Brain regions that were more active during the task and that showed more interindividual variation in activation were better at capturing individual differences in performance on the task, but this was only true for activations when successfully inhibiting. Cortical regions outperformed subcortical areas in explaining individual differences but the two hemispheres performed equally well. These results demonstrate that the detection of reproducible links between brain function and performance can be improved with multivariate approaches and give insight into a number of brain systems contributing to individual differences in this fundamental cognitive control process.The stop‐signal task (SST) is one of the most common fMRI tasks of response inhibition, and its performance measure, the stop‐signal reaction‐time (SSRT), is broadly used as a measure of cognitive control processes. The neurobiology underlying individual or clinical differences in response inhibition remain unclear, consistent with the general pattern of quite modest brain–behavior associations that have been recently reported in well‐powered large‐sample studies. Here, we investigated the potential of multivariate, machine learning (ML) methods to improve the estimation of individual differences in SSRT with multimodal structural and functional region of interest‐level neuroimaging data from 9‐ to 11‐year‐olds children in the ABCD Study. Six ML algorithms were assessed across modalities and fMRI tasks. We verified that SST activation performed best in predicting SSRT among multiple modalities including morphological MRI , diffusion tensor imaging, and fMRI task activations, and then showed that SST activation explained 12% of the variance in SSRT using cross‐validation and out‐of‐sample lockbox data sets  methods to improve the estimation of individual differences in stop‐signal reaction time (SSRT) with region of interest‐level multimodal structural and functional neuroimaging data from the ABCD Study. Six ML algorithms were assessed across modalities and fMRI tasks. As expected, SSRT was best explained by stop‐signal task activation with 12% of individual differences in SSRT explained in cross‐validation and out‐of‐sample lockbox data sets. Brain regions that were more active during the task and that showed more interindividual variation in activation were better at capturing individual differences in performance on the task, but this was only true for activations when successfully inhibiting. By segregating a large sample into independent model development and model testing subsets, ML methods can identify potential novel associations in an exploratory fashion and then, importantly, assess if these associations are generalizable. Regression‐based methods have been a popular approach for multivariate analyses, such as linear, ridge, least absolute shrinkage and selection operator (LASSO), elastic net, support vector machine  in predicting SSRT and to identify important brain regions in predicting SSRT. In this study, we (1) tested whether SST brain activation would best estimate SSRT, compared with other imaging modalities and across different ML algorithms, (2) evaluated the relative predictive power of left and right hemisphere activation, cortical and subcortical activation, and the different activation contrasts derived from the SST, and (3) identified the most important features for explaining SSRT for each of the multivariate models.22.1Predictive models were generated with data from the ABCD study, an ongoing longitudinal project that includes 11,875 youths recruited for their baseline assessment at ages 9 and 10 years  on stop trials , and fMRI for resting state, the stop‐signal task (SST), Monetary Incentive Delay (MID) task (assessing reward and loss anticipation and receipt), and an EN‐Back working memory task that contained place stimuli and face stimuli evincing different emotions and contained both 0‐Back and 2‐Back conditions  in the Destrieux atlas , incorrect stop versus correct go (stop fail contrast), and correct go versus fixation (go success contrast); from the MID, large loss versus neutral anticipation, large reward versus neutral anticipation, loss positive versus negative feedback, and reward positive versus negative feedback; from the EN‐Back, 2‐back versus fixation and 2‐Back versus 0‐Back. Although SST brain activations were anticipated to be most strongly related to SSRT, we sought to determine whether reward processing (MID) and working memory (EN‐Back) were also linked to inhibitory control. Previous work suggests that reward and punishment can have distinct effects on go and stop performance  of the time series of BOLD signals  features coefficients of similar magnitude.Standard linear regression performs poorly in a situation with multicollinearity. An alternative is regularized regression, wherein a constraint is placed on the model coefficients to be regressed. A few methods exist to shrink the magnitude of regression coefficients  to the closest points in either class. However, SVM algorithms have been developed for a numeric prediction  that share many of the properties encountered in the classification case: they produce a model that can usually be expressed in terms of a few support vectors and can be applied to nonlinear problems using kernel functions. It is commonly applied with different kernel functions to map the original input variables into high‐dimensional feature space which allows the incorporation of nonlinearity in the solution. Given a set of data points, an SVM regression model relates the output variable and the input variables as:ψX corresponds to the features mapped from the original input variable with a kernel function. SVM extension to SVR is realized by introducing an ε‐insensitive region around the function, which is referred to as the ε‐tube that best approximates the regression function  of total features would be considered as split candidates, as using only a subset of the predictors can eliminate the correlations among all N trees. The variable m is generally recommended as the square root of the total number of predictors. Subsequently, bagging essentially averages all N trees to reduce the overall model variance  at each split point is calculated for predicted values and observed values and the minimum SSE is selected for the node. Overall, it operates by constructing a multitude of decision trees at training time and outputting the class that is the mean SSRT of the individual trees.Random forest model is an ensemble method that generates a number of decision trees in parallel in order to balance between underfitting and overfitting to produce a generalizable model. Training a random forest model mainly involves two statistical techniques: bootstrapping and bagging. First, N bootstrapped sample sets are obtained from the entire training set by random sampling with replacement. Each set is then used to construct a regression tree without limiting the number of decision trees in order to enable the tree to reach its best performance of predicting the target. Instead of considering all features, only a smaller and fixed number  and lockbox (20%) subsets. The training subset was used solely for the learning procedure of the predictive model algorithm and the lockbox subset was used to estimate the performance of the trained predictive model algorithm. Thus, the lockbox sample was not made available in the learning stage and was separated from the training data and served to determine whether overfitting had occurred in the model construction.2.5.2K‐fold cross‐validation was used to assess each model's generalizability. The training data were first divided into K subsets, one of which was used as a validation set, and models were trained on the remaining K‐1 subsets. Model errors were averaged over K iterations. In this study, fivefold cross‐validation was selected to randomly split the training data set.R2), was used to assess the deviation between the predicted and actual SSRT. R2 represents the proportion of variance of actual SSRT values that a model's SSRT predictions explain and compares the fit of the model with that of a sample mean null hypothesis. If the chosen model has higher error on the validation set than the sample mean, then R2 is negative. In ML studies, it is not uncommon for models to have negative validation R2, which indicates that the model does not generalize.In order to evaluate the accuracy of each model, the coefficient of determination, R‐squared score  brain predictors which would affect our primary goal to quantify the maximum extent to which brain activation can explain SSRT. That said, analyses were run separately for males and females and returned comparable effect sizes Table , neither33.1R2 values  while among the structural measures, the DTI data performed better than the morphological measures . The increased sample size improved model performance for all algorithms except random forest . Ridge regression and SVM regression both had notably strong performance with R2 of 11.86 and 11.70%, respectively.With confirmation of SST activation data being the strongest predictor of SSRT, we focused next on an in‐depth exploration of this data set using all participants with SST data that passed quality control and all exclusion criteria n = 798. The it = 23.53, p < .01) with cortical (max R2 = 11.09%) compared to subcortical features (max R2 = 3.84%). Similarly, performance restricted to left hemisphere features was slightly better than performance with right hemisphere features but the differences for the elastic net model were modest .To identify subsets of brain features particularly valuable for capturing individual differences in SSRT, we examined a number of meaningful feature groups  across the different ML algorithms. Since the elastic net model performed best on predicting SSRT , both of which were significantly greater than the Go response contrast . This pattern held across all algorithms , we evaluated its performance on the set‐aside 20% lockbox set and examined the features that contributed to its performance . This indicates that the magnitude of each feature's contribution in the predictive model is tightly coupled with how closely linked that feature, in isolation, is with individual differences in SSRT. Next, the correlation between corr/SSRT and the mean activation of each feature is significant for stop success  but not for stop fail  nor go trial . This result demonstrates that brain regions that are most activated by the task are also more related to individual differences in performance on the task, but this is only true for activations when successfully inhibiting. Finally, the standard deviations of activation levels of regions were positively correlated with corr/SSRT for stop success , uncorrelated for stop fail , and negatively, but not significantly, correlated for go trials . This pattern suggests that regions with more interindividual variation when successfully inhibiting were better at explaining interindividual variation in SSRT. Conversely, the more interindividual variation a region showed on go trials, the worse it was in capturing interindividual variation in SSRT.In order to understand the characteristics of the final model's features (regions), we plotted the relationships between each feature's correlation with SSRT (corr/SSRT) and its coefficient in the final predictive model, its mean activation, and its standard deviation of activations  in Figure p < .01) correlation with SSRT.There were a number of brain features with notable properties. As shown in Figure r = −.84, p = .016) between their activation levels and their correlation with SSRT indicating that the more active an insular region was on error trials, the better it performed in explaining individual differences in SSRT.As noted in Figure 44.1In this study, we investigated the relationship between stop task performance and structural and functional neuroimaging measures. Exploring multiple ML algorithms enabled us to assess any advantages to be obtained by combining feature elimination and feature coefficient reduction and doing so in the current analysis resulted in a model that was able to predict 12.54% of the variance in SSRT in the set‐aside 20% lockbox.Elastic net regressions yielded the most accurate predictions but both ridge regression and SVM both generated comparable estimations. There was no significant difference between SVM and the elastic net regression model, indicating that linear associations provided sufficient information for the predictive models. We also evaluated adding a first‐step univariate feature (ROI) selection approach Table . Althoug4.2Although some concerns regarding the SST implemented in the ABCD study have been suggested , the present results underscore the importance of multivariate analysis for capturing individual differences.In general, coefficients of the predictive model were highly correlated with the pairwise correlations between brain activations and SSRT , this result indicates that the magnitude of activation in these visual regions contributes very substantially to how closely coupled that region is to individual differences in motor inhibition.Visual regions were prominent among stop success activations for having both relatively high levels of activation as well as being relatively highly correlated with SSRT. The results suggest that the visual processing of the task stimuli, including the possibility of their top‐down attentional modulation, may contribute in important ways to task performance at this age. Across the cortical visual regions in the final model, the association between activation levels and the magnitude of the correlation between the region's activation and SSRT was A number of cingulate regions were activated on stop fail trials and showed negative correlations between activation levels and SSRT. A large body of research links the dACC to cognitive control, especially regarding the monitoring of suboptimal performance  are related to increased occipital cortex activity  or incorporation of other relevant features such as inter‐regional task connectivity offer potential. Importantly, determining how intraindividual and interindividual differences develop throughout adolescence as the ABCD sample ages will be key to understanding how this neurocognitive process influences, and is influenced by, a multitude of psychosocial factors. There are several limitations to this study that should be highlighted. First, the study was restricted by the limited number of SST contrasts that probe only the main aspects of the task (go and stop success and failure), although current studies suggest that other cognitive processes such as proactive control of motor responses are relevant to SSRT  is the final number of participants excluded for any of the exclusion‐types listed in the table.TABLE S2 Proportion of variance in SSRT explained by each model averaged across the fivefold cross‐validation analyses.TABLE S3R2 (%) of predicting SSRT for each model on SST data set as well as three different contrastsTABLE S4 T‐statistics between each pair of modalities across 5 folds R‐square performance TABLE S5 T‐statistics between each pair of algorithm across 5 folds R‐square performance predicting SSRT with SST TABLE S6 Characteristics of Stop success occipital regionsTABLE S7 Top 10 negative corr/SSRT with stop failTABLE S8R2 (%) when predicting SSRT separately for males and females. One‐tailed t‐test comparing male and female model results across the six algorithms was not statistically significant t(5) = 1.514, p = .08.TABLE S9R2 (%) for predicting SSRT across numerous algorithms and with 10 different univariate feature selection groups.Click here for additional data file.


