In the original article, we mistakenly reported on a secondary outcome for the Towbin et al. (2020) study, specifically the Children's Depression Rating Scale—Irritability item rather than the primary outcome, the Clinical Global Impression's for Chronic Severe Irritability. We have updated this information to Pharmacological Interventions” section, at the end of paragraph 2:An update has been made to the “d = 0.69, p < 0.001); however, a non-significant difference was observed between the citalopram and placebo groups for change in irritability during the subsequent blinded randomized trial phase (18)” has been replaced by “For their primary outcome of the Clinical Global Impression's for Chronic Severe Irritability, a significant decrease in irritability was found (d = 0.60), with the estimated response differing between the citalopram (35%) and placebo (6%) groups  (18).”The original text “A significant decrease in irritability was observed from baseline to the end of the 5-week open trial on MPH  and one utilizing youth self-report on The Irritability Scale (19). In contrast, five of the studies created irritability composites based on ODD items from rating scales for disruptive behavior, including the Swanson Nolan and Pelham version IV scale, the Disruptive Behavior Disorder Rating Scale, and the ADHD Symptom Checklist . Further, two utilized measures of depression or mania to assess irritability including the Children's Depression Rating Scale and Young Mania Rating Scale . Surprisingly, none employed the Affective Reactivity Index (ARI) (36), which has well established psychometrics in pediatric populations. As such, we recommend that future research assessing interventions for irritability among youth with ADHD utilize well-validated measures, such as the ABC-I and ARI, as well as consider integrating multiple informants vs. sole reliance on parent report. Relatedly, several studies were excluded from the review due to only examining irritability as a side effect/adverse event .” has been replaced by “Specifically, only six of the included studies utilized an established scale for irritability, with four using the ABC-I subscale , one utilizing the Clinical Global Impression focused on irritability which was generated based on a semi-structured diagnostic interview (18), and one utilizing youth self-report on The Irritability Scale (19). In contrast, five of the studies created irritability composites based on ODD items from rating scales for disruptive behavior, including the Swanson Nolan and Pelham version IV scale, the Disruptive Behavior Disorder Rating Scale, and the ADHD Symptom Checklist . Further, one utilized a measure of mania to assess irritability, specifically the Young Mania Rating Scale (25). Surprisingly, none employed the Affective Reactivity Index (ARI) (36), which has well-established psychometrics in pediatric populations. As such, we recommend that future research assessing interventions for irritability among youth with ADHD utilize well-validated measures, such as the ABC-I and ARI, as well as consider integrating multiple informants vs. sole reliance on parent report. Such an approach was only used in one of the included studies in this review (18). Relatedly, several studies were excluded from the review due to only examining irritability as a side effect/adverse event .”The original text “This mini review addresses an important gap in the literature by discussing the existing pharmacological and psychosocial interventions targeting irritability among youth with ADHD. Findings suggest that CNS stimulants used alone or in combination with behavioral therapy  are effective at reducing irritability in youth with ADHD only or comorbid ADHD and DMDD/SMD. Less evidence was found for the efficacy of alpha agonists and atomoxetine, with existing studies focusing on youth with comorbid ASD. Parent training alone or in combination with atomoxetine was found to be effective at reducing irritability in youth with comorbid ADHD and ASD. Future research assessing the efficacy of other psychosocial interventions, particularly CBT is necessary, as are randomized trials with large samples using well validated scales designed to measure irritability that assess intervention sequencing and intensity among youth with ADHD.” has been replaced by “This mini review addresses an important gap in the literature by discussing the existing pharmacological and psychosocial interventions targeting irritability among youth with ADHD. Findings suggest that CNS stimulants used alone or in combination with behavioral therapy  are effective at reducing irritability in youth with ADHD only or comorbid ADHD and DMDD/SMD. Less evidence was found for the efficacy of alpha agonists and atomoxetine, with existing studies focusing on youth with comorbid ASD. One study found evidence for benefits of adding a selective serotonin reuptake inhibitor  to MPH for reducing irritability in youth with ADHD and DMDD/SMD. Parent training alone or in combination with atomoxetine was found to be effective at reducing irritability in youth with comorbid ADHD and ASD. Future research assessing the efficacy of other psychosocial interventions, particularly CBT is necessary, as are randomized trials with large samples using well-validated scales designed to measure irritability that assess intervention sequencing and intensity among youth with ADHD.”The authors apologize for this error; beyond these minor changes, this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Natural killer/T-cell lymphoma (NKTL) is a rare type of aggressive and heterogeneous non-Hodgkin's lymphoma (NHL) with a poor prognosis and limited therapeutic options. Therefore, there is an urgent need to exploit potential novel therapeutic targets for the treatment of NKTL. Histone deacetylase (HDAC) inhibitor chidamide was recently approved for treating relapsed/refractory peripheral T-cell lymphoma (PTCL) patients. However, its therapeutic efficacy in NKTL remains unclear.We performed a phase II clinical trial to evaluate the efficacy of chidamide in 28 relapsed/refractory NKTL patients. Integrative transcriptomic, chromatin profiling analysis and functional studies were performed to identify potential predictive biomarkers and unravel the mechanisms of resistance to chidamide. Immunohistochemistry (IHC) was used to validate the predictive biomarkers in tumors from the clinical trial.We demonstrated that chidamide is effective in treating relapsed/refractory NKTL patients, achieving an overall response and complete response rate of 39 and 18%, respectively. In vitro studies showed that hyperactivity of JAK-STAT signaling in NKTL cell lines was associated with the resistance to chidamide. Mechanistically, our results revealed that aberrant JAK-STAT signaling remodels the chromatin and confers resistance to chidamide. Subsequently, inhibition of JAK-STAT activity could overcome resistance to chidamide by reprogramming the chromatin from a resistant to sensitive state, leading to synergistic anti-tumor effect in vitro and in vivo. More importantly, our clinical data demonstrated that combinatorial therapy with chidamide and JAK inhibitor ruxolitinib is effective against chidamide-resistant NKTL. In addition, we identified TNFRSF8 (CD30), a downstream target of the JAK-STAT pathway, as a potential biomarker that could predict NKTL sensitivity to chidamide.Our study suggests that chidamide, in combination with JAK-STAT inhibitors, can be a novel targeted therapy in the standard of care for NKTL.Trial registration: ClinicalTrials.gov, NCT02878278. Registered 25 August 2016, https://clinicaltrials.gov/ct2/show/NCT02878278The online version contains supplementary material available at 10.1186/s13148-023-01436-6. Natural killer/T-cell lymphoma (NKTL) is an aggressive Epstein–Barr virus-associated T-cell lymphoma (TCL) that accounts for 10% of all TCLs . It is mEP300, MLL2, ARID1A and BCOR [Histone acetylation is an important mode of epigenetic regulation controlled by the antagonistic actions of two classes of enzymes: histone acetyltransferases (HATs) and histone deacetylases (HDACs). HATs acetylate histones, leading to open chromatin regions that promote gene transcription. On the other hand, HDACs deacetylate histones, resulting in chromatin condensation, which induces transcriptional repression . Frequenand BCOR , 11. Abeand BCOR . InhibitMultiple HDAC inhibitors, such as vorinostat (SAHA), romidepsin (FK-228) and belinostat, have shown promising clinical efficacy in TCL patients . ApproxiIn this study, we evaluated the therapeutic effects of chidamide in 28 relapsed/refractory NKTL patients, and a remarkable ORR of 39% was achieved. We identified potential biomarkers to predict the clinical response to chidamide. Moreover, we deciphered the underlying mechanisms of primary resistance to chidamide mediated by aberrant JAK-STAT signaling and found that JAK-STAT inhibitors can effectively overcome this resistance in the treatment of NKTL.A phase II clinical trial to evaluate chidamide monotherapy in relapsed/refractory NKTL was conducted at Sun Yat-sen University Cancer Center (SYSUCC). The study was approved by the institutional review board of SYSUCC in accordance with the Declaration of Helsinki, and applicable regulations according to Good Clinical Practice guidelines. Written informed consent was obtained from all patients before enrollment in the trial. This trial was registered at ClinicalTrials.gov, number NCT02878278. Research protocols for tumor collection and analysis were approved by the ethical committees of SYSUCC. The details for the patients and cell lines are described in the Supplementary Methods. We routinely tested mycoplasma contamination in cell culture using Myco-Blue® Mycoplasma Detector (Vazyme) monthly.Chidamide was provided by Shenzhen Chipscreen Biosciences Ltd. SAHA was obtained from Sigma–Aldrich. Trichostatin A (TSA), belinostat and tofacitinib were purchased from Selleckchem. Romidepsin was obtained from MedChemExpress. Ruxolitinib and Stattic were purchased from Axon Medchem and Millipore, respectively.A total of 2000 cells were seeded in 96-well plates in triplicate and treated with the indicated drugs at various concentrations or not treated for 96 h. Cell viability was measured by