For generations, the rhizomes of Drynaria ferns have been used as traditional medicine in Asia. Despite this, the bioactivities of Drynaria rhizomes and leaves have rarely been studied scientifically.Drynaria quercifolia, Drynaria rigidula and Drynaria sparsisora. The phenolic and flavonoid contents of the samples were respectively quantified with the total phenolic content (TPC) and total flavonoid content (TFC) assays, while the antioxidant activities were determined via measuring the DPPH radical scavenging activity (FRS), ferric reducing power (FRP), ferrous ion chelating (FIC) activity and lipid peroxidation inhibition (LPI). The tyrosinase inhibition activity of all three species was also reported.This study evaluates the antioxidant properties of the methanolic extracts of the fertile fronds and rhizomes from three species in this genus: D. quercifolia were found to exhibit the highest overall TPC (2939 ± 469 mg GAE/100 g) and antioxidant activity amongst all the samples, and the fertile fronds of D. quercifolia and D. rigidula exhibited superior TPC and FRP compared to their rhizomes, despite only the latter being widely used in traditional medicine. The fronds of D. quercifolia had high tyrosinase inhibition activity (56.6 ± 5.0 %), but most of the Drynaria extracts showed unexpected tyrosinase enhancement instead, particularly for D. sparsisora’s fronds.The fertile fronds of The high bioactivity of the fertile fronds in the fern species indicate that there is value in further research on the fronds of ferns which are commonly used mostly, or only, for their rhizomes. Interest in phenolic compounds has been on the rise due to their potential human health benefits . They arDrynaria quercifolia (L.) J. Smith is one of the best-known members of this genus, commonly used in Ayurvedic medicine (“Ashwakatri”) [Neisseria gonorrhoeae and Streptococcus-β-haemolyticus [D. quercifolia’s rhizomes, it has been the emphasis of phytochemical-centric research on the species. In comparison, little research has been conducted on the fertile fronds (leaves), which are sometimes used in conjunction with the rhizome to treat tuberculosis and throat infections [The rhizomes of the Drynaria genus of ferns have a long history of being used as traditional medicine, particularly in India , China  and Soutakatri”)  where itakatri”) , diarrheakatri”) . Additioolyticus . Due to fections , or as afections .Drynaria rigidula (Sw.) Beddome and Drynaria sparsisora (Desv.) T. Moore have also been claimed to treat similar diseases as the rhizomes of D. quercifolia, such as diarrhea and gonorrhea [D. quercifolia, there have been no reports on the bioactivity of the fertile fronds of both species. D. rigidula is an endangered species in many locations [D. sparsisora is nearly identical to D. quercifolia in physical appearance, but with shorter fertile fronds [D. rigidula and D. sparsisora fertile fronds and rhizomes have ever been reported, and the comparison between the antioxidant activity of fertile fronds and rhizomes in a fern remains a rarely-explored avenue in most literature. This is also the first time that the tyrosinase inhibition activity for any of these species has ever been reported.The rhizomes of onorrhea . Howeverocations , while De fronds  and a dae fronds . This stD. quercifolia and D. rigidula were obtained from the Putrajaya Botanical Garden, Kuala Lumpur, while the leaves and rhizomes of D. sparsisora were obtained from Sunway, Petaling Jaya. The ferns were identified by plant taxonomist Anthonysamy S., formerly from University Putra Malaysia.The fertile fronds and rhizomes of 18H32O2 , Tween 40 ; tyrosinase inhibition activity: 3, 4-dihydroxy-l-phenylalanine C9H11NO4 , dimethyl sulfoxide analytical grade , kojic acid C6H6O4 , tyrosinase  3400 units/mg solid .The various reagents used throughout this project were purchased from suppliers as follows. TPC analysis: Folin–Ciocalteu’s phenol reagent , gallic acid , anhydrous sodium carbonate ; total flavonoid content (TFC) analysis: aluminium chloride , potassium acetate , quercetin ; diphenyl-2-picrylhydrazyl (DPPH·) assay: 1,1-diphenyl-2-picrylhydrazyl ; ferric reducing power (FRP) assay: ferric chloride hexa-hydrate , potassium ferricyanide , trichloroacetic acid , potassium dihydrogen orthophosphate , dipotassium hydrogen phosphate , iron chloride ; ferrous ion chelating (FIC) assay: ferrozine , ferrous sulphate hepta-hydrate , ethylenediaminetetraacetic acid (EDTA) ; lipid peroxidation inhibition (LPI): β-carotene , chloroform , linoleic acid, CFresh fertile fronds were extracted at a ratio of 50 mL 70 % methanol to 1 g of leaf material after liquid nitrogen-aided crushing. For the rhizomes, 1.0 g of de-skinned rhizome was extracted at a ratio of 50 mL 50 % methanol to 1 g of rhizome. The methanol concentrations chosen for extraction were based on preliminary extraction efficiency screening, where 70 and 50 % methanol were found to be more efficient at extracting the fertile fronds and rhizomes respectively. The extracts were then filtered with a Buchner funnel. The methanolic extracts were stored at −20 °C when not in use.2CO3) solution. The test tubes were then left to stand for 30 min in the dark at room temperature before the absorbance values were measured at 765 nm. The total phenolic content was expressed as mg gallic acid equivalent per 100 g of sample (mg GAE/100 g).The determination of the total phenolic content of the samples was done using a procedure modified from Kähkönen et al.  utilizinFlavonoid content in the extract was determined with the aluminium chloride colorimetric method as described in Chang et al. . Equal v50 of AA used for calculation of FRS was 0.00387 mg/mL.The DPPH· assay was based on the procedures described in Leong and Shui  and Mili3+) to ferrous ions (Fe2+). One mL of sample extract of different concentration (diluted with methanol) was added with 2.5 mL of 0.2 M phosphate buffer (pH 6.7) and the same volume of 1 % (w/v) potassium ferricyanide. The solutions were mixed and incubated in 50 °C water bath for 20 min. Subsequently, 2.5 mL of 10 % trichloroacetic acid was added to stop the reaction. Then, the solution in each test tube was separated into aliquots of 2.5 mL, added with 2.5 mL of Milli-Q water and 0.5 mL of 0.1 % FeCl3. The solutions were mixed and left on bench for 30 min before the absorbance was measured at 700 nm. FRP was expressed as mg gallic acid equivalent per gram of sample, mg GAE/g.The reducing power of the extracts was determined using potassium hexacyanoferrate(III) as described in the procedure described by Tan and Chan . The FRP4 was added to 1 mL of sample of different concentrations , followed by 1 mL of 0.25 mM ferrozine. The mixtures were incubated at room temperature for 10 min before the absorbance was measured at 562 nm. It was expressed as the percentage of iron chelating activity. EDTA (0.017–0.067 mg/mL) was used as a positive control.The determination of ferrous ion chelating strength of the extract was based on the procedures described in Mau et al. , and SinLipid peroxidation inhibition was adapted from Kumazawa et al.  based onl-DOPA and a 96-well microplate for screening of multiple samples simultaneously. For testing of the sample extract, the wells in triplicate were added with 80 μL of buffer , 40 μL of tyrosinase (1 mg/mL of tyrosinase diluted 50-folds with Milli-Q water), and 40 μL of sample (1 mg of freeze-dried extract in 400 μL of 50 % DMSO), and lastly 40 μL of l-DOPA. For the control, the same reagents were added to the wells except that the sample was substituted with 40 μL of DMSO. The blank contained 120 μL of buffer, 40 μL of tyrosinase, and 40 μL of sample. The mixture in each well was mixed and left on the bench for 30 min (after the addition of l-DOPA). Subsequently, the microplate was measured using BIOTEK PowerWave XS Microplate Scanning Spectrophotometer at the absorbance of 475 nm, with reference wavelength at 700 nm. The percentage tyrosinase inhibition activity was calculated using equation below:The measurement of tyrosinase inhibition activity of extracts was based on the method used by Masuda et al. , with slp < 0.05 was considered to be