The COVID‐19 pandemic necessitated unprecedented changes in alcohol availability, including closures, curfews and restrictions. We draw on new data from three UK studies exploring these issues to identify implications for premises licensing and wider policy.n = 17) with licensing stakeholders in Scotland and England reporting how COVID‐19 has reshaped local licensing and alcohol‐related harms; (ii) semi‐structured interviews (n = 15) with ambulance clinicians reporting experiences with alcohol during the pandemic; and (iii) descriptive and time series analyses of alcohol‐related ambulance callouts in Scotland before and during the first UK lockdown (1 January 2019 to 30 June 2020).(i) Semi‐structured interviews .COVID‐19 restrictions  affected on‐trade premises only and licensing stakeholders highlighted the relaxation of some laws  and a rise in home drinking as having long‐term risks for public health. Ambulance clinicians described a welcome break from pre‐pandemic mass public intoxication and huge reductions in alcohol‐related callouts at night‐time. They also highlighted potential long‐term risks of increased home drinking. The national lockdown was associated with an absolute fall of 2.14 percentage points [95% confidence interval (CI) −3.54, −0.74; COVID‐19 gave rise to both restrictions on premises and relaxations of licensing, with initial reductions in alcohol‐related ambulance callouts, a rise in home drinking and diverse impacts on businesses. Policies which may protect on‐trade businesses, while reshaping the night‐time economy away from alcohol‐related harms, could offer a ‘win–win’ for policymakers and health advocates. The COVID‐19 pandemic necessitated unprecedented changes in alcohol availability with closures, curfews and operating restrictions in many countries. Complete sale bans were introduced in several countries , whereasOther pandemic control measures also have implications for where, when and how much alcohol is drunk. It may be ‘easier’ to drink alcohol where people work more from home with less need to drive and lower visibility . GreaterAlongside changes in alcohol consumption, a general decline in healthcare utilisation for non‐COVID‐19 issues was observed early in the pandemic, with decreases across wide ranging conditions , 23, 24.Much UK public discourse has centred on hardships for hospitality businesses arising from public health measures with surprisingly little discussion of reduced pressures on emergency services , 29. TheMeanwhile government policy has moved on apace in many countries to considering post‐pandemic recovery. While some public health experts have drawn attention to the need for stronger alcohol controls to protect health services and public health , 36, theHow might the pandemic have reshaped regulation of alcohol sales via the local premises licensing system in England and Scotland?What are ambulance clinicians' experiences and views of alcohol‐related ambulance callouts during the pandemic?How did alcohol‐related ambulance callouts in Scotland change in volume and timing during the first UK national lockdown compared to non‐alcohol callouts?Together, these sources can contribute to an understanding of the potential implications of changes in the alcohol regulatory environment and related behaviours during the pandemic, for public health and emergency service utilisation in the short and medium term.In this paper, we: (i) review primary qualitative data from interviews which took place during the pandemic for two separate studies not originally focused on COVID‐19; and (ii) analyse secondary data on alcohol‐related ambulance call‐outs during the first UK national lockdown. Using this data, we seek to inform discussions of the following three questions:First, we report in‐depth interview data from a study on the role of public health in licensing  in whichIn response to the pandemic, the UK Government, in tandem with the Scottish Government, initiated a national lockdown requiring all ‘on‐licence’ premises , including bars, restaurants and nightclubs, to stop trading from 20 March 2020. In Scotland, such premises remained fully closed until 19 June, when they were permitted to re‐open to serve alcohol in outdoor spaces (e.g. beer gardens) only, with indoor spaces opening in Scotland from 15 July. From early July in England, premises gradually re‐opened with social distancing and other requirements in place, albeit implemented to varying degrees . PrevailSampling: As part of the wider ExILEnS study  now can't go out and they sit in the house all day drinking, and they drink and drink and drink and drink and drink’ and added their view that isolation from friends affects the mental health of these patients.Alongside this reduction, domestic alcohol‐related incident numbers appeared to have gone up, though ‘‘… alcohol intake is definitely increased in the house. My thoughts on when this is all finished, the people that used to go socializing, might not… I still think the younger generation will go back out to the nightclubs and the public houses and that, probably forty year old people upwards…I think they might get used to being in the house drinking, why get dressed and go out, when I can just sit here and watch TV and do what I've been doing for the last seven months’ (#18)‘All these people can't stay at home drinking without a consequence at some point.’ (#23)While some ambulance clinicians expressed a hope that people might be less inclined to call an ambulance for minor issues post‐pandemic, others described fears about longer‐term consequences arising from new alcohol consumption habits formed during the pandemic that might persist.Ambulance callouts of all causes from March to June 2020 decreased in number compared to the previous year, with falls of 1.1% in March, 12.3% in April, 11.5% in May and 10.9% in June  was 4.06, but fell to 3.74 in the 3 months of lockdown (20 March–30 June 2020), reaching a low point in the first month (21 March–20 April 2020) of 3.36. Levels appeared to be returning to pre‐lockdown volumes from the end of May 2020 .P=0.003). In relative terms, this corresponds to a 13% reduction in the proportion of callouts that were alcohol‐related compared to the period before the lockdown (1 January 2018–20 March 2020). This association was immediate and then lessened over time with a significant daily increase (post lockdown announcement) in the percentage of alcohol related callouts of 0.03% . The analysis finds that the proportion of alcohol‐related call‐outs compared to total call‐outs came back to pre‐lockdown levels after approximately 2 months, at the end of May 2020  showed an absolute reduction of 2.14 percentage points  more visible, including to children and people in recovery from alcohol problems who may be passing by. The pandemic may have contributed to expansions in availability as licence applications received less public health scrutiny, but may have had some benefits in building relationships between stakeholders seen as important for facilitating successful public health engagement in licensing .mass intoxication’ and ‘battlefield environment’ on city streets, which they described. As premises re‐open and especially in Scotland where the ambulance service is under intense pressure in 2021 [Second, qualitative data from the Scottish Ambulance Service indicate that the pandemic period, during which premises were either closed completely or not open late at night, was associated with a substantial reduction in demand for ambulances arising from alcohol consumption. The quantitative data shows that total alcohol‐related callouts fell even more than overall ambulance call‐outs in the first months of lockdown, but rebounded after 2 months. The reduction was particularly acute at weekend night‐times however, and call‐outs at these times remained lower than usual through to the third month after lockdown. Considering the reports of ambulance clinicians, it seems clear that reductions in call‐outs were linked to the closure of licensed premises and the night‐time economy (NTE), more than ‘stay at home’ measures introduced at the same time, even though these probably reduced socialising in people's homes. Exceptionally good weather in May 2020 may have mitigated the latter effect, perhaps explaining a rebound in weekend callouts that month even with premises still closed; that disappeared in June 2020. They reported clearly that, in normal times, many ambulance callouts are associated with people in or around pubs/bars/clubs in the NTE and that these were greatly reduced. The views expressed by paramedics are powerful and give pause for thought about whether business recovery post‐COVID has to mean a return to the ‘ in 2021 , authori in 2021  and viol in 2021  and coulIt is timely to consider whether economic prosperity in the NTE must rely on alcohol and whether there is a third way or ‘sweet spot’ approach via policies which transform and build the NTE to prioritise other forms of entertainment, food, music or more family‐friendly environments. The nature of such policies and their feasibility and acceptability to communities , 49 and The third issue highlighted by both licensing stakeholders and ambulance clinicians, is that the pandemic shifted alcohol sales and drinking from the on‐ to off‐trade, exacerbating existing trends. Reportedly driven by increases in home delivery of alcohol coupled with closures of premises and ‘stay at home’ orders, interviewees' perceptions of increases in home drinking are supported by sales and survey data from other sources , 9, 54. Overall, it seems likely that the closure of licensed premises led to net reductions in the burden of alcohol‐related harm on emergency services, even with a shift to home drinking, but that this was relatively short‐lived. Total closures or prohibitions are neither practical nor desirable in a liberal society however and our data support the suggestions of others that an associated shift to home drinking may result in mid‐ and long‐term harms , 33. In A final issue raised by our data is that the pandemic is likely to have longer term impacts on the alcohol sector . SmallerThis paper provides timely, relevant data to inform current debates around how to support businesses to recover from the COVID‐19 pandemic, while still protecting public health and health services. It makes pragmatic use of new data arising spontaneously in two studies not designed to focus on COVID‐19 and reports new analyses of data on ambulance call‐outs during the pandemic. Drawing together data from three studies allows for triangulation across the datasets and we found broad agreement on issues raised by the different sources, allowing us to draw questions and hypotheses from the findings.There are however, some important limitations to bear in mind. The ExILEnS study was not focused on the pandemic and questions regarding the issues reported were not explored with all interviewees. Furthermore, relevant data were naturally only available from interviews conducted after the start of the first UK lockdown in March 2020, during which time only two public health stakeholders were interviewed. The views reported in the ExILEnS study are mostly those of others without a specific public health remit —a larger sample of public health stakeholders may broaden and strengthen findings relating to alcohol harm implications of the pandemic. In contrast, in the IMPAACT study, a strength is the varied sample of interviewees in terms of region, role and length of service and that questions about the pandemic were included in the topic guide. Both studies capture professional perspectives but there is also a need for in‐depth qualitative research to understand consumer experiences, including their view of possible long‐term outcomes of changes in alcohol consumption. Considering the quantitative ambulance callout data, while the algorithm used to identify alcohol‐related callouts has been found to be highly accurate , it may Drawing on new data from three studies, we report potential impacts of the COVID‐19 pandemic that have implications for public health and alco hol policy. Licensing stakeholders reported the liberalisation of licensing laws that may increase availability of alcohol and the possible reshaping of the on‐trade sector as different premises types and businesses are more or less negatively affected by the pandemic. There were large short‐term reductions in alcohol‐related ambulance callouts while public health measures closed or severely restricted on‐trade premises, especially at weekend night‐times and reports from both licensing and ambulance interviewees of rises in home drinking with potential long‐term consequences. Paramedics gave stark reports about the volume of late night alcohol‐related callouts prior to the pandemic. We argue that these reports should give pause for thought about the wisdom of a return to ‘normal’ in the night‐time economy and tentatively draw on prior literature to suggest possible approaches to both reduce harm and support business recovery. At a time when policymakers are reluctant to be seen to hurt already suffering hospitality sectors, but also wish to protect health services, it is going to be vital for politicians, advocates and lobbyists alike to find win–win’ policies that can do both.The authors have no conflicts of interest.