the CellTiter-Glo Luminescent Cell Viability Assay (Promega) according to the manufacturer’s instructions, and luminescence was read using an Infinite M200 plate reader . Half maximal inhibitory concentration (IC50) values were calculated, and proliferation curves were plotted using GraphPad Prism.CI values were determined by the inhibition rate of the cells treated with drugs and computed using CalcuSyn . CI valu5 cells were treated with indicated drugs for 72 h. Cells were washed with cold PBS and fixed with 70% ethanol for more than 1 h. Fixed cells were washed with cold PBS twice and treated with 100 μl 100 μg/ml ribonuclease A  for 5 min. The cells were then stained with 400 μl 50 μg/ml propidium iodide  for 40 min in the dark and analyzed by flow cytometry .2 × 105 cells were treated with indicated drugs for 72 h. Cells were washed with ice-cold PBS and stained with Annexin V-FITC Apoptosis Detection Kit (Vazyme) following the manufacturer’s instructions. The stained cells were detected using flow cytometry .2 × 10H&E staining, IHC for TNFRSF8 (CD30) and Epstein–Barr encoding region in situ hybridization (EBER ISH) were performed with formalin-fixed, paraffin-embedded (FFPE) tissues from NKTL patients using standard procedures , 21 withThe primer sequences used for ChIP-qPCR are listed in Additional File The primer sequences used for RT–qPCR are listed in Additional File In vivo studies were conducted in compliance with animal protocols approved by the SingHealth Institutional Animal Care and Use Committee. Five- to eight-week-old female NOD/SCID/IL2rγnull (NSG) mice were kept under standard laboratory conditions according to the National Advisory Committee for Laboratory Animal Research guidelines. The details are provided in the Supplementary Methods.t test, while significant differences among multiple groups were analyzed by two-way ANOVA with Bonferroni’s correction using GraphPad Prism 7. Fisher’s exact test was used to assess the association between protein expression and clinical response. p < 0.05 was considered statistically significant.Graphs include results that are expressed as the mean ± SD of three independent experiments. Statistical differences between the two groups were evaluated by a two-tailed Student’s p < 0.0001, log-rank test; median OS, 39.7 months vs. 2.9 months, p = 0.0003, log-rank test) . The patients were treated with chidamide until disease progression or intolerance. The demographic and clinical characteristics of the 28 patients are summarized in Additional File To assess the in vitro efficacy of chidamide in NKTL, we evaluated the IC50 of the drug in 11 NKTL cell lines Fig. A and divn = 28,727) revealed that the H3K27ac signals were significantly different between resistant and sensitive cells  was found  for H3K27ac and H3K4me3 in two resistant cell lines HANK1 and SNK6) and two sensitive cell lines (KHYG1 and MEC04). Epigenetic changes in H3K27ac and H3K4me3, which are associated with gene enhancers and promoters, respectively, have been shown to modulate gene activities and confer resistance to drug treatment , 24. Uns and SNK6TNFRSF8 and TNFRSF9. Increased H3K27ac signal intensities at the enhancers of the TNFRSF8 and TNFRSF9 gene loci were observed in HANK1 and SNK6, but not in KHYG1 and MEC04 cells  and two chidamide-sensitive (KHYG1 and MEC04) cell lines. Integrative transcriptomic and chromatin profiling analyses revealed that the differential H3K27ac peaks were positively correlated with gene expression in NKTL cells  [p = 0.0272, Fig. As the JAK-STAT pathway was found to be implicated in resistance to chidamide, we examined the activity of its effector molecules, STAT3 and STAT5 in both sensitive and resistant NKTL cell lines. High levels of phospho-STAT3 and STAT5 were detected in resistant cell lines compared to sensitive cell lines Fig. A. Aberrad MEC04) . UpregulWe hypothesized that targeting the JAK-STAT pathway could reverse chidamide resistance in NKTL cell lines. To test this hypothesis, we treated the chidamide-resistant NKTL cell lines HANK1 and SNK6) with tofacitinib (JAK inhibitor), chidamide or a combination of both drugs for 96 h and determined the combination index Fig. A. Combin and SNK6To understand how JAK inhibitors reverse chidamide resistance in NKTL cells, we performed RNA-seq of HANK1 cells treated with DMSO, tofacitinib or ruxolitinib. Compared to the control treatment, tofacitinib and ruxolitinib treatment remarkably upregulated 131 common genes and downregulated 141 common genes in HANK1 cells Fig. B. TofaciWe next assessed whether JAK-STAT inhibition could be a potential therapeutic strategy to overcome resistance to chidamide in the clinic. A 48-year-old Chinese male patient who was initially diagnosed with stage IIA extranodal NKTL of the nasal type was recruited. He was refractory to first-line asparaginase-based chemotherapy. Salvage therapy with a combination of chidamide  and the anti-programmed death protein 1 (PD-1) antibody sintilimab  was given, but the disease progressed with multiple new lesions accompanied by a dramatic increase in the peripheral blood Epstein–Barr virus (EBV) viral load after four 21-day cycles Fig. A. The JAIn the current management of NKTL, a substantial proportion of patients still experience treatment failures, and salvage options are severely limited. Increasing evidence suggests that PTCL is characteristically an epigenetic disorder, with several HDAC inhibitors receiving regulatory approvals for the treatment of relapsed/refractory disease. Here, we conducted the first focused trial of chidamide, an oral HDAC inhibitor, in relapsed/refractory NKTL. Chidamide induced durable tumor regression, with a complete response rate of 18% and a partial response rate of 21%. Compared with other HDAC inhibitors, chidamide is the first selective oral HDAC inhibitor to show effective and increased immunomodulatory effects and convenient administration , 32. AltSTAT3 activating mutations in 21% of patients with this disease [STAT3 activating mutations, which were not found in the three chidamide-sensitive cell lines , suggesting that mutations might be one of the underlying mechanisms leading to activation of the JAK/STAT pathway in NKTL, which confers resistance to chidamide. Our study shows that hyperactivation of JAK-STAT signaling confers resistance to chidamide in NKTL, suggesting that the activity of JAK-STAT may serve as a predictive biomarker of the response to chidamide.We further found that aberrant JAK-STAT signaling contributes to primary resistance to chidamide, and inhibiting this oncogenic pathway can resensitize resistant NKTL cells to chidamide. We have previously demonstrated the prevalence of JAK-STAT pathway alterations in NKTL and discovered frequent Interestingly, we found that JAK-STAT signaling mediates resistance to chidamide through chromatin remodeling of enhancer elements. We demonstrated that drug resistance to chidamide was correlated with chromatin remodeling alterations, which could be reversed by targeting the JAK-STAT pathway. Enhancer but not promoter alterations were significantly correlated with the response to chidamide in NKTL. In addition, STAT motifs were enriched at gained enhancers over lost enhancers, which was consistent with the enrichment of JAK-STAT target genes in resistant cells. Previous studies have demonstrated that the JAK-STAT pathway can regulate chromatin accessibility and influence the chromatin landscape through different mechanisms. For example, JAK2 can directly phosphorylate Tyr41 in histone H3 (H3Y41), preventing the binding of heterochromatin protein 1α (HP1α) with chromatin and destabilizing heterochromatin in hematopoietic cells . The carTNFRSF8 and TNFRSF9. Consequently, inhibition of JAK-STAT overcomes resistance to chidamide. A recent study showed that massive enhancer remodeling plays a key role in acquired resistance to anaplastic lymphoma kinase (ALK) inhibitors and that panobinostat can induce chromatin remodeling in resistant cells to overcome acquired resistance to ALK inhibitors in ALK-positive lung cancer [Importantly, chromatin-mediated drug resistance is often reversible, as demonstrated by JAK-STAT inhibitor reprogramming of the chromatin state from a resistant to a sensitive state. JAK-STAT inhibition suppresses H3K27ac marks of JAK-STAT target genes, such as g cancer . Knoecheg cancer . Moreoveg cancer . ConsistFurthermore, JAK-STAT target genes can also serve as predictive or therapeutic biomarkers of chidamide resistance. Frequent overexpression of TNFRSF8 (CD30) in NKTL has been reported previously , 44. CD3In summary, our study shows favorable clinical outcomes of chidamide in treating relapsed/refractory NKTL patients. JAK-STAT inhibitors may further expand the clinical utility of HDAC inhibitors, including chidamide, romidepsin and belinostat, by remodeling the resistant enhancer landscape toward a sensitive state. The combined use of JAK-STAT and HDAC inhibitors is a promising new treatment option for relapsed/refractory NKTL patients. We believe that our study could promote the clinical application of HDAC inhibitor chidamide in NKTL, provide personalized and precision treatments for NKTL patients and improve their survival rate.Additional file 1. Supplementary Methods and Figures S1–2. Figure S1. H3K4me3 landscapes in chidamide-resistant and chidamide-sensitive NKTL. Figure S2. JAK inhibitors overcome the resistance of SAHA/TSA in NKTL cells.Additional file 2. Tables S1-10. Table S1. List of primers used for ChIP-qPCR. Table S2. List of primers used for RT–qPCR. Table S3. Demographic and clinical characteristics of the 28 relapsed/refractory NKTL patients. Table S4. Patient characteristics at baseline. Table S5. List of GAIN and LOSS regions identified in both resistant and sensitive samples. Table S6. List of motifs in GAIN regions. Table S7. List of GAIN and LOSS regions overlapping with nearest genes identified in both resistant and sensitive samples. Table S8. List of differentially expressed genes overlapping with GAIN and LOSS regions in sensitive versus resistant samples. Table S9. Differentially expressed genes from JAK-STAT pathway in sensitive versus resistant samples. Table S10. Differentially expressed genes in HANK1 treated with DMSO, tofacitinib or ruxolitinib.