significantly different.Statistical analysis was carried out with one-way ANOVA and the Tukey HSD test was used to identify any significance between the TPC values of the samples. D. quercifolia fertile fronds exhibited a very high TPC of 2939 ± 469 mg GAE/100 g—consistent with previous findings reported by our research group where D. quercifolia fertile fronds ranked second highest in TPC amongst the fifteen ferns screened with an average TPC exceeding 2500 mg GAE/100 g [D. quercifolia had the highest TPC of the three species screened: approximately three times higher than the fronds of D. rigidula and nearly ten-fold higher than the fronds of D. sparsisora; a surprising result given the similar physical appearance of the D. quercifolia and D. sparsisora fronds. The results for the FRS activity (IC50 and AEAC) reflected a similar correlation, with D. quercifolia fronds exhibiting the highest activity, followed by D. rigidula and D. sparsisora. However, despite the higher TPC and FRS in D. rigidula fronds compared to D. sparsisora fronds, the FRP between the fronds of both species were similar thus indicating that phenolic compounds may not necessarily be the only reducing agents present in the fronds of D. sparsisora.The TPC, FRS, FRP and TFC results reported in Table AE/100 g . D. querD. quercifolia and D. rigidula exhibited lower primary antioxidant activity, with lower TPC and weaker FRP activity  activity Fig. , measureD. rigidula showing the highest chelating activity at approximately 45 % at 7 mg/mL  activity is a measure of secondary antioxidant activity—the ability to prevent the formation of radicals, such as those generated by the Fenton reaction in the presence of free ferrous ions . Interes/mL Fig. . This apactivity . The pheD. quercifolia fronds showed a high tyrosinase inhibition activity (exceeding 50 %) [d-glucopyranosyl-(1→2)-β-d-glucopyranosyl-quercetin-3-O-β-d-glucopyranosyl-(1→4)-β-d-glucopyranoside [D. sparsisora, which showed remarkable tyrosinase enhancement activity.As shown in Table ng 50 %) , and mayng 50 %) . Althougranoside . Macrocyranoside . Tyrosinranoside , and as ranoside . It can ranoside . The exaD. quercifolia fertile fronds exhibited the highest TPC, FRS, FRP and LPI. The fertile fronds of D. quercifolia and D. rigidula showed significantly higher TPC and FRP when compared to their rhizomes, with D. quercifolia fronds also exhibiting significantly higher FRS. D. sparsisora rhizomes on the other hand showed similar antioxidant activity with the fronds, with the exception of the rhizome’s FRS being significantly higher than that of the fronds. Interestingly, regardless of the differences in other antioxidant properties, the LPI between the fronds and rhizomes of all three species were comparable at the highest concentration studied. The fronds of D. quercifolia showed good tyrosinase inhibition, while the fronds of D. sparsisora showed remarkable tyrosinase enhancement. These findings may hopefully provide further impetus into looking at the fronds of other ferns which are commonly used mostly for their rhizomes.Of the three fern species screened,


Optimal stimulation theory and moderate brain arousal (MBA) model hypothesize that extra-task stimulation (e.g. white noise) could improve cognitive functions of children with attention-deficit/hyperactivity disorder (ADHD). We investigate benefits of white noise on attention and inhibition in children with and without ADHD (7–12 years old), both at behavioral and at neurophysiological levels.Thirty children with and without ADHD performed a visual cued Go/Nogo task in two conditions (white noise or no-noise exposure), in which behavioral and P300 (mean amplitudes) data were analyzed. Spontaneous eye-blink rates were also recorded and participants went through neuropsychological assessment. Two separate analyses were conducted with each child separately assigned into two groups (1) ADHD or typically developing children (TDC), and (2) noise beneficiaries or non-beneficiaries according to the observed performance during the experiment. This latest categorization, based on a new index we called “Noise Benefits Index” (NBI), was proposed to determine a neuropsychological profile positively sensitive to noise.Noise exposure reduced omission rate in children with ADHD, who were no longer different from TDC. Eye-blink rate was higher in children with ADHD but was not modulated by white noise. NBI indicated a significant relationship between ADHD and noise benefit. Strong correlations were observed between noise benefit and neuropsychological weaknesses in vigilance and inhibition. Participants who benefited from noise had an increased Go P300 in the noise condition.The improvement of children with ADHD with white noise supports both optimal stimulation theory and MBA model. However, eye-blink rate results question the dopaminergic hypothesis in the latter. The NBI evidenced a profile positively sensitive to noise, related with ADHD, and associated with weaker cognitive control. Attention-deficit/hyperactivity disorder (ADHD) is a highly prevalent developmental disorder that affects about 5 % of school-aged children and adolescents –4. TheseWhile stimulant medication has been shown to improve behavioral symptoms  and schoSeveral models, including the optimal stimulation theory , the cogOne possible explanation for why adding stimulation might be beneficial lies in the stochastic resonance (SR) phenomenon . SR is aThe MBA model  is a neuMBA model is corroborated by three studies using a long-term memory task , 26, 40,To our knowledge, the potential effect of a white noise exposure on a visual Go/Nogo task has not been previously examined in children with ADHD, nor at behavioral or at neurophysiological levels. The P300 is an event-related potential (ERP) component that allows different processes (see below) to be studied and is often used as an interest marker in ADHD –47. ERP In this study, the first objective was to compare noise benefit between children with ADHD and TDC in a visual cued Go/Nogo task using both behavioral and neurophysiological measures. This task, in which a non-informative cue precedes each Go or Nogo trial, enables the separate examination of P300 evoked by: (1) preparation (Cue P300); (2) inhibition (Nogo P300); (3) attention and orienting processes . The CuChildren with ADHD were recruited and assessed according to DSM IV-TR criteria  by a mulAt first, 36 children (7–12 years old) were recruited for the study. Three children were excluded because of a too poor signal-to-noise ratio after EEG qualitative observation. Two children were excluded because their estimated IQ was below 80, and one child asked to stop the experiment because it was too long for him. The two groups (ADHD and TDC) consisted then of 13 children with ADHD  and 17 TDC . TDC were recruited from primary schools.Chi square test of independence to assess whether the gender ratio was different between the groups. It showed that the gender ratio was not statistically different between the groups .In addition to the neuropsychological assessment see 2.1.1.), each child performed subtests of the Wechsler Intelligence Scale for Children . Estimated IQ was computed based on two perceptual processing subtests  and two verbal comprehension subtests  of the WISC-IV. Intellectual assessment was aimed at excluding children who presented an intellectual weakness but was not used for group comparison as it is known to be influenced by attentional and executive factors , 56. Par.1., eachInformed consents were obtained from all subjects and from their parents with the prior approval of the Ethics Committee of the Queen Fabiola Children’s University Hospital , of the Erasme Hospital  and of the Faculty of Psychology and Education .Data was acquired during two separate sessions: a neuropsychological assessment and an experimental session.During this session, we used the computerized TAP battery , 60 and IQ testing as well as the completion of the CBCL questionnaire and the informed consent was performed during this session.In the visual cued Go/Nogo task, the child was submitted to three kinds of 3 × 3 cm stimuli briefly displayed 150 ms), in