No enrichment was observed for activity-regulated cytoskeleton-associated protein (P=0.23) or N-methyl-D-aspartate receptor (P=0.74) post-synaptic signalling gene sets previously implicated in schizophrenia. Enrichment of ADHD CNV hits for genes impacted by autism de novo SNVs (P=0.019 for non-synonymous SNV genes) did not survive Bonferroni correction. Hypothesis-free testing yielded several highly significantly enriched biological pathways, including ion channel pathways. Enrichment findings were robust to multiple testing corrections and to sensitivity analyses that excluded the most significant sample. The findings reveal that CNVs in ADHD converge on biologically meaningful gene clusters, including ones now established as conferring risk of other neurodevelopmental disorders.A strong motivation for undertaking psychiatric gene discovery studies is to provide novel insights into unknown biology. Although attention-deficit hyperactivity disorder (ADHD) is highly heritable, and large, rare copy number variants (CNVs) contribute to risk, little is known about its pathogenesis and it remains commonly misunderstood. We assembled and pooled five ADHD and control CNV data sets from the United Kingdom, Ireland, United States of America, Northern Europe and Canada. Our aim was to test for enrichment of neurodevelopmental gene sets, implicated by recent exome-sequencing studies of (a) schizophrenia and (b) autism as a means of testing the hypothesis that common pathogenic mechanisms underlie ADHD and these other neurodevelopmental disorders. We also undertook hypothesis-free testing of all biological pathways. We observed significant enrichment of individual genes previously found to harbour schizophrenia  The knowledge gap on pathophysiology is not straightforward to address given practical limitations to directly assess biological and molecular systems in those who are affected. Genetic findings offer one non-invasive approach to providing clues about the biology and pathogenesis of neuropsychiatric disorders.7Attention-deficit hyperactivity disorder (ADHD), a childhood-onset neurodevelopmental disorder,12 Common genetic variants also contribute to ADHD when considered ‘en masse' as a composite risk.19 One important observation from the original ADHD CNV studies was that the CNVs spanned chromosomal and gene regions that overlapped with schizophrenia and autism CNV loci.11 Since then, exome-sequencing investigations of schizophrenia and autism have been published and have highlighted additional novel genomic regions that harbour neurodevelopmental disorder risk variants .23 Exome-sequencing results for ADHD are awaited. Unlike CNVs, which generally span multiple genes, SNVs are located in individual genes and thus offer improved resolution for testing cross-disorder pathogenic mechanisms.In the last 5 years, one class of genetic variant ) has been found to contribute to ADHD risk across multiple studies.24 and in schizophrenia, genes involved in glutamatergic post-synaptic processes: ARC (activity-regulated cytoskeleton-associated protein) and NMDAR (N-methyl-D-aspartate receptor) complexes.25 Additional pathways have been reported to be enriched for SNVs , but such findings are not yet replicated across multiple studies and research designs.Schizophrenia and autism genetic findings have also indicated specific biological mechanisms; these involve targets of the Fragile X Mental Retardation protein (FMRP) in schizophrenia and autism10 Our aim was to test the hypothesis that ADHD-associated CNVs show enrichment for gene sets that have been: (a) implicated by SNVs in the most recent, largest schizophrenia and autism de novo exome-sequencing studies23 and (b) selected for function involving ARC, NMDAR and FMRP targets. We also undertook a hypothesis-free meta-analysis of all biological pathways across the five ADHD data sets.In the current study, we assemble five ADHD and control CNV data sets from the United Kingdom, Ireland, Northern Europe, United States of America and Canada.10 Case–-control CNV data sets were provided by the CNV lead for each study . The five data sets came from: (a) Canada,11 (b) Cardiff, UK,17 (c) the Children's Hospital of Philadelphia, USA (CHOP),8 (d) the International Multi-Center ADHD Genetics (IMAGE) 2 Project16 and (e) the Pfizer-funded study from UCLA, Washington University and Massachusetts General Hospital (PUWMa).26 All affected subjects from each of the five studies were children aged 5–18 years, had an IQ⩾70, were of European descent, free of psychosis, epilepsy, serious neurological impairment and met Diagnostic and Statistical Manual of Mental Disorders (DSM)-III-R or DSM-IV criteria for ADHD as confirmed by semi-structured research diagnostic interviews. Collection and analysis of case–control data had been approved at each site by each Institutional Review Board or ethics committee. Case data were collected with informed consent from parents and assent from children.The recruitment, assessment processes and clinical description of ADHD case subjects and controls have been described in detail previously.27 A total of 692 ADHD cases from PUWMa and 1101 controls were genotyped on the Illumina 1M BeadChip.After quality control exclusions (see later), the Canadian study included 247 DSM-IV ADHD cases and 2357 comparison subjects genotyped on the Affymetrix 6.0 array . The UK Cardiff study (excluding those that were included in IMAGE 2) included 603 DSM-IV ADHD cases genotyped on Illumina Human 660Q-Quad Beadchip  and 1047 comparison subjects genotyped on HumanHap550Beadchip. The CHOP study included 1013 participants with DSM-IV ADHD and 4105 comparison children genotyped on the Illumina Infinium HumanHap550K Beadchip. IMAGE 2 included data from 732 affected children collected in the United Kingdom, Ireland, Germany, Switzerland, the Netherlands and the United States. Control data came from 2010 subjects collected for a genome-wide association study of schizophrenia, described elsewhere.et al.10 for a summary). CNVs were filtered for frequency (<1%), and also for overlapping (>50% length) sites of known common CNVs defined by the Genome Structural Variation Consortium (http://projects.tcag.ca/variation/ng42m_cnv.php) or segmental duplications present in the March, 2006, human reference sequence . To allow for differences in CNV detection between the arrays, we used controls genotyped on platforms that matched the platforms used for the cases. The primary analyses used all CNVs >500 kb as these are the most reliably called across different genotyping platforms, have consistently been found to show increased burden in ADHD cases vs controls and have withstood experimental validation.10 Pathways highlighted by the primary analysis were also tested for enrichment with all CNVs >100 kb (results available from last author).Quality control and procedures for CNV calling are described in each of the original manuscripts The following two logistic regression models were fitted to the sample of CNVs:χ2 on one degree of freedom.29 CNV length was fitted in the model because long CNVs are more likely to hit any set of genes than small ones, and CNV length may differ systematically between cases and controls. The ‘number of genes hit outside pathway' was fitted to allow for case CNVs influencing disease status by hitting genes other than those in the pathway being tested. A binary variable (yes/no) is used for whether a CNV ‘hits gene(s) in a pathway' rather than the number of genes in the pathway hit by the CNV  to allow for some pathways having several genes that are physically close together . The same analysis method was used to determine gene-specific enrichments, by defining the ‘pathway' as the gene.and the deviances of the two models compared. If there is no enrichment of case CNV hits on pathway genes, then the difference in deviances should be distributed asymptotically as a 10 we tested for differential strengths of enrichment by comparing case duplications to case deletions in the logistic regression framework described above.For all analyses, we tested pathways showing significant enrichment in the combined data set of five studies and then undertook tests of sensitivity by examining each of the five data sets separately and rerunning analysis after excluding the most significant single sample. Additional analyses were stratified by CNV type (deletions and duplications). As the number of duplications was greater than the number of deletions,de novo SNVs in schizophrenia were taken from the most recent published de novo exome-sequencing study of schizophrenia, which also catalogued and annotated all SNVs in previous studies in a consistent way.20 There were a total of 611 genes containing at least one non-synonymous de novo SNV, with 87 of these containing a de novo loss-of-function SNV. We also took sets of genes containing: (c) non-synonymous and (d) loss-of-function de novo SNVs from the two largest, recent exome-sequencing studies of autism.23 Combining the autism SNV sets from these studies, there were 2726 unique genes containing at least one non-synonymous de novo SNV, of which 538 contained a loss-of-function de novo SNV.Four sets of genes were defined: (a) non-synonymous and (b) loss-of-function 30 and the ARC (28 genes) and NMDAR (61 genes) complexes, as defined by Kirov et al.25 Bonferroni corrections were used to adjust for testing seven gene sets.Next, we examined a further three gene sets by function, selecting those implicated in schizophrenia and autism by CNV and more recent exome-sequencing studies. These were genes involved in FMRP targets (840 genes) as defined previously Gene