The B chromosome (B) is a dispensable component of the genome in many species. To evaluate the impact of Bs on the transcriptome of the standard A chromosomes (A), comparative RNA-seq analyses of rye and wheat anthers with and without additional rye Bs were conducted. In both species, 5–6% of the A-derived transcripts across the entire genomes were differentially expressed in the presence of  2Bs. The GO term enrichment analysis revealed that Bs influence A chromosome encoded processes like “gene silencing”; “DNA methylation or demethylation”; “chromatin silencing”; “negative regulation of gene expression, epigenetic”; “post-embryonic development”; and “chromosome organization.” 244 B chromosome responsive A-located genes in + 2B rye and + B wheat shared the same biological function. Positively correlated with the number of Bs, 939 and 1391 B-specific transcripts were identified in + 2B and + 4B wheat samples, respectively. 85% of B-transcripts in + 2B were also found in + 4B transcriptomes. 297 B-specific transcripts were identified in + 2B rye, and 27% were common to the B-derived transcripts identified in + B wheat. Bs encode mobile elements and housekeeping genes, but most B-transcripts were without detectable similarity to known genes. Some of these genes are involved in cell division-related functions like Nuf2 and might indicate their importance in maintaining Bs. The transcriptome analysis provides new insights into the complex interrelationship between standard A chromosomes and supernumerary B chromosomes.The online version contains supplementary material available at 10.1007/s10577-022-09704-6. The B chromosome (B) is one of the most interesting nonessential components of the genome in a wide range of taxa, from fungi to plants and animals. Although dependent on the species, Bs may vary in copy number, size, and structure; they share features that make them distinguishable from other types of chromosome polymorphisms like, for example, aneuploidy  tolerates up to eight Bs . Sequence alignment of ScAGO4B showed that compared to the A-originated transcript, the B-transcript had minor changes, and none of the point mutations caused a non-sense mutation. In vitro analysis of the A and B chromosome encoded ScAGO4 variants demonstrated that both proteins retain RNA slicer activity  with and without additional rye B chromosomes  at the IPK, Gatersleben (Germany). The B chromosome of the wheat-rye B addition line was initially introduced into a Nepalese strain of wheat from a spring rye variety from Transbaikal, Siberia  plants of the Japanese JNK strain , according to Klemme et al. (http://www.invitrogen.com).Rye  according to the manufacturer’s manual. RNA samples were treated with DNA-free DNase before cDNA synthesis following the on-column DNA removal protocol . The RNA concentration was determined using a Qubit device . RNA Integrity Number (RIN) was verified using a 2100 Bioanalyser . Library preparation (Illumina TruSeq RNA Sample Preparation Kit) and sequencing-by-synthesis using the Illumina HiSeq 2500 device  involved standard protocols from the manufacturer . The libraries were quantified by qPCR  genome using the HISAT2 Galaxy tool v2.0.3 with default settings  were aligned against S. cereale genome  using HISAT2 v2.1.0 with default settings. The generated by HISAT2 aligned mapping reads were proceeded further with the pseudo-alignment Kallisto (v0.44.0) genome using the HISAT2 Galaxy tool v2.0.3  were catenated into one dataset and further processed by CD-HIT (CD-HIT-EST) v4.6.8 (sequence identity threshold 0.90)  values were generated and “PtR” of the Trinity pipeline was used to examine the data quality based on Pearson’s correlation. The final FASTA file with contigs starting with 2B and 4B prefixes was used as a reference file in mapping of rye samples and identification of B-specific reads (see below). B-specific transcripts were filtered based on a minimal expression of 0.1 TPM in all replicates representing 2B and 4B conditions, respectively, and a maximal expression of 0.09 TPM in all replicates representing 0B condition. Additionally, only transcripts with prefixes 2B and 4B were considered. The principal component analysis was done in DESeq2  which did not align against the 2-transformed counts-per-million (CPM) values were generated, and “PtR” of the Trinity pipeline was used to examine the data quality based on Pearson’s correlation   was used to predict open reading frames (ORFs) in the B-specific transcriptome, which were annotated by Interproscan Galaxy v5.0.2 mkh. Additionally, sequences were BLASTed against the wheat genome IWGSC Ref.Seq.1.1 using E-values < 1.0E − 3 in at least 100 amino acids.Transdecoder v5.3.0  against the wheat genome IWGSC Ref.Seq.1.1 using a bit score at least 50 in at least 100 amino acids using the first best hit criterion. The annotated DETs were used in downstream GO term enrichment analysis.2(fold change) |≥ 0.5 and adjusted P < 0.05 were selected. Gene-specific P values were obtained from the hypergeometric test and were further corrected via Hochberg FDR option. A FDR < 0.05 cut-off has been set for detecting significantly enriched groups. For the GO term enrichment analysis, the separated analyses of upregulated and downregulated transcripts were performed. It was demonstrated that the analysis of all DE genes together could greatly reduce the power to identify significant pathways due to the imbalance between the upregulated and downregulated genes. In wheat and rye, biological processes (BP), molecular functions (MF), and cellular components (CC) were assessed using T. aestivum (“Chinese Spring”) as a background reference since the wheat is better annotated and described than rye. GO term enrichment analysis results were visualized with REVIGO  as well as the coordinates of (i) upregulated and downregulated DETs and (ii) all protein-coding features. Overlaps were identified and counted using the bedops suite (v2.4.19)  under the study accession number ID PRJEB46034, available at To study whether rye B chromosomes influence the transcriptome of rye and wheat standard A chromosomes, first, we identified 0B and + B containing plants by FISH using the repeat D1100  ≥ 1). Among highly DETs, 864 were activated in the presence of + 2B and 424 strongly decreased their expression , including 1366 upregulated and 854 downregulated transcripts  were computed. A total of 244 in rye identified DEGs are shared with the DEGs of the wheat host genome Fig. . A fractTo explore the regulatory pathways of DETs identified as a host response, a GO enrichment analysis was performed with rye and wheat datasets with wheat as a background reference. Separate studies of upregulated and downregulated transcripts were carried out, and different enriched GO terms were found. The dataset of upregulated transcripts in + 2B rye was represented with 578 annotated genes. Biological processes (BP) were described with 110 GO terms. Among them, “negative regulation of gene expression, epigenetic”; “chromatin silencing”; and “chromosome organization” were the most statistically significant. Other overrepresented GO terms included “methylation,” “cell cycle process,” “chromatin organization,” and “mitotic cytokinesis.” In the molecular function (MF) category, there were 18 overrepresented terms with the most statistically significant term “choline dehydrogenase activity.” The cellular component (CC) category was represented with 50 terms. Among them, there were “chromosome,” “chromatin,” “microtubule,” and “chromosome, centromeric region.”The dataset of downregulated rye A chromosome transcripts in the presence of 2B was represented with 363 annotated genes. The BP category was characterized with 89 terms; MF and CC categories included eight and 38 terms, respectively. The enriched GO terms of the BP category were related to metabolism, photosynthesis, and several kinds of responses. The most statistically significant terms of the MF were “catalytic activity” and “hydrolase activity.” The CC category included terms related to chloroplast and plastids  genome. To identify rye B-specific transcripts, RNA-seq data from wheat with and without Bs were compared. The de novo transcriptome assembly followed by filtering and clustering led to the final FASTA file with 8611 transcript sequences specific to + 2B and/or + 4B transcriptomes  confirmed biological replications  in the background of wheat was used as a reference to map B chromosome-specific reads in rye  were highly differentially expressed, while in wheat, the highest proportion of DETs included those with log2 FC < 1) B-responsive transcription in hexaploid wheat. (II) The rye B in the background of wheat could be considered as an alien chromosome, while the B of rye is part of the rye pangenome. During species evolution, the interaction between A and B chromosomes of rye likely coadapted. It is likely that Bs may bring about various epigenetic effects, including the differential regulation of A-localized transposable elements through mechanisms such as homology-dependent RNA interference pathways. Since the rye B likely originated from the rye A chromosome complement, it seems reasonable to suggest that the transcription alterations of A-located sequences are caused by homology-dependent mechanisms, as has been proposed for the remodelling of gene activities after chromosome segment duplications  the genome size and ploidy differ (diploid versus hexaploid) between both species. As suggested by Blavet et al. , the effLilium amabile plants, recent RNA-seq analysis from leaf tissues of 0B and + 1B chromosomes revealed that 5.1% of the total transcripts were differentially expressed (threshold log2FC >|2|)  and rye (297 transcripts in + 2B) identified in our study compared to 1569 transcripts in Ma et al. Supplemental Dataset 2 PCA plots based on the “PtR” of the Trinity pipeline. Pearson’s replicate correlation heatmap. (DOCX 88 KB)Supplemental Dataset 3 Correlation of four biological replicates of rye B specific transcripts derived from wheat +2B plants. Correlation of four biological replicates of rye B specific transcripts derived from wheat +4B plants. PCA based on four biological replicates of rye B specific transcripts derived from wheat +2B and wheat +4B plants. (DOCX 73 KB)Supplemental Dataset 4 GO term enrichment analysis of up-regulated and down-regulated DETs of rye in the presence of +2B chromosomes. T. aestivum is the background reference. (XLSX 29 KB)Supplemental Dataset 5 GO term enrichment analysis of up-regulated and down-regulated DETs of wheat in the presence of +2B chromosomes. T. aestivum is the background reference. (XLSX 121 KB)Supplemental Dataset 6 Common overrepresented GO terms identified in up-regulated and down-regulated DETs in rye and wheat in the presence of +2B chromosomes. T. aestivum is the background reference. (XLSX 30 KB)Supplemental Dataset 7 Rye B specific transcripts identified in wheat and rye and annotated based on the Interproscan and BLASTP against T. aestivum. (XLSX 2556 KB)Supplementary file8 (DOCX 36 KB)Supplementary file9 (PNG 31 KB)Supplementary file10 (PNG 38 KB)Supplementary file11 (PPTX 620 KB)Supplementary file12 (PPTX 71 KB)Supplemental Table 1 RNA sequencing and read mapping onto the T. aestivum reference genome. (XLSX 11 KB)Supplemental Table 2 Differentially expressed transcripts identified in rye in the presence of +2B chromosomes. (XLSX 9 KB)Supplemental Table 3 Annotated differentially expressed transcripts in rye in the presence of +2B chromosomes. (XLSX 182 KB)Supplemental Table 4 Differentially expressed transcripts identified in wheat in the presence of +2B chromosomes. (XLSX 10 KB)Supplemental Table 5 Annotated differentially expressed genes identified in wheat in the presence of +2B chromosomes based on the DESeq2. (XLSX 1270 KB)Below is the link to the electronic supplementary material.