black, one by one on a grey background. A square  always preceded Go (“×”) or Nogo (“+”) stimuli. Go and Nogo stimuli each had a 50 % of probability of following a cue and were pseudo-randomly displayed  from 14 channels , embedded in a waveguard cap (10–20 system) and all referred to the mean of the two mastoids. Horizontal and vertical eye movements were monitored using two bipolar recordings: one between each outer eye canthus and one between a supraorbital electrode and an electrode positioned just below the lower eyelid on the left side. The ground was placed on the left wrist. All impedances were kept below 10 kΩ. After amplification (×20) and online filtering , the inAfter the experimental task, children were asked to keep their eyes open to record eye-blinking rate, during two blocks of 2 min, one with and one without white noise exposure .Independent samples t tests were used to assess group differences (ADHD vs. TDC) with regard to IQ, CBCL questionnaire as well as scores from the TAP and from the Counting Stroop.In the TAP subtests, subject’s median RT was chosen as a measure of response latencies because it is less sensitive than the mean to the enhanced intra-individual variability in response time usually observed in the ADHD population . AnotherTwo variables were computed to investigate the classical interference effect in the Counting Stroop, i.e. the difference scores  betweenDependent variables were omissions , false alarms , RTs and the RT variability.Subjects were assigned to two kinds of groups. First, they were assigned according to their diagnosis status: “ADHD” or “TDC”. Second, they were assigned according to their benefit from noise or not during the task. Therefore, we computed a NBI that calculated the difference of omissions, for each subject, in noise vs. no-noise conditions (i.e. the percentage of hits in the noise condition minus the percentage of hits in the no-noise condition). Children with a higher NBI index (i.e. more hits/less omissions in the noise condition) were grouped as “noise beneficiaries” (n = 12) and the others (same or less hits/more omissions in the noise condition) were assigned to the “non-beneficiaries” group (n = 18). Mean and standard deviation (SD) for the NBI scores in each group are: ADHD ; TDC , noise beneficiaries ; noise non-beneficiaries .Chi square test of independence to assess whether the gender ratio was different between the groups (“noise-beneficiaries” and “non-beneficiaries”). It showed that the gender ratio was not statistically different between the groups . Age did not statistically differ between these groups .A median split, another possible way to create two groups regarding noise benefit, might have assigned participants who made more omissions in the noise condition to the “noise beneficiaries” group. Therefore, the former method, although not perfect, was considered more appropriate. As gender influences symptom expression and the cognitive profile , we perfFor all dependent variables, two mixed ANOVA’s were performed with between Group factors: (1) ADHD vs. TDC and (2) “noise beneficiaries” vs. “non-beneficiaries”. Within-subjects factors were block (first vs. second block) and condition (noise vs. no-noise).Chi square test of independence was applied to examine the potential relation between ADHD and benefiting from noise. Aside from the Chi square test, an independent samples t test was used to assess the difference in the mean of the NBI score between ADHD and TDC groups. Pearson correlations were used to examine relationships between the NBI and neuropsychological scores.Continuous EEG was segmented in 1200 ms epochs including a 200 ms pre-stimulus onset baseline. Averaged waveforms were computed for each subject and then across groups for each of the following trials: “Go” (“×”), only when the subject had responded, “Nogo” (“+”), only when the child did not respond and “cue” (square).Blinks were corrected with the SOBI algorithm  to avoidFor each “Type of P300” , two mixed ANOVA’s were performed for mean amplitude with Groups: (1) ADHD vs. TDC and (2) “noise beneficiaries” vs. “noise non-beneficiaries” as between-subjects factors. “Site”  and condition (noise vs. no noise) were within-subject factors.Spontaneous eye-blink rates (after the task) were analyzed with Matlab 2012. A free Matlab script, “peakdet” ( cited inWe chose to register spontaneous eye-blinks after the main experiment (and not during) because our goal was to make an indirect inference between dopaminergic functioning and white noise. While eye-blinks during a task seem to reflect the transition of activation between different neural networks , spontanA repeated measures ANOVA was performed for eye-blink rates with Groups: (ADHD vs. TDC) as between subject factor and condition (noise vs. no noise) as within-subjects factor.All statistical analyses were performed using “Statistica 8.0”. When necessary, post hoc Tukey tests were applied.As illustrated in Table In the Alertness TAP subtest, analyses disclosed larger mean median RTs in ADHD than in the TDC group in tonic  and phasic alertness condition . Coefficient of variation was also higher in ADHD for tonic  and phasic alertness . ADHD children had less correct responses  and made more anticipations  than the TDC group.In the Go/Nogo TAP subtest, coefficient of variation  and the number of errors  were larger in the ADHD than in the TDC group.In the Counting Stroop task, analyses disclosed significantly larger time interference  and errors difference indices  in ADHD than in TDC.All results of the TAP tests and the Counting Stroop are shown in Table Relevant behavioral and electrophysiological data are presented according to the comparison between ADHD and TDC groups in Section 1 and according to the comparison between “noise-beneficiaries” and “non-beneficiaries” groups in Section 2. Block factor was included in each behavioral analysis but will not be presented, as the results were not relevant in this context.Behavioral measuresOmissionsChildren with ADHD made more omission errors than TDC . There was no main effect of Condition , but a significant Group*Condition interaction  = 9.98, p = .004] an more variable  than TDC. All other factors did not reach significance and did not interact with each other .FA cueChildren with ADHD committed more FA cues than TDC . No other factor reached significance and there was no interaction .FA nogoNo factor reached significance and there was no interaction .Electrophysiological measuresCue P300Cue P300 amplitudes were similar regardless of the Group, Condition or Site and there was no interaction .Go P300Go P300 amplitudes were similar regardless of the Group, Condition or Site and there was no interaction .Nogo P300There was a Group*Site interaction  and a three-way interaction Group*Condition*Site , indicating that ADHD had a significant higher Nogo P300 than TDC but only at Pz in the noise condition (p = .05). No other factor reached significance and there was no interaction .In this section, children who benefited from noise (noise-beneficiaries) were compared to those who did not (non-beneficiaries), independently of their diagnostic status. Given that we split our groups according to the fact that children benefited from noise on omissions errors, analyzing behavioral effects on these omissions would be redundant and was not included. Here we present analyses on electrophysiological because no relevant behavioral analysis reached significance. The clinical status * noise benefit * block * condition four-way mixed ANOVA would have allowed quantifying the interaction effect of clinical status and noise benefit on our dependent variables. However, we did not follow this approach because the two types of group assignment yielded an imbalanced design with cells with low sample sizes  = 9.62, p = .005] indicating, as illustrated in Fig. There was no relevant result for Group, Condition or Site factors as regards Cue and Nogo P300.All significant correlations between NBI and neuropsychological tests (TAP and Counting Stroop) are presented in Table These correlations indicate a relationship between benefitting from noise and lower attention and