Emotion dysregulation and functional brain network topology in adult attention-decit/hyperactivity disorder

Emotion dysregulation is common in attention-decit/hyperactivity disorder (ADHD). It is highly prevalent in adult ADHD and related to reduced well-being and social impairments. Neuroimaging studies reported neural activity changes in ADHD in brain regions associated with emotion processing and regulation. It is however unknown whether decit in emotion regulation relate to changes in functional brain network topology in these regions. We used a combination of graph analysis and structural equation modelling (SEM) to analyze resting-state functional connectivity in 147 well-characterized young adults with ADHD and age-matched healthy controls from the NeuroMAGE database. Emotion dysregulation was gauged with four scales obtained from questionnaires and operationalized through a latent variable derived from SEM. Graph analysis was applied to resting-state data and network topology measures were entered into SEM models to identify brain regions whose local network integration and connectedness differed between subjects and predicted emotion dysregulation. The latent variable of emotion dysregulation was characterized by scales gauging emotional distress, emotional symptoms, conduct symptoms, and emotional lability. In individuals with ADHD characterized by prominent hyperactivity-impulsivity, the latent emotion dysregulation variable was related to an increased clustering and local eciency of the right insula. Thus, in the presence of hyperactivity-impulsivity, clustered network formation of the right insula may underpin emotion dysregulation in adult ADHD.


Introduction
Attention-de cit/hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by core symptoms of inattention and/or hyperactivity-impulsivity that may persist well into adulthood 1 . Emotion dysregulation, although not a core symptom, is a frequently co-occurring clinical problem 2,3 . Emotion dysregulation refers to the inability to adequately modulate and control emotions 4 . Its prevalence within the ADHD population changes with age, from 25-45% in children to 30-70% in adults, and its cooccurrence is associated with reduced well-being, risky behavior, and social impairments 2 . Especially individuals with ADHD and impulsivity-hyperactivity symptoms often suffer from emotion dysregulation 5 . However, the neural roots of ADHD associated emotion dysregulation remain unclear.
In emotion dysregulation, a distinction between explicit and implicit emotion regulation has been made.
Explicit emotion regulation requires conscious effort and is commonly achieved by applying cognitive control and reappraisal strategies. While cognitive control and reappraisal are particularly associated with activity in structures of the ventral attention and frontoparietal network 6 , it is precisely these structures in which deviant -often reduced -activity is frequently found in childhood and adult ADHD studies 1 . Implicit emotion regulation, on the other hand, is an unconscious stimulus-driven process based on experiencebased reward estimations. The ventromedial prefrontal cortex and the anterior cingulate cortex have been linked to implicit emotion regulation 6 . With regard to ADHD, it appears that not only structures related to explicit emotion regulation, i.e. structures for cognitive control and reappraisal, are affected, but also those associated with implicit emotion regulation and rather fundamental emotion reactivity processes 7 .
Functional connectivity studies consistently showed deviations in structures of the limbic system including the orbitofrontal, ventromedial prefrontal and anterior cingulate cortex in patients with ADHD [8][9][10][11][12][13][14] . Also, task-based fMRI studies using emotion perception and processing tasks in individuals with ADHD found evidence for functional abnormalities in the amygdala and insula, possibly indicating increased bottom-up emotional reactivity 15,16 . Task-based fMRI studies related to implicit emotion regulation, i.e., using fear extinction via habituation or emotional Stroop paradigms, which controlled for differences in cognitive control, found ADHD-speci c differences in the ventral anterior cingulate and ventromedial prefrontal cortex [17][18][19] . Given the heterogeneity of ADHD, existing neuroimaging studies, and postulated neurocognitive models, e.g., the dual-pathway model 20 , one might expect ADHDassociated emotion dysregulation to be similarly complex, with both explicit and implicit regulatory processes accounting for it.
While several studies, using task-based as well as resting state fMRI, reported brain activity deviations in structures commonly associated with emotion processing and emotion regulation, few have attempted to directly correlate corresponding activation patterns with emotion dysregulation and none has investigated changes speci c to ADHS subtypes. Two childhood ADHD studies used seed-based connectivity approaches focusing on the amygdala. They reported associations between high emotion dysregulation scores and reduced negative connectivity with the insula and frontoparietal structures as well as increased positive connectivity with the anterior cingulate cortex 21,22 . Connectivity changes beyond the amygdala have, however, not been investigated, and it remains uncertain whether the reported associations are indeed ADHD-or possibly even ADHD subtype-speci c.