Ontology KEGGhttp://www.pantherdb.org/pathway/)PANTHER pathways version 3.1 Mouse Genome Informatics databasehttp://www.biocarta.com, accessed on June 2013)BioCarta Reactome NCIThis final analysis was undertaken using a large, unbiased, general set of pathways comprising:P-values described above and to reduce the chance of a small pathway being falsely declared to be enriched based on a few gene hits, analysis was restricted to pathways with at least 10 gene hits in the total sample . Correction for multiple testing of pathways was performed by calculating q-values.36Pathways containing between 3 and 1500 genes were used in the analysis . To increase the accuracy of the asymptotic 37de novo SNVs20 were significantly enriched for case ADHD CNV hits  but not for deletions (P=0.37). However, there was no significant difference between ADHD duplication and deletions in terms of the rate of hits for genes previously found to carry non-synonymous schizophrenia SNVs (P=0.142). For sample-specific enrichments, see P-values. Restricting analysis to genes hit by loss-of-function de novo schizophrenia SNVs showed no significant enrichment for case CNV hits.The genes containing non-synonymous schizophrenia de novo SNV genes (non-synonymous or loss-of-function), although these do not survive Bonferroni correction.P=0.0018), which remained significant after Bonferroni correction (see P=0.005) but not for deletions (P=0.18), although there was no significant difference between the rate of case duplication and deletion hits for FMRP target genes (P=0.247). See There was significant enrichment of ADHD case CNVs in FMRP targets .de novo SNVs. Previous work demonstrated overlap of ADHD CNVs with schizophrenia and autism CNVs;11 however, because we were now able to define our gene sets using SNVs, which impact on single genes rather than chromosomal regions encompassing multiple genes, the current findings more precisely suggest overlap at the level of genes. These findings therefore extend, refine and are independent of previous studies that suggest biological overlap across these clinically very different disorders.In these analyses of ADHD case–control CNV data, the largest to date, we found highly significant enrichment of CNVs in many, although not all, hypothesised neurodevelopmental gene sets. Hypothesis-free testing revealed additional biological pathways enriched for CNVs in those with ADHD. Genes spanned by the ADHD CNVs were enriched for those that have recently been found to harbour schizophrenia-associated 38 Although previous studies, using some of the data sets in the present paper, found ADHD and autism overlap at the level of CNVs11 and CNV-associated biological pathways,39 we did not observe this extended to autism de novo SNV genes.The clinical co-morbidity and co-heritability of ADHD is much more strongly established with autism than with schizophrenia.23 Complete expression failure of FMRP itself, which characterises Fragile X syndrome, is known to be associated with elevated rates of ADHD, autism and other neurodevelopmental disorders.40 Indeed, the majority of males with Fragile X syndrome show ADHD. Previous work has shown that the protein FMRP regulates activity of 842 biological targets and suggested that some of these likely underlie the manifestation of autistic features in Fragile X syndrome.30 Our findings show that genes encoding FMRP targets are also enriched in ADHD CNVs, which could help explain why children with Fragile X syndrome show such high rates of ADHD. The results suggest that FMRP-mediated biology may be relevant across multiple neuropsychiatric disorders that include ADHD, as well as autism and schizophrenia. Although a recent report suggests that what has been considered as the specific contribution of FMRP targets to the pathogenesis of autism might simply reflect involvement of long, highly brain-expressed genes,41 here large gene size is controlled for. Nevertheless, further work is needed to understand how FMR1 is involved in the pathogenesis of different neurodevelopmental phenotypes, that is, ADHD, autism and schizophrenia, each of which has very different phenotype features and treatments.Targets of the FMRP have been implicated previously in both schizophrenia and autism.23 we did not observe that to be the case for ADHD. Nor did we replicate prior pathway analysis implicating neurite outgrowth at synapses in ADHD, although previous analyses have used SNP or candidate gene data or smaller data sets.45 We cannot be certain if this is a genuine point of difference or whether larger studies of ADHD, which consider additional types of mutations, for example, through exome sequencing, will yield a different pattern of findings.Although schizophrenia and autism genetic findings have all strongly implicated the involvement of synaptic functions in pathogenesis,The hypothesis-free analysis of all ADHD CNV pathways yielded strongly significant enrichment for multiple biological pathways. Many but not all of these were robust to analysis after excluding the most significant study. As expected for this final, hypothesis-free set of analyses, the significance of the enrichment was reduced by the removal of the most significant study, even when there was no significant difference in the strength of the enrichment between that study and the remainder of the sample, as was the case for most of the pathways highlighted here  and especially involvement of 45 However, the biology and validity of ADHD remain poorly understood and it is still widely considered to be primarily a catecholaminergic disorder.43 Findings from the present study highlight the consistent and robust neurodevelopmental nature of ADHD and provide novel insights about its biological underpinnings.Although our study involves analysis of the largest ADHD CNV data set to date, there are several limitations. First, analyses of this type for any disorder are restricted by the quality of gene and pathway annotations and the fact that the same genes belong to multiple different functional gene sets. Second, genotyped ADHD sample sizes have markedly lagged behind those of many other neurodevelopmental and psychiatric disorders and results of exome sequencing are awaited. Third, ADHD-associated CNVs do not capture all possible forms of genetic risk, or risk loci; however, previous findings from smaller, individual studies do suggest biological convergence of CNVs and common gene variants.In conclusion, in an international, cross-centre analysis of ADHD CNV data, we find evidence of biological overlap with schizophrenia at multiple levels; previous findings of overlap with schizophrenia CNVs have now been extended to SNVs. Furthermore, FMRP target enrichment appears to characterise ADHD as well as schizophrenia and autism. Ion channel pathway involvement in ADHD was robust to type of CNV and across samples. The findings reveal that CNVs in children with ADHD, from across multiple centres, converge on biologically meaningful gene clusters that are now robustly established as involved in neurodevelopmental disorder risk.


The declaration represents a coordinate effort to control or eliminate ten of the neglected diseases by 2020, and has already led to important coordination and partnership, and mobilised considerable resources. Although the progress made so far is to be celebrated, now is the time to count down to 2020 and start monitoring the progress and trends towards achieving the control and elimination targets.The London Declaration on Neglected Tropical Diseases2 Efforts are being made to measure the progress and forecast future trends, but most efforts are disease-specific and occur at a global scale.3 Therefore, comprehensive measurements to track the goal of the declaration are needed. Country-specific case studies, considering different aspects of policy, programming, and intervention would provide the whole picture of the analysis needed for the countdown to 2020. Focus on assessment from a multidimensional aspect and pinpointing of bottlenecks will help to identify the gaps in implementation and point out areas that need improvement.First, progress towards the 2020 targets should be measured through collation, presentation, and analysis of standardised core metrics that track country progress. Elimination and control targets in the WHO Roadmap could serve as initial standardised metrics.Second, development of standardised methods to measure the progress towards reaching the targets is crucial. Modelling methods, which use the available data to forecast future trends will be important for understanding how countries are progressing towards achieving the goals of the London declaration. These methods will enable comparison of the outputs of such an analysis, and several sources of data would be needed to provide continuous high-quality data for analysis. Collaboration with national surveys, such as the demographic and health surveys and several indicators surveys, would be crucial for achieving this goal.4 The group would have an important role in the definition of the core methods to monitor progress and develop a framework for country assessment mechanisms. This approach will help to further improve the quality of country-level reports.Third, establishment of a global independent monitoring group would be essential to assess country-specific reports and set up an accountability framework.As we go further down the road, understanding if we are on track to achieve the targets is necessary. Standardised metrics, continued monitoring, and comprehensive country-specific case studies will be crucial to advance the goal of the declaration.