inhibition (motor and cognitive) in the neuropsychological tests.The Chi square test highlighted a significant relationship between the Group factor (ADHD or TDC) and benefitting from noise . There was no other significant effect or interaction .In this study, we first aimed to evaluate the potential cognitive benefit of white noise exposure during a visual cued Go/Nogo task in children with ADHD and TDC. The second objective was to examine, through the use of a new index (NBI), whether potential correlations could be observed between noise benefit in the visual cued Go/Nogo task and individual neuropsychological profiles. The third objective was to investigate whether this NBI had a neurophysiological correlate, i.e. whether differences in the P300 component can be observed between “noise-beneficiaries” and “non-beneficiaries”. The fourth objective was to discuss, through the use of spontaneous eye-blink rates, the Moderate Brain Arousal model according to which white noise modulates dopaminergic functioning .Regarding participant characteristics’, ADHD and TDC differed in the CBCL only in the ADHD problems subscale, indicating a limited influence of comorbidities on results. Results from the attentional and executive assessment showed significant differences between children with ADHD and TDC. Children with ADHD were slower and had higher RT variability on all neuropsychological tasks , confirming previous observations –73. TheySimilar results were obtained in the experimental task. Children with ADHD made more omissions, more impulsive errors (i.e. FA cue) and had slower and more variable RTs than TDC. However, exposure to white noise during the experimental task showed a positive impact on cognitive performance in children with ADHD. To the best of our knowledge, this is the first demonstration of a beneficial effect of white noise in the Go/Nogo paradigm in a study comparing children with and without ADHD. Positive effects of white noise have already been documented on memory  and on GElectrophysiological data showed a significant Group*Condition*Site interaction with a higher Nogo P300 amplitude in ADHD than in TDC at Pz in the noise condition. This result might indicate enhanced inhibitory processes in children with ADHD in the noise condition. However, behavioral results did not show a similar effect, as we did not observe a difference between groups according to noise exposure. Possibly, this Nogo P300 underlines a positive neurophysiological effect of white noise that is not observed at the performance level (see e.g. ). More sWhen groups were assigned according to their noise benefit (noise-beneficiaries and non-beneficiaries), we demonstrated a significant relationship between group classification (ADHD or TDC) and benefiting (or not) from noise. In addition, we found significant correlations between benefitting from noise and markers of vigilance  and motor/cognitive inhibition (anticipation errors and interference errors) in Stroop and TAP tasks . These markers have been identified as core cognitive symptoms in ADHD .Furthermore, noise-beneficiaries had a marginally larger Go P300 mean amplitude in the noise condition compared to the no-noise condition. This was not found in participants who didn’t benefit from noise. Interestingly, white noise modulates only, at an electrophysiological level, the Go P300 mean amplitude (for noise-beneficiaries), which is associated with attentional processes (like vigilance); and white noise modulates only vigilance at a behavioral level  in children with ADHD. We propose two possible complementary explanations: (1) White noise, perceived as a potential distractor, could make noise beneficiaries (prone to distraction regarding their correlations with markers of vigilance) gathering up more attentional resources not to be distracted. (2) The perceptual load hypothesis  proposesFinally, noise benefit on cognitive functioning supports the MBA model. However, it is unclear whether this noise improves performance through dopamine system modulation, as suggested by Sikstrom, Soderlund . We addrSome limitations of our study have to be mentioned including, first, the small number of participants. Second (probably related to the first), the three-way interaction showing a larger Go P300 for the noise beneficiaries in the noise condition was only a trend. Third, in ERP studies on cognition, it is recommended to use two-year groupings over the age of 8 years because of significant ERP changes over a short time period . Due to The finding of a white noise benefit in children with ADHD during a Go/Nogo task validates the optimal stimulation theory. However, the benefits of white noise are not to be generalized to all analyzed functions within this task and seem more specifically associated with vigilance improvement. The NBI allowed us to characterize a neuropsychological profile, related to that of ADHD, positively sensitive to noise at both behavioral and electrophysiological levels. We also provide neurophysiological correlates of noise benefit. The eye-blink rate investigation distinguishes ADHD between TDC groups, but was not sensitive to white noise, questioning the influence on dopaminergic functioning suggested by the MBA model. With regard to the literature, the type of extra-task stimulation has a different impact on cognitive functioning according to task requirements and interpersonal differences , 86. Fut


The characterization of samples were carried out using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and ultraviolet–visible absorption spectroscopy (UV-vis). The results show that the Bi2S3 layer is uniformly formed on the surface of WO3 nanoplates and less interfacial defects were observed in the interface between the Bi2S3 and WO3. More importantly, the Bi2S3/WO3 films as photoanodes for photoelectrochemical (PEC) cells display the enhanced PEC performance compared with the Bi2S3/WO3 films prepared by a sequential ionic layer adsorption reaction (SILAR) method. In order to understand the reason for the enhanced PEC properties, the electron transport properties of the photoelectrodes were studied by using the transient photocurrent spectroscopy and intensity modulated photocurrent spectroscopy (IMPS). The Bi2S3/WO3 films prepared via an in situ approach have a greater transient time constant and higher electron transit rate. This is most likely due to less interfacial defects for the Bi2S3/WO3 films prepared via an in situ approach, resulting in a lower resistance and faster carrier transport in the interface between WO3 and Bi2S3.In this study, Bi Photoelectrochemical (PEC) and photocatalytic (PC) water splitting could be two potential approaches to counter these challenges because solar energy is a clean and inexhaustible energy source133) with a large band gap (2.6 ~ 3.0 eV) has been extensively investigated, because of its excellent photocatalytic activity, high electron mobility and nontoxic nature8343 platelike films show an excellent PEC performance and have attracted intensive attention363738404142As a well-known photocatalyst, tungsten trioxide (WO2S3) is a small band gap semiconductor (~1.3 eV) and has attracted great interest as a sensitizer for PEC and photovoltaic cells recently due to its large absorption coefficient and reasonable IPCE2744452S3 is more negative than that of WO3472S3/WO3 heterojunction photoelectrode. Recently, we have reported on the synthesis of Bi2S3/WO3 photoelectrode by a sequential ionic layer adsorption reaction (SILAR) process and the photoelectrodes exhibit an excellent PEC activityin-situ growth method is considered as a good way to prepare ideal heterojunction with perfect interfacial contact50512WO6 synthetized from WO3 can be used as interim product and Bi2S3 can be obtained from Bi2WO6 by a hydrothermal process. Thus, it is feasible to the formation of Bi2S3/WO3 heterojunction by in-situ growth method.Bismuth trisulfide  dissolved in 30 mL of deionized water at room temperature. Then, 10 mL of 3 M HCl was added to the solution under constant stirring, followed by the addition of 0.2 g of ammonium oxalate ((NH4)2C2O4). After several minutes of stirring, 30 mL of deionized water was added