In a baseline condition, participants simply responded to the sacrificial moral dilemmas as is typically done in previous research. In an “emotion‐salient” condition, participants had to reason about the emotions of another person after solving moral dilemmas (deliberative processing). In the “emotion‐ambiguous” condition, participants saw images of people in distress, after solving moral dilemmas, but did not have to explicitly reason about such emotions (spontaneous processing). The four PCL‐R facets predicted distinct interference effects depending on spontaneous versus deliberative processing of hypothetical victim's emotions. The findings suggest that the use of a multi‐faceted approach to account for cognitive and moral correlates of psychopathy may help address previously mixed results. Implications and future directions for theory and research are discussed.Recent research has documented a small but significant correlation between psychopathic capacities and utilitarian moral judgment, although the findings are generally inconsistent and unclear. We propose that one way to make sense of mixed findings is to consider variation in perspective‐taking capacities of psychopathic individuals. With this in mind, we had criminal offenders ( Their a1.1Despite psychopathic individuals' antisocial behavior, a recent meta‐analysis found only a small relationship between non‐standard moral judgment and psychopathic traits, suggesting that perhaps psychopathic individuals may only exhibit subtle differences in moral judgment in comparison to less psychopathic individuals . While both scenarios, the trolley and the footbridge dilemma, involve the sacrifice of one life for many, most people experience a moral difference between the two dilemma types  emotional processing may shed light on the discrepant research findings on psychopathy and moral decisions. In the current study, we incorporated the distinction between spontaneous versus deliberate emotion processing of empathy‐related content into the investigation of behavioral moral actions. Based on the research discussed, we hypothesized that elevated psychopathic traits would be associated with more utilitarian behavioral choices in sacrificial moral dilemmas. With the incorporation of spontaneous versus deliberate emotion feedback, however, psychopathic traits were expected to be less likely to be associated with utilitarian choices when deliberately processing a potential victim's emotions, compared to a spontaneous processing condition.More specifically, based on findings that psychopathy is associated with more endorsement of utilitarian choices rather than utilitarian judgment  from several German correctional and forensic‐psychiatric institutions. Inclusion criteria were defined as broadly as possible with fluent German language skills and the offender status. We included only male offenders for reasons of generalizability  with lower secondary education (41.7%) or general secondary education (15%). About 16.7% of the participants had completed an apprenticeship, 11.7% were qualified with a university‐entrance diploma, and 3.3% had a University degree. No education was stated by 11.7% of the sample.For this study, we drew 17 out of the 60 participants from a former study, with previously collected Psychopathy Checklist‐Revised (PCL‐R) scores  and the Affective facet  load on a superordinate Factor 1 mapping psychopathic core personality traits  and the Antisocial facet  load on a superordinate Factor 2 that maps social deviance.The German version of the PCL‐R , and a range between 4 and 37.90, covering almost the whole spectrum of PCL‐R scores. Half of the participants in our sample had a PCL‐R score of 25 or more, which indicates a high degree of psychopathic traits according to the German manual of the PCL‐R , which is lower than an average IQ in the general population but representative for a forensic sample considering the lower education levels in prison samples  with INQUISIT 5 . After a5.60″ witbaseline condition, six moral dilemmas were each directly followed by a set of questions. Based on the findings by Pletti et al. , and then how likely they would engage in such behavior from 0 to 7 (choice of action). Following the tradition of sacrificial moral dilemma research, utilitarian responses were coded positively . Participants were then required to rate their arousal from extremely calm (0) to extremely aroused (7) and how pleasant this decision felt (0 indicating extremely unpleasant and 7 indicating extremely pleasant) based on the Arousal and Valence scales of the Self‐Assessment Manikin by Lang et al. (In the first (1), the i et al. , we distg et al. . Throughspontaneous emotion processing condition, eight new moral dilemmas were presented to the participants that were each again followed by the set of questions described previously. Additionally, after the choice of action, a female and a male face were shown that expressed one of the six basic emotions  along with the question “do you think your victim in the dilemma before was a female or a male?” Pictures with facial expressions were drawn from the Radboud database  within‐subjects task, the deliberate emotion processing condition, instead of guessing the gender of the victim, participants saw two distinct emotions in a person and were asked to respond to the question “what emotion did you likely cause in the victim of the dilemma?” The emotions again matched the participant's response in the choice of action question and were displayed as text clicks below the pictures. The task structure was tested for feasibility and timing in a pilot study, prior to conducting the study within correctional and forensic‐psychiatric institutions with strict time limits for visitors.Following the same task structure, in the third task (3), the 3.4Bivariate correlations were calculated in IBM SPSS statistics version 23 (2015) to assess associations between the key variables. Path models were conducted with Mplus , (2), and (3) are represented by moral judgments ratings with higher scores indicating utilitarian judgment (“it is acceptable to sacrifice one for many”) in path model 1, and in model 5 for choice of action ratings . For an overview of the models run, see Table , (2), anTo test our hypotheses of distinct decision outcomes based on spontaneous or deliberate emotion processing, we used difference score criterion variables between task conditions. Thus, we ran separate models including the task condition difference scores (2‐1), (3‐2), and (3‐1) as dependent variables . With regard to the hypothesized change in moral decisions based on deliberate emotion processing, differences in decisions between the deliberate (3) and the spontaneous (2) were of particular interest.We pursued the same procedure for valence and arousal ratings, by mapping the distinct ratings per condition onto the separate facets (models 10 and 14) and included the differences between valence (models 11–13) and arousal (models 15–17) ratings into the models. Total moral judgment ratings and choices of action were included in supplemental analyses, to test the hypothesized distinction between judging and choosing (model 9).χ2 test. If the χ2 test is not significant , it is assumed that the difference between the reproduced covariance matrix of the model and the observed covariance matrix of the sample is not significantly different from zero and thus, that the hypothesized model accounts for the sample data sufficiently well. The Comparative Fit Index (CFI) and the Tucker Lewis Index (TLI) are reported for incremental fit, with values of 0.95 or above signifying good model fit. Also, absolute fit was evaluated with the Standardized Root Mean Square Residual (SRMR) and the Root Mean Square Error of Approximation (RMSEA) to gauge how well the path models accounted for the data. This is assumed with a recommended value of ≤0.08 for the SRMR, and ≤0.05 for the RMSEA. Path modeling analyses were conducted with Mplus 7.0  in model 1 and choice of action ratings in model 5  in the spontaneous emotion feedback condition.Utilitarian decisions in the three experimental conditions were analyzed in separate path models for moral judgment  and comparably, choices of action . The Antisocial facet on the other hand predicted more acceptance ratings of utilitarian judgments in the presence of spontaneous emotion feedback  as well as the willingness to engage in such behavior . The Interpersonal facet predicted utilitarian moral judgments at trend level  and choices of action significantly  in the spontaneous emotion processing condition. Results of total moral judgments and choices taken together across conditions (model 9) revealed a significant prediction of the Interpersonal facet for more utilitarian choices  but not judgments . The Affective facet was highly predictive for less utilitarian decisions in both judgments  and choices . The Antisocial facet was associated with utilitarian responses at trend level .The Affective facet also predicted deontological moral judgments in the deliberate condition . The Affective facet predicted more utilitarian judgments in the deliberate (3) compared to the spontaneous (2) condition at trend level . Also at trend level, higher IQ scores were associated with less utilitarian judgments between these (3‐2) conditions . Difference scores in choice of action were not, however, significantly related to the PCL‐R facets, age, or IQ.The inclusion of difference scores between the separate task conditions along with age and IQ as covariates in model 3  in model 10, indicating that higher age led to a more unpleasant experience in moral decisions. Age was unrelated to arousal ratings, however, in all conditions (model 14). The Lifestyle facet predicted an unpleasant experience  and excitement  during decisions in the spontaneous condition only . The Affective facet predicted pleasantly experienced decision‐making in the deliberate emotion processing condition at trend level  but did not reveal a significant relation to reported excitement during decisions.The models fit the data well  and more excitement  experienced with spontaneous emotion feedback compared to the baseline condition (2‐1). The Antisocial facet predicted in the other direction with a more pleasant experience at trend level  and highly significant less excitement . The Interpersonal facet was negatively related to the difference score (3‐2) for valence  but unrelated to arousal, indicating that higher Interpersonal facet scores predicted a more unpleasant decision‐making process when deliberately processing emotions of the proposed victims compared to a spontaneous emotion processing with no difference in excitement. The Affective facet was related to the opposite trend, with higher scores predicting a more pleasant decision‐making process with the deliberate compared to no emotion feedback , but no relation was found between the Affective facet and arousal difference scores (3‐1). The Lifestyle facet though, was associated with more reported excitement during decisions in the deliberate emotion processing condition compared to the baseline , but was unrelated to how pleasant the decision‐making was experienced.When including difference scores into the models, the Lifestyle facet predicted significantly less pleasant decision‐making  was linked to divergent associations with PCL‐R facets. Also, individual psychopathic traits were only related to utilitarian