To investigate the relationship between emotional dysregulation and functional brain network organization, we used graph theory to analyze fMRI resting-state data from healthy individuals and individuals with ADHD with and without hyperactivity-impulsivity symptoms. We captured the centrality of each brain network node and analyzed whether nodes were highly integrated and connected, either locally towards their direct neighboring nodes or globally with the entire network. Graph theory-based methods have previously been used in ADHD research 11,12,14 to describe changes in network topology of functional connectivity. These studies show increased local connectivity and e ciency and decreased global integration. It is however unclear how such changes in information processing properties of brain networks relate to emotional de cits. We gauged emotional dysregulation through structural equation modelling (SEM), using a combination of several self and informant scales assessing emotional problems, emotional lability, and conduct problems as well as one experimental task of emotion recognition. SEM is particularly well suited for testing the signi cance of certain assumed (groupspeci c) relations while simultaneously estimating latent variables embedded within the relational model. We focused our analysis on nodal topology measures and aimed to identify those brain regions whose local, functional brain network integration speci cally contributes to emotion dysregulation in predominately inattentive ADHD (ADHD-I) and ADHD with symptoms of hyperactivity-impulsivity (ADHD-C/H).
We hypothesized that an ADHD-speci c association exists between the functional brain network measures for local network integration and emotion regulation. Thereby, we focused on measures of network topology within limbic brain regions that have previously been associated with emotion processing. We assumed that the strongest relation would be found in individuals with ADHD and prominent hyperactivity-impulsivity symptoms.

Sample characteristics
Age and sex did not signi cantly differ between the ADHD and control group. Participants with ADHD showed higher scores on all four measures of emotional problems and dysregulation. Further, compared to controls, participants with ADHD had a signi cantly lower IQ, partially showed oppositional de ant disorder (ODD) or conduct disorder (CD) comorbid diagnoses and, in general, more often used stimulant medication. ADHD types did not signi cantly differ with regard to age, sex, IQ, the four measures of emotion dysregulation, or stimulant use. The demographic details of the sample are given in Table 1.

Structural equation models
To identify brain regions in which emotion dysregulation is speci cally linked with functional brain network topology as a function of ADHD subtype we combined graph analysis with SEM.
Three-group structural equation model. Second, to identify group dependent differences between network topology and the latent variable gauging emotion dysregulation, we used three-group SEM and estimated regression parameters for the group-speci c relationship between the latent emotion dysregulation variable and measures of nodal network topology. Pooling over different density levels of connectivity, the SEM revealed group-speci c differences in the relation between clustering coe cient as well as local e ciency measures, respectively, and the latent variable for emotion dysregulation in the right insula (see Figure 1a). For participants with ADHD-C/H the regression parameter estimates of the SEM were greatest, while they were smallest for participants with ADHD-I (see Figure 1b). Table 2 displays the group-speci c parameter estimates for the relation between the latent variable and the right insula's clustering and local e ciency measures. It also shows p-values (uncorrected), and estimates of the completely standardized solution. Further, Table 2 gives model-speci c χ 2 , degrees of freedom, p-values of the models, goodnessof-t measures (CFI, SRMR, & RMSEA), p-values that are the result of the χ 2 difference tests between the main models and the corresponding models with xed regression parameters, and signi cant pairs of the post-hoc two-group SEM.
Only in the ADHD-C/H group were the topology measures of the insula signi cantly related to emotion dysregulation. The respective models showed satisfactory goodness-of-t measures. Results were highly signi cant for the density-integrated models and signi cant group-speci c differences were found across multiple density thresholds below 25% connectivity (see Figure 1c & d). Results for SEM with clustering coe cients and local e ciency were very similar, as the measures themselves are very similar. Correspondingly, both measures, at the integrated level, showed a correlation of 0.913 for the right insula.
Post-hoc two-group comparisons revealed signi cant group differences between the participants with ADHD-I and ADHD-C/H as well as between participants with ADHD-C/H and controls. While not surviving FDR-controlling procedures, further group-speci c differences in the relation between topology measures and emotion dysregulation occurred for measures of nodes in the right orbitofrontal cortex (OFC), frontal pole, right insula, right anterior mid-cingulate cortex, and left insula (all uncorrected p-values <0.01).