into it with continual stirring for 0.5 h. The as-prepared precursor was transferred into a 100 mL of Teflon-lined stainless autoclave. The FTO substrates with the conducting side facing down were immersed and leaned against the wall of the Teflon-vessel. The hydrothermal synthesis was carried out at 140 °C for 3 h. The as-prepared films were calcined at 500 °C for 1 h.All chemicals were analytical grade. WO2WO6/WO3 films were prepared through a simple soaking process. The prepared WO3 films were soaked in 0.2 M glacial acetic acid solution of Bi(NO3)3·5H2O for 12 h. To ensure the same thickness Bi2WO6 layer for all samples, the WO3 film was pulled from the solution with a pulling rate of 3 mm/s, not rinsing. Next, the films were dried in 25 °C for 1 h and then calcined at 520 °C in air for 4 h.The Bi2S3/WO3 films could be obtained from hydrothermal Bi2WO6/WO3 film. 0.1 g thiourea as the sulphur source was dissolved in 50 mL of DI water at room temperature. Then, 100 μL of 2 M HCl was added to the solution under constant stirring. Next, the solution was transferred into an 80 mL of Teflon-lined stainless autoclave. The Bi2WO6/WO3 film was placed into the autoclave. The hydrothermal synthesis was carried out at 140 °C for 4 h. The as-prepared films were dried at 200 °C for 2 h. For comparison, we also prepared another kind of Bi2S3/WO3 and Bi2S3 films by the SILAR method according to our previous work2S3/WO3 films prepared by the SILAR method have been annealed at low-temperature  after the Bi2S3 nanoparticles deposition.The Bi3 and Bi2WO6/WO3 film and the Vis-NIR diffuse reflection spectra of Bi2S3/WO3 film were obtained using the UV-Vis 2450 (Shimadzu) and Vis-NIR U-4100 (Hitachi) spectrophotometer, respectively. The diffuse reflection spectra could convert into the absorption spectra by Kubelka-Munk function. Scanning electron microscope  was used to observe the surface morphology of samples. Transmission electron microscopy  was operated at an accelerating voltage 200 kV to investigate the microstructure and the crystallinity of samples. On account of the nonuniformity of Bi2S3 thickness in s-Bi2S3/WO3 film . The samples with 2 cm2 were soaked in 5 ml of 1 M HNO3 solution for 12 h and the Bi2S3 would be dissolved completely. The amount of Bi2S3 in the film was obtained to characterize the concentration of Bi3+ using ICP-MS. The concentration for Bi2S3/WO3 films prepared via an in situ approach and the SILAR method is 12.26 and 10.78 mg L−1, respectively. The PEC properties of samples were investigated in a typical three-electrode electrochemical cell using an electrochemical analyzer . The synthesized films were employed as the working electrode and a platinum foil and an Ag/AgCl/satd. KCl electrode were employed as the counter and reference electrodes, respectively. All electrochemical tests were conducted in an aqueous solution containing 0.1 M Na2S and 0.1 M Na2SO3 (pH ≈ 9). The illumination source was a 150 W xenon lamp  with a 400 nm cutoff filter to remove UV irradiation. IPCE measurements were carried out using a xenon lamp  with an AM 1.5 filter and a monochromator with a bandwidth of 5 nm. Intensity modulated photocurrent spectroscopy (IMPS) were recorded by using a Zahner CIMPS-2 system. A white light lamp emitting diode (λ = 540 nm) driven by a PP210 was used as lamp and the light intensity is adjusted to 50 mW cm−2. Light intensity modulation was driven by current modulation with a depth of 10%. The bias potential is at −0.4 V vs. Ag/AgCl during the IMPS measurement.The crystalline phase of the sample was characterized by X-ray powder diffraction . The UV-Vis diffuse reflection spectra of WOWO3 film , the amo2S3/WO3 films by in-situ growth method and surface morphology change of sample for each step are shown in 3 arrays films were obtained via a hydrothermal process. The SEM images of platelike WO3 film in 2WO6 was formed on WO3 nanoplate by a simple soaking and calcined process. The SEM images of Bi2WO6/WO3 film shown in 2WO6/WO3 is different from that of the pristine WO3. Compared with the pristine WO3, the surface of Bi2WO6/WO3 nanoplates became rough and the surface textures reduce obviously. It may because the formation of the bigger molecule Bi2WO6 lead to the surface swell. Third, Bi2S3/WO3 films can be obtained from hydrothermal Bi2WO6/WO3 film. During the hydrothermal process, the S2− can be generated by the decomposition of thiourea, and then the Bi2WO6 will react with the S2− to form the Bi2S3 on the surface of WO3 nanoplates2S3/WO3 nanoplates is rougher than that of Bi2WO6/WO3.The depiction of the fabrication process for Bi3 nanoplate is clearly shown in the low-resolution TEM image (3 (3 (JCPDS 83-0950). The HR-TEM image recorded on the rim of a Bi2WO6/WO3 nanoplate was shown in 2WO6. In addition, the lattice fringes with the spacing of 0.384 and 0.335 nm are in good agreement with the (002) and (120) planes of WO3, respectively. The TEM results for the Bi2WO6/WO3 nanoplate can confirm that Bi2WO6 layer can be formed on the surface of WO3 plate via an in situ approach. The HR-TEM image of the interface between Bi2S3 and WO3 is shown in 3 (JCPDS 83–0950) and the lattice spacing of 0.312 nm is assigned to the (121) plane of Bi2S3 (JCPDS No. 84–0279). The two corresponding fast Fourier transform patterns  was used to identify the elaborate structure of the composite nanoplates. Typical WOEM image . The uniimage . The three small diffraction peaks of the Bi2WO6/WO3 sample can be observed at 28.38, 33.05 and 47.16o, attributed to the characteristic diffraction peak of Bi2WO6 (JCPDS No. 73–2020). In addition, many weak diffraction peaks observed from the Bi2S3/WO3 sample, are identical to Bi2S3 (JCPDS No. 84–0279). The result indicates the formation of Bi2S3 on the surface of WO3 plate. The optical behavior of the prepared films was evaluated by using UV-vis and Vis-NIR absorption spectroscopy. 3, Bi2WO6/WO3 and Bi2S3/WO3 film. For the pristine WO3, a clear absorption edge about 460 nm can be observed, corresponding to its indirect band gap energy , the i-Bi2S3/WO3 photoelectrodes show a higher photocurrent density (8.0 mA cm−2 at −0.1 V vs Ag/AgCl). In addition, the stability of i-Bi2S3/WO3 photoelectrode is tested by performing long-duration PEC experiments. The testing experiment is carried out at the −0.1 V vs Ag/AgCl under continuous illumination and lots of H2 bubbles can be found on the surface of Pt electrode throughout the entire test. The result is shown in The PEC measurements were implemented in a three-electrode PEC cell using the prepared films as photoanodes under illumination of visible light. Linear sweep voltammetry (LSV) was employed to evaluate the PEC performance of films. For comparison, the BiTo investigate the quantitative correlation between the wavelength of the incident light and the PEC activity, incident photon-to-current conversion efficiency (IPCE) measurement was performed at a bias of −0.5 V vs. Ag/AgCl. The IPCE were estimated by the following relation15I is the photocurrent density, λ is the incident light wavelength, and Jlight is the incident light power density. The IPCE of the WO3, s-Bi2S3/WO3 and i-Bi2S3/WO3 photoelectrodes are shown in 3 photoelectrode exhibits the photoresponse only at the wavelength range of ~460 nm, because of the large band gap of WO3 (≥2.6 eV). Relative to the pristine WO3 photoelectrode, the photoelectrode decorated by Bi2S3 shows enhanced IPCE in the entire testing wavelength region due to the increased absorption by the Bi2S3. Importantly, the i-Bi2S3/WO3 photoelectrode shows the highest IPCE value, which is in accordance with the LSV result.Where 2S3/WO3 photoelectrode has an enhanced PEC property, the electron transport properties of the photoelectrodes were investigated by using intensity modulated photocurrent spectroscopy (IMPS) analysis. 