decisions  and deontological decisions  with the additional presence of emotion feedback. Earlier findings have suggested that psychopathic offenders compared with non‐psychopathic offenders draw on cognitive compensation strategies when confronted with moral tasks, which helps them to process emotional information despite neural abnormalities  psychopathy trait suggests that low empathy is linked to enjoyment in hurting more people, respectively letting more people die. Therefore, beyond a lack of concern that implies indifference toward the harm of others, the reported pleasantness during deontological decisions associated with low empathic traits implies joy in experiencing the harm of others. Recent research has shown that there is indeed a psychopathic domain that pertains to enjoyment in doing harm  compared to the second condition . This finding is in contrast to earlier studies that reported deliberate thinking patterns to be associated with more utilitarian judgments, which was linked to higher working memory capacities (Moore et al., 5.4Although we controlled for mean differences in conflict ratings of moral dilemmas between the separate conditions in our pilot study sample, the comparison of distinct dilemmas in the three conditions is a possible confounding factor for attributing the within‐subject differences to the emotion interference. Moreover, moral dilemmas were not randomized among experimental conditions to avoid randomization as confounding factor (Saint‐Mont, The implementation of spontaneous versus deliberate emotion processing in imagined, theoretical sacrificial moral dilemma scenarios remain delicate by the very nature of these tasks. For obvious ethical reasons, it is unthinkable to involve real victims in a task that would allow the participants to cause suffering in order to manipulate the deliberation of the perpetuators' emotional responsiveness. One recent approach of real‐life dilemmas was conducted with monetary rewards (Bostyn et al., 6Studies on psychopathy and morality using sacrificial dilemmas have resulted in heterogeneous findings. While some researchers have claimed utilitarian judgment to be characteristic for psychopathic individuals, others found no such relation. In our study, the four psychopathy facets of the PCL‐R revealed contrasting associations to moral decisions. It therefore adds to the current knowledge by reuniting the formerly contradictory findings and suggests that the heterogeneity of other studies is related to the multi‐faceted structure of the psychopathy construct. Indeed, several studies reported distinct associations of the psychopathy facets to moral correlates (Gao & Tang, Moreover, in this study, we paid tribute to the growing evidence referring to the dimensional character of the psychopathic construct with a path analytic approach Patrick, . Our modTo our knowledge, our study is the first that included the differentiation between spontaneous and deliberate emotion processing of psychopathic individuals in the moral context of social decisions. Our findings encourage further research to take notice of these distinct processes to the better understanding of emotional, moral, and behavioral features of the psychopathic personality.supplementary Material 1Click here for additional data file.


Human‐centered, active‐learning approaches can help students develop core competencies in biology and other STEM fields, including the ability to conduct research, use quantitative reasoning, communicate across disciplinary boundaries, and connect science education to pressing social and environmental challenges. Promising approaches for incorporating active learning into biology courses include the use of course‐based research, community engagement, and international experiences. Disruption to higher education due to the COVID‐19 pandemic made each of these approaches more challenging or impossible to execute. Here, we describe a scalable course‐based undergraduate research experience (CURE) for an animal behavior course that integrates research and community engagement in a remote international experience. Students in courses at two U.S. universities worked with community partners to analyze the behavior of African goats grazing near informal settlements in Western Cape, South Africa. Partners established a relationship with goat herders, and then created 2‐min videos of individual goats that differed in criteria (goat sex and time of day) specified by students. Students worked in small groups to choose dependent variables, and then compared goat behavior across criteria using a factorial design. In postcourse surveys, students from both universities indicated overall enthusiasm for the experience. In general, students indicated that the laboratory provided them with “somewhat more” of a research‐based experience compared with biology laboratories they had taken of similar length, and “somewhat more” to “much more” of a community‐engagement and international experience. Educational benefits were complemented by the fact that international educational partners facing economic hardship due to the pandemic received payment for services. Future iterations of the CURE can focus on goat behavior differences across ecological conditions to help herders increase production in the face of continued environmental and social challenges. More generally, applying the structure of this CURE could facilitate mutually beneficial collaborations with residents of under‐resourced areas around the world. Course‐based research, community engagement, and international experiences can enhance undergraduate biology education, but each were made more challenging due to the COVID‐19 pandemic. Here, we describe a scalable course‐based undergraduate research experience focusing on African goat behavior that integrates research and community engagement in a remote international experience. Educational benefits for students were complemented by the fact that international educational partners facing economic hardship due to the pandemic could receive payment for services. A key cOne main approach for realizing V&C recommendations is through research‐based instruction. Developing opportunities for authentic research experiences for undergraduates is essential for capturing the imagination of students by exposing them to the creativity of STEM disciplines PCAST, . One appA second promising approach for creating active‐learning experiences is through community‐engaged education Enos, . CommuniA third approach for creating active‐learning experiences for biology students is through international education  at the University of St. Thomas  that benefited students but also provided income for people in Cape Town facing economic hardship. This CURE was then replicated in a Behavioral Ecology course at Metropolitan State University  in Summer 2021.The design of the Goat CURE is described in Figure In an initial laboratory period (Week 1), students viewed the panoramic videos and conducted a literature search to learn more about African goats and goat behavior  and tasked with determining what might serve as independent variables that could facilitate hypothesis testing using short videos of individual goats. These suggestions were then filtered by all of the authors, who ultimately decided to create videos that would allow students to contrast goat sex  and time of day (morning vs. late afternoon).Between weeks 2 and 3, LB conducted all filming over the course of 1 day. Each video was ~2 min and focused on one individual  they would focus on. They were informed that variables would have to be something they could quantify using only observations of the videos. Examples of dependent variables chosen by students included % time spent feeding, % time spent scanning, and % time with tail raised. Then, each group of students watched all 32 videos and quantified their chosen variable(s). Students were asked to have each video independently evaluated by two group members to test for possible observer effects.In Week 4, students analyzed their results using statistical training from previous laboratory periods. After analysis, each group made a short oral presentation to their peers and prepared a final written report to be shared with the community partners.The educational and community outcomes we envisioned for this CURE are also presented in Figure 4We surveyed students from St. Thomas and Metro State in December 2021, 7 and 4 months, respectively, after the courses were completed. All survey work was first approved by the Institutional Review Boards at St. Thomas and Metro State. Student participation in the survey was voluntary, and no individual identifiers were collected. We sent the survey to the 47 students who had taken either course and received 10 responses from St. Thomas and nine responses from Metro State students . Students received no compensation for responding.We asked students seven multiple‐choice questions and four written‐response questions. The multiple‐choice questions asked students to compare their experience in the African Goat laboratory to “labs of similar length that you have had in other biology classes” Table . Answer F2,54 = 3.076, p = .0543).Multiple‐choice responses suggest that, overall, students valued the research, community engagement, and international aspects of the laboratory Table . Mean reWritten response questions showed that students identified and valued the learning objective targets but struggled somewhat with the open‐ended nature of the laboratory . We adjusted to this lack of a true control by asking students to compare their experiences to those in other biology laboratories. Third, survey questions asked students to compare this experience to their previous experiences rather than an objective standard. As a result, survey responses may differ if the same laboratory is taught by different instructors or at other institutions with more (or fewer) research, community engagement, or international experiences incorporated into the curriculum. Fourth, our survey relies on students' opinions rather than learning outcomes. However, our main objective was to increase students' engagement, so the enthusiasm expressed in their responses provided a reasonable measure of whether we achieved this objective. Finally, our survey was administered several months after the experience. Regardless of these limitations, our survey responses suggest that at least respondents viewed the laboratory as offering a “somewhat” to “much” better research, community engagement, and international experience compared to alternative opportunities in their biology programs.5A main value of CUREs is the possibility of engaging many students in relevant research that is iterative. Video‐based CUREs focused on the behavior of goats or other domesticated animals are ideally suited for iteration given the relative ease with which short videos of individual animals can be taken. The possibility of iteration as described in this Goat CURE can also build and reinforce relationships with individuals and communities that are remote from one another.Several students' comments suggested opportunities for iteration focused on additional temporal comparisons . Such comparisons have been made previously in direct observation studies. For example, Bakare and Chimonyo  found thOther iterations are possible by creating videos that allow for local or regional spatial comparisons. Examples include creating opportunities for students to assess behavioral differences across habitats or rangeland quality. Snapshot samples of forage conditions could also allow students to assess some ecological variables . Community partners could be hired to make some of these ecological measurements, creating additional employment opportunities.A critical opportunity for iteration is to create extended dialogue among students, community partners, and community members (including goat herders). Successive iterations of the Goat CURE could allow for students to share findings with herders, who then could provide feedback on whether students' observations were consistent with herder knowledge and practices. Such exchange could deepen students' understanding of goat behavior and help them see the useful but potentially incomplete knowledge that comes from scientific studies.One other opportunity for improvement that emerged from students' comments is students' apparent