This study compared the sociodemographic characteristics, comorbidities, healthcare resource utilization, and work productivity among Japanese adults who reported being diagnosed with attention-deficit/hyperactivity disorder (ADHD) to those of a non-ADHD control population.Data for this study were captured from an online survey of adults in Japan conducted by Kantar Health using consumer panels. A total of 84 survey participants reported they had received a diagnosis of ADHD from a physician. Survey responses pertaining to functional status and resource utilization from this ADHD group were compared to those from a non-ADHD control group of 100 participants. Comparisons between the ADHD and non-ADHD groups were made using chi-square tests for categorical variables and t-tests for continuous variables.Participants in the ADHD group were on average slightly younger with a higher proportion of males. ADHD respondents reported significantly more comorbid depression, sleep difficulties, headaches, and anxiety than non-ADHD controls. Over the previous 6 months, the ADHD group made more visits to healthcare providers and the emergency room, and had more hospitalizations than non-ADHD controls. The ADHD group also rated their overall health status lower than the non-ADHD control group. Respondents with ADHD reported a significantly higher degree of health-related work impairment compared to non-ADHD, with greater absenteeism and decreased work productivity. The ADHD group indicated their symptoms negatively impacted relationships, self-esteem, and regular daily activities.Japanese adults with ADHD face a substantial burden of illness, including lower overall health status, increased number of comorbidities, greater healthcare utilization, and significant health-related occupational impairment compared to those without ADHD. Additional research is needed to develop a better understanding of both the consequences and treatment approaches for Japanese adults with ADHD. Attention-deficit/hyperactivity disorder (ADHD) is characterized as a pattern of inattention and/or impulsivity-hyperactivity, starting in early childhood, that is more persistent, frequent, and severe than typically observed at a similar stage of development in others . This diADHD is a prevalent disorder that affects an estimated 4% to 7% of children worldwide . While AOutside of Japan, reports indicate that adults with ADHD frequently present with impairment at school, in social environments, and in the workplace ,4,9,10. In the workplace, adult ADHD has been shown to impair and reduce productivity, and also to lead to behavioral issues as a result of irritability and low frustration tolerance . In addiAn increased risk of serious vehicular accidents in adults with ADHD has also been reported . In one In addition to the related impairments in social and occupational functioning, ADHD in adults is also associated with an increased burden on the healthcare system. A study conducted in the United States comprehensively reviewed the literature for studies reporting incremental costs related to ADHD over a 20-year period . This stIn Japan, adult ADHD had been less recognized in the field of psychiatry, but this has recently started to change with the approval of medication used to treat this disorder . While iData for this study were captured from the National Health and Wellness Survey (NHWS), administered in 2012 by Kantar Health to the adult population of Japan. The NHWS is a self-reported, Internet-based survey that assessed information on approximately 160 different conditions. These data were drawn from a larger study that also collected survey data across Europe, Australia, and the United States . All resTwo cohorts were defined for comparison; an ADHD group and a non-ADHD control group. Respondents from Japan were included in the ADHD group if they indicated they had received a diagnosis of ADHD from a physician (n = 84). The control group was comprised of 100 participants from Japan, drawn randomly from the entire sample of respondents, who indicated they did not have experience with ADHD and had not received a diagnosis of ADHD (n = 100).Measures were included to assess domains pertaining to comorbidities, general health, functioning, work productivity, general activity and use of healthcare services for both cohorts. Comorbidities were assessed based on self-report. Participants were asked to select which health conditions they had experienced in the past 12 months, from a list provided. The general health of participants was assessed with a single question from the Medical Outcomes Survey–Short Form– 12 Version 2 (SF-12v2): “In general, would you say your health is: excellent, very good, good, fair, or poor” .For the ADHD group, the effect of ADHD on functioning was evaluated using a series of 6 questions, rated on a 5-point scale ranging from strongly disagree (1) to strongly agree (5). The 6 questions were as follows. ADHD has negatively impacted my: (1) ability to achieve success in my career/work life, (2) relationship with my spouse or significant other, (3) relationships with other people in my life, (4) self-esteem, (5) overall physical health, and (6) overall mental health.The impact of health problems  on work productivity and general activity was measured using the Work Productivity and Activity Impairment Questionnaire (WPAI) . The WPAHealth care resource use was assessed based on self-report. Participants were asked how many times they had visited various types of healthcare providers in the past 6 months. In addition, participants were asked how many times they had been to the emergency room or hospital for their medical condition in the past 6 months.Comparisons between the ADHD and non-ADHD control groups were made using chi-square tests for categorical variables and t-tests for continuous variables. All analyses were completed using SAS version 9.2 .A total 184 individuals from Japan were included in the analysis; 84 individuals from the online survey met inclusion criteria for the ADHD group and 100 individuals were included in the non-ADHD control cohort. Demographic characteristics were similar between the two groups, with the exception of age and gender. Participants in the ADHD group were on average slightly younger and had higher proportion of males .Over the past 12 months, comorbid conditions reported significantly more frequently in the ADHD group than in the control group included depression, sleep difficulty, and anxiety-related disorders . The most common comorbid condition reported by the ADHD group was depression . SpecifiParticipants in the ADHD group also utilized significantly more healthcare resources over the previous 6 months, as measured by total healthcare provider visits, emergency room visits, and hospitalizations .Consistent with the greater frequency of visits to a healthcare provider, the ADHD group also rated their health status lower than the control group. The mean self-rated health status for the ADHD group and control group was 3.6 (standard deviation [SD] 1.2) and 3.0 (SD 0.8), respectively (p < 0.05), with a higher score indicating worse overall health. The modal rating for the ADHD group was “Fair” while the modal rating for the control group was “Good” . In the With regard to occupational functioning, as assessed by the WPAI, participants in the ADHD group reported a higher rate of absenteeism over the past 7 days when compared to the control group . In addition, health problems affected work productivity in the ADHD group to a greater extent than the control group as measured by the presenteeism scale . Taking into account both absenteeism and presenteeism, the mean work impairment score was significantly greater for the ADHD group  than for the control group .According to the responses on the WPAI, the impact of health problems was not limited to work productivity. In the same time period, participants in the ADHD group also reported that their health problems affected their ability to engage in regular daily activities to a greater degree than reported by the control group. The activity impairment score was 59.2% for the ADHD group and 18.3% for the control group (p < 0.05).Specific to the ADHD group, respondents reported that their ADHD symptoms negatively impacted their relationships and self-esteem . The majThe results from this study underscore the significant impact of adult ADHD on individuals in Japan with this condition in terms of overall health, as well as functioning in social and occupational settings. Japanese adults with ADHD reported a significantly greater number of comorbid central nervous system (CNS)-related conditions. In the presence of ADHD, depression and general anxiety occurred at particularly high rates that were approximately 60- and 40-fold greater than non-ADHD controls, respectively . The ratFurthermore, individuals in the ADHD group utilized substantially more healthcare resources, as measured by visits to a physician, emergency room visits, and hospitalizations. In fact, the ADHD group reported visits to physicians at a rate that was 10 times higher than the control group, and approximately 3 times more visits to the emergency room and hospitalizations . Given tSignificant impairments in occupational and social functioning were also attributed to the symptoms of ADHD. Individuals with ADHD missed more work, and reported being less productive while at work, than individuals in the control group. In fact, the mean overall work impairment score for the ADHD group was over 3 times higher compared to the control group. Furthermore, nearly 75% of participants in the ADHD group either “agreed” or “strongly agreed” that ADHD had a negative impact across relationships with their spouse, coworkers, and others.The negative impact associated with adult ADHD found in this sub-analysis of Japanese patients is consistent with the overall results from the larger study that included Europe, Australia, and the United States . Adults In regard to comorbidities of CNS-related disorders, higher rates of Substance Use Disorders (SUD) in ADHD patients (12.5%) have been reported previously in the literature . The ratIn the present study, the disruption associated with ADHD in occupational and social settings reflects the findings from other parts of the world as well. A consistent pattern starts to emerge regarding lost productivity and difficulties in relationships with family, friends, and co-workers. Küpper and his colleagues reviewed the literature (limited to articles published in English or German) to assess the negative effects of ADHD in adults on work productivity and occupational health . The autThe results from this study add to, and are consistent with, the one known study regarding the consequences of ADHD adult in Japan in which ADHD was associated with increased stress and more frequent visits to a healthcare provider .ADHD in adults is consistently associated with negative outcomes regarding relationships, work productivity, and healthcare utilization. Therefore, it is not surprising that they also tend to suffer from depression and anxiety. Proper diagnosis, and identifying the appropriate treatment and support are essential to help adults with ADHD improve their lives and also offset the costs associated with lost productivity and overall healthcare utilization.Information from this study was obtained through self-report by online survey respondents, rather than from clinical or experimental data. Therefore, it was not possible to verify demographic and behavioral information provided by participants, nor corroborate the diagnosis of ADHD (or the absence thereof). In addition, the ADHD and control groups were not matched on age and gender.When compared to adults not suffering from ADHD, adults from Japan with ADHD face a substantial burden of illness including lower overall health status, increased number of comorbidities, greater healthcare utilization, and significant health-related occupational and social impairment. The results from this study add to the limited information available regarding the impact of adult ADHD specific to Japan. Additional research is needed to develop a better understanding of both the consequences and treatment approaches for Japanese adults with ADHD.