2S3/WO3 and i-Bi2S3/WO3 photoelectrodes at −0.4 V vs. Ag/AgCl. The IMPS is popularly used to characterize the electron transport of PEC cells54τd) is the average time photogenerated electrons need to reach the back contact and can be obtained from the IMPS result by the following formula192856In order to understand the reason that the i-Bifmin is the frequency at the imaginary minimum. τd of the s-Bi2S3/WO3 and i-Bi2S3/WO3 photoelectrode are 15.87 and 3.80 ms, respectively. The results suggest the electron transit rate in i-Bi2S3/WO3 photoelectrode is faster than in the s-Bi2S3/WO3 photoelectrode. It may be attributed to the stronger contact and lower resistance in the interface between Bi2S3 and WO3 for the i-Bi2S3/WO3 photoelectrode. To further verify the IMPS results, we also measured IMPS at −0.6 and −0.2 V vs. Ag/AgCl and the results are shown in τd of i-Bi2S3/WO3 photoelectrodes are smaller than that of s-Bi2S3/WO3 photoelectrodes at the same potential due to the higher fmin.where τt) of photoelectrodes can be calculated and obtained from the transient photocurrent plots by using the kinetic equations as follows585960To further support the above ideas, the transient photocurrent plots were measured at −0.7 V and −0.5 V vs. Ag/AgCl and were shown in D is defined asWhere I is the current, i and f are related to the initial and final steady states, and t denotes time, respectively. The transient time constant, τt, can be defined as the time at ln D = −1. Generally, a longer transient time constant implies a smaller extent of recombination and the transient time may be treated as a lifetime of the photogenerated carriers. As shown in τt values of the s-Bi2S3/WO3 and i-Bi2S3/WO3 photoelectrode are shown in 2S3/WO3 photoelectrode has longer the transient time than that of the s-Bi2S3/WO3 photoelectrode, suggesting a smaller extent of recombination for the i-Bi2S3/WO3 photoelectrode. This may be because of the faster carries transit rate for the i-Bi2S3/WO3 photoelectrode, resulting in the reducing of the recombination of the photogenerated carriers.Where 3 and Bi2S3 to their conduction band (CB). The accumulated electrons at the CB of Bi2S3 easily move to the CB of WO3 due to the more negative CB of Bi2S3, and then are collected by the FTO conductor3 and Bi2S3 will be transferred to the semiconductor/electrolyte interface in the opposite direction and react with S2− to avoid the photocorrosion of Bi2S3. The electrons in the FTO are migrated to the Pt electrode/electrolyte interface by the external bias voltage and will reduce the H2O to H2. It should be noted that the PEC cells discussed here need the presence of Na2SO3 and Na2S as the sacrificial reductants to steadily generate H2, which has been widely reported15243133To better understand the charge separation and transfer process in the PEC cells, the schematic illustration of the charge transfer of the photoelectrode is illustrated in 2S3 sensitized WO3 nanoplate arrays films via an in situ approach. The film characterization results suggest that the Bi2S3 layer was uniformly formed on the surface of WO3 nanoplates. The prepared films were used as photoanodes and the PEC performance was studied. The Bi2S3/WO3 films prepared via an in situ approach have a higher photocurrent density (8.0 mA cm−2 at −0.1 V vs Ag/AgCl) than that of the Bi2S3/WO3 films prepared by SILAR method. This may be due to the stronger contact interface for the Bi2S3/WO3 films prepared via an in situ approach, resulting in a higher electron transit rate and the reduced photogenerated carrier recombination. This versatile preparation method has potential to be applied in the synthesis of other hybrid films with less interfacial defects.In conclusion, we have synthesized BiHow to cite this article: Liu, C. et al. In situ synthesis of Bi2S3 sensitized WO3 nanoplate arrays with less interfacial defects and enhanced photoelectrochemical performance. Sci. Rep.6, 23451; doi: 10.1038/srep23451 (2016).


Attention-deficit hyperactivity disorder (ADHD) is a common neurodevelopmental disorder that affects the child and adolescent population. It is characterized by impairment in attention/concentration, hyperactivity, and impulsivity, all of which can impact performance of athletes. ADHD treatment within the athletic population is a unique challenge. The research in this field has been relatively limited. The National Collegiate Athletic Association and International Olympic Committee both regulate the use of psychostimulants for treatment of ADHD due to their performance-enhancing effects. In this article, authors have discussed the screening methods, pharmacological treatment, side effects, and behavioral approaches for the treatment of ADHD in adolescent athletes. Attention-deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder that affects about 6–7% of the child and adolescent population . ReportsFor this article, we searched the following databases for the period of January 1985, to December 2013: Medline, PsychINFO, Ovid, PubMed, and Cochrane Library. We used the following search terms: “ADHD,” “adolescents,” “athletes,” and “sports.” Key articles were selected on the basis of topic covered, and emphasis was placed on articles published after 2000. Reference sections of identified publications were also searched for relevant information. The current article is supplemented with the authors’ clinical knowledge, and the references list was modified on the basis of suggestions from peer reviewers.Diagnostic and Statistical Manual of Mental Disorders-III (DSM-III) was introduced. ADHD is a chronic disorder that affects multiple areas of life including education, work, family, and social functioning. If left untreated, ADHD can have long-lasting detrimental effects on the individual’s development and psychosocial functioning.Descriptions of ADHD in the literature go back to the beginning of the twentieth century. This disorder was further conceptualized and refined at the time that the Diagnostic and Statistical Manual of Mental Disorders-IV-TR identifies inattention, hyperactivity, and impulsivity as the three cardinal symptoms of this disorder  and International Olympic Committee (IOC). The NCAA updated their drug testing medical exception procedures in 2008 and now requires more stringent documentation for ADHD diagnosed athletes. It allows for stimulant use by athletes with adequate documentation of ADHD diagnosis and evidence of continued follow up . Stimulant use by Olympic athletes for ADHD and narcolepsy has been approved recently under the TUE. Stringent rules apply and approval by two independent consultants is required.Athletes on continued stimulant use must obtain a TUE from the World Anti-Doping Agency (WADA) prior to competing internationally . AthleteThe true prevalence of ADHD in athletes is not known, as among the athletes who have this condition, few are open about it. Despite the risks, a few athletes have come forward and discussed their condition openly: Michael Phelps, the multiple Olympic gold medalist, was diagnosed with ADHD at 9 years of age; Terry Bradshaw is a pro football Hall-of-Famer who quarterbacked the Pittsburgh Steelers to four Super Bowl victories in the 1970s; Pete Rose became a World Series MVP in 1975; Cammi Granato scored more goals than any other player in the history of U.S. women’s hockey and led her team to a gold medal at the 1998 Winter Olympics in Nagano, Japan. These are just a few examples of the many professional athletes who have excelled despite their disabilities related to ADHD .As noted above, stimulant medications have the tendency to increase heart rate and blood pressure . ExercisDuring the last few years, concerns have been raised about the effects of stimulant medications on the cardiovascular system. In 2005, Health Canada suspended the sale of Adderall XR following reports of sudden deaths in children taking this medication. The U.S. Food and Drug Administration (FDA) later issued a warning about the use of these medications in children with pre-existing heart disease. The effects of stimulants on the heart and their association with sudden cardiac death are still being debated, but in 2008, the American Heart Association (AHA) published monitoring guidelines that are currently being followed by most physicians .For varied reasons, the use of stimulant medications in athletes has been an area of concern. It is known that ADHD diagnosed athletes can benefit from these medications, but because of the above-mentioned concerns, a thorough cardiac evaluation needs