need for more explicit instruction. Several students mentioned that they would have liked more guidance about which behaviors to study and exactly how they should quantify them. Given that a main value of CUREs is to provide students with an opportunity for self‐directed, original discovery, this lack of guidance is actually an essential part of the experience. What we learned from this experience is that we need to communicate more effectively about the CURE's objectives and their rationale, and the general value of CUREs for students. The implementation of CUREs in more courses, especially early in the curriculum, should also help in making students more comfortable with the open‐ended nature of the scientific process.6Our analysis of the Goat CURE suggests standardized video segments taken by international community partners can provide substrate for original student‐led research while exposing students to distant social and environmental challenges. From student surveys and community partner testimonials, we can point to advantages of this approach and opportunities for improvement and expansion.Our work builds upon previous published activities that have incorporated video footage into animal behavior research and teaching , provides an efficient way for students to ask distinct research questions on dependent variables of their choice. Although this approach obviously does not include every aspect of the scientific process , it provides students with significant opportunity for asking their own questions. The Goat CURE also provides clear opportunities for collaboration and discovery given that students worked in small groups and chose their own dependent variables to study original video segments. Collaboration opportunities also existed through repeated interactions with community partners. Iteration opportunities exist as described above. Future iterations may help to identify goat behavior differences across time and space that can help researchers understand constraints on African goat rearing in the understudied, semi‐intensive production system, and potentially provide herders with information about herd condition. These broader comparisons can increase the extent to which this CURE gives students the opportunity to produce useful knowledge. Overall, students indicated that they viewed the Goat CURE to have “somewhat more” opportunity to conduct original research  relative to “labs of similar length that you have had in other biology classes.” Additional footage across different conditions will likely enhance students' impressions that they are making important contributions to an original research effort.The first goal of this work was to help students participate in course‐based original research, characterized by five features: use of scientific practice, collaboration, discovery, iteration, and the opportunity to produce useful knowledge ; formal analysis ; funding acquisition (lead); methodology ; project administration ; writing – original draft (lead); writing – review and editing . Zach Lager: Conceptualization ; investigation ; methodology ; project administration ; writing – original draft (supporting); writing – review and editing . Litha Bhebheza: Conceptualization ; methodology ; project administration ; writing – review and editing . Justa L. Heinen‐Kay: Conceptualization ; formal analysis ; methodology ; project administration ; writing – original draft (supporting); writing – review and editing .The authors have no financial or nonfinancial interests that are directly or indirectly related to the work submitted for publication.Video S1Click here for additional data file.


Despite the relationship between food insecurity and poor mental health, food insecurity in young people attending mental health services in Australia remains understudied. This study aimed to determine the occurrence and predictors of food insecurity, and the relationship with dietary factors in young people attending a mental health service.A cross‐sectional online survey was conducted in a sample of young people (15–25 years) who attended a mental health service in Launceston, Australia. The survey utilised a single‐item food insecurity screening tool and eight demographic, health and service use questions. Five questions determined self‐reported intake of fruit, vegetables, breakfast, water, sugar‐sweetened beverages and takeaway foods. Binary logistic regression determined predictors of food insecurity. Cross‐tabulations determined differences in dietary intake according to food security.n = 48; 68% female), 40% (n = 19) were food insecure. Respondents living out of home or in unstable accommodation were at significantly higher risk of food insecurity  compared to those living with their parents. Those receiving government financial assistance  were also at significantly higher risk of food insecurity. Regardless of food security status, self‐reported intake of fruits, vegetables and breakfast were low, and respondents regularly consumed takeaway foods and sugar‐sweetened beverages.Of survey respondents (There was a high occurrence of food insecurity and poor dietary intake in young people attending a youth mental health service demonstrating that initiatives to support access to healthy food in this group should be a priority, with potential benefits for mental health outcomes. Secondly, we hypothesised that there will be a relationship between food security status and poor dietary habits, namely lower intakes of fruits, vegetables as well as the consumption of breakfast, and/or higher intakes of takeaway and sugar‐sweetened beverages.Although a great deal of research exists on the burden of food insecurity in many vulnerable groups within Australia,2headspace Launceston (Cornerstone Youth Services) is a youth mental health service based in the regional city of Launceston in Tasmania, an island state of Australia sitting south of the mainland. Tasmania has a population size of nearly 542 000 peopleThe headspace Launceston provides early intervention mental health and wellbeing services to young people aged 12–25 years including counselling, psychology and mental health services delivered by a range of practitioners including mental health workers, general practitioners and psychiatrists. Clients are referred to external dietetic support upon request from clients, with limited funding to support this service internally for their clients.There are several services in Tasmania providing support, counselling, and therapeutic interventions for young people experiencing poor mental health.headspace Launceston to improve nutrition and physical activity behaviours in their clients. Ethical approval to conduct this study was obtained from the Tasmania Social Science Human Research Ethics Committee (H0023475) and the study was conducted in accordance with the Declaration of Helsinki. Compliance to the STrengthening the Reporting of OBservational studies in Epidemiology – Nutritional Epidemiology (STROBE‐nut) tool has been addressed.This study is a sub‐analysis of a larger project,headspace management and staff, members of headspace Launceston's Youth Reference Group , and a high school teacher. The survey was also edited using Grammarly (2021 © Grammarly Inc) to ensure the reading level for the survey was no higher than the age of 12 years, to suit the known literacy level in the area.A quantitative, online survey was developed by the project team for the purpose of this study. Only the relevant questions for the analyses underpinning this manuscript are described. For a full list of the survey outcomes please see the published report.The survey measured food security status using a single item food security screening tool, which is the most commonly applied tool in Australia, including in national health surveys.headspace over the past 12 months (continuous).Demographic characteristics were collected including age (continuous variable), gender , living situation , postcode and town name, whether they were of Aboriginal and/or Torres Strait Islander background (yes or no), whether they were receiving government financial assistance (yes or no) and the number of visits to Brief nutrition‐related questions were based on recommendations made in a review of short questions to assess the food consumption of children in Australia.headspace Launceston within the past 12 months. Young people aged 12–14 years and attending headspace were not eligible due to ethical concerns about providing assent. Data collection was undertaken between February and June 2021 through LimeSurvey . Participants were recruited through headspace, where clinicians distributed printed flyers promoting the study to clients attending the service. The flyers had a link and QR code to enter the survey. Due to the low uptake via this method, a 5‐week paid advertisement was run on social media platforms Facebook and Instagram with the parameters set to target 15–25 year olds living in Launceston or surrounds (within a 50 km radius). Over this period, the survey post reached 26 253 young people, with 621 survey link clicks that led to the information sheet that specified eligibility, and the opportunity to review the screening questions to enter the survey. Of this, there were 140 attempts at the screening questions that were employed to determine informed assent (those aged 15–17 years) and consent (those aged 18–25). Participants had to correctly confirm that (i) they were not under a child protection order; (ii) that no one would know they have done the survey; (iii) and, that they understood if they started the survey, they could stop at any time. Of the 140 attempts, 53 completed and answered all screening questions correctly. There were several instances of duplicate IP addresses being used within a matter of minutes with a high level of homogeneity between survey answers, indicating a lack of authenticity in these survey responses. Therefore, duplicate surveys received within 10 min from the same IP addresses were deleted, leaving a final sample size of 48.Participants were young people aged between 15 and 25 years who self‐identified as having contact with p < 0.05.Data were exported from the online software into Microsoft Excel , then transferred into IBM SPSS Statistics for Windows, version 26.0  for analysis. All available survey data were used in the analyses, with surveys excluded if a response to the food security question were not provided. If data were missing for other demographic characteristics, they were only excluded for the relevant univariate analyses or cross‐tabulations to retain as much data as possible. The significance level for all analyses was set at headspace were coded into 1–3 visits or 4 or more visits using a data‐driven approach. Living situation options ‘living in short term/unstable accommodation’ and ‘homeless’ were merged into one group due to low cell counts. Postcodes and town names were used to classify people according to levels of rurality using the Modified Monash ModelFood security status was determined by classifying an affirmative response to the single item question as food insecure and a negative response as food secure. To assist in comparative analyses and due to the distribution of the data, binary variables were generated, with age coded into those aged 15–17 years or 18–25 years, and the number of visits to Intake of fruits and vegetables was interpreted against the Australian Guide to HealthyCross‐tabulations with chi‐square statistics were used to compare the proportions of food secure and food‐insecure respondents, according to their demographic characteristics, and to compare dietary intake variables by food security status. Binary logistic univariate regression was used to determine the demographic predictors of food security status in the sample.3n = 48) were predominantly classified as food secure  and 40% (n = 19) were classified as food insecure (Table headspace more than three times over the past 12 months (Table headspace 4 or more times in the past 12 months were food insecure (46%) compared to those who attended 3 times or less (30%) (Table Survey respondents providing a response to the food security screening question ( 40% n = 9 were cl 40% n = 9 were cl 40% n = 9 were cl 40% n = 9 were cl 40% n = 9 were clThe univariate regression Table  identifiMost respondents (70%) consumed fruit daily and around half (52%) reported consuming vegetables