Recently, the master equations for the interaction of two-mode photons with a three-level Λ-type atom are exactly solved for the coherence terms. In this paper the exact absorption spectrum is applied for the presentation of a non-demolition photon counting method, for a few number of coupling photons, and its benefits are discussed. The exact scheme is also applied where the coupling photons are squeezed and the photon counting method is also developed for the measurement of the squeezing parameter of the coupling photons. Electromagnetically induced transparency (EIT) has been theoretically introduced by Kocharovskaya and Khanin  and expeMoreover, profile of the absorption and dispersion spectra are shown to depend on the number of coupling photons so that the number of coupling photons could be measured using the absorption spectrum of the probe photons. This scheme is applied for the presentation of a non-demolition photon counting method. The present method is applied to the squeezed coupling photons. Straightforwardly, it is shown that the exact absorption and dispersion spectra drastically depended on squeezing parameter of the coupling photons. This scheme is also applied for presenting measurement of the squeezing parameter.In “In this section a review on the exact model of a three-level Λ-type atom interacting with two quantized electromagnetic field  interact with a three-level Λ-type atom see Fig. a. The inAssume the system is initially in the ground state entclass1pt{minimaAn ensemble of cold three-level atoms is prepared by an optical pumping initially in the state entclass1pt{minimaThe master Eqs. –7) are  are 7) aThe real and imaginary parts of entclass1pt{minimaIn the next section, a photon counting method based on the exact form of absorption spectrum of the probe field, which is derived from Eq. , is presΛ-type atoms.In this section, a non-demolition photon counting method is presented for measuring the number of coupling photons which are trapped in a quantum cavity and interact with an ensemble of three-level It is worthwhile to estimate the number of probe photons which is required to determine the probe absorption peak. As an example, the cavity field decay rate can be estimated as It is clear in Eqs. –17) and and17) aAlthough Fig. A considerable difference between the plots in Fig. r and Another application of the full-quantum interaction of two-mode photons with three-level Λ-type atoms is in measurement of squeezing parameter of coupling photons. Supposed the trapped coupling photons are squeezed In this case, the interaction Hamiltonian in the interaction picture is given by “” 19\docuEquation  is similentclass1pt{minima in Eqs.  and, aftr, which leads to applying the photon counting method for a squeezing measurement by measuring the DAPs or DDPs.The real and imaginary parts of the probe coherence term  correspor. Furthermore, they are more sensitive to the squeezing parameter for a larger number of photons. Of course, it is important to note that n2 is not the average of coupling photons; but, it can be easily derived by r. Therefore, number of coupling photons and their squeezing parameter can be obtained by measuring DAP and DDP of the absorption and dispersion spectra simultaneously. Some typical values of DAP and DDP are shown in Table By taking a derivative of the imaginary and real part of Eq.  and settmentclass2pt{minimmentclass2pt{minimIn this paper, the master equations of Λ-type three-level atom interacting with two-mode quantized electromagnetic field and its exact coherence term are applied to obtain the squeezed and non-squeezed coupling photons. The following results were obtained: (1) The method was applied for presenting a photon counting and a squeezing measurement method by measuring the absorption spectrum of the probe photons. (2) The difference between the exact and WFA photon counting methods and benefits of the exact method  were demonstrated. This sensitivity increased for the larger number of coupling photons by increasing the squeezing parameter. (3) It was shown that the photon counting method was more sensitive for the smaller number of coupling photons. (4) The present method for the measurement of squeezing was more sensitive for larger values of squeezing parameters. (5) The number of coupling photons and their squeezing parameter can be obtained simultaneously by measuring the DAP and DDP. (6) The photon counting method was non-demolition for the strong coupling photons.


Apripona germari, comprehensive metabolomic changes of poplar bark and xylem in response to A. germari infection were examined by gas chromatography time-of-flight mass spectrometry (GC–TOF/MS). It was found that, four days after feeding (stage I), A. germari infection brought about changes in various metabolites, such as phenolics, amino acids and sugars in both bark and xylem. Quinic acid, epicatechin, epigallocatechin and salicin might play a role in resistance response in bark, while coniferyl alcohol, ferulic acid and salicin contribute resistance in xylem. At feeding stages II when the larvae fed for more than one month, fewer defensive metabolites were induced, but levels of many intermediates of glycolysis and the tricarboxylic acid (TCA) cycle were reduced, especially in xylem. These results suggested that the defense strategies against A. germari might depend mainly on the early defense responses in poplar. In addition, it was found that bark and xylem in infected trees accumulated higher levels of salicylic acid and 4-aminobutyric acid, respectively, these tissues displaying a direct and systemic reaction against A. germari. However, the actual role of the two metabolites in A. germari-induced defense in poplar requires further investigation.Plants have developed biochemical responses to adapt to biotic stress. To characterize the resistance mechanisms in poplar tree against  Populus spp.), a fast-growing species with high timber yields, play important roles in wood production and ecological maintenance, and are widely distributed in China [Apriona germari (Hope), is a destructive stem-boring pest in China. The main damage associated with A. germari is caused by the larvae, which create long tunnels in the trunk. This affects the growth of the trees, increases the chances of windbreak and decreases the quantity and quality of the timber. Notably, during population outbreaks, the insects are capable of causing extensive mortality and severe damage to poplar plantations. Breeding for resistance is considered as an economical and environment-friendly approach to manage A. germari [The poplar tree (in China . However germari . KnowledPhyllocnistis populiella) induced increased levels of tremulacin and salicortin in aspen leaves [Malacosoma disstria) and leaf beetle (Chrysomela confluens) larvae significantly increased condensed tannin concentrations in poplar leaves [Anoplophora glabripennis [It is well known that plants have evolved a variety of strategies for defending themselves against insects. Chemical defense is one of the most effective strategies for insect control because metabolites play important roles in the resistance reaction . Comparin leaves . Feedingr leaves . Howeverripennis . HoweverA. germari-induced defense response in the poplar tree will be helpful for exploring underlying defense mechanisms.Metabolomics, a post-genomics tool, can elucidate the host biochemical responses to exogenous elicitors by comprehensively analysis of all metabolites in plant. A systematic analysis on plant metabolism upon insect challenge allows the identification of defensive compounds, revealing novel pathways and discrimination the mechanistic responses to different elicitors ,10. In fA. germari induced metabolic responses were investigated in poplar clone DH (P. deltoids cv. “Danhong”), which have been widely planted in China. Non-targeted metabolic profiling of poplar bark and xylem, with and without A. germari infection, were analyzed by gas chromatography time-of-flight mass spectrometry (GC–TOF/MS). The results revealed that upon infecting the tree, the insect induced metabolic changes in the tree to satisfy nutritional requirements; conversely, there was a shift in the tree’s metabolism to defend against attack. The levels of phenolics, amino acids and sugars were significantly disturbed by A. germari in both bark and xylem. The defense strategies against A. germari might depend on early defense responses in poplar because more defensive metabolites were induced at earlier feeding stages. Quinic acid, epicatechin, epigallocatechin and salicin might play a role in resistance response in bark, while coniferyl alcohol, ferulic acid and salicin contributed to resistance in the xylem. The results will be helpful for identifying metabolic biomarkers for inductive resistance to the insect and may help us to gain an improved understanding of the molecular mechanisms underlying resistance of trees to A. germari.In the present study, A. germari induced metabolic responses in poplar, metabolic profiles of bark and xylem tissues with and without infection, were analyzed by GC–TOF/MS. In bark tissues, 1055 peaks were obtained, in which 412 and 643 peaks were identified in the polar and lipophilic phases, respectively. In xylem tissues, 784 peaks were measured, with 305 peaks being identified in the polar phase and 305 peaks in the lipophilic phase. A library search using the NIST mass spectral data base showed that the metabolites in the lipophilic phases included phenolic acids, benzenoids, sterols, fatty acids and their derivatives, while those in polar phases mainly included amino acids, sugars, organic acids and their derivatives. Metabolic profiles of the mock- and A. germari-infected samples were differentiated by PLS-DA model in SIMCA-P 11.5 . As shown in A. germari infection. At feeding stage I, four days after the larva feeding, the infected bark tissues were clearly separated from their mock controls, but the two groups of the xylem tissues were only just distinguishable, indicating that more metabolic changes occurred in the bark than the xylem. At feeding stage II when the larvae fed for more than one month, separations were observed between the infected bark and the xylem tissues with their corresponding controls  were down-regulated (O-phosphorylethanolamine) were increased at 20 days but decreased at 60 days. Pathway enrichment and topological analyses revealed that A. germari induced metabolites in bark were mainly assigned to biosynthetic pathways for unsaturated fatty acids, fatty acid biosynthesis, cyanoamino acid metabolism, methane metabolism, sphingolipid metabolism and nitrogen metabolism  and later stage (stage II) were investigated using comprehensive untargeted chemical analysis. Metabolic profiles of bark and xylem near the infection sites were analyzed, respectively, giving more detailed information on metabolic changes triggered by A. germari in poplar.In this study, the metabolic responses of poplar to A. germari. Especially, the amounts of some sugars, amino acids and fatty acids were significantly changed both in the bark and the xylem. Sugars and amino acids are important nutritional components assimilated by the insect from the plant, while amino acids and fatty acids are precursors of some defensive metabolites. The alteration in the metabolism of these metabolites might be associated with a reallocation of nutrients and provision of precursors for the biosynthesis of defensive metabolites and indicate that the insect might induce some egocentric changes in the host to satisfy nutritional requirements, and conversely, the tree may switch the metabolism to synthesize defensive metabolites, such as tannins in bark and phenolics involved in lignin synthesis in xylem, thus attempting to defend itself from the insect [A. germari indicating that this metabolite might be also involved in the induced resistance to stem borers. In contrast to salicin, some secondary metabolites were induced by A. germari in specific tissue possibly resulting from different chemical components in bark and xylem. For