity. Indeed, synaptic competition during assembly of neural circuits has emerged as an important component of brain development. Although activity-dependent synaptic competition is widely studied, how plasticity of inputs from distinct neuronal classes are regulated and the role of post-synaptic cells in this process is still being elucidated. Much of the work in this area has focused on the interplay between excitatory and inhibitory inputs that control overall output of a circuit, along with the neurodevelopmental disorders that occur when the process is disrupted . Among iTonic and phasic neuronal subtypes were initially defined in vertebrates based on distinct excitability properties, with tonic neurons firing in a sustained manner and phasic neurons displaying burst properties with rapid adaptation. Studies of invertebrate locomotion revealed motoneuron subclasses with similar differences in excitability properties. Although initial studies of phasic and tonic motoneurons where described in amphibians ,b, studiAlthough several differences in crustacean and Drosophila neuromuscular junctions (NMJs) have been described , the latA receptors are located in both synaptic and extrasynaptic membranes in the brain and represent the primary receptors for inhibition. Synaptic GABAA receptors mediate phasic inhibition, while extrasynaptic GABAA receptors mediate tonic inhibition. Dysfunction in tonic or phasic inhibition has been associated with epilepsy, depression, and anxiety , tonic and phasic dopaminergic neurons contribute to behavioral flexibility associated with task switching vs. maintaining a learned behavior , 2015. T anxiety . As such2+ and represents the primary driver for contraction  at single AZs, indicating individual release sites have on average higher Pr at phasic Is terminals  to a smaller fraction (∼10%) of high Pr AZs (0.2–0.7) depending on extracellular [Ca2+] (Pr has been mapped to increased accumulation of presynaptic Ca2+ channels and late AZ scaffolding proteins like Bruchpilot (BRP) at high Pr AZs in Ib terminals  were also ruled out as potential mechanisms. Indeed, tonic synapses were found to have a larger pool of docked SVs by EM (11 vs. 4) and a bigger RRP pool measured by electrophysiology (130 vs. 60) compared to phasic synapses  in Pr are currently unknown. Defining these mechanisms would be highly informative and potentially suggest molecular changes that might be harnessed for other forms of presynaptic plasticity throughout the nervous system , appears to alter Ca2+ buffering and ATP production between the two terminal types. Indeed, PMCA plays a key role in Ca2+ extrusion at Ib terminals, with lower activity at Is synapses  that controls SSR development  with large infoldings of the muscle post-synaptic membrane that develop around the boutons over the course of larval development . Mature elopment . The difelopment . Drosophelopment . Whetherelopment  and thatelopment  suggest Neuronal plasticity occurs through multiple mechanisms across invertebrate and vertebrate species and is often associated with changes in synaptic strength or synapse number . For exaSynaptic competition is a widespread form of plasticity that leads to robust synaptic inputs being strengthened and weaker inputs undergoing elimination, occurring in both the CNS and peripheral nervous system (PNS) of vertebrates . SynaptiNeuromuscular junctions are not classically considered highly plastic synapses, but mutants in many of the genes required for learning and memory in the Drosophila brain also show defects in synaptic function or morphology at the NMJ . As a glPr and RRP size, tonic Ib synapses typically display short-term facilitation while phasic Is synapses undergo short-term depression during high frequency trains  appears to have both shared and distinct roles in the two neuronal populations . In additeolysis . Definin2+ sensitivity of release between the two classes. It remains to be seen what parallels at the molecular level are present between invertebrate tonic and phasic neurons and vertebrate neurons with similar properties. Once the underlying mechanisms are identified in a genetically approachable system like Drosophila, it will provide a robust set of hypotheses that can be tested in mammalian models. In conclusion, further dissection of the pathways that generate the unique properties of invertebrate tonic and phasic neurons will no doubt continue to provide important insights into neuronal diversity mechanisms that contribute to biophysical properties, synaptic target choice, synapse structure and function, and differential synaptic plasticity pathways.In addition to changes in the transcriptome of tonic and phasic motoneurons, post-translational modifications to ion channels and other synaptic proteins may alter their location or function differentially within the two neuronal classes. Altered phosphorylation of key components of the SV fusion machinery would be an interesting target for the dynamic differences in CaAll authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Dear Editor:Tharakan et al. found a trend of increased mortality with poor temperature control in severely ill COVID-19 patients and suggest further studies to determine if controlling high temperature might alleviate the inflammatory response and improve outcomes [ClinicalTrials.gov Identifier: NCT02706275) has recently completed enrollment.Multiple aspects of both humoral and cellular immunity -induced α4 integrin activation and signaling, and macrophage function) are boosted by elevated temperature, and numerous studies have found no benefit to treating fever of infectious origin . After aWith the results of the Induced Hypothermia in Patients With Septic Shock and Respiratory Failure (CASS) randomized, controlled study, as well as a more recent but smaller randomized study of fever reduction in sepsis showing potential harm , we woulSerena Tharakan and Kiyotake IshikawaWe thank Drewry et al. for their interest in our Research Letter. As the authors pointed out, we agree