to be done before the start of medications and before participation in sports , 68. TabAtomoxetine is a non-stimulant medication that is approved by the FDA for the treatment of ADHD in children. It can be an alternative medication in competitive sports where stimulants are completely banned despite the clinical necessity. Strattera’s side effect profile does not increase the chance of cardiac problems. On the other hand, in 2005, the FDA issued a boxed warning for this drug because of reports of increased suicidality in individuals taking this medication . AtomoxeClonidine and guanfacine are two other alternatives to stimulants for the treatment of ADHD. Clonidine is the more sedating of the two, but both can cause this undesirable side effect for athletes. Cardiovascular parameters including blood pressure, heart rate, and EKG need to be evaluated before the start of these medications . These mFor one reason or an other, neither stimulant nor non-stimulant medications may be the initial choice for adolescent athletes. This is especially true for adolescents with aspirations to compete on the national or international level. In these individuals, implementation of a dedicated behavioral program can help to improve mental capabilities and to overcome ADHD symptoms, namely inattention and impulsivity . ProblemMild brain stimulation techniques and neurofeedback (NF) improve brain wave imbalances, facilitating a calmer mental state and therefore better communication throughout the brain . NF, whiAttention-deficit hyperactivity disorder is a common disorder that affects millions of children all over the world. These children suffer from inattention, hyperactivity, impulsivity, poor self-esteem, and academic and social problems. Because of these symptoms and deficits in motor control, their performance in sports is also impaired . StimulaThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Plantago virginica L. has invaded many lawn ecosystems in the Eastern part of China. The invasion has incurred an economic cost to remove them. In order to prevent the invasion, it is critical to understand the invasive mechanisms of this species. However, few studies have been conducted on the allelopathic mechanisms of its invasion. In this study, we examined allelopathic effects of P. virginica on germination of seeds and growth of seedlings of four widely used lawn species. We found extensive allelopathic potential of P. virginica on other lawn species, which varied with species and developmental stage. While most effects of the extracts of P. virginica were inhibitory, some variables in some species were promoted by the addition of the extracts. The extracts of P. virginica significantly inhibited seed germination of Agrostis matsumurae. While the overall differences in seed germination rate of Poa annua were significant among treatments, difference between control and any of the treatments was not significant. The height of seedlings of A. matsumurae and Cynodon dactylon was significantly lower under the treatments of adding extracts of P. virginica. In contrast, growth of seedlings of Festuca elata and P. annua did not show significant differences among treatments. The root length of A. matsumurae, C. dactylon and P. annua was suppressed by the extracts of P. virginica whereas root length of F. elata was not affected. Aboveground biomass of A. matsumurae and F. elata was significantly higher than control, except for F. elata at the concentration of 50mg/mL, whereas aboveground biomass of C. dactylon and P. annua was reduced at higher concentrations of the extracts. Except for A. matsumurae, root biomass of the other three lawn species declined under the treatments with the extracts of P. virginica. Our results revealed that P. virginica had allelopathic potential on four lawn species and supported the theory of “novel weapons hypothesis”. Invasion by P. virginica in lawn can be moderated by selecting those species that are not affected or promotionally affected by it. Biological invasion has become a great concern in the world. Many studies had expatiated the impacts of invasive species on native species, ecological environments, and incalculable economic loss from different aspects. Invasion threatens the biodiversity of the world , 2, 3, 4Solidago canadensis in its introduced range has been proven a key mechanism for its successful invasion [et al. found Phragmites australis, one of the most aggressive invaders, had significant negative effects on germination and growth of a native species Melaleuca ericifolia [Acacia dealbata produced an allelopathic substance, which influenced soil microbes, with bacteria being more sensitive to it than fungi [Ambrosia artemisiifolia had allelopathic effects on some but not all native species [Lonicera maackii interacted with biotic and abiotic conditions [Alliaria petiolata that impeded seed germination of Brassica rapa was in seeds [Carduus nutans and Carduus acanthoides that inhibited growth of lettuce, was found only in roots [Successful invasion by a species may be a result of interaction between the invasive species and native species , 24. Allinvasion . Uddin eicifolia . Acacia an fungi . Ambrosi species . Allelopnditions . Allelopin seeds , but aplin roots .Plantago virginica L. is a native annual species in North America and its introduction to China might be unintentionally [P. virginica was first found in Jiangxi province of China in 1951 [P. virginica in lawns has obvious effects on the landscape and possible profound effects on the ecosystems. As a result of invasion by P. virginica, large amounts of money and labor are needed to remove them.Invasion by alien species will remain a significant challenge in the future due to the globalization of human society . Therefotionally . P. virg in 1951 , but now in 1951 , 41. It P. virginica, some studies have been conducted on this invasive species. Most of the researches were focusing on its biological or physiological characteristics, population dynamic and reproductive ecology [P. virginica could be replaced after some time of establishment if no more disturbances happened [P. virginica has an r-strategy of reproduction with more investment into reproductive organs [P. asiatica . P. virginica allocated more resource to reproductive growth than P. asiatica under aluminium stress [aluminium stress than P. asiatica [P. virginica [With the growing concerns on  ecology , 43. Amohappened . P. virge organs , resultim stress  and chloasiatica . Warmingirginica .P. virginica has caused economic loss. Here we conducted a study to investigate possible mechanism for successful invasion of P. virginica in lawn ecosystems. Our hypothesis is that P. virginica may have allelopathic potential on other species in lawns and this helps its establishment and becoming the dominant species in the lawns. Our study is also aimed to give some managerial suggestions to control P. virginica and maintain landscape.Other possible mechanisms on how this species becomes successfully invaded is rarely studied and therefore poorly understood. It is crucial to thoroughly understand the mechanisms that facilitate its invasion to prevent or remove them, especially in lawn ecosystems where the invasion by P. virginica including roots in the lawn of Shanghai Institute of Technology  in May, 2013. No specific permission was required for those locations where we took the plant samples. And the field work did not involve endangered or protected species at this location. Plants were washed, dried, milled and sieved with 40–80 mesh. 200g milled powder was put into a 5000mL wide-mouth bottle, and 2000 mL of 50% ethanol solution were added into the bottle. After fully stirring, the sample was kept at room temperature for 48hrs to extract substance from P. virginica powder. The extraction solution was filtered by Buchner funnel. The filtrate was collected and concentrated with rotary evaporator . After ethanol was completely evaporated, the remaining extracted substance was re-dissolved in distilled water to a total volume of 2000mL. The extracts were stored in a 4°C refrigerator before application. The original concentration of the extracts was 