Additional analyses. Robustness of the results was controlled by evaluating the group-speci c differences in overall connectivity strength of the individual networks. The analysis of overall connectivity did not reveal signi cant group differences. Finally, the multi-group SEM analysis was repeated using an alternative parcellation scheme to investigate the robustness of the results with respect to the chosen parcellation. The repetition of the main analysis with an alternative parcellation scheme revealed a groupspeci c difference in the relation between the clustering or local e ciency, respectively, and emotion dysregulation in the right insula. Further signi cant differences were found for the left insula and the right OFC and frontal pole. The latter also showed for the density-integrated values and after applying FDRcontrolling procedures (clustering coe cient: p=0.047, FDR-corrected; local e ciency: p=0.015, uncorrected). The signi cant right insula result, however, was not revealed using threshold-integrated values but only with a density threshold of 25% connectivity and not after FDR-controlling procedures (clustering coe cient: p=0.033, uncorrected; local e ciency: p=0.030, uncorrected).

Discussion
Emotion dysregulation is a key component of many psychiatric conditions, including adult ADHD. We identi ed properties of functional brain network topology related to a latent measure of emotion dysregulation by using a combination of graph theoretical methods with structural equation modelling. Our results provide evidence that especially in individuals with ADHD-C/H emotion dysregulation is accompanied by increases in local e ciency and clustering of the right insula.
Local e ciency and clustering measure how interconnected a node's neighbors are to each other.
Increases imply stronger functional connections between structures directly connected to the respective brain region. Here the insula was identi ed as showing increased connections to its nearest neighbors contributing to emotion dysregulation in ADHD-C/H. The insula integrates interoceptive states with emotional information via connections to other limbic regions such as the amygdala 23 . It plays an essential role in implicit emotion regulation via reciprocal connections with the ventromedial prefrontal cortex and other regions particularly associated with experience-based reward estimations 24 . Throughout brain maturation, functional network formation is characterized by speci c patterns of integrative and segregating processes 25 . These processes are also re ected in measures such as the clustering coe cient and local e ciency. Local e ciency in particular has previously been used as a measure of local brain segregation and it was suggested that increased local e ciency goes along with a loss of global brain network integration. For example, it was shown that higher local e ciency is associated with lower global e ciency and reduced performance during cognitive tasks 26 . Prior research consistently provides evidence of delays in the neural maturation of individuals with ADHD 8,27,28 . Increases in the insula's clustering and local e ciency, i.e. functional connectivity increases between neighboring nodes, may indeed, originate from reduced or delayed network-forming processes. This may negatively affect the e ciency with which the insula performs its integrative tasks and helps to appropriately evaluate emotional stimuli or to facilitate emotion regulation. Our results may re ect ADHD-subtype speci c de ciencies in functional network forming processes. At the behavioral level, this could lead to inappropriate behavior and some of the commonly observed emotional problems in ADHD.
The positive association between the functional connectivity of structures directly connected to the insula, i.e. local e ciency and clustering, and emotional functioning in ADHD-C/H was neither found in healthy controls nor participants with ADHD-I. This is consistent with expectations, given that this subgroup was shown to be most severely affected by co-occurring emotion dysregulation 3 . Previously, it was observed that deviations of functional connectivity in ADHD-I occur most clearly in areas of the frontoparietal network, especially the dorsolateral prefrontal cortex, whereas for ADHD-C deviations are most pronounced in areas of the default mode network, i.e. in areas associated with motivation and emotion processing. Since ADHD-I is predominantly characterized by inattention symptoms, it has been suggested that problems in top-down control systems are more likely to underlie this subtype, whereas combined type ADHD appears to be more clearly associated with motivation and emotion processing networks 29 . Interestingly, in the comparison of the different ADHD subtypes, the combined type shows local functional hyperconnectivity in regions associated with emotion processing and implicit emotion regulation, namely the vmPFC 30 . Taken together, the clinical distinction between ADHD subtypes with and without hyperactivity-impulsivity symptoms and the associated prevalence differences commonly observed in co-occurring problems such as emotion dysregulation may be re ected in differential deviations of the underlying functional organization of the brain.