example, quinic acid, epicatechin and epigallocatechin were increased in infected bark, while ferulic acid and coniferyl alcohol were up-regulated in infected xylem. Quinic acid, epicatechin, epigallocatechin are low molecular weight tannins and have been shown to be toxic to several herbivores [A. germari induced accumulation of the two group metabolites might be important resistance factors of poplar clone DH against this insect in bark and xylem, respectively.Before ovipositing, the female adult bit through the bark and gnawed a nidus within which an egg was laid in the xylem. Therefore, early metabolic changes in infected trees might be associated with the damage caused by the adult and feeding of larvae. At stage I in this study, metabolic analysis of samples revealed that primary and secondary metabolisms in the tree were significantly disturbed by e insect . Salicine insect . The prerbivores ,15. Ferurbivores . Lignin rbivores ,18. Furtrbivores ,20. TherA. germari fed in the pith. The metabolic changes in bark were decreased while those in xylem were increased. Less defensive metabolites were induced by the infection both in bark and xylem. The metabolites that were perturbed were mainly sugars, amino acids, organic acids and fatty acids which were associated with primary metabolism in the tree. As shown in After feeding for more than one month, the larvae of A. germari induced metabolic responses in the two tissues were somewhat different. Specifically, some tissue-specific metabolic changes were found in the present study. For example, in the bark, salicylic acid (SA) was significantly up-regulated by A. germari infection at all sampling stages. Meanwhile, the amount of linolenic acid, the precursor of jasmonic acid (JA), decreased in the infected bark although several fatty acids had increased. SA and JA usually act as signaling molecules in the regulation of induced responses to biotic stress. Generally, the former regulates the responses to pathogens, while the latter plays a role in responses to chewing herbivores [A. germari induced responses in poplar has to be corroborated by further studies. In the infected xylem at feeding stages I and II, the contents of glutamic acid and 4-aminobutyric acid (GABA), which is a non-protein amino acid derived from glutamic acid, were all significantly induced by A. germari. GABA accumulation can be stimulated by feeding of leaf eating larvae and thus affect larval performance possibly as the ingested GABA interferes with the normal development of the insects [A. germari induced accumulation of GABA in xylem suggested that GABA might also be involved in plant defense against stem-boring insects.Pathway enrichment and topology analyses revealed that the perturbed metabolites enriched pathways in bark tissues were not the same as those in xylem indicating that rbivores ,25,26,27rbivores ,29. Addirbivores . Indeed, insects ,32,33,34 insects . Thus, AP. deltoids cv. “Danhong”), which is widely planted in China, was chosen to investigate the A. germari induced metabolic changes in poplar. Tissues of DH were obtained by vegetative propagation using cuttings which were grown in a greenhouse . When the height reached 20–35 cm, the poplars were transferred to an open-air garden. We selected for bioassay and metabolic profiling 16 two-year-old trees  that lacked any signs or symptoms of pathogen or insect attack (eight corresponding to A. germari infected and eight mock infected). In early July, the female and male A. germari adults were captured from paper mulberry (Broussonetia papyrifera) and were placed in wire cages  which were fixed in the trees at 2 m above the ground. One male and 1 female were placed in each cage. Fresh branches of paper mulberry tree were supplied to the adults and were replaced every 2 days. The mock-infected trees received fixed wire cages but without insects.The poplar clone DH  and 3-mm-thick outer xylem tissues near the dejection site under which the larvae hid in the infected trees and those in the corresponding altitude of the mock infected trees were sampled. Hereafter, this sampling stage is referred to as stage I. Forty days later, the larvae bored into the pith of the tree, with small frass expulsion holes made through the bark. Tissues of infected bark and xylem and their corresponding mock controls were also sampled. The second sampling stage is referred to as stage II. After removal from the tree, the bark and the xylem were separated and then frozen immediately in liquid nitrogen.et al. and Qu et al. with some modifications [g, 10 min) and 1.0 mL supernatant was collected. Five hundred microliters distilled water and 400 μL chloroform were mixed with the supernatant and then centrifuged . Finally, five hundred microlitres polar extracts and 200 μL lipophilic extracts were taken out and dried using a nitrogen gas stream in a vacuum rotary evaporator without heating. Fifty microliters of methoxyamine hydrochloride  dissolved in pyridine (ACS grade) were added to the dried extracts and incubated for 2 h (30 °C). Then, 100 μL of MSTFA (N-Methyl-N-trifluoroacetamide) was mixed with the above solution and incubated for 30 min (37 °C).Metabolites in bark and xylem tissues were extracted according to the protocol of Weckwerth ications ,37. The m/z 80–500 at 20 scans per second. The detector voltage was set to 1800 V.The derivatized samples were analyzed using a LECO Pegasus IV GC-TOF/MS system. An Agilent 6890 gas chromatograph  was used with a 30 m long, 0.25 mm i.d. (inside diameter) fused silica capillary column with 0.25 μm DB-5 MS stationary phase . One μL was injected in splitless mode and 10:1 split ratio for lipophilic and polar extracts, respectively. Chromatography was performed at a flow rate of 1.5 mL/min. For the lipophilic metabolites, the initial column temperature was set as 100 °C and maintained for 2 min, 100 to 200 °C at 8 °C/min and held for 1 min, 200 to 260 °C at 3 °C/min and held for 1 min, and then 260 to 320 °C at 8 °C/min and held for 4 min. For the polar metabolites, the initial column temperature was set as 100 °C and maintained for 2 min, 100 to 260 °C at 4 °C/min and held for 1 min, 260 to 320 °C at 20 °C/min and held for 3 min. The column’s effluent was then directed to the ion source of a Leco Pegasus IV time off light mass (TOF) spectrometer with an ion source temperature of 200 °C, transferline temperature of 250 °C, and electron ionization at −70 eV. Mass spectra were obtained from p < 0.01). Compounds were tentatively identified using NIST standard mass spectral databases (similarity ≥ 80%). Heat maps derived from the log-transformed normalized data were obtained using the cluster program and visualized using the Mev software v4.9 [Arabidopsis thaliana pathway library.ChromaTOF version 4.0  was used for the raw data processing. Weights of the samples were normalized to the maximum value to minimize the discrepancy caused by sample weights. Then, the data was normalized using min-max normalization. After normalization, the missing values for each metabolite were replaced with the observed minimum value. Multivariate and univariate statistical techniques were used to analyze the data. The multivariate analyses were performed by the software package SIMCA-P 12.0 . Using the autofit function, PLS-DA model was fitted on mean centered unit variance scaled data. Potential differential metabolites were extracted according to the variable importance plot (VIP) values (VIP > 1). Meanwhile, the non-parametric Mann–Whitney test  was used to distinguish differential metabolites (are v4.9 . A red/gare v4.9 . The outA. germari at earlier and later stages (stages I and II) of feeding. It was observed that the primary and secondary metabolisms in bark and xylem tissues in infected trees were significantly altered by A. germari. More defensive metabolites were induced at the earlier feeding stage (stage I) than the later stage (stage II) when the energy production of the tree was disrupted. The defensive compounds induced in bark were quinic acid, epicatechin, epigallocatechin and salicin, while those in xylem include coniferyl alcohol, ferulic acid, salicin and GABA. The present results may prove to be useful in identifying the metabolic biomarkers for A. germari resistance in poplar and to gain a better understanding of the mechanistic basis for resistance against stem-boring pest in trees.This study traced the metabolic responses in bark and xylem of poplar trees against infection by


Although atomoxetine demonstrates efficacy in individuals with attention-deficit hyperactivity disorder, its treatment effects on brain resting-state functional connectivity remain unknown. Therefore, we aimed to investigate major brain functional networks in medication-naïve adults with attention-deficit hyperactivity disorder and the efficacy of atomoxetine treatment on resting-state functional connectivity.After collecting baseline resting-state functional MRI scans from 24 adults with attention-deficit hyperactivity disorder (aged 18–52 years) and 24 healthy controls (matched in demographic characteristics), the participants with attention-deficit hyperactivity disorder were randomly assigned to atomoxetine (n=12) and placebo (n=12) arms in an 8-week, double-blind, placebo-controlled trial. The primary outcome was functional connectivity assessed by a resting-state functional MRI. Seed-based functional connectivity was calculated and compared for the affective, attention, default, and cognitive control networks.At baseline, we found atypical cross talk between the default, cognitive control, and dorsal attention networks and hypoconnectivity within the dorsal attention and default networks in adults with attention-deficit hyperactivity disorder. Our first-ever placebo-controlled clinical trial incorporating resting-state functional MRI showed that treatment with atomoxetine strengthened an anticorrelated relationship between the default and task-positive networks and modulated all major brain networks. The strengthened anticorrelations were associated with improving clinical symptoms in the atomoxetine-treated adults.Our results support the idea that atypical default mode network task-positive network interaction plays an important role in the pathophysiology of adult attention-deficit hyperactivity disorder. Strengthening this atypical relationship following atomoxetine treatment suggests an important pathway to treat attention-deficit hyperactivity disorder. Attention-deficit hyperactivity disorder (ADHD) is an early-onset neurodevelopmental disorder with clinical symptoms often persisting into adulthood . Given cIntrinsic resting-state functional connectivity (RSFC), represented by the correlation of low-frequency  spontaneous fluctuations in neural activity measured by a resting-state functional MRI (rs-fMRI) BOLD signal, can reliably characterize the functional organization of the brain at a systems level . AberranPharmacological treatment is recommended as the first-line treatment for adults with ADHD . AtomoxeOnly 2 studies have investigated atomoxetine treatment effects on neural activity and its relationship with clinical improvement. After an 8-week treatment with atomoxetine, adults with ADHD had increased fronto-parietal activation during interference . In a 6-supplementary Material for the rationales of selecting these networks). Second, we investigated how an 8-week treatment with atomoxetine affected RSFC in medication-naïve adults with ADHD. We further probed how posttreatment changes in RSFC were related to clinical and neuropsychological performances in the atomoxetine-treated group. Based on earlier studies, we hypothesized that medication-naïve adults with ADHD would exhibit reduced anticorrelated relationships between the DMN and task-positive networks (attention and control networks), as well as decreased RSFC in the DMN, compared with controls. Further decreased connections in the cognitive control and attention network and increased RSFC in the affective network would be found in the patient group relative to the control group. These changes might