This cohort study assesses trends in incidence of attention-deficit/hyperactivity disorder (ADHD) among children, adolescents, and adults in Japan from 2010 to 2019. Demographic and diagnostic information was collected from the NDB, which includes all electronic health insurance claims from April 2009 to March 2020. The NDB is described on the Ministry of Health, Labour and Welfare website4 and in our previous study.5This cohort study was approved by the ethics committee of Shinshu University School of Medicine. Informed consent was not required because anonymous data were used. This study followed the ICD-10 codes F90.0, F90.1, and F98.8) between fiscal years (FY) 2010 and 2019 were retrieved. Extracted information were sex and the year and age group at diagnosis. Incidence in each age group was calculated by dividing the number of ADHD diagnoses by the total population in the age group. Microsoft Excel 2016 was used for analysis. Details are given in the eMethods in the Data on individuals newly diagnosed with ADHD  were female and 97 986 (80.8%) were male. Of 381 753 individuals diagnosed at age 7 to 19 years, 91 891 (24.1%) were female and 289 862 (76.0%) were male. Of 335 234 individuals diagnosed after age 19 years, 160 239 (47.8%) were female and 174 995 (52.2%) were male.The Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition) (DSM-5) in 2013, which allows combined diagnosis of ADHD and autism spectrum disorder (ASD). Although the ICD-10 precludes a dual diagnosis of ADHD and ASD, the release of the DSM-5 may have prompted physicians to give an additional diagnosis of ADHD to individuals with ASD to prescribe ADHD medications.The incidence of newly diagnosed ADHD in Japan increased from 2010 to 2019. One reason may be the release of the 6 Thus, the sensitivity to detect ADHD in the adult population seems to have reached a standard in Japan.The increase was most prominent among adults between 2012 and 2017. This may be explained by increased sensitivity of diagnosis, especially of adult ADHD. The approval of the first medication in Japan in 2012 for adults with ADHD may have contributed to the expanding awareness of the disorder. The annual incidence of adult ADHD seems to have leveled after reaching a peak of 6 per 10 000 person-years in 2018. This figure is comparable to the adult ADHD incidence reported for Asian individuals in the US in 2016 (6.88 per 10 000 person-years).Findings of this study are limited in that the available demographic data were restricted to sex and age group. Nevertheless, nonetiological factors could explain at least part of the apparent increase in ADHD incidence. Further investigation is warranted to elucidate how etiological and nonetiological factors contribute to the changing incidence of ADHD in Japan.


To meet the high demand for white Guinea yam, there is a need to develop and release improved varieties to farmers. Unfortunately, low rate of adoption of most of the improved yam varieties by both producers and consumers was observed. Information regarding agronomic characteristics and food qualities of popular white Guinea yam landraces with high market value are not available to establish minimum standards to be considered by breeding programs. To fill this gap, surveys using rural appraisal tools were carried out in 20 villages and 16 markets throughout Benin. Data on the agronomic performance suggested that for an improved variety to be adopted by Beninese farmers it should have a minimum yield of 4.16 ± 0.15 kg per mound, and average number of marketable tubers of 1.23 ± 0.05, a mean tuber length of 36.41 ± 1.22 cm, and a minimum diameter of 25.44 ± 1.16 cm. The sensorial attributes for boiled and pounded tubers of this improved variety should have minimum score of 3.16 for texture, 0.75 for softness, 3.75 for elasticity, and 1.34 for colour during the sensory evaluation. The improved variety must also have a minimum average severity score of 1.1 for yam mosaic virus disease, 1.33 for anthracnose and 1 for nematodes. Landraces Amoula, Laboko, and Djilaadja should be considered as the standard for yield, sensory attributes, and tolerance to pest and diseases while landraces Danwari, Kodjewe, Mondji, and Gnidou should be characterized as possessing good flowering and fruit setting capacities for breeding programs. Dioscorea spp.) is a climbing crop species that produces edible underground tubers, which contribute to the food security and poverty alleviation in many developing countries [Dioscorea rotundata Poir.) remains the most produced and preferred [Yam  identify the popular white Guinea yam landraces with high market value in the study area; (ii) assess the on-farm agronomic performance of these elite yam landraces across farmers’ fields and production zones; (iii) evaluate the sensory quality of the boiled and pounded yam of these elite yam landraces; (iv) establish the basic standards for yam breeding and selection programs in the Republic of Benin.The study was carried out in the intensive yam-growing areas of four regions  located in the central (latitude 7° 45’ and 8° 40’ North and longitude 2° 20’ and 2° 35’ East) and northern (latitude 9° 00 ’and 12° 30’ North and longitude 1° and 30° 40’ East) regions of Benin. In Benin, yam is not produced in the far north because of drought stress and in the far south due to ignorance of cultural practices [Twenty villages were selected based on the yam production statistics collected from the agriculture and rural development office of each department. Sixteen yam local markets close to the chosen villages were also surveyed. Surveyed villages covered sixteen ethnic groups. These are Idaatcha, Tchabè, Nago, Mahi and Yoruba in central Benin; Yom, Ani, Lokpa, Natimba, Kotocoli, Gnindé, Ditamari, Wama and Berba in the north west; Bariba and Boko in the north east.Surveys were conducted using participatory research appraisal tools and techniques, such as group discussions and yam field observations . In eachDuring focus group discussions, information on the agronomic, technological, and culinary performance of each yam landrace listed in each village were recorded. Fourteen variables , resistance to anthracnose, susceptibility to weeds, stay-green characteristic, relative productivity, storage aptitude of the fresh tuber, quality of chips, storage aptitude of the chips, quality of the pounded yam, and quality of the boiled yam) were used to assess the performance of each landrace. According to Loko et al. , yam lanSixteen closest rural markets to the surveyed villages were selected for the estimation of the market value of the identified popular yam landraces. In each market, 5 to 12 yam wholesalers and retailers of yam were randomly selected for the survey. A total of 128 traders were surveyed in the study area. In the markets, yams were sold by retailers in small lots of 3 to 5 tubers per landrace, while the wholesalers sold them in lots of 50 to 100 tubers. It was observed in some markets such as Glazoué market that some traders were selling yam in kilograms using baskets and an adapted weighing scale. The market value of tubers of each yam landrace was estimated regarding the price per kg, the number of sellers and available quantity (in tons). For each yam landrace, five representative lots of tubers were selected and weighed to provide the peak price per kilogram for both wholesale and retail sales. The average price per kg of tubers of all popular yam landraces found in the markets was used as a standard to identify those considered to have high prices. A popular white Guinea yam landrace was considered as sold by many sellers when, Sc ≥ Ns/2 . A popular white Guinea yam landrace sold by many sellers with high price and available in large quantities (≥ 1 ton) were considered as having high market value.Scutellonema bradys Steiner and LeHew) damage on harvested yam tubers were assessed using the standard descriptor of Asfaw [In each surveyed village, five yam producers were chosen for field visits with the help of the village chief and/or heads of farmer associations. These farmers were selected on the basis of similarity in their farming practices , and in order to include all popular yam landraces with high market values identified at the village level. Three visits were made in each field at different yam development stages. During the first visit (yam germination stage), five plants of each identified popular and high market value landraces were randomly selected and tagged with white tape for data collection. During the second visit, at the vegetative yam development stage (10 to 12 weeks after planting), the vigour, flowering intensity and fruiting intensity were recorded according to the yam descriptors . In addiof Asfaw .Fresh matured tubers of each popular white Guinea yam landrace with high market value were collected from the surveyed villages. The yam tubers were washed with clean water to remove adhering soil and other undesirable materials in the laboratory of the Biotechnology, Genetic Resources and Animal and Plant Breeding (BIORAVE). The yam samples were sorted, hand-peeled using kitchen knives and then cut into slices of about 10 cm each. About 2 kg of yam tuber slices from each landrace were cooked (temperature of 100°C) with 200 ml of water. Cooking times were recorded as they varied from one yam landrace to the other. Qasa yam pounding machine (Model QYP-6000) was used for the pounding of the boiled yam samples for 3 min.Four randomly selected boiled and pounded yam samples from each landrace were served to consumers to determine consumer acceptability and assess sensory characteristics. The yam dishes prepared from each landrace was coded. To determine the consumer acceptability of the dishes prepared from each landrace, 60 untrained consumer panellists from the students and staff members of the National High School of Applied Biosciences and Biotechnologies (ENSBBA) were selected as panellist based on their willingness to participate. A 5-level hedonic scale ranging from very unpleasant (1) to very pleasant (5) were used for the evaluation . SubsequThe research protocol was approved by the ethic committee of the National University of Sciences, Technologies, Engineering and Mathematics (UNSTIM). Interviews were carried out in accordance with the guidelines of the Declaration of Helsinki. Written informed consent was obtained from all participants prior to the interviews.hclust function was used to construct a dendrogram. Analysis of variance (ANOVA) was performed using the IBM SPSS software version 23. Before the ANOVA test, the data normality was tested using Levene’s test. To achieve normality and homogeneity of variances, yield and yield component data were log-transformed (log (x + 1)). Significant differences between means were separated using the Student Newman Keuls test (p < 0.05). Principal Component Analysis (PCA) was performed to describe the relationship between the popular yam landraces and their yield and yield components. Pearson correlation analysis was used to measure the degree of association between the total number of tubers harvested, yield in kg per mound, length, diameter and weight of marketable tubers per mound.Descriptive  and inferential statistics were used to analyse the data collected. The Hierarchical Cluster Analysis (HCA) was used to establish the relationship between the popular yam landraces with high market value and their agronomic and culinary characteristics as perceived by the surveyed farmers. The data analysis was performed using the R software  with theTo classify and characterize the popular yam landraces with high market value, the data collected including agro-morphological and sensorial attributes  were conDI) and severity index (MS) were calculated using the following formula [Disease incidence , number of harvested marketable tubers per mound (CV = 52.4%), marketable tuber weight per mound (CV = 38.7%), marketable tuber length (CV = 31.4%), and width (CV = 30.3%) were observed among the popular white Guinea yam landraces . The yieResults of the principal component analysis showed