Signi cant results were found for the right insula only. While ADHD is not understood as a generally rightlateralized disorder, some functions are more strongly associated with right hemisphere processing 31 .
Indeed, the right hemisphere is assumed to take a dominant role in emotion processing. Regarding the regulation of emotions, ndings are not as conclusive. However, evidence concerning the insula suggests that processes for integrating interoceptive and emotional/motivational information are rather right lateralized 32 . Note that the left insula yielded -like the right OFC, frontal pole and cingulate cortexsimilar but weaker results, which did, however, not survive correction for multiple comparisons.
The latent variable for emotion dysregulation encompassed self-reported emotional distress, emotional and conduct symptoms and informant-reported emotional lability. Callous-unemotional traits and emotion recognition did not contribute signi cantly. Their relation to the other emotional measures (i.e., emotional distress & lability) appeared to be rather negligible. This is not surprising as callousunemotional traits are characterized by de cient affect and lack of empathy, which is a conceptually different aspect of emotion 33 . While emotion recognition is an important prerequisite for the regulation of emotions, adolescents with and without emotion dysregulation have shown similar abilities in recognizing emotions, suggesting again a conceptual difference 34 . Nevertheless, de cits in both are frequent in ADHD, a common occurrence is, however, not imperative 35 .
The novel combination of SEM and graph theory presented here enabled us to reliably assess the functional network topology underlying emotion dysregulation. By using a SEM approach, it was possible to simultaneously determine emotional dysregulation as a latent variable of several emotion questionnaires and assess the group-speci c relationships of emotion dysregulation with different measures of nodal brain topology. Previous approaches were limited in focusing on seed based functional connectivity of the amygdala only 21,22 and using only unidimensional emotional measures. Compared to prior neuroimaging studies, we used a relatively large and well characterized sample of adult ADHD patients and healthy controls. Also, cases with missing values were excluded from the data set before performing the analyses. Nevertheless, even larger sample sizes may be needed to obtain appropriate power and avoid parameter biases in SEM Note however that simple models, as used here, require smaller samples (e.g., about 30 cases for a simple CFA with four indicator variables) and larger samples are only necessary if missing values exist 36 . The robustness of the ndings was further tested by conducting an additional analysis with an alternative parcellation scheme. Even though we were able to replicate the increased local e ciency and clustering in core regions of the implicit emotion regulation network, effects in the insula were weaker and those in orbitofrontal cortex and frontal pole more pronounced.
In conclusion, the present study shows a positive relation of the right insula's clustering and local e ciency with emotion dysregulation in adult individuals with ADHD-C/H. A similarly strong connection was not found for individuals with ADHD-I or healthy controls. The results suggest ADHD-type speci c de ciencies in network forming processes that are associated with emotion processing and its implicit regulation. The commonly observed emotional problems in ADHD may partially be linked to the present ndings. Given that emotion dysregulation is present in many other psychiatric disorders it is interesting to note that several of those, i.e. major depressive disorder, bipolar disorder, anxiety disorders and schizophrenia are also associated with changes in the structural and functional connectivity of the insula 37 , which may represent a worthwhile target area for future treatment efforts.

Participants and procedures
Data were taken from NeuroIMAGE II, the third wave of an integrated genetics-cognition-MRI-phenotype project on ADHD 38 . It includes resting-state fMRI data of 249 individuals with ADHD as well as age-and sex-matched healthy controls. Initial recruitment criteria for ADHD participants were an ADHD combined type diagnosis, availability of one or more siblings, age between 6-18 years and availability of subject, sibling, and at least one biological parent for DNA collection. Exclusion criteria (for all participants) were IQ (as measured by Wechsler Intelligence Scale for Children/Wechsler Adult Intelligence Scale) < 70, diagnoses of autism or schizophrenia, and neurological disorders. For controls, it was required that neither they nor any of their rst-degree relatives had a prior ADHD diagnosis.

Reassessment of the ADHD diagnosis was established by combining information from the Kiddie
Schedule for Affective Disorders (K-SADS) 39 and parent, teacher, and self-report versions of the Conners' rating scale (CPRS-R:L, CTRS-R:L & CAARS-R:L) [40][41][42] . All diagnostic and phenotypic data was acquired on the same day as the fMRI data. A detailed description of the diagnostic procedures is given by von Rhein et al. 38 .