demonstrate some patterns distinct from the report of The aim of the present study was thus 2-fold. First, we used rs-fMRI to explore ADHD-related RSFC differences between medication-naïve adults with ADHD and healthy controls in the 5 predefined neural networks . All participants provided written informed consent.DSM-IV-TR criteria for childhood and current ADHD diagnosed by the corresponding author and further confirmed with the semistructured Conners’ Adult ADHD Diagnostic Interview as described in the DSM-IV    for currms Inc.) , 2013b. Twenty-four healthy adult controls without any lifetime diagnosis of ADHD based on the same clinical and psychiatric assessments as the ADHD group were recruited from the community according to the same age, sex, intelligence, and handedness of the ADHD group.All participants who had any systemic medical illness; a history of bipolar disorder, psychosis, major depression, substance use disorder, pervasive developmental disorder; or currently had depressive or anxiety symptoms or suicidal ideations; had been treated with any psychotropic agents, including medications for ADHD; or IQ <80 as assessed by the Wechsler Adult Intelligence Scale-Revised were excluded from this study.supplementary Material.All enrolled participants underwent clinical, neuropsychological, and imaging assessments. Control participants completed a single MRI scan and the Rapid Visual Information Processing (RVP) of the Cambridge Neuropsychological Test Automated Battery ; adults Medication-naïve adults with ADHD were randomly assigned to double-blind treatment with atomoxetine (n=12) or placebo (n=12) according to computer-generated random sequencing. Participants with ADHD were initially administered atomoxetine 0.5mg/kg/d in the morning at baseline and would titrate the drug dosage at week 2 , and at week 4 depending on clinical response and adverse effects (maximum daily dosage of atomoxetine=1.2mg/kg) depending on clinical response and adverse effects. Only nonspecific supportive psychotherapy and counseling were implemented as needed during the follow-up. All ADHD participants completed the pre- and posttreatment MRI scans and other required assessments without any missing data .supplementary Material for imaging parameters and preprocessing details).Data were obtained on a 3T scanner (Siemens Magnetom Tim Trio) with a 32-channel phased-arrayed head coil. All participants were verbally instructed to remain still with their eyes closed to complete a 6-minute rs-fMRI scan . WakefulFollowing preprocessing, whole-brain functional connectivity was calculated by correlating the seed time-series with the time course of all other voxels using REST toolbox . The resP<.05. One-way ANOVA was used to determine differences in clinical symptoms, neuropsychological performances, vital signs, and measures of in-scanner head motion at baseline. One-way repeated-measures ANOVA was used to compare differences within subjects between weeks 8 to 10 and baseline in each treatment arm of the clinical trial. Mixed-effect ANOVA was used to test for treatment × time interaction effect.All behavioral data analysis was conducted using IBM SPSS Statistics for Macintosh . The alpha value was preselected at P<.01 and a spatial extent threshold (corrected for nonstationarity) that ensures a cluster-wise FWE at P<.05  was used to localize the significant clusters and the related Brodmann area (BA). The results were visualized using BrainNet Viewer  calculated based on random field theory implemented in SPM8, using a cluster-forming voxel-level height threshold of at P<.05 . xjView8t Viewer .t tests to determine significant differences in RSFC between ADHD and controls. For treatment effects, we entered each ADHD participant’s seed-based connectivity map into a 2×2 repeated-measure factorial model using SPM8. We treated time as a repeated measure with 2 levels: pretreatment and posttreatment scans. We used treatment as a between-group factor with 2 levels: atomoxetine and placebo. We isolated a time×treatment term to determine differential treatment effects on connectivity, followed by conducting a posthoc pairwise t test for seed-regions of interest connectivity to determine the nature of the interaction. As suggested by supplementary Table 1).For baseline comparisons, we analyzed rs-fMRI data in SPM8 using 2-sample F test on the functional connectivity maps for covarying mean FD, with an uncorrected P<.05. We then created a mask that excluded any voxels, in which there were baseline RSFC differences across the 2 treatment arms, from subsequent analyses. Owing to the finite spatial coverage of the EPI scan (102mm thickness), we further excluded the cerebellum in the analysis by subtracting the cerebellar regions derived from the Automated Anatomical Labeling template =19.53, P=.001) and hyperactivity-impulsivity =15.01, P=.003) assessed by the Chinese ASRS and increased total correct hits in RVP =8.8, P=.013), while ADHD participants treated with the placebo had significant improvement only in inattention symptoms from baseline to week 8 =13.45, P=.004) (supplementary Table 2).At baseline, there were no group differences  in demographics, intelligence, or RVP performances except that adults with ADHD had higher clinical symptom severity compared with control participants . After 8F=4.52, P=.045). After treatment, the postplacebo group had higher mean FD compared with the postatomoxetine group =6.06, P=.022), while there was no significant difference between the postplacebo and postatomoxetine groups in maximum head displacement and jerky movement .The 2 comparison groups  were separately matched on the amount of composite movement in terms of mean FD, maximum FD, and number of outliers, in the baseline scans, except that the preplacebo group had higher mean maximum FD compared with the preatomoxetine group , between the right FEF and right parahippocampal gyurs/fusiform (P=.032), and between the right FEF and right middle frontal gyrus . The cognitive control network displayed reduced negative connectivity between the DLPFC and PRE/PCC for the ADHD group relative to the control group (P<.001). Adults with ADHD, compared with the controls, had weaker positive connections in the DMN between the left PRE and right middle temporal (MTG)/fusiform gyrus (P=.02). There was no hypoconnectivity in the affective network and ventral attention network for adults with ADHD compared with the controls .For adults with ADHD, relative to the controls, weaker positive connections were found in the dorsal attention network, between the left FEF and right fusiform/inferior temporal gyrus (P=.02), alongside the left middle frontal gyrus , respectively, adults with ADHD displayed increased negative connections relative to the controls . Adults with ADHD, relative to the controls, did not have significant hyperconnectivity in the cognitive control, ventral attention, affective, and DMNs .In the dorsal attention network between the right FEF and left MTG/angular gyrus ; in the DMN between the left PRE and right middle frontal gyrus, lateral part , and between the PCC and left inferolateral temporal lobe (P=.024); and in the dorsal attention network between the bilateral FEF (P=.008) and orbitofrontal cortex/mPFC (P=.002), respectively. After treatment with atomoxetine, the posthoc analyses demonstrated increased connectivity strength in the DMN between the mPFC and right middle occipital/temporal gyrus (P<.001); in the affective network between the left subgenual ACC and right inferior temporal/middle occipital gyrus (P=.005); in the cognitive network between the left DLPFC and right hippocampus (P=.008); and in the right ventral attention network between the right TPJ and left middle occipital gyrus (P=.037) .Significant time × treatment interactions were extensively found throughout all major brain networks. After treatment with atomoxetine, we detected stronger negative connections in the cognitive control network between the left DLPFC and left superior frontal gyrus, medial part , between the left VFC and left TPJ (P=.001), between the right VFC and left MTG (P=.046), and between the right VFC and left TPJ (P=.017) in the ventral attention network. As hyperactivity/impulsivity improved, increased RSFC was observed in the ventral attention network between the left VFC and TPJ (P=.014) and in the DMN between the PCC and left middle/inferior occipital gyrus (P=.007). We identified negative correlations between symptom improvement and changes in RSFC between the right IPS and PRE in the dorsal attention network . Regarding neuropsychological performances, we observed positive correlations of RVP total hits with increased RSFC between the left TPJ and left middle frontal gyrus/VFC in the ventral attention network (P<.001); between the left DLPFC and right TPJ (P=.019), as well as right precentral gyrus (P=.002), respectively; and between the right DLPFC and left mid-cingulate cortex (P=.001) in the cognitive control network. With improving RVP performances, we identified less connectivity between the PCC and middle occipital/calcarine in the DMN (P=.003). We observed no significant associations between changes in behaviors and RSFC in the affective network in the atomoxetine-treated adults.As shown in To our best knowledge, this is the first study to incorporate rs-fMRI into a randomized clinical trial of atomoxetine in medication-naïve adults with ADHD. We are the first to display causal relationships between atomoxetine use and a strengthening anticorrelated relationship between the task-positive networks  and DMN in adults with ADHD. Moreover, we provide some evidence to support a modulating effect of atomoxetine on all major neural networks in adults with ADHD.Consistent with prior literature , our resP=.053) in adults with ADHD . First, the postplacebo group had higher mean FD than the postatomoxetine group. It may arise from atomoxetine effects on ADHD symptoms. Despite that several strategies were applied to minimize in-scanner head motion impacts, including motion-censure exclusion criteria , component-based denoising method (Several methodological limitations of rs-fMRI should be acknowledged (see the g method , alongsig method , even a g method . Also, tg method  despite g method . Anotherg method . Nonetheg method .Earlier studies employing different preprocessing techniques  suggest Owing to the finite spatial coverage, we excluded the cerebellar regions from the rs-fMRI analyses. However, the cerebellum is structurally and functionally connected with prefrontal and striatal circuits implicated in ADHD . IndividAnother limitation of this study is selection bias of our sample. To increase the internal validity of the sample and explore disease-specific alterations, only participants who were medication naïve and comorbidity free were recruited in the present study. This makes generalization of the current findings to the “real-world” clinical settings indistinct, given the high comorbidity and medication exposure rates in an adult cohort of ADHD . Lastly,This study shows the atypical natures of relationships between functional brain networks and of connectivity within dorsal attention network and DMN in medication-naïve adults with ADHD. We further provide evidence for a mechanism by which atomoxetine therapy strengthens an anticorrelated relationship between the task-positive networks and DMN and modulates RSFC across all major brain networks in medication-naïve adults with ADHD. Our results support the idea that atypical DMN task-positive networks cross talk may contribute to the pathophysiology of ADHD. Strengthening this relationship following atomoxetine treatment suggests an important pathway through which atomoxetine may improve ADHD.Dr. Lin has received grant or research support from National Taiwan University Hospital. Dr. Gau has received grant or research support from the National Science Council, Taiwan, the National Health Research Institute, Taiwan, and National Taiwan University Hospital. All authors declare no biomedical financial interests or potential conflicts of interest.