that the first axis explained 85.24% of the total variability. The length (LtubL), diameter (DtubL), and weight (PtubL) of tubers and yields were found to be positively correlated with the first axis, while the number of harvested tubers per mound (Ntub) was negatively correlated with the first axis . The priTwenty-nine elite yam landraces with high market values had low flowering capacity. Of these yam landraces, 24 yam landraces had flowering capacity but no fruit production, whereas five landraces were found to be completely sterile . Two popS. bradys infestation.Under on-farm conditions, several of the 44 popular yam landraces were found to be susceptible to anthracnose, virus and nematodes and had relatively high incidence of infections, ranging from 10 to 92%, 10 to 100% and 0 to 66%, respectively . Very feMajority (81.8%) of popular yam landraces with high market values were preferred in both boiled and pounded forms. Only 6.8% and 11.4% landraces were preferred in the boiled and pounded forms, respectively. Ahimon, Baniouré, Déba, Douroubayésirou, and Kratchi landraces were most preferred in the pounded form while, Adoro, Ossoukpana, and Singou landraces were the most preferred in the boiled form. The hedonic tests of boiled and pounded yam of the 44 popular white Guinean landraces with high market value showed that more than half of them were classified as pleasant (95.6%), and 4.5% as very pleasant by the tasters. Boiled and pounded form of Laboko and Djiladja landraces were considered as very pleasant by all the taste panellists. A significant variability in the scores obtained for pounded tubers of the different yam landraces was observed in terms of colour (CV = 29.8%), sweetness (CV = 33.1%), hardness (CV = 34.6%), aroma (CV = 168.1%), presence of lumps (CV = 199.1%) and elasticity (CV = 30.6%). The sweetness of Laboko and Tchee landraces were most appreciated by the panellists, while Tamsam and Baniouré landraces were the least appreciated for this attribute. Okogan, Tamsam, and Gnarambo landraces were considered by panellists as producing pounded yam with good texture attributes. In terms of the elasticity of pounded yam, Laboko had the highest score by the panellists . Deba, DThe multiple factor analysis showed that the first factor (Dim 1) explained 13.3% of the total variation among the yam landraces, the second factor (Dim 2) 12.4% while the first two axis explained 25.7% of the total variation among the yam landraces . The perOur results showed that, subject to local yam landraces only 44 of the 116 popular white Guinea yam landraces recorded in the study area had a high market value. Yam production in the study area is mainly intended for family consumption . ConsequIn general, farmers perceived most of the identified popular yam landraces with high market value as having high agronomic and culinary performances. Indeed, according to Zannou et al. , the priOur results showed that, the yield of the popular yam landraces with high market value varied significantly from one mound to another. This variation of yam yield could be due to an important interaction between the environment and the expression of their phenotypic potential . In addiMost of popular yam landraces with high market value recorded in this study showed poor flowering and fructification capacities. Indeed, it is known that natural pollination of white Guinea yam occurs very rarely in the field , 44, andOur results have revealed that the identified popular yam landraces with high market value in the Republic of Benin offer enormous potential for yam breeding programs. In fact, the recorded yam landraces that showed tolerance to diseases and nematodes, and had good agronomic and culinary characteristics should be prioritized for demonstration and use in yam breeding programs , 46. LabOur results suggested that a minimum yield of 4.16 ± 0.15 kg/mound, and tubers of minimum length and width of 36.41 ± 1.22 cm, and 25.44 ± 1.16 cm respectively must be the minimum standards that an improved variety of yam must have to be accepted by Beninese farmers. Furthermore, in addition to resistance to various biotic and abiotic stresses the improved variety should reveal minimum scores of 3.16 for texture, 0.75 for softness, 3.75 for elasticity, and 1.34 for colour preference during sensory evaluation to be acceptable by consumers. In the framework of a varietal improvement program for early maturity yam landraces, the MFA showed that landraces in cluster 1 would be good parents while the white Guinea yam landraces in cluster 2 would be ideal parents for a late-maturing yam for a breeding program.Our study identified 44 popular white Guinea yam landraces with high market value and agronomic and culinary performance that could be very useful for market-oriented breeding programs. Among them, Danwari, Kodjewe, Mondji, and Gnidou landraces should be integrated in the national breeding programs because of their high flowering intensity, and fruit setting capacity. Amoula, Laboko, and Djilaadja landraces should be considered the standard landraces for high yield, good sensory attributes, and resistance to diseases and nematodes, respectively. This study allowed to identify basic breeding standards, which used by breeders could increase the acceptability of improved white Guinea yam varieties by farmers and consumers in the Republic of Benin.S1 Table(DOCX)Click here for additional data file.


Attention Deficit/Hyperactivity disorder (ADHD) is one of the most common disorders in school-age children. In addition to learning difficulties associated with the disorder’s core symptoms of inattention and hyperactivity, children with ADHD display substantial social impairments. Methylphenidate (MPH) in formulations such as Ritalin or Concerta mitigates inattention and hyperactivity, but the effects of the therapy on social behavior in children with ADHD are not clear. This review aims to determine the effectiveness of short term (up to 6 months) MPH treatment on three domains of social skills in children aged 6–14 with ADHD: (i) Recognition of nonverbal emotional expressions, which are a marker of inherent (unlearned) social understanding, (ii) theory of mind (ToM) components that relate to learned cognition and social communication, and (iii) social competence in everyday environments. 15 relevant studies were identified based on inclusion/exclusion criteria. The results show mixed effects: the overall social performance as evaluated by parents, teachers or peers, and some components of ToM, were found to improve following a weeks-long course of MPH treatment. However, the effects of the medication are less clear when evaluating momentary/nonverbal social responses such as reactions to emotional facial expressions. While the findings of this review indicate that an MPH medication regime of order weeks to months could improve, to a degree, social impairment in children with ADHD, more studies are required to identify the medications’ mechanism and confirm such a conclusion. Attention-Deficit/Hyperactivity disorder (ADHD) affects an estimated 2–12% of school-age children worldwide . IncreaThe disorder is defined by a range of inattention and/or hyperactivity–impulsivity symptoms , 10 that© or Concerta© has been found to be highly effective in the treatment of ADHD symptoms, alone or in combination with behavioral therapy,  in formulations such as Ritalinexample, , 44). Asexample, –49. MPH example, –52).Appropriate social functioning is a complex decision-making process based on the understanding of emotional and motivational states in others, the application of prior learning of social norms to the choice of behavior, and the ability to act appropriately . As a reThe aim of this review is to examine the effects of MPH on specific components of social performance in children 6–14 years old with ADHD. We chose this age range based on several considerations. First, diagnosis of preschoolers with ADHD is complicated due to other aspects of their development . As a rOur cutoff at the older age range of our sample is 11–13, namely, pre-adolescence . ADHD chRecognition of emotional inputs, as measured through accurate identification of emotions: communication of emotions is a core, essential step in social functioning. Individuals that are impaired in their ability to recognize emotions in others cannot perform socially in a competent manner , 23, 30.‘Theory of mind’ (ToM): ToM is a components of social cognition that relates to the skills required to manage social communication and relationships . In typiSocial competence : social competence is generally defined as the effective functioning of individuals within social context, and may be divided into three aspects: attainment of society-defined accomplishments, global  judgment of social competences, and evaluation of peer acceptance . AssessmThree domains of social skills are examined, as representatives of the different steps in the execution of social performance:Better understanding of the effects of MPH on these three domains will enable answering questions important for the treatment of children with ADHD, such as: does medication with MPH improve social deficits in children with ADHD? If it does, are the effects short term and ‘in the moment’, or long term? Answers might help design appropriate medication regimes to address social deficits in children with ADHD.We follow a framework adapted from Whittemore and Knafl  for the The search included papers published between 1980 and 2021 in APA PsycNet, MEDLINE, PubMed, ISI Web of Science, Scopus.- non-reviewed publications , papers published in languages other than English, or review papers.- studies in which the average age of the participants was not in the target range of 6–14 years.- case studies .- studies where the participants were not consistently diagnosed with ADHD based on DSM III , DSM IV - studies where a substantial fraction (40% or more) of the ADHD group had additional psychological disorders, namely, comorbidities, such as oppositional defiant disorder (ODD), conduct disorder (CD), or learning disability (LD).- studies where the ADHD group was also medicated with drugs other than MPH.- studies whose focus was on comparing different formulations or dosages of MPH.- studies where the effects of medication were evaluated after more than 12 months of a medication regime: Social cognition in children changes relatively rapidly, so that such long-term studies may include effects not directly related to the medication.Studies were eligible for the review if they were peer reviewed and published in the English language. Inclusion criteria were based on the following Boolean descriptors: (ADHD OR “attention deficit disorder” OR “attention deficit hyperactivity disorder” OR “attention-deficit/hyperactivity disorder”) AND  AND (methylphenidate OR MPH or stimulant) AND (children OR youth). Exclusion criteria included:The abstracts were assessed by the authors to evaluate compliance with the inclusion/exclusion criteria. Papers that complied with the criteria were then read in total. The authors extracted the relevant data including testing methodology, tested traits/behaviors, and outcomes.Database Boolean search results yielded 626 papers. These were examined individually by the authors to exclude those that did not comply with the exclusion criteria listed above. The process is described in Fig. Table The Diagnostic Analysis of Nonverbal Accuracy (DANVA) was used by Hall et al.  to examiKobayashi et al.  investigIn a functional Magnetic Resonance Imaging (fMRI) study, Bottelier et al.  examinedA somewhat different methodology was used by Williams et al.  and GumuGumustas et al.  found thThe ‘reading the mind in the eyes’ test (RMET) examines ToM by using photos of the eye region, where participant are asked to identify the emotion portrayed. Unlike the facial expression pictures  where emotions are clearly defined, RMET illustrates complex mental states such as ‘‘embarrassed’’ and require application of ToM concepts of inferences.Golubchik et al.  tested tDemirci et al.  tested tOther aspects