daily Table . However4This study is the first in Australia to screen for the occurrence, and determine the demographic predictors, of food insecurity in a convenience sample of young people attending a youth mental health service. The study also assessed the relationship between food security status and dietary intake in this group. The results suggest that the occurrence of food insecurity in this vulnerable group is much higher than the wider Australian population, and that young people living out of home or living in unstable accommodation, and those receiving government financial assistance, are at the highest risk of food insecurity. The results are not surprising, given the relationship between food security status and social disadvantage and low incomes that persists across all age groups in Australia, but particularly in young people.In the current study, 40% of survey respondents were food insecure, which is very similar to the 38% of university students experiencing food insecurity in the same state of Tasmania, Australia,Critically, the current study found that young people living independently or in unstable accommodation, and those receiving government financial assistance were at substantially higher risk of food insecurity. It is well established that people or households with low economic resources are at high risk of food insecurity, and that the primary reason for food insecurity in Australia is material hardship and inadequate financial resources.The current study indicated that a higher proportion of respondents who were food insecure attended the mental health service more frequently and had received a mental health diagnosis compared to those who were food secure, but this was not significantly different between groups. This is most likely due to the small study sample resulting in underpowered subgroup analyses. In international research, food insecurity has been associated with higher mental health service use,In the current study, food security was not related to lower dietary intakes of fruit, vegetables or breakfast. However, most young people in our study did not meet the Australian Government's dietary recommendations for these foods regardless of their food security status. In 2017–2018, it was reported that of young Australians aged 15–24 years, 18% did not consume a whole serving of fruit daily, and 9% did not consume a serving of vegetables daily.Most participants in the current study, regardless of food security status, reported consuming sugar‐sweetened beverages and takeaway foods at least once a week, indicating that these are potentially contributing substantially to overall dietary patterns. In addition, the frequency of consumption of water as a beverage was higher in food secure young people in our study. This may relate to the relatively better dietary habits or better access to fresh water in this group. Lastly, in the current study, only a third of respondents reported consuming breakfast daily regardless of food security status, which is lower than in other studies of young people in Australia showing 92% consume breakfast daily.headspace Launceston in over the previous 12 months was approximately 1100, the exact number of eligible participants (aged 15–25 years) is unknown due to limitations in accessing this data from the headspace referral system. However, the current study employed multiple recruitment strategies to reach this at‐risk group, highlighting challenges in sampling this population that should be considered in future research. In addition, the regional location of the study setting further limits the generalisability of the results to other populations and geographical areas. Lastly, the sample contained a high proportion of female respondents (68%) which, while not uncommon in research related to food security,While this study presents some of the first results of food insecurity in young people attending a mental health service, the study should be considered in the context of some limitations. First, as is common for this cohort of vulnerable young people, the study has a small sample size. This may lead to Type II errors in the identification of risk factors and does not allow for subgroup analyses. While the total number of young people (aged 12–25 years) who were engaged in at least one appointment with In conclusion, this research found a high occurrence of food insecurity in young people attending a youth mental health service, with those living independently or in unstable housing, or receiving government financial assistance at the highest risk indicating that financial access to food is a major contributor to food insecurity in this group. Given the results of the study, there is a clear need for additional research in Australia to explore food insecurity in a representative sample of young people, including the main demographic drivers and relationship with mental and physical health outcomes. This should be a priority given the long‐term impacts on physical healthConceptualization, KK, SM, TM, HB; methodology, all authors; formal analysis, KK and DV; investigation, TM, HB; resources, HB; data curation, TM and KK; writing—original draft preparation, KK; writing—review and editing, all authors.; project administration, HB; funding acquisition, HB. All authors have read and agreed to the published version of the manuscript.The authors declare no conflicts of interest.


We are living in the era of genome editing. Nowadays, targeted editing of the plant nuclear DNA is prevalent in basic biological research and crop improvement since its first establishment a decade ago. However, achieving the same accomplishment for the plant mitochondrial genome has long been deemed impossible. Recently, the pioneer studies on editing plant mitogenome have been done using the mitochondria‐targeted transcription activator‐like effector nucleases (mitoTALENs) in rice, rapeseed, and Arabidopsis. It is well documented that mitochondria play essential roles in plant development and stress tolerance, particularly, in cytoplasmic male sterility widely used in production of hybrids. The success of mitochondrial genome editing enables studying the fundamentals of mitochondrial genome. Furthermore, mitochondrial RNA editing (mostly by nuclear‐encoded pentatricopeptide repeat (PPR) proteins) in a sequence‐specific manner can simultaneously change the production of translatable mitochondrial mRNA. Moreover, direct editing of the nuclear‐encoding mitochondria‐targeted factors required for plant mitochondrial genome dynamics and recombination may facilitate genetic manipulation of plant mitochondria. Here, the present state of knowledge on editing the plant mitochondrial genome is reviewed. Mitochondria play essential roles in plant development and stress tolerance, particularly, in cytoplasmic male sterility widely used in production of hybrids. Precise editing of mitochondrial genomic DNA using the mitoTALENs, mitochondrial RNA editing mostly by nuclear‐encoded PPR proteins and editing of the nuclear‐encoding mitochondria‐targeted factors required for mitochondrial genome dynamics and recombination enable better understanding of the fundamentals of mitochondria. Agronomists and breeders are putting enormous resources into crop improvement to encounter these challenges. But until recently, such efforts are mainly have been made by the conventional breeding strategies. The application of the CRISPR/Cas9  system in plants in recent years has paved a new avenue for crop improvement. Although nuclear genome editing accounts for most majority of the gene editing attempts in plants, mitochondrial gene editing tools are also developed and applied in the crop improvement, as such tools have their merits. In this article, we will walk the authors through the history, molecular basis, and application of major mitochondrial gene editing approaches and their possible roles in crop improvement for future agriculture. Mitochondria are the hub of energy supply chains, biosynthesis center for different types of lipids, and play important roles in the programmed cell death (apoptosis), calcium and redox homeostasis, the reaction to stressors, and nucleic gene expression regulation. Mitochondria genomes underwent many changes during the endosymbiosis process, but a reduced set of protein‐coding genes remains and are replicated, transcribed and translated independently from the nuclear genome. In general, mitochondrial genomes are double‐stranded DNA molecules but range in size and differ in architecture  in different species.Mitochondria are specialized organelles derived from endosymbiosis in eukaryotic cells.1 Mito The long repeat sequences involved in homologous recombination give rise to the dynamics of mitochondrial genome, with a potential to generate subgenomic forms and extensive genomic variation even within the same species. Such changes in genome structure are responsible for the rapid evolution of plant mitochondrial DNA and for the variants associated with cytoplasmic male sterility and abnormal growth phenotypes. In contrast to the variations in mtDNA sizes, the number of mitochondrial genes is relatively conserved in the plant kingdom, with about 60–70 genes found in the mitogenomes of different terrestrial plant species. Most of mitochondrial DNA is non‐protein coding. However, most mitochondria encoded genes are critical for the mitochondria functions and the normal development in plants.Mitochondrial genomes in flowering plants differ in size from about 200 kb to 11 Mb, and their circular, linear, and branched structures are distinct from those in mammalian and yeast cells.5, 6To better understand the functions of mitochondrial genes, the mitochondrial and nuclear interaction, and the evolution of mitochondrial genome, the intensive efforts have been put into the development of methodologies for stable plant mitochondria genome editing. Here, we review the exciting efforts and the most recent advancement in the mitochondrial genome editing research. We cover three major areas: precise editing of mitochondrial genomic DNA, mitochondrial RNA editing and editing of the nuclear‐encoded genes targeting mitochondria.2 While it is fairly easy to introduce the targeted modification to the nuclear genome by using CRISPR/Cas9, targeted mitochondrial genome editing in plants remains very challenging. The application of the CRISPR/Cas9 system in mitochondrial genome editing is complicated by the double membrane of mitochondria, and it is very difficult for the guide RNA to cross the inner mitochondrial membrane due to the strong electrochemical potential present across the membrane. Stable transformation of mitochondrial genomes using the mitoCRISPR‐Cas approach was not achieved until recently. Hussain et al. were able to perform gene editing in the mitochondrial DNA by using the CRISPR/Cas9 system; they reduced the expression of the mitochondrial‐encoded gene ND4 (NADH‐ubiquinone oxidoreductase chain 4) by appending an ND4‐targeting guide RNA to an RNA transport‐derived stem loop element (RP‐loop) and expressing the Cas9 enzyme with a preceding mitochondrial localization sequence in the murine cell cultures.Genomic DNA editing has been widely used in plants for characterizing gene function and crop improvement, especially in recent years with the application of the CRISPR/Cas9 system in plants.10 Whi Subsequently, stable genomic editing of the mitochondrial genome using mitoTALENs was established in model plant Arabidopsis. Applying mtDNA editing in Arabidopsis is difficult because most mitochondria‐encoded genes in Arabidopsis are the single‐copy and essential genes, and knocking out any of them would cause embryo lethality. To circumvent this issue, Arimura et al chose to work on ATP synthase 6 , a protein encoded by two gene copies, ATP6‐1 and ATP6‐2, in Arabidopsis thaliana Columbia‐0 (Col‐0). They successfully knocked out either ATP6‐1 or ATP6‐2 by using mitoTALEN vectors via floral dipping. They also pointed out the importance of avoiding the influence of nuclear copies of mitochondrial DNA (Numts) when designing the confirmation primers after the transformation.The first case of directed double‐strand breaks in plant mitochondrial genomes using mitochondria‐targeted TALENs in cytoplasmic male sterile rice and rapeseed was achieved by Arimura and his colleagues.13 Sub However, the precise gene editing of mitochondrial