and noted in our letter that we were limited by our inability to adjust for disease severity due to the lack of relevant parameters in our dataset. Still, the correlation between maximum body temperature and mortality from COVID-19 was obvious and showed a clear trend. Thus, we believe our conclusion remains solid: maximum body temperature during the course of COVID-19 is a good and easily obtainable predictor of worse outcomes.Drewry et al. discussed the potential benefit of elevated body temperature based on the results of clinical trials that primarily included patients with bacterial sepsis. However, there is a lack of evidence to suggest that sepsis due to SARS-CoV-2 and bacterial sepsis have the same pathophysiology or can be treated in the same way. There are notable differences between sepsis due to virus and sepsis due to bacteria . Remy etElevated body temperatures may augment the immune response to SARS-CoV-2 and potentially inhibit viral replication. However, it is important to consider this argument in the context of the widespread damage that high body temperature might cause on top of virally induced systemic inflammation. Fever increases metabolic demand and oxygen consumption of many organs, which may exacerbate tissue injury caused by systemic inflammation. For example, it is known that COVID-19 is associated with increased risk of myocardial injury . Fever cGiven the observed high mortality in patients with elevated temperatures and the paucity of data on temperature management for COVID-19, it is important to explore every potential approach to identify optimal temperature management strategies. This includes warming studies as Drewry et al. mention, as well as maintaining normothermia and hypothermia.


Successfully preparing for and conducting on-animal ectoparasiticidal evaluations is key in providing accurate results and inferences on product performance. However, the procedures associated with designing sound-reliable research projects while using animal test subjects can become complex. The current manuscript offers insights towards the characterization of different evaluation types highlighting key considerations and potential problematic barriers that may otherwise be overlooked by researchers new to the area of on-animal product evaluation. Furthermore, recommendations on reporting inferences from findings based on various study designs are discussed. The authors of the current manuscript offer these considerations in the hopes of maintaining harmony in future reports used to develop and evaluate on-animal ectoparsiticidal products in the field of veterinary entomology. The complex parasitic interactions that drive large economic losses in agricultural animal production continue to provide a broad range of challenges for producers and researchers alike . In respParasiticidal intervention has long been an effective tool in combating the detrimental effects associated with a broad range of economically important animal pests . UnfortuThe purpose of the current manuscript is to offer general guidelines that can be used to ensure consistent and reliable product efficacy determinations for both marketed and experimental products aimed at on-animal dosing and exposures. An all-inclusive effort which would transcend multiple pest species and even commodities is beyond the scope of the current effort. Moreover, available products, application types, and formulations across animal production systems are as vast as the species labeled for control. However, many of the concepts presented may be applicable on a case-by-case basis. For purposes of simplicity, the following guidelines are intentionally structured broadly around key approaches, concepts and considerations for future researchers attempting to undertake on-animal product testing. The authors in no way, challenge existing guidelines or recommendations. Rather, we offer insights within existing procedures that may be useful to novice researchers in the field of ectoparsiticide evaluations.On-animal product assessments, as the name would imply, include the use of animal test subjects. Prior to any work being conducted on animal, appropriate regulatory  approval is necessary which includes the justification for use of live animals. Furthermore, the experimental use of agricultural animals generally falls under United States Department of Agriculture guidelines. Under no circumstances should animals be used for research purposes prior to or without the appropriate authoritative approvals. In fact, approvals are often required for publication in most scientific journals. There is a plethora of resources available to ensure animal welfare and established guidelines for certain species that are already in place .The first step to any on-animal product evaluation must begin by identifying specific research objectives. Many off-animal or animal alternative models might be more suitable for certain projects and should be considered. For instance, parasiticidal responses of given pest population can be assessed using laboratory based toxicological evaluations . Such teAnimals will constitute experimental units driving statistical assessments of findings. As with all scientific investigations, the number of experimental units used to conduct a study should provide statistical power and reliability in support of the study design. Proper identification of the experimental units for any on-animal product testing is key. Typically, in field applications animals are treated individually but housed as a herd. In this case, the herd should serve as the experimental unit . TherefoLess commonly, researchers may be afforded the ability to utilize individual animals as the experimental unit. In this case, a combination of housing, treatment, infestation, and assessments are made at the animal level. Typically, while animal numbers can be decreased in this scenario, labor associated with conducting these types of projects are increased. Regardless of the role of the animals used in on-animal product