of ToM were examined by Maoz et al. who used a ‘faux pas’ (FPR) , 107 tesMore recently, Levi-Shachar et al.  examinedA related concept to ToM is empathy, which is defined as an affective response based on an assessment of another person’s emotional state. This definition encompasses the cognitive awareness of the inner state of another person—that is, the emotions, thoughts, perspectives, and intentions of the other . GumustaA number of studies followed the effect of MPH on measures of overall social performance in children with ADHD, using self, peer, parental or teacher evaluations.Shang et al.  examinedGolubchik et al. assessed the effect of a 12 week long MPH treatment regime on the Emotional Quotient (EQ)  and on sSomewhat similar findings after a 12 week long MPH regimen applied to mostly drug-naïve children were observed by Kim et al. . ResultsLonger-term effects of MPH on children with ADHD were studied by Abikoff et al. , 96, whoOverall social skills can also be tested in laboratory settings. Pelham et al.  examinedADHD has been linked to social impairment in children , 27–30. Core emotional facial expressions such as happy or angry are a universal method of emotion communication that is largely independent of social learning –88. CommThe results of studies examining the effect of MPH on emotion recognition in children with ADHD are somewhat contradictory. Both Gumustas et al.  and HallThe answer to this question is partially given in studies of brain activity: Bottelier et al.  used fMRThe next type of studies examined aspects of ToM. Meta-analysis shows that children with ADHD have impaired ToM when compared to their TD counterparts . As noteRegarding the short-term effect of MPH on ToM, Golubchik et al.  did not Most evaluations of how MPH affects global aspects of social performance in children with ADHD focused on long-term medication regimes. Whether through self, parental or teacher assessments, it is agreed that medication with MPH for a period of 8 weeks or more improves social skills in children with ADHD , 104. InSocial performance in children is a multi-facet and complex process. For example, Dodge’s Social Information Processing (SIP) proposes six steps  that inc


Attention‐deficit/hyperactive disorder (ADHD) has substantial heterogeneity in clinical presentation. A potentially important clue may be variation in brain microstructure. Using fractional anisotropy (FA), previous studies have produced equivocal results in relation to ADHD. This may be due to insufficient consideration of possible sex differences and ADHD's multi‐componential nature.n = 234; 32% female youth) and non‐ADHD , case‐control cohort (ages 7–12). Sex‐specific effects were tested.Using whole‐brain analyses, we investigated the association between FA and both ADHD diagnosis and ADHD symptom domains in a well‐characterized, ADHD , after controlling for overlap with hyperactivity‐impulsivity. Hyperactivity‐impulsivity symptoms were associated with higher FA in the SLF, and lower FA in the superior cerebellar peduncles, after control for overlap with inattention. Meanwhile, both categorical and dimensional analyses revealed ADHD‐by‐sex interactions (voxel‐wise p < .01). Girls with ADHD had higher FA, but boys with ADHD had lower FA (or no effect), compared to their same‐sex peers, in the bilateral anterior corona radiata. Further, higher ADHD symptom severity was associated with higher FA in girls, but lower FA in boys, in the anterior and posterior corona radiata and cerebral peduncles.No ADHD group differences were found using categorical assessment of ADHD without consideration of moderators. However, dimensional analyses found total symptoms were associated with higher FA in the superior corona radiata. Further, ADHD symptom domains appear to be differentially related to white matter microstructure, highlighting the multi‐componential nature of the disorder. Further, sex differences will be crucial to consider in future studies characterizing ADHD‐related differences in white matter microstructure. Sex differences in ADHD have not been adequately considered, particularly for studies of fractional anisotropy (FA), a measure of white matter microstructural development. Results here demonstrate that ADHD symptom domains are differentially related to FA, highlighting the multi‐componential nature of the disorder. Males and females showed opposing patterns of FA, and studies of ADHD and white matter microstructure should consider symptom heterogeneity and sex to develop neurobiologically plausible models of ADHD brain development. Previous investigations into the relationship between attention‐deficit/hyperactive disorder (ADHD) and white matter microstructure have produced equivocal findings due to possible sex‐specific effects and ADHD's multi‐componential nature.Exploratory whole‐brain analyses in one of the largest pre‐adolescent case‐control cohorts to‐date confirmed sex‐specific effects of ADHD on white matter microstructure, particularly in the prefrontal cortex. Boys with ADHD showed consistently lower fractional anisotropy (FA) and girls with ADHD showed consistently higher FA than their typically‐developing peers.Dimensional analyses of separate symptom domains reveal unique, and sometimes opposing, associations in white matter microstructure.Future studies of white matter microstructure in ADHD are encouraged to include examinations of symptom domains and sex differences.Attention‐deficit/hyperactive disorder (ADHD) is one of the most common and costly neurodevelopmental psychiatric disorders , estimated using diffusion tensor imaging (DTI). The most common white matter microstructural metric, FA has historically been thought to reflect microstructural components of white matter, such as fiber density, axonal diameter, and myelination , internal capsule, forceps minor and cerebellum, regions that may subserve fronto‐striatal‐cerebellar neurocircuitry and relate to cognitive and motor functions in ADHD  and in severity of symptoms (diagnostic criteria are met with 6–18 symptoms in childhood). Studies using total ADHD symptom severity composites have also identified both higher   categorical ADHD diagnosis will be associated with widespread n = 618) ages 7–12 were recruited from the local community. After screening for sufficient data, motion contamination during imaging, and restriction to tight criteria for ADHD and non‐ADHD cases (see Supporting Information n = 36) was reported previously , substance use disorder, prior diagnosis of autism spectrum disorder or psychosis, current use of non‐stimulant psychoactive medications, estimated IQ < 70, or MRI contraindications. Participants prescribed stimulant medication were asked to wash‐out for at least five medication half‐lives (24–48 h), prior to scanning.ADHD diagnoses were established in a multi‐step process using standardized, nationally‐normed rating scales from parents and teachers, parent semi‐structured clinical interviews, child intellectual testing, and clinical observations  with gradient‐encoding pulses in 30 directions  and six images collected with a b‐value of 0 s/mm2. Participants received either two  or three  DWI runs. A diffusion field map was also acquired .Participants were scanned on a 3T Siemens Magnetom Tim Trio with a 12‐channel head coil. Diffusion‐weighted imaging (DWI) scans were collected using a whole‐brain, high‐angular resolution, echo‐planer imaging (EPI) sequence  practices, as described previously , both symptom domains were modeled simultaneously in dimensional analyses to identify regions unique to each domain. Main effects and interactions from each model were identified voxel‐wise (p < .01) and FWE‐corrected  FA ∼ diagnosis + sex + age(2)FA ∼ diagnosis + sex + diagnosis × sex + age(3)FA ∼ total ADHD symptoms + sex + age(4)FA ∼ total ADHD symptoms + sex + total ADHD symptoms × sex + age(5)FA ∼ inattention symptoms + hyperactivity‐impulsivity symptoms + sex + age(6)FA ∼ inattention symptoms + hyperactivity‐impulsivity symptoms + sex + inattention symptoms × sex + hyperactivity‐impulsivity symptoms × sex + ageVoxel‐wise analyses were carried out using the Analysis of Functional Neuroimages' 3dMVM to examine categorical (Hypothesis 1) and dimensional (Hypothesis 2) effects of ADHD on FA, as well as differences by sex (Hypothesis 3), controlling for age. Though inattention and hyperactivity‐impulsivity were highly correlated , inattention (p = .96) and hyperactivity‐impulsivity (p = .98) ADHD symptoms severity scores did not differ by sex, after controlling for ADHD status. The group with ADHD also had more associated psychiatric conditions and lower estimated IQ. IQ was not included as a covariate in DTI analyses because covarying features that reflect population‐level differences is not generally appropriate  and girls with ADHD had higher FA  compared to controls   and higher FA in girls   Figure . There w) Figure .higher FA in the body and splenium of the corpus callosum (CC) and left ansa lenticularis (AL), and lower FA in the right SLF   but not boys (p > .05)  Figure . There w) Figure .ps < .01) in post‐hoc models that included covariates for lifetime mood, anxiety, oppositional defiant disorders, current and lifetime stimulant medication prescription, signal‐to‐noise ratio, number of useable diffusion volumes, and familial relatedness in a random‐effects structure  the largest case‐control sample to date. Further, it adds new information regarding symptom‐dimension specific associations and sex‐specific effects, which help to clarify prior contradictions in the literature.higher FA, while boys with ADHD showed lower FA (or no differences) compared to their peers in the bilateral anterior and left PCR and right CP. The neuromechanistic phenomena behind this sex‐specific effect is yet unclear; as FA may be related to alterations in fiber density, myelination, glial cells swelling, neuronal excitotoxicity, and neuronal branching at the gray‐white matter boundary  limits our ability to carry out more advanced analytic techniques. However, tensor metrics are fairly limited in their interpretability and future studies should utilizing more advanced multi‐shell sequences, will be better equipped to handle issues of crossing fibers  was an epidemiological sample that included a small proportion of children with ADHD (Albaugh et al., The issue of sample size also remains important. The size of ADHD groups in all but one (van Ewijk et al., Overall, two major conclusions emerge. First, while categorical findings were not present, more sensitive dimensional analyses using separate symptom domains showed unique FA correlates and, at times, opposite patterns of correlation, highlighting the multi‐componential nature of the disorder. Second, results strongly support sex‐specific effects in which girls with ADHD had higher FA, but boys with ADHD had lower FA, than typically‐developing peers of the same sex, particularly in the prefrontal cortex. These results underscore the importance of examining symptom domains and sex differences in studies of white matter in ADHD.Scott A. Jones: Conceptualization; Data curation; Formal analysis; Methodology; Visualization; Writing – original draft; Writing – review & editing. Bonnie J. Nagel: Conceptualization; Data curation; Funding acquisition; Methodology; Project administration; Writing – review & editing. Joel T. Nigg: Conceptualization; Data curation; Funding acquisition; Methodology; Project administration; Supervision; Writing – review & editing. Sarah L. Karalunas: Conceptualization; Data curation; Methodology; Project administration; Supervision; Writing – original draft; Writing – review & editing.Advances Editorial Advisory Board. The remaining authors have declared that they have no competing or potential conflicts of interest.Joel T. Nigg serves on the JCPP All study procedures were approved by the Oregon Health & Science University Institutional Review Board. During recruitment, parents/legal guardians and children provided written consent and assent, respectively.Supporting Information S1Click here for additional data file.