DNA (mtDNA) has been reported recently. This new method utilizes a novel bacterial cytidine deaminase, named “dsDNA deaminase toxin A”, or DddA, which specifically convert cytosines to thymines (C→T) present in double‐strand DNA. This is achieved by adding DddA to the MTS‐TALEN construct, forming MTS‐TALEN‐DddA fusion proteins, named “DddA‐derived cytosine base editor” (DdCBE). The Mok group tested different combinations of MTS‐TALEN‐DddA fusion vectors and precisely edited at least six different mitochondrial genes in the human cell lines, with efficiencies ranging from 4.6% to 49%. This breakthrough is going to accelerate the mitochondrial genome research in the animal model systems. The usage of MTS‐TALEN‐DddA has been tested in the plants as well. Recently, Kang et al. have assembled 16 DdCBE plasmids and used the resulting DdCBEs to efficiently introduced point mutagenesis in the mitochondria and chloroplasts of lettuce or rapeseed calli, we can expect this system will serve as a valuable resource for future organelle DNA function research and potential application of mitochondrial DNA editing in the crop improvement. As mitochondria play a critical role in the plant biotic and abiotic stress response, redox homeostasis, and phytohormone regulation. Targeted mitochondrial gene editing could create specific retrograding signaling and trigger downstream gene expression changes, and eventually create novel phenotypes.It is exciting to see the mitoTALEN approach eventually used in the model plants, but, this system has its own limits. The linearized mtDNA generated by fusion degrades rapidly; consequently, the modified mtDNA copies will eventually be outnumbered by the uncut version. The other issue is the mitoTALENs or mitoZFNs approaches cannot introduce specific nucleotide changes.17 How3 It is a process that changes the bases of RNA at the post‐transcriptional level. So far, at least 400 editing sites have been discovered in Arabidopsis mitochondrial RNAs, with most of them located in the protein‐coding gene regions and only a few sites in the non‐coding regions. In plants, natural RNA editing is carried out by a protein complex called editosome, a complex that contains multiple protein factors with the PPR proteins as its core components. The PPR proteins carry a tandem array of 35‐aminoacid helix‐turn‐helix motifs that can be classified into two subgroups: PPR‐P and PPR‐PLS. These two subgroups are distinguished by their structures and the functions they carry out. The PPR‐P subclass plays a crucial role in a wide range of organellar RNA metabolism, whereas the PPR‐PLS subclass mainly involves C‐U RNA editing. Many of the PLS proteins contain three types of highly conserved protein domain modules  at their carboxy terminus. The DYW domain named after the conserved amino acids aspartate (D), tyrosine (Y), and tryptophan (W), contains sequences similar to a conserved cytidine/deoxycytidylate deaminase motif. The sequence specificity of RNA editing is achieved by the PPR array recognizing the target RNA, and the DYW domain catalyzing the C‐to‐U conversion.In addition to mitochondrial DNA editing, it is also possible to introduce or reverse mitochondria‐associated traits by mitochondrial RNA editing, which can be achieved by RNA‐binding pentatricopeptide repeat (PPR) proteins to target editing sites in a sequence‐specific manner. RNA editing is based on intrinsic biological machinery that exists in many eukaryotic species.22 It  It was demonstrated for the first time in an individual moss (Physcomitrella patens) that a DYW‐type PPR protein alone can introduce efficient C‐to‐U editing in Escherichia coli and reproduce the moss mitochondrial editing. Using this straightforward E. coli system, different types of designed DYW‐type PPRs from different species were tested. They expressed the P. patens editing factor PPR79 and successfully edited all target sites in the Arabidopsis cwm1 mutant background; the homolog of PPR79 in Arabidopsis is CWM1. 2) P‐type PPR engineering. Precise mitochondrial RNA editing might also be achieved by P‐type PPR (non‐editing factors) protein engineering. The first example of a targeted block in the expression of a specific mitochondrial transcript by a custom‐designed PPR protein was reported by Colas des Francs‐Small et al, whereby they used a modified PPR protein RNA PROCESSING FACTOR 2 (RPF2) to eliminate the expression of the mitochondrial NAD6 gene in A. thaliana. 3), RNA editing using CRISPR‐Cas system. Although designed RNA editing can also be achieved by using the CRISPR‐Cas system, so far in plants, no successful targeted mitochondrial RNA editing via CRISPR‐Cas has been reported. Just as in DNA editing via the mitoCRISPR‐Cas, the challenge that CRISPR‐Cas‐mediated mitochondrial RNA editing faces is to introduce an efficient mean to deliver the gRNA to the mitochondrial matrix. The development of bioinformatics also fueled the research on the targeted mitochondrial RNA editing. A complete list of bioinformatics tools and databases that can be used for RNA editing study and engineering is included in the Giudice's comprehensive review. Considering all of these developments in methodologies for targeted mitochondrial RNA editing, and a wide range of important plant traits for RNA editing, we believe the targeted mitochondrial RNA editing approach opens a novel avenue for reverse genetics studies of mitochondrial gene function and will eventually have great applications in crop breeding.Based the molecular basis of natural mitochondrial RNA editing, So far, at least three approaches have been developed to achieve precise designed mitochondrial RNA editing: 1) RNA editing factors (PLS‐type with DYW domains) engineering. The sequence specificity and the one‐to‐one relationship between the DYW‐type PPRs and their targets enables the designed precise C‐to‐U modification in the targeted plant mitochondrial transcripts.32 It 4MSH1, RecA2, RecA3, WHY2, OSB1, and RECG1.As mentioned, the plant mtDNAs have quite complicated and dynamic structures and can form a set of subgenomic forms that are permutable by recombination. Therefore, surveillance machinery is developed to control mtDNA mutation rates and to maintain genome stability. In plants, this surveillance is carried out by a group of proteins involved in DNA replication and mismatch repair. Some examples are  It functions in mismatch repair as an inhibitor of strand invasion during homologous recombination. The MSH1 has been shown to limit ectopic recombination in plant cytoplasmic genomes and maintain the stability of the mitochondria genome in higher plants. It was reported that disrupting the MSH1 (but not the other mtDNA repair genes) leads to massive increases in the frequency of point mutations and small indels (approximately 10‐ to 1000‐fold) and changes to the mutation spectrum in mitochondrial DNA. Disruption in the MSH1 gene resulted in enhanced mitochondrial genome recombination at numerous repeated sequences, and a range of distinct phenotypes in five species including Arabidopsis, sorghum, tobacco, tomato, and pearl millet. Further investigation revealed that the mitochondrial genome rearrangements associated with the MSH1 depletion in mitochondria resulted in enhanced heat tolerance and the curly and wrinkled leaf phenotypes observed in the msh1 mutant. Hemi‐complementation of msh1 mutants with MSH1 specifically targeted to mitochondria completely restored mtDNA stability and the curly or wrinkled leaf phenotypes. In Brassica juncea, the MSH1 mediated higher substoichiometric shifting (SSS) activity and increased in the ORF220 copy number to induce male sterility and fertility reversion. Plant mitochondria possess two eubacteria‐type RecA proteins that are core components of the mitochondrial repair mechanisms. In Arabidopsis, the recA2 and recA3 mutations trigger increased ectopic mitochondrial DNA recombination. The recA2 is lethal at the early seedling stage, whereas the recA3 first‐generation mutant plants are normal in most cases, but develop more severe phenotypes and became hypersensitive to genotoxic treatments in the later mutant generations. These results suggest it is possible to introduce changes to the mitochondrial genome via editing the nuclear‐encoded mitochondria‐targeted genes and eventually create new traits in plants.The MSH1, MUTS HOMOLOG 1 is dual targeted to both chloroplasts and mitochondria. It was the first protein described as an mtDNA mutator in Arabidopsis.39 It 5 Nevertheless, the development of CRISPR tools for mitochondrial genome editing has been described in mammalian cells, with controversial results because of the efficiency of import into mitochondria. It was demonstrated that gRNA could be imported efficiently into the mitochondria of mammalian cells with an additional 20‐bp stem‐loop element of nuclear RNase P. More recently, an RNA‐free base editor has been developed for human cells by fusing a non‐toxic half of an interbacterial toxin with the transcription activator‐like effector array proteins and an uracil glycosylase inhibitor to edit precisely the mitochondrial genome. This raises the possibility of precisely manipulating mitochondrial genomes in plants. As the MTS‐TALEN‐DddA DNA editing already been tested in plants, we would expect decent amount of research using this technology to study mitochondrial gene function. In addition, it would be exciting to see the mitoCRISPR‐Cas DNA editing get applied in plants in the future. The development of more mitochondrial DNAs editing approaches will help us reach a better understanding of the cross‐talk between the nuclear and mitochondrial genome, and to eventually get applied to the creation of novel germplasms for crop improvement. More specifically, we expect mitochondrial DNA editing approaches will play an important role in creating abiotic and biotic stress tolerant plant germplasms in the future. We can also expect that the designed mitochondrial‐targeted PPR proteins will be used more frequently and efficiently in mitochondrial gene function studies. The feasibility of designed PPR proteins to bind specifically to a wide array of mitochondrial RNA sequences will be enhanced. The range of species this technology can be applied to will expand.Although much is known about the genomic sequences of plant mitochondria, the attempts to directly edit the mitochondrial genome using CRISPR/Cas systems have been hindered by the challenge of transporting the gRNA into the mitochondrial matrix.49 Nev Moreover, the fertility reversion rate of the CMS line in Brassica juncea can be altered by manipulating the MSH1 expression; In both cases, the mitochondrial genome‐modified materials can facilitate the utilization of heterosis, several studies reported that msh1 mutation derived epigenetic variation can be selected for crop improvement, produce plant materials with growth vigor and abiotic tolerance in tomato, soybean, and sorghum. Although to what extent mitochondria retrograde signaling contribute to these epigenetic variations still remain unclear, these findings suggest that editing the nuclear‐encoded mitochondria‐targeting genes could have agricultural applications. We believe the ongoing improvement in the mitochondrial genome editing methodologies will eventually lead to more advances in crop improvement, as the mitochondria genome manipulation used to introduce novel traits, or increase the plant resilience to emerging agricultural challenges. One thing is for sure, this is going to be an exciting era for scientists studying mitochondrial biology in plants (Figure The development of mitochondrial genome editing methodologies will benefit the plant breeding efforts; actually, we have already started to see such examples. Kazama et al managed to cure cytoplasmic male sterility in rice and rapeseed by using mitoTALEN to knock out the CMS‐associated genes in the CMS line.54 MorThe authors declare no conflicts interest.J.Y.: Conceptualization; writing‐original draft; writing‐review and editing. X.Y.: Conceptualization; writing‐original draft; writing‐review and editing. T.S.: Writing‐review and editing. Z.H.: Writing‐review and editing. M.Z.: Supervision; writing‐review and editing.The peer review history for this article is available in the Supporting Information for this article.Supplementary Information: Record of Transparent Peer ReviewClick here for additional data file.