Attention deficit hyperactivity disorder (ADHD) is a developmental neuropsychiatric disorder characterized by inappropriate levels of inattention, impulsivity and hyperactivity. The cause of ADHD is unknown, but may involve both genetic and environmental factors. It has been suggested that exposure to inflammation in infancy may increase the risk for ADHD in later life. Familial Mediterranean Fever (FMF) is the most common inherited autoinflammatory disorder. In many FMF patients the inflammation persists in attack-free periods. The prevalence of ADHD among FMF patients has not been studied previously.To explore the prevalence of ADHD among FMF patients and to examine the relationship between FMF characteristics and ADHD.The cohort consisted of 103 consecutive children with FMF, followed in a single referral center. Clinical manifestations, demographic and genetic data were abstracted from the patients' medical records, supplemented by information obtained by interviews conducted during routine follow up visits. The presence of ADHD was assessed using the Diagnostic and Statistical Manual of Mental Disorders questionnaire .ADHD was diagnosed in 33 (32.4%) FMF patients, a rate significantly higher that known in our local unselected population (about 8%). The distribution of ADHD subtypes in our patients was similar to the general population: 10 children had predominantly inattentive type (9.8%), 6 hyperactive-impulsive (5.9%) and 17 combined type (16.7%). FMF patients diagnosed with ADHD had a higher rate of arthritis and family history of FMF than patients without ADHD.The high prevalence of ADHD in children with FMF may support the neuroimmune hypothesis, in which inflammatory conditions increase the risk for ADHD. Furthermore, our findings suggest that physicians should be alert to the possible presence of ADHD among FMF patients.


That some of these signatures of coherence in an atom-based system extend to ambient temperature suggests a novel mechanism that could be a synchronized, dynamical, disproportionation excitation, possibly via the solid state analog of a Feshbach resonance that promotes the coherence. Such a mechanism would demonstrate that the use of ultra-low temperatures to establish the BEC energy distribution is a convenience rather than a necessity, with the actual requirement for the particles being in the same state that is not necessarily the ground state attainable by other means. A macroscopic quantum object created by chemical doping that can persist to ambient temperature and resides in a bulk solid would be revolutionary in a number of scientific and technological fields.Bose-Einstein condensates (BECs) composed of polarons would be an advance because they would combine coherently charge, spin, and a crystal lattice. Following our earlier report of unique structural and spectroscopic properties, we now identify potentially definitive evidence for polaronic BECs in photo- and chemically doped UO Because of their formation from the ground states of simple or composite bosons, atomic Bose-Einstein condensates12457910115131617181920f Mott insulator UO2(+x) that we interpreted as the signatures of superfluid droplets of aggregated polarons that self-organized as coherent bosonic states232526282931323334352+x systemWe have previously described structural and both ultrafast optical pump-probe and static x-ray absorption spectroscopy experiments on the chemically and photo-doped, partly filled 5f character of excitations across the Mott gap may cause other free electron “in gap” states (IGS)384245464751525355572+xf-6d electron manifold. By influencing the dynamics of excited states40Partly filled Mott insulators exhibit a variety of fascinating properties. In actinide dioxides in particular, the 5Optical pump-TTDS probe experiments on UO2. The defining property of a condensate is its coherence, spatially over dozens to hundreds of μm and temporally, in conventional BECs, msec. This is manifested statically as evidence that the 103–106 constituent particles are synchronized by, e.g., exhibiting position and momentum distributions much narrower than the expected thermal ones12357146061626364652.00 would be the IGS and/or collective states. On pumping at both 3.14 eV (5f→5f) and 1.57 eV (5f→IGS) the THz spectra display a peak around 1.8 THz  ion will give an EPR spectrum when the U site is distorted either statically to give a symmetric signal or dynamically to give an asymmetric one with widths around 100 G and g values up to 4.6 at 77 K772.16 spectra around 30 K resemble the small regions of those that were reported earlier2 at 10 K. Some spectral features occur in all three samples while others vary with the U:O ratio. In contrast to only the two phases found by diffraction, susceptibility, and other bulk spin probes, the microwave absorbance is consistent with the EXAFS, another local probe, in showing a number of distinct states77X-ray diffraction patterns show the known phase separation between the UOns712.00  matches 2.00 at g = 2.1 for T ≥ 90 K. These exhibit a flat baseline and a signal that is similar in shape to that expected from a geometrically distorted S = 1/2 system. This could be from the individual polarons, whose increasing amplitude with temperature implies that they are forming from a lower energy EPR-silent state. Like most of the features in these spectra, its width is far too large for simple dipolar broadening2.08 and UO2.16 (B/U (257 K), 2.9 μB/U (185 K), and 2.7 μB/U (140 K) for x = 0, 0.08 and 0.16, respectively. The AFM transition occurs at the same temperature for all three compounds. However, the direction is inverted for the doped ones, although the magnitudes of the step are comparable and that of UO2.16 less than UO2.08. It is also suppressed at high fields, being almost eliminated for UO2.08. Since the susceptibility shows that these materials are not ferromagnetic in the normal sense the inversion in direction could signify a change in the canting angle of the U spin vector in the UO2 domains. The canting induced by addition of extra oxygen would be minimal, θ ≈ 5.6 × 10–4/1.74. The abrupt roll over of χ(T) at 6–7 K in lower fields could be another reorientation of the ordered moment. However, this type of flattening or rollover in the magnetization at low T following an abrupt rise was found in TiCuCl3, where it contributed to the interpretation of a magnon BEC in this system2.16 the dip near 3400 G is still present, reaching its greatest amplitude at 180 K similar to the UO2.00 spectra from 90–140 K. However, the rising portion at lower field of this particular signal is absent in the UO2.16 spectrum. This signal in the UO2.08 spectrum appears as a mixture of the two, reflecting the UO2+δ-U4O9–δ phase separation as seen in the diffraction pattern. Two other very broad and complex signals are observed below 25 K in all three samples, albeit in somewhat modified form, at 0–2500 and 3400–5000 G.The spectra that are possibly recognizable are those of UOd UO2.16  there ap.7 μB/U 10 K for x2.08 and UO2.16  exhibit a difference in amplitude throughout the entire sweep that is even superimposed on other features – like cuprates – but with the return downfield sweep higher in amplitude than the upfield one – unlike cuprates. The collective microwave dissipation in UO2+x is therefore of a novel type. The extension of the differential absorption to higher field as the spectral range is increased is a special and possibly unique property of cuprates where the flux line/vortex dis- and reordering from superconducting domains is continuous in energy. The opposite pattern, no change in the field where the hysteresis loop closes so that it is relatively symmetric around zero is characteristic of ferromagnets where the reversal of the magnetization occurs at a well defined energy232428292+x spectra are also notable in not exhibiting any features coincident with the AFM ordering transition. It is therefore unsurprising that these spins or other phenomena that give the spectra are also decoupled from those giving the anisotropy in the thermal conductivity2.00 domains despite the UO2.00 spins being affected by their presence. A notable feature of this separation, however, is that the UO2.00 spin system and the condensate domains are presumably intermingled within the crystal on the scale of the domain sizes. In addition, the many features of the spectra and their complicated temperature dependence are analogous to the RF absorbance of the ultracold fermionic systems, where the different states including those from defects give very similar kinds of behaviors31323334352.00 signal, may therefore originate in precisely the type of phenomena observed in fermionic condensates.Following initial cooling of the UO2+x and the extremely long lifetimes of the collective phases of the excited states and their coherent oscillations in photodoped material2(+x) hosts polaronic condensates. The condensate emerges from individual or aggregated quasiparticles whose collective interactions are enabled by dynamically expanded wavefunctions extending into the UO2 host. This interpretation of the unique oscillation observed in the IGS pump-TTDS probe experiment