The novel brain-penetrant peroxisome proliferator-activated receptor gamma agonist leriglitazone, previously validated for other rare neurodegenerative diseases, is a small molecule that acts as a regulator of mitochondrial function and exerts neuroprotective, anti-oxidative and anti-inflammatory effects. Herein, we tested whether leriglitazone can be effective in ameliorating the mitochondrial defects that characterize an hiPS-derived model of Pantothenate kinase-2 associated Neurodegeneration (PKAN). PKAN is caused by a genetic alteration in the mitochondrial enzyme pantothenate kinase-2, whose function is to catalyze the first reaction of the CoA biosynthetic pathway, and for which no effective cure is available. The PKAN hiPS-derived astrocytes are characterized by mitochondrial dysfunction, cytosolic iron deposition, oxidative stress and neurotoxicity. We monitored the effect of leriglitazone in comparison with CoA on hiPS-derived astrocytes from three healthy subjects and three PKAN patients. The treatment with leriglitazone did not affect the differentiation of the neuronal precursor cells into astrocytes, and it improved the viability of PKAN cells and their respiratory activity, while diminishing the iron accumulation similarly or even better than CoA. The data suggest that leriglitazone is well tolerated in this cellular model and could be considered a beneficial therapeutic approach in the treatment of PKAN. Neurodegeneration associated with pantothenate kinase 2 deficiency  is an autosomal recessive disease which belongs to the group of disorders named Neurodegeneration with Brain Iron Accumulation (NBIA) ethoxy] phenyl]methyl]-1,3-thiazolidine-2,4-dione hydrochloride) is the hydrochloride salt of the active metabolite M4 (M-IV) of pioglitazone  in comparison with CoA. We noted that the treatment with leriglitazone did not affect the differentiation of neuronal precursors into astrocytes. Most importantly, it improved the viability of PKAN cells, increased the mitochondrial respiration, and, at the same time, significantly reduced iron accumulation.The hiPS clones for controls and PKAN patients were previously generated and fully characterized . SubsequAstrocytes were treated with leriglitazone  in the range from 3 nM to 3 μM for three days after they reached maturation to determine the optimal dose of the drug. In all the other experiments, astrocytes were differentiated in the presence or absence of 100 nM of leriglitazone or 25 μM of CoA for more than 45 days with fresh medium and drugs changed every 2–3 days.Cells grown on coverslips were fixed in 4% paraformaldehyde and processed for immunofluorescence as described in . Immunof4 astrocytes/well in 96-well plates were grown in the appropriate medium at 37 °C for 18 h and then incubated with 10 μL of MTT solution  for 2 h at 37 °C. The color absorbance was read at 570 nm on an iMark Microplate Reader .A total of 2 × 10The oxygen-consumption rate (OCR) of astrocytes was measured with an XF96 Extracellular Flux Analyzer  as previously described (Orellana).Astrocytes were differentiated for more than 70 days in the presence or absence of 100 nM leriglitazone or 25 μM CoA and then stained for iron content with Perls reaction by incubation for one hour in 1% potassium ferrocyanide, 1% hydrochloric acid in distilled water. Cells were counterstained with nuclear fast red (Sigma). Images were taken on Zeiss AxioImager M2m equipped with AxioCam MRc5 using 40× objective.® software  was used togenerate the graphs and to perform statistical analysis. The results are expressed as mean ± SD or ±SEM. The statistical analysis performed was indicated in the corresponding figure legend. Significant p-values: * p < 0.05; ** p <0.01; *** p < 0.001; **** p < 0.0001 were considered.GraphPad PRISMStarting from the previously obtained hiPS clones , we diffThe differentiation method, previously set up in  and descTo determine the optimal dose for leriglitazone treatment, we added to the growth medium increasing concentrations of the drug ranging from 3 nM to 3 μM and maintained the mature d-astrocytes culture of one control and one patient for 3 days. At the end, we analyzed the d-astrocytes’ viability and respiration, two features affected in PKAN neuronal models ,24, to vBased on these results, we decided to use a concentration of 100 nM of MIN-102 and extend the time of treatment to assess the effect of longer treatment with the drug and directly compare the results with the already proven beneficial effect of 25 μM CoA addition ,26,32, fMaintaining these identified conditions of treatment, we investigated the ability of leriglitazone in the amelioration of the pathological phenotypes. The estimated cell vitality revealed that all the untreated PKAN d-astrocytes were less viable than controls, as expected A. The adRespiratory parameters of PKAN d-astrocytes were also examined by measuring the oxygen consumption rate (OCR) and comparing it with the controls. The results showed that all patient’s cells had significantly lower values of respiratory parameters than controls, showing impaired mitochondrial functionality . Herein,The peculiar feature of this PKAN astrocyte model is the presence of intracellular iron deposits  that areTreatment with CoA was already proven effective in lowering iron aggregation , and herLeriglitazone was recently evaluated in in vitro and in vivo preclinical models of neurodegenerative disorders and clinically tested in healthy volunteers in a phase 1 study ,30 and cHerein, we checked whether this could be confirmed in our PKAN models, which are characterized by mitochondrial dysfunction. Indeed, our data indicated that the treatment of PKAN d-astrocytes during their differentiation with leriglitazone does not affect their proliferation. This could be probably explained by the recovery of the mitochondrial functionality as demonstrated by the rescue of respiratory activity. Interestingly, in two patients, the respiratory activity resulted in higher improvement after leriglitazone treatment compared to CoA treatment, suggesting a similar efficacy of the leriglitazone in rescuing mitochondrial functionality. The positive effect of the leriglitazone treatment cannot be ascribed to the increased amount of the PANK2 enzyme in these cells as seen by the absence of PANK2 peptide in untreated or treated cells. More intriguingly, the phenotype of iron deposition appeared strongly reduced after leriglitazone treatment with a slightly greater efficacy of leriglitazone as compared to CoA in reducing the percentage of cells containing iron deposits. This results is aligned with the data reported recently in FRDA patients treated with leriglitazone in the FRAMES clinical trial, showing that iron accumulation in the dentate nucleus, as assessed by QSM, was greater in the placebo arm . We cannCoA is a key metabolic cofactor for many biochemical functions; among them, it is a vehicle of the acyl group, and it is able to activate the carbonyl groups in many reactions taking place within cells, including the tricarboxylic acid cycle and metabolism of fatty acids . FurtherFurther studies will be needed to define this point; however, these results are interesting from a therapeutic point of view. Until now, PKAN patients have been treated with drugs that alleviate their symptoms. These therapies are based on the usage of dopaminergic drugs, anticholinergics, tetrabenazine, baclofen and botulinum toxin. Other current therapies include the administration of iron chelating agents, such as deferiprone. In recent clinical trials, deferiprone’s long-term effect has been confirmed by the reduction of iron accumulation in the globus pallidus and a stabilization of motor symptoms ,41,42. OC.V., L.R.-P., M.M. and P.P. are employees of Minoryx and have stock options and/or patents with Minoryx.