Participants with a subthreshold ADHD diagnosis (2-4 ADHD-speci c symptoms), left-handedness, excessive movement during the scanning, or insu cient quality of rs-fMRI or questionnaire data were excluded. 147 participants were used for the nal analysis, including 31 participants with ADHD but without hyperactivity-impulsivity symptoms, i.e., predominantly inattentive ADHD (ADHD-I), 25 participants with ADHD and hyperactivity-impulsivity symptoms (ADHD-C/H), i.e., 21 with combined type ADHD and 4 with predominantly hyperactive-impulsive ADHD, and 91 healthy controls. Demographic information are given in Table 1.
Forty-eight hours prior to testing, stimulant medication use was discontinued. Data acquisition took place at the Donders Institute for Cognitive Neuroimaging, Radboud University Nijmegen, Netherlands. Participants (and their parents when < 18 years old) gave written informed consent for participation.

Graph analysis
For graph analysis we used Python 3.5 with NetworkX 46 . Parcellation of preprocessed time series data was realized using a hemisphere-speci c functional brain template with 268 parcels 47 . It was created using graph-theoretically based parcellation that ensures functional homogeneity within the parcels of the atlas, even across different individuals. The atlas thus minimizes the likelihood that different functional areas lie within a single parcel 47 . For 221 relevant parcels, covered by the MR measurement, subject-speci c average intensity time series were calculated. Correlation matrices were created by computing pairwise Pearson's correlations between the extracted time series. Matrices were Fisher's ztransformed and absolute values were taken. Absolute value transformation was performed as preprocessing of the present data did not involve global signal regression and as anti-correlations are thought to be functionally relevant 48 . Due to the naturally low density of negative correlations, we refrained from performing speci c analysis for positive and negative correlations. To distinguish differences in network density from those of network topology 49 , matrices were binarized based on seven equally spaced density thresholds with a minimum density of 0.10 and maximum density of 0.40 50 . In this range of low to medium network densities, previous studies found signi cant associations between network topology and ADHD symptoms 11,12,14 . Thus, seven threshold-speci c graphs for each subject were investigated in the following graph analysis. Nodal topology measures were calculated for 46 of the 221 nodes. These 46 nodes were chosen based on their association with parcels overlapping with the orbitofrontal cortex, ventromedial prefrontal cortex, anterior cingulate cortex, insula, ventral striatum, amygdala, and hippocampus as de ned by the Harvard-Oxford Brain Atlas by more than 30%. These brain regions have been previously documented to be involved in emotion processing, its implicit regulation, and brain dysfunctions in ADHD 7 . Since we focused on implicit regulation, regions more strongly related to explicit processes, such as cognitive reappraisal, (e.g., frontoparietal network) were not considered.
Here, we captured the centrality of each node and analyzed whether nodes were highly integrated and connected, either locally towards their direct neighboring nodes or globally with the entire network. Thus, our focus is on six nodal measures, that is betweenness, closeness, eigenvalue centrality, clustering coe cient, nodal e ciency, and local e ciency, which were used in previous studies on ADHD and showed ADHD-speci c deviations 11,12,14 . Betweenness, closeness, and eigen-value centrality are measures that describe the centrality of a node within a network. While betweenness describes how often a node is part of the shortest path between two other nodes, closeness describes how many of the theoretically possible direct connections to other nodes actually exist. Eigenvalue centrality also considers the centrality of the node's direct neighbors. The clustering coe cient of a node describes how strongly its neighboring nodes are interconnected. E ciency values indicate how directly nodes can be reached from other nodes of the network. In the case of nodal e ciency, this refers to the shortest connection from a particular node to all other nodes, while in the case of local e ciency it refers to the e ciency amongst the nodes adjacent to a particular node of interest. All measures entered into separate SEM models described in 2.4.
Density-integrated topology measures were calculated 49 . Differences between populations of weighted networks may be due to the networks' wiring costs and not the targeted topological features. Densityintegration of measures from binarized networks, however, can eliminate cost-related differences and also allow the assessment of topology measures under different density-thresholds. Figure 2 summarizes the functional connectivity and network analysis.

Statistical analysis and structural equation modeling
Variable Selection. Statistical analyses were conducted with R software 51 . Lavaan was used for SEM 52 .