is corroborated by the presence of distinct mesoscale domains of a coherent quasiparticle fluid, as derived from our prior structural measurements and observed for other quasiparticle condensates5These results validate the conjecture, based on the observed superfluid atom tunneling and broadening of the electronic states in UO2 occurs in its [111] plane832 (111)-type acoustic phonon in which the cubic U(IV/V) “neutron” configuration is more stable when the [111] U planes are on the compressed and the layered U(IV/VI) “x-ray” conformation on the expanded sides, respectively, of the vibrational motion ↔U(IV)+U(VI). This can be combined with the finding that stress relief in UOl motion . Concert2.00 via metal-to-metal charge transfer reactions to produce the required U(V). Another is the formation of the vortices and other states of the system2+x is one more example of the valence-structure-coupled polaron dynamics, but an extreme one whose non-adiabatic spin-charge-lattice fluctuations synchronize to become coherent oscillations of the condensate. The persistence of the condensate to high temperature would therefore occur because this reaction in UO2(+x) is a non-degenerate one to an excited state originating in unique aspects of the structural chemistry of U0.6Se0.416192(+x) the BEC is composed of electrons and not atoms, it is a demonstration that BECs are extensible to the fundamental components of condensed matter212+x relative to UO2.00f band that would be at the Fermi level would produce much more complicated Fermi surface topology than the relatively flat band in UO2.00 before doping and also contribute to forming and stabilizing a condensate via exchange. These results supporting BEC-formation in UO2(+x) are the next step on this path. To the extent that the proposed mechanism is correct, we note that the Feshbach resonance occurs in the crossover or pseudogap regime, which would suggest that the UO2(+x) condensate could be tuned in both the BEC and BCS directions.The coherence and subsequent condensate have additional consequences. One is the superfluidity of the atoms across the oscillation that allows the requisite rapid rearrangement of the O atoms without a barrier. A second is the stability of the condensate that appears to cause the formation of O-enriched domains on irradiation84UO2+x. The UO2 crystal for the TTDS measurements was the same as used earlier and the UO2+x powder samples were prepared identically2 sample was located within this arrangement, but was removed when the original THz pulse was to be measured. The THz pulse was finally focused between our detection slits. Meanwhile, the remaining portion of the original laser pulse passed through a 50/50 beamsplitter. The transmitted half of the beam passed through a delay stage, was rotated to p-polarization, and was eventually focused between the detection slits and combined with the THz pulse. The total pulse energy of this beam could be as high as 300 μJ although this was usually further attenuated with a variable ND filter. The slits had a 500 Hz, 2 kV bias applied and it was to this signal that a lock-in amplifier was locked to. A weak second harmonic field was generated between the slits and modulated by the mixing of the two beams. The second harmonic signal then passed through a 400 nm bandpass filter and was detected with a photo multiplier tube. Together, the THz and 800 nm laser beams allowed a time-domain measurement of the THz waveform with the air biased coherent detection (ABCD) method.In our experiment, we investigated 200 μm thick highly polished uranium dioxide samples. The sample was fixed in a rotational mount to allow the sample orientation to be varied with respect to the laser polarization. A 6 mJ, 40 fs laser pulse was first split with a 90/10 beamsplitter. The stronger portion of the beam was sent through a half-wave plate and then focused with a 12.5 cm focal length lens into an argon filled gas cell for the broadband THz pulse generation .The cell was kept at a pressure of ~700 Torr which was chosen to maximize the THz pulse energy and bandwidth. Also located inside the gas cell was a barium borate (BBO) crystal. This was placed ~2 centimeters before the laser focus to avoid damage to the crystal while also producing a sufficient amount of second harmonic for the THz pulse production. The half-wave plate and the BBO orientation were chosen to provide the maximum THz pulse energy along with a linear p-polarized pulse after the gas cell. Typically we estimated the THz pulse energy at nearly 1 μJ. This was measured by directing the THz photons onto a pyroelectric detector and calibrating the response with a helium-neon laser pulse of known energy. The end of the gas cell contained a silicon filter to remove the residual 800 nm and 400 nm laser light while passing as much of the THz radiation as possible. From there, the THz pulse passed through an 8-f optical arrangement of 2 inch, gold-coated, parabolic mirrors. The UO2 sample. This served as the pump pulse for our experiments. Within this portion of the beam path was also located a half-wave plate, variable ND filter, and another BBO crystal for possible 3.1 eV pumping of the UO2 sample in the future. Additionally, a reflection is seen at ~6 ps resulting from a Fabry-Perot effect from the sample itself. All subsequent data was truncated in the time domain before the reflection to avoid artificial spectral modulations after converting the time-domain signal to the frequency domain with an FFT. Reducing the time window to ~5 ps after time zero reduced the low frequency resolution below 1 THz which was inconsequential to our data accumulation. The pump beam was not collinear with the THz probe beam. The sample was pumped at an angle that could not be captured by the collection optics for the THz, preventing the signal from originating in THz emission stimulated by the pump beam.The remaining portion reflected from the 50/50 beamsplitter also passed through a delay stage and was focused onto the UO2 through the direct frozen-phonon method using the Vienna Ab-initio Simulation Package (VASP) combined with the phonopy package on supercells consisting of 4 × 4 × 4 primitive cells (192 atoms). To obtain a good description of the electronic structure of this highly correlated system, we have used density functional theory including an additional Hubbard term (DFT+U). The gradient generalized approximation (GGA) was chosen for the DFT exchange-correlation functional. In the employed GGA + U approach, the Hubbard and exchange parameters, U and J, respectively, are introduced to account for the on-site Coulomb correlations between the uranium 5f electrons; the +U term also helps to remove the self-interaction error. We have chosen a Hubbard U value of 4.5 eV and an exchange parameter J value of 0.54 eV, which have been shown to provide good results. To deal with the problem of occurring metastable states when using the DFT+U method, we have used the occupation matrix control (OMC) technique proposed by Dorado et al.s, 7s, 6p, 6d, 5f) and  states were treated as valence states. A plane-wave basis with an energy cutoff of 510 eV was used to expand the electronic wave functions. The Brillouin zone integrations were performed on a special k-point mesh generated by the Monkhorst Pack scheme (6 × 6 × 6 in the bulk and 2 × 2 × 2 for the phonon supercells), and a Gaussian smearing of σ = 0.1 eV was used. The electronic minimization algorithm used for static total-energy calculations was a blocked Davidson algorithm.Phonon calculations were performed for UOIn the phonon calculations we corrected the long range macroscopic electric field generated by collective ionic motions near the gamma point by adding a non-analytical term to the dynamical matrix.2+x was sandwiched between two plastic discs whose susceptibility was determined independently and subtracted from the magnetic response of the sample plus discs. In plots shown, χ is defined as the magnetic moment of the sample divided by applied field, M/H.Magnetic measurements were performed in a Quantum Design SQUID magnetometer over the temperature range 2 ≤ T ≤ 350 K and in various fixed magnetic fields between 1 and 50 kOe. For these measurements, a powder of UOAll samples for EPR spectroscopy were prepared in an inert nitrogen atmosphere glove box equipped with a liquid nitrogen fill port to enable sample freezing to 77 K within the glove box. EPR samples were prepared in 4 mM OD thin wall precision Suprasil quartz screw cap EPR tubes from Wilmad Labglass. All samples for EPR spectroscopy were prepared in the solid state. X-band EPR spectra were recorded on a Bruker EMXplus spectrometer equipped with a 4119HS cavity and an Oxford ESR-900 helium flow cryostat. The instrument parameters employed for all samples were as follows: 2 mW power; time constant 41 ms; modulation amplitude 12 G; 9.38 GHz (10K spectra)/9.83 GHz (298K spectra); modulation frequency 100 kHz.How to cite this article: Conradson, S. D. et al. Possible Demonstration of a Polaronic Bose-Einstein(-Mott) Condensate in UO2(+x) by Ultrafast THz Spectroscopy and Microwave Dissipation. Sci. Rep.5, 15278; doi: 10.1038/srep15278 (2015).