Methylphenidate (MPH) is the recommended first-line treatment for attention-deficit/hyperactivity disorder (ADHD). While MPH’s mechanism of action as a dopamine and noradrenaline transporter blocker is well known, how this translates to ADHD-related symptom mitigation is still unclear. As functional connectivity is reliably altered in ADHD, with recent literature indicating dysfunctional connectivity dynamics as well, one possible mechanism is through altering brain network dynamics. In a double-blind, placebo-controlled MPH crossover trial, 19 medication-naïve children with ADHD underwent two functional MRI scanning sessions (one on MPH and one on placebo) that included a resting state scan and two inhibitory control tasks; 27 typically developing (TD) children completed the same protocol without medication. Network control theory, which quantifies how brain activity reacts to system inputs based on underlying connectivity, was used to assess differences in average and modal functional controllability during rest and both tasks between TD children and children with ADHD (on and off MPH) and between children with ADHD on and off MPH. Children with ADHD on placebo exhibited higher average controllability and lower modal controllability of attention, reward, and somatomotor networks than TD children. Children with ADHD on MPH were statistically indistinguishable from TD children on almost all controllability metrics. These findings suggest that MPH may stabilize functional network dynamics in children with ADHD, both reducing reactivity of brain organization and making it easier to achieve brain states necessary for cognitively demanding tasks. Neurobiologically, it has been proposed that attention-deficit/hyperactivity disorder (ADHD) emerges from disruptions to cognitive control and motivation systems , 2, bothNetwork control theory (NCT) provides such an approach. NCT is a mathematical framework that provides tools to connect the topology of a network with the underlying dynamics of the complex system that the network describes , 22. ConThus, in a preliminary study, we used NCT to assess how the controllability of the functional connectome differs between medication-naïve children with ADHD and TD children across cognitive contexts, and how those differences change after MPH administration in children with ADHD. This study seeks to extend prior literature using static FC approaches in a single context  to assess differences in children with ADHD and TD children, as well as the effects of MPH, on the functional dynamics of the whole brain across three cognitive contexts (rest and two response inhibition tasks).Recent reviews suggest that functional connectivity within and between the default mode, fronto-parietal, salience, and attention networks is altered in children with ADHD, however the direction and magnitude of these alterations are inconsistent between studies , 26. A rn = 12; see MRI data acquisition and processing section below for details), (2) inability to swallow the MPH/placebo pills (n = 1), and (3) diagnosis of autism spectrum disorder (n = 1). Five TD children were excluded for insufficient scan quality. Therefore, 23 medication-naïve children with ADHD  and 27 TD children  were included in these analyses. Children with ADHD and TD children included in analyses were not statistically different from the overall sample with regard to age, sex, race, or ADHD symptom severity. Sample characteristics are provided in Table Thirty-seven medication-naïve children with ADHD  and 32 TD children  between the ages 8 and 12 years participated in this study. Children with ADHD participated in a randomized, double-blind, placebo-controlled crossover trial with MPH, while TD children completed two identical sessions without any drug or placebo administration. Of these participants, a total of 14 children with ADHD were excluded for the following reasons: (1) insufficient scan quality  on MPH relative to placebo. This criterion was selected for two primary reasons. First, prior evidence demonstrates that in-scanner head motion and ADHD symptoms are significantly correlated, perhaps due to common genetic influences [acute behavioral response to MPH, given that our goal was to assess possible mechanisms underlying the behavioral effects of MPH. The participants categorized as acute behavioral responders were different for each scan context. Thus, this criterion was applied to each scan context separately. Table acute behavioral response criteria for all three scan types. When restricting analyses to this subset of participants, results were consistent with the findings reported in the main text .Prior evidence suggests that ~70% of children with ADHD respond favorably to MPH , 38, 39,fluences . In suppfluences , 43. Secp-values > 0.29; see Supplemental Information, Tables Finally, we tested whether in-scanner head motion  was differentially related to different ADHD symptom domains to determine whether our results were driven by a single subgroup in the children with ADHD. We found that neither absolute head motion nor the change in head motion between placebo and MPH sessions was significantly related to either symptoms of hyperactivity/impulsivity or symptoms of inattention . MPH or placebo was administered to the participants with ADHD in a counterbalanced order that was randomly assigned and double blinded. MPH dose was 0.3 mg/kg (rounded up to the nearest 5 mg). Given the MPH half-life  and peakFor the resting state runs, participants saw a white crosshair on a gray background and were asked to remain awake and focused on the crosshair.For the go/no-go task, eight sports balls were separated into six ‘go’ stimuli and two ‘no-go’ stimuli; no-go stimuli were determined randomly for each participant. During each trial one sports ball was presented for 600 ms followed by a jittered intertrial interval sampled from a uniform distribution . Participants were instructed to respond on go trials by pressing a button with the index finger of their right hand, and to withhold a response on no-go trials. Each of two runs consisted of 128 trials with stimuli presented in pseudorandom order such that go trials were presented in blocks of two to four, followed by a no-go trial. The rewarded go/no-go task was identical to the go/no-go task, with the addition of rewards for performance. Participants were informed that correct go responses faster than 650 ms and correct no-go non-responses would be rewarded with one cent and five cents respectively. Feedback following each trial was presented for 600 ms, and the interval between response and feedback was jittered identically to that of the intertrial interval. There were 64 trials in each of four runs for the rewarded go/no-go task. Participants were provided the amount of money earned at the end of each session.Scanning was performed on a 3.0-T Siemens Prisma Scanner using a 32-channel head coil at the Biomedical Research Imaging Center at the University of Carolina at Chapel Hill. A T1-weighted multiecho MPRAGE was acquired for coregistration with fMRI images . BOLD signal during functional runs was acquired using a gradient-echo T2*-weighted EPI sequence . Before each scan, five images were acquired and discarded to allow for magnetization to reach equilibrium. 150 volumes were acquired for each resting state scan, 195 for each go/no-go scan, and 185 for each rewarded go/no-go scan.Participants section, these criteria resulted in the exclusion of 12 children with ADHD and 5 TD children from analyses. Importantly, we conducted a set of linear regression models assessing whether children with ADHD who were included in analyses (n = 23) differed in terms of symptom severity (inattention or hyperactivity/impulsivity) or task performance (go/no-go or rewarded go/no-go d’) from children with ADHD who were excluded from analyses for scan quality (n = 12). We did not observe any significant differences  was used to examine the differences in network-level average and modal controllability between TD children (using both scan sessions coded as placebo) and children with ADHD (both on MPH and on placebo). A second set of mixed effects models (one for each scan context for each network) was used to examine the effects of MPH on average and modal controllability in children with ADHD (on-off MPH). The two model sets were estimated separately since the study design was not fully crossed, as TD children were not administered MPH. Both model types controlled for age, biological sex, and in-scanner motion . They additionally used a random intercept at the level of the participant to account for participant-specific differences . This resulted in a total of 156 models (2 model sets x 2 metrics x 3 scan contexts x 13 networks). All models were fit using the lme4 R package . We chosp-values > 0.07; Fig. First, we quantified average controllability of individual functional networks, or how reactive the whole brain was to inputs to each network. During rest, children with ADHD on placebo exhibited increased average controllability in the somatomotor dorsal network relative to TD children , reward  networks relative to TD children. These group differences were not present when children with ADHD were on MPH . There were corresponding significant within-participant reductions in average controllability associated with MPH for the visual . In both cases there was a significant within-participant effect of average controllability associated with MPH . Additionally, there was a corresponding significant increase in modal controllability in children with ADHD on MPH compared to on placebo for the visual . There was a corresponding significant within ADHD effect of MPH for the somatomotor dorsal and somatomotor ventral networks . Additionally, there was a significant increase in modal controllability in children with ADHD on MPH compared to on placebo of the parietal memory network p-values > 0.05). All other effects were non-significant  task context, group differences in controllability were more readily observable. Prior literature has observed larger group differences , 34, 35 Consistent with our hypotheses, MPH administration generally eliminated differences in controllability between ADHD and TD groups, with the only significant difference in the controllability metrics of children with ADHD on MPH compared to TD children localized to the dorsal attention network during rest. This is consistent with prior literature that MPH reduces differences between children with ADHD and TD children for brain activity and FC of specific connections during cognitive tasks , 66, 67., the analyses we performed are likely underpowered to detect group differences, and the pattern of effect sizes suggest that the included participants had slightly lower hyperactivity/impulsivity severity and slightly better performance on both go/no-go tasks. Induced bias due to exclusion criteria is particularly important to consider in high motion populations such as the one under study, and it is likely that our included participants have less severe symptoms than the entire population of children with ADHD. It should be noted, however, that having less severe symptoms is likely to attenuate any effects, rather than lead to upwardly biased estimates. Given these limitations, although the observed networks in which we observed group differences are consistent with prior literature, our results should be taken as exploratory, rather than confirmatory. Future research is needed to confirm the specific networks that show differences in functional controllability as related to ADHD, especially in larger samples and with prolonged treatment to better assess MPH response.There are several limitations to the present study. First, the small sample size, particularly of the children with ADHD (18 or 19 depending on scan context), limits our power to detect group differences, as does the reduction in number of timepoints due to the censoring of high motion volumes. Related to this, differences in findings between scan contexts could in part be due to differences in the number of timepoints going into each task . Second, the study was designed to evaluate acute response to MPH administration, rather than longer term behavioral response to MPH treatment, which curtailed our ability to identify MPH responders using typically applied criteria . This limitation, combined with our small sample size, led us to define acute behavioral response on a task-by-task basis to balance sample size with the behavioral response criteria, which in turn led to some participants being considered responders for one task and non-responders for other tasks. We tested a stricter behavioral response criterion, defining responders as those with reductions in head motion across all three tasks, which reduced the sample size to 13. These models In summary, our findings suggest that the functional dynamics of children with ADHD are characterized by instability in a context of high cognitive demands. Children with ADHD were less likely to enter difficult-to-reach states that may underlie successful response inhibition , while they were more likely to be “pushed out” of those states by non-task-related stimuli (higher average controllability). Furthermore, we identified a possible network-level mechanism through which MPH, a dopamine and noradrenaline transporter blocker, may reduce symptom severity in ADHD: by increasing stability and the ability to enter states necessary for cognitive control. This is consistent with recent work reporting that increased dopamine D1 receptor gene expression was related to increased stability of brain states related to cognitive control, in that case working memory representations . FinallySupplemental Materials