evaluations, individual animal descriptions are suggested to establish clarity of the procedures used for the reader. Physiological state, age, sex, and breed should be detailed for any experiment using animals. When using individual animals as the experimental unit, the authors suggest allocating treatments based on pretreatment population assessments and/or body weights of the animals.In general, the authors have identified two types of assessments that are heavily relied upon when conducting on-animal product testing; 1) General confirmational studies which include marketed product evaluation and assessments on product performance and 2) experimental evaluations of off-label or non-marketed products. When evaluating existing marketed products through confirmational studies, label directions will dictate procedures and use of the products as results and inferences should be structured around applicable product directions. Researchers might want to use these confirmational studies to provide insight on regional variations of product performance which would offer more detailed insight towards future recommendations. Additionally, confirmational studies can be structured to evaluate various rotational programs, integrated pest management assessments, and/or offer early detection of the onset of pest population resistance to various parasiticidal products.The authors classify any off-label product use as an experimental evaluation. When researchers are conducting experimental evaluations as described above, characteristics of the procedures employed will need to be scientifically just and approved by regulatory groups with jurisdiction over specific research groups. However, specific research objectives can be dramatically broadened as the application and subsequent findings with regard to performance are no longer bound by label claims. General on-animal product development employs the use of experimental evaluations to screen novel compounds and/or formulations and explore dosage rates leading to cost effective large scale development. For any experimental evaluation, appropriate animal disposition should be clarified and approved prior to initiation.Another common factor to consider when designing on-animal product evaluations would be animal housing and testing location. Modifications of existing facility structures to enhance on-animal product evaluation capabilities will rarely, if ever, be justified in budgetary requests. As such, many researchers interested in assessing product performance must design product assessment studies within the confines of specific environmental capabilities and limitations. Oftentimes, in the case of confirmational studies, collaborations with local animal producers become beneficial. In this case, researchers should ensure that the facility chosen for an on-animal product assessment provides the necessary working environment that would facilitate both animal and worker safety. In many cases, producer collaborations extend research potentials in areas where on-animal product evaluations would otherwise not be possible. The authors implore researchers working in this capacity to nurture these relationships by exhibiting professionalism in their interactions as they not only represent themselves, and their institution, but the entire field of researchers utilizing similar resources. Collaborative producers will continue to be valuable contributors to on-animal product evaluations and will most likely be the first to adopt novel approaches for wide-scale implementation.A final key consideration when designing a study evaluating on-animal product performance is labor. Effective, safe, and appropriate animal husbandry during trials can become rather laborious. Under no circumstances, should an experiment be conducted using animals that will not be provided the most suitable methods to alleviate any undue pain or suffering. Ensuring animal and researcher safety often will require multiple individuals to be listed as study participants. Research leaders should properly plan for experienced personnel to be available or in attendance during each animal handling event.Following the identification of key objectives and establishment of an appropriate study design, the authors suggest the development of a calendar of events structured around treatment application and other key data collection and animal handling events. This document should detail each activity and provide insight to potential scheduling conflicts with key personnel that could jeopardize successful completion of said activities or more importantly, compromise animal health.A properly prepared on-animal product assessment protocol will ensure a successfully conducted research project. For purposes of the current document, no standard protocols or procedures are presented. Rather, an arguably over simplification of commonalities across a wide application of scenarios is presented.Pretreatment population assessments are key in determining population reductions in response to product application or performance. When making claims or inferences on a products ability to effectively reduce an existing population associated with an animal(s), pretreatment population assessments are necessary. Guidelines for estimating population densities vary from pest to pest and techniques utilized by the researchers evaluating product performance should be validated or commonly accepted for the given pest of interest. Pretreatment population assessments should be conducted at a time prior to treatment application. It may be common and in the best interest of the researcher to conduct such an estimate on the same day as treatment application but prior to administration. If researchers are estimating populations from a subset of animals within a herd, effort should be made to sample from the same animals throughout the trial to minimize inter-animal variation in population densities.The procedures used for posttreatment population assessments should mimic those used to establish pretreatment populations. Sampling