Anthropogenic climate change has resulted in warming temperatures and reduced oxygen concentrations in the global oceans. Much remains unknown on the impacts of reduced oxygen concentrations on the biology and distribution of marine fishes. In the Southern California Channel Islands, visual fish surveys were conducted frequently in a manned submersible at three rocky reefs between 1995 and 2009. This area is characterized by a steep bathymetric gradient, with the surveyed sites Anacapa Passage, Footprint and Piggy Bank corresponding to depths near 50, 150 and 300 m. Poisson models were developed for each fish species observed consistently in this network of rocky reefs to determine the impact of depth and year on fish peak distribution. The interaction of depth and year was significant in 23 fish types, with 19 of the modelled peak distributions shifting to a shallower depth over the surveyed time period. Across the 23 fish types, the peak distribution shoaled at an average rate of 8.7 m of vertical depth per decade. Many of the species included in the study, including California sheephead, copper rockfish and blue rockfish, are targeted by commercial and recreational fisheries. CalCOFI hydrographic samples are used to demonstrate significant declines in dissolved oxygen at stations near the survey sites which are forced by a combination of natural multidecadal oscillations and anthropogenic climate change. This study demonstrates in situ fish depth distribution shifts over a 15‐year period concurrent with oxygen decline. Climate‐driven distribution shifts in response to deoxygenation have important implications for fisheries management, including habitat reduction, habitat compression, novel trophic dynamics and reduced body condition. Continued efforts to predict the formation and severity of hypoxic zones and their impact on fisheries dynamics will be essential to guiding effective placement of protected areas and fisheries regulations. This study examined changes in species distributions along a steep bathymetric gradient from 1995 to 2009 to determine whether declining concentrations of dissolved oxygen has triggered redistributions in marine ecosystems. One third of fish species observed on Southern California rocky reefs exhibited a significant shift in peak distribution to shallower habitat concurrent with shoaling hypoxic waters. As anthropogenic warming and ocean deoxygenation intensify, the rapid redistribution of marine species to more suitable habitats and the extirpation of populations that are unable to relocate will have unprecedented consequences on marine ecosystem structure and the provisioning of ecosystem resources As the ocean surface layer becomes more buoyant, transport of highly oxygenated surface waters into the ocean interior is reduced  from central California that were exposed to hypoxic conditions (DO concentration of 3.15 ml/L) exhibited increased metabolic costs, exploration behaviour and predation mortality compared to normoxic controls  and black rockfish (Sebastes melanops) from northern California decreased in hypoxic conditions  experienced elevated mortality rates  stock, the population has since recovered steadily and the stock was declared rebuilt by the Pacific Fishery Management Council in 2019  or severely hypoxic (DO <0.5 ml/L) environments .Oxygen concentration data were pulled from the California Cooperative Oceanic Fisheries Investigations program  hydrographic bottle data set. Seawater measurements are derived from discrete bottle sample depths collected on CTD/rosette casts during quarterly CalCOFI cruises at 66 stations. While oxygen concentrations are primarily derived from bottle samples, concurrent CTD sensor measurements may be used for corrections and interpolations. Detailed information about the CalCOFI program and methodologies can be found in CalCOFI cruise data reports  and non‐YOY. YOY were defined for each taxon as individuals with a total length less than the average length at 1 year, as predicted by the von Bertalanffy growth function using taxon‐specific growth parameters following Claisse et al.  Table S.N is the number of fish within a specific taxon, t, and life‐history stage, s, counted on a transect; D is the mean depth of the transect; Y is the year of the survey; D*Y is the interaction between depth and year; and L is the transect length. The depth squared term was added so that fish count estimates were not forced to monotonically increase or decrease across the large depth range observed in this study. This allows the models more flexibility to define the relationship between fish abundance and depth, creating a more realistic fit for species that may have a peak in distribution in this range. All Poisson models were tested for overdispersion using the AER library in R with alpha set at α = 0.05 , and had observation years spanning at least a decade. Fish abundance was modelled using Poisson regression as follows:Model predictions were generated for each survey year across the range of depths that a species was observed. The interaction between depth and year provides information on whether the modelled fish type is moving deeper or shallower over time. We define the annual peak distribution as the depth of maximum predicted fish abundance for a given survey year calculated from the model coefficients. Cases were excluded if a modelled peak in distribution across the three reefs did not occur within the taxon's observed depth range during any of the years within our time series.The model‐estimated change in the depth of peak distribution over time was calculated for each fish type. This depth change is defined as the difference between the peak depth distribution of the fish type in its first observed year and its final observed year, divided by the span of observation years. This method may result in an underestimate of the change in peak distribution depth because our surveys were limited to depths between 44 m and 365 m. Due to this limitation, we were not able to observe or model peaks in fish densities that may have moved shallower than 44 m over the 15‐year observation span.3p < 0.001; Figure 2/kg per year. This simple model was a better fit than the more complex model which included the interaction between year and depth category (ΔAIC = 8.75), indicating that the decline in oxygen over time was relatively consistent between depth categories. The same linear oxygen concentration model was also run during the restricted time period 1995–2009 corresponding to the timing of the fish surveys with a significant decline in oxygen at a rate of 0.70 μmol O2/kg per year , one of the most valuable commercial fishery species in the region  under hypoxic conditions in the laboratory  shifting deeper . This State Marine Reserve overlaps with most of our study area, including about half of the surveys in Anacapa Passage and all of the Footprint and Piggy Bank survey sites. ROV surveys monitoring the first 5 years after the establishment of nearby Channel Islands state marine reserves show higher abundances of rocky reef fishes inside the no‐take reserves relative to paired sites outside the reserves  are both commercially valuable with >60,000 lbs each landed in the Santa Barbara Channel region in 2019  and starry rockfish (Sebastes constellatus) are all valuable to recreational fishers, with more than 25,000 individual landings documented for each of these species in the Santa Barbara Channel region in 2019 (RecFIN, It is essential to document and predict distribution changes in fish species that are actively managed to monitor sustainable fishing practices and promote stock recovery. Among the fish species that exhibited a statistically significant distribution shift into shallower waters, California sheephead and copper rockfish ( RecFIN, . The 33  RecFIN,  may be lThe identification of hypoxic environments, tracking their spatial and temporal dynamics, and predicting the response of fish species to this environmental degradation is critical to supporting meaningful fishery management. Oxygen stress will emerge in nearly half of the global no‐take marine protected areas by 2050 under the business‐as‐usual climate projection RCP8.5 (Bruno et al., Declining oxygen will exceed the range of natural variability in most of the global ocean by 2052 (Henson et al., Fig S1Click here for additional data file.Table S1Click here for additional data file.Table S3Click here for additional data file.Table S4Click here for additional data file.Supplementary MaterialClick here for additional data file.