Prior to SEM, multi-group con rmatory factor analysis was used to identify data suitable for calculation of the latent emotion dysregulation variable. NeuroIMAGE II includes 6 tasks, questionnaires, or questionnaire subscales, respectively, that gauge emotional problems, emotional lability, and associated features: the Kessler Psychological distress scale, NESDA version of K-10 53 with 10 items for the assessment of emotional problems including anxiety and depression, the strength and di culties questionnaire subscales (SDQ; ve items about anxieties, worries, happiness, and physical symptoms of emotional stress for the emotional symptoms subscale & ve items about temper tantrums, compliance, quarrelsomeness, stealing, and lying for the conduct symptoms subscales) 54 , the emotional lability subscale of the Conners' parent rating scale (CPRS-R:L consisting of three items for unpredictable mood changes, temper tantrums, and tearfulness) 42 , the Inventory of Callous-Unemotional traits (ICU) 55 (24 items with a callousness and unemotional traits score), and the MINDS Testmanager's gradual emotion recognition task (GERT) 56 (accuracy of correct emotion classi cation).
Mediation structural equation model. The latent variable was used to investigate the relationship between functional brain network activity, emotion dysregulation, and ADHD. In the initial, preregistered analysis plan, we intended to use SEM mediation models in which the relationship between topological measures and the emotional latent variable was mediated by ADHD scores (CPRS-R:L) without taking into account the different ADHD subtypes. However, those models did not produce good model-t (as measured by the standard goodness-of-t measures described below) and had no signi cant results. One reason may be that the ADHD scores were derived from a parents' questionnaire for children's ADHD symptoms (CPRS-R:L) which may be less valid than a clinical diagnosis.
Three-group structural equation model. Alternatively, we chose a multi-group SEM approach using the clinical diagnoses. The diagnoses re ect all available diagnostic information and may thus provide more accurate models. We investigated the group dependent differences (i.e., healthy controls, ADHD-I participants, & ADHD-C/H participants) in the association between topology measures and the latent variable gauging emotion dysregulation. For each node of interest and each density-speci c topology measure, one multi-group model was built. Models consisted of the latent variable, questionnaire scores, associated parameter estimates and variances as well as the nodal topology measure variable with its regression parameter for the latent variable (see Fig. 3). Factor loadings of the latent variable were xed across groups, latent variable variance was standardized, and to account for between-group mean differences, group-speci c intercepts were added. Models were compared with almost identical models, in which however the regression parameters between the network topology variable and the latent emotion dysregulation variable were xed across groups. Model estimation was conducted using maximum likelihood procedures. To evaluate the signi cance of the group effects, model-speci c χ 2 -values were compared between the two models (χ 2 -difference-test). Following the approach of Ginestet et al. 49 , a model was considered signi cant if it revealed a signi cant effect on the density-integrated level (averaged over densities). Subsequently, signi cance was tested on each density threshold to provide detailed information on whether effects were found with stronger or weaker network connections.
Post-hoc analysis, multiple comparison procedures and goodness-of-t. We used post-hoc two-group SEMs with Bonferroni-corrected p-values to investigate pairwise between-group differences. These twogroup models resembled the three-group models described above, except that each only considered participants from two of the three groups. Alpha in ation due to multiple comparisons was controlled by the Benjamini-Hochberg false discovery rate (FDR) procedure 57 . The comparative t index (CFI), standardized root mean square residual (SRMR), and root mean square error of approximation (RMSEA) were calculated to evaluate the models' goodness-of-t. Acceptable goodness-of-t measures should at least be above 0.95 for the CFI, below 0.08 for the SRMR, and below 0.10 for the RMSEA 58 .
Additional analyses. To further control the robustness of the results, we evaluated whether group-speci c differences in overall connectivity strength of the underlying adjacency matrices exist 59 . Furthermore, an alternative parcellation scheme was used to investigate the robustness of the results with respect to the chosen parcellation approach. For this, we used the AAL atlas 60 , further subdivided with K-mean clustering to adapt the size and number of parcels to that of Finn et al. (2015). Procedures for calculating topology measures and SEM analyses were conducted as described above.

Data availability statement
The data are property of the Donders Institute for Cognitive Neuroimaging and are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.
Further information about the NeuroIMAGE project may be requested via the project leader Jan K. Buitelaar.