times will be dictated by individual pest species biology and general constructs of the study. Additional data, such as animal performance, behavior or health can be incorporated in protocols in an attempt to demonstrate the positive effects as a result of effective pest population reduction.Documentation of key events such as, treatment times, application rates, and population assessments is critical when justifying claims of performance for all types of on-animal product testing. This becomes particularly important when experimental evaluations are aimed at product registration or general label inclusions. If researchers are working in this capacity, strict adherence to Good Laboratory Practice GLP;  or Good For simplicity sake, the authors will address two common metrics of determining effective product performance. The first, population reduction, is in regards to an animal(s) posttreatment pest population assessments as a percentage of the pretreatment population and is commonly used when evaluating repellent compounds . InferenAlternatively, general efficacy is another common metric of product performance. In this case, the use of and access to a control animal(s) is essential as efficacy calculations require relatable control population estimations . InferenControl populations should also be scrutinized by researchers. Efficacy calculations can be misleading in cases where control performance is less than optimal and fail to accurately represent naturally occurring population levels. As such, statistical comparisons may be used in addition to efficacy calculations to highlight control performance. Application of appropriate statistical comparisons may highlight variation between samples that is overlooked when using efficacy calculations alone. Specific statistical models should be structured to account for study design. Statistical comparisons may help solidify efficacies but must also provide biological soundness.It should be noted that researchers may become limited in their ability to design statistically sound approaches to evaluating efficacy in response to limited animal availability. Modifications of ideal study designs including animals serving as their own control and adjusting treatment schedules across time may be implemented to maximize available animals for testing purposes.A variety of environmental influences may impact findings and inferences regarding on-animal product performance . ResearcWhen conducting on-animal product evaluations, researchers should incorporate scheduled health evaluations in an attempt to maximize animal welfare. If adverse events occur, effort should be made to determine whether or not the event was due to the test product.On-animal product evaluations can become costly for a number of reasons. The following discussion draws from the authors own experiences conducting on-animal experimental evaluations. Dependent upon commodity, budgetary expenses associated with animal purchases used in experimental evaluations may account for as much as 60% of allocated budgets. The remainder of budgetary allocations are devoted to supplies (~10%) and labor (~30%). Furthermore, off-animal models of product performance  can be conducted for less than 1/10th of those conducted directly on-animal . Budgetary expenses will undoubtedly vary between researchers and locations. However, when preparing to conduct an on-animal product evaluation, researchers should ensure that the cost associated with safely and accurately conducting the study is reflective in appropriate funding requests.The results of any on-animal product assessments can take many different shapes depending upon variables of interest and related research objectives. Therefore, comprehensive recommendations for providing research findings is beyond the scope of the current effort. Rather, general guidelines for considerations are provided below to in an attempt to capture a broad range of measurements taken.Pest population descriptions are key in product assessments and require special attention when inferring product performance based on findings. Pretreatment pest populations should always be reported to detail natural occurring densities. Such assessments are particularly important when attempting to demonstrate population reductions in response to product application without the use of a control. In cases such as these, researchers may report reductions but must always disclose the reasons for control exclusion. Furthermore, inferences regarding the duration of reductions or product to product comparisons using observed reductions must be done with great caution.The use of efficacy calculations and subsequent inferences on product performance in terms of pest population management should be restricted to research studies which utilize controls. Inferences between and within labeled on-animal products should additionally be presented in relation to labeled claims , pretreatment population reductions, and duration of efficacies based on suitable control performance can be reported.There are a number of reasons for researchers to pursue on-animal product evaluations which make providing specific guidelines difficult. In the present manuscript we provide generalized concepts, not in an attempt to displace existing guidelines established for individual pests or commodities, but to offer additional considerations for new researchers pursuing objectives related to on-animal product evaluations. The utilization of on-animal investigations to evaluate ectoparasiticidal compounds will continue to benefit long-term integrated pest management options for animal producers. Researchers in this area must take care to ensure that any animal use is justified and will provide contributions to further understanding the plethora of host-pest interactions that drive economic distress in animal production. It is our hope that the provided considerations along with existing recommendations will assist future researchers by offering these insights largely based on personal experiences.