100mg (milled powder)/mL. We further diluted the extracts to create five gradients of concentration: 0, 25, 50, 75 and 100mg/mL in our experiment treatments. Compounds contained in the extracts were analyzed with Agilent 7890–5975 gas chromatograph-mass spectrometer (GC-MS) system  after dissolved in trichloromethane. The compounds in the extracts were shown in We took samples of whole individuals of We didn’t use activated carbon in this study because it has been proven that activated carbon can also absorb substance other than allelochemicals, and change soil properties and soil microbial population structure , 52, 53.Agrostis matsumurae Hack. ex Honda, Cynodon dactylon (Linn.) Pers., Festuca elata Keng ex E. Alexeev and Poa annua L. were used in this study to investigate the allelopathic potential of P. virginica on these species.Four commonly used lawn species, P. virginica of five different concentrations, 0mg/mL, 25mg/mL, 50mg/mL, 75mg/mL and 100mg/mL, were added to the filter paper. The Petri dishes were kept in an incubator for seeds to germinate. The temperature was 24°C and the humidity was 80%, with light turned on for 12 hrs per day. Each concentration treatment of each species had 3 replicates, making a total of 60 samples (four species * five concentration * three replicates). We calculated the germination percentage when no new germination would occur.100 seeds of each species were put onto the filter paper in a Petri dish that was 10cm in diameter. The extracts of P. virginica that were same as in the germination experiment were applied into soils. Meanwhile the soils were watered with the same amount of tap water to keep the soils at a water content that was favorable for seedlings to grow. There were also 60 pots in total (four species * five concentration * three replicates). The pots were held in a greenhouse with a controlled temperature of 24°C and humidity of 80%. After growing for 2 weeks, the height of plants was measured. Then plants were removed from the soils and washed for measurements of root length. The plants were then dried at 60°C to a constant weight to determine the aboveground and root biomass.Seeds of each species were sowed into sandy soil in a pot of 10cm in diameter. After germinating, 40 individuals were kept in the pot and the rest of individuals were removed. Five concentrations of the extracts of One-way analysis of variance (ANOVA) and post-hoc Fisher tests were used to analyze the significance of the differences in germination rate, growth and biomass of seedlings among treatments. We tested the normality and homogeneity of variance before conducting ANOVA analysis. All of the statistical analysis was performed with Statistic 6.0.P. virginica had significant effects on the germination rates of A. matsumurae and P. annua, but not on those of the rest two species  is (are) responsible for the inhibitory or promotional effects because we used the extracts of whole plants of P. virginica. Actually potential allelopathic substance is diverse. It could be only one compound or several compounds together to inhibit the germination of seeds or growth of seedlings of the other species. Silva et al. found that it was a kind of secondary metabolites in Carduus, aplotaxene that had allelopathic effects [Centaurea maculosa by Inderjit et al., (±)-catechin was responsible for allelopathic potential [Lonicera maackii that functioned as allelopathic substance [While we have found some allelopathic potential of invasive  effects . In a stotential . Howeverubstance .et al. found that ethyl phydroxybenzoate, p-hydroxybenzaldehyde, dhurrin, and apigenin of Sorghum halepense in invaded area in China had allelopathic effects, but their concentrations have changed a lot compared with those reported by Czarnota et al. in 2003 [P. virginica between invaded area and its original habitat (unpublished data). For example, with the rutin as a standard substance, we analyzed the content of total flavonoids in the leaves of the native and invasive P. virginica. The content of total flavonoids of native and invasive P. virginica was 1.51% and 2.56%, respectively. Another possibility is that invasive species can release a totally new substance that does not exist when grown in its original environment. Further studies are needed to separate the specific substance that causes allelopathic potential of P. virginica.It is also possible that the content of some extant substance has changed largely after a species invaded into a new area and it is the increased substance that acts as allelopathic chemical. Liu  in 2003 , 57 in t in 2003 , 59. We P. virginica. In addition, so far most noticeable invasion by P. virginica occurs in lawn ecosystems. This provides an opportunity for the managers of the lawn engineering to select those lawn species that are insensitive to the extracts of P. virginica in order to avoid invaded by P. virginica. For example, from our study, at earlier stage of growth, i.e., seed germination, C. dactylon and F. elata would not been affected by P. virginica. During later stage of seedling growth, F. elata was less affected by P. virginica. Both seedling height and root length of F. elata remained unchanged under treatments of extract addition, but its biomass was significantly affected. Considering the biomass, A. matsumurae would be better choice for lawns because root biomass was not influenced and aboveground biomass was stimulated by the extracts of P. virginica. However, as we can see from the above recommendations, different conclusions may be drawn when different variables are taken into account. A comprehensive evaluation is necessary to select a better species for lawns from the perspective of preventing invasion by P. virginica.As shown above, some variables of some species were not affected or even promoted by use of the extracts of P. virginica on other species is inhibitory but we did find some promotional effects. Fox example, aboveground biomass of A. matsumurae and F. elata was higher under the treatments with addition of the extracts of P. virginica. This suggests that some native species derive benefits from the invasion of P. virginica [Most allelopathic potential of invasive irginica . We may P. virginica is known as a pioneer plant in its native habitats [P. virginica has changed the microenvironment. The altered micro-environment, in turn, was more favorite for its successful invasion.habitats . Pioneerhabitats , 62, 63,habitats , 65. OurPlantago in the U.S., P. lanceolata, was native in Europe. Recently its seeds were developed as health products. In China, many species of the same genus have historically and extensively been used as materials of Chinese medicine due to its medicinal functions [P. asiatica [P. major and P. lanceolata. However, there is no report about active compounds in P. virginica. Therefore, we should amplify research in this area. After we know more about its active compounds, it is possible to utilize P. virginica for medical purpose or developing health products. At the same time we utilize it, we can also control its invasion.One species of unctions , 67. Lotasiatica , 72, 73,S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.


The present study investigated the relationship between different types of childhood maltreatment  and the symptoms of attention deficit hyperactivity disorder (ADHD) in young adulthood.Data were collected from a Danish national study conducted by The Danish National Centre for Social Research in 2008 and 2009. A sample of 4,718 young adults (24 years of age) were randomly selected using the total birth cohort of children born in 1984. Structured interviews were conducted with a response rate of 63%, equating to a total sample size of 2,980 participants.Chi-square analyses revealed significant relationships between child maltreatment groups and a probable diagnosis of ADHD using the Adult ADHD Self-Report Scale (ASRS). Binary logistic regression analysis showed that the overall abuse class was more strongly associated with probable ADHD (OR=5.08), followed by emotional abuse (OR=3.09) and sexual abuse (OR=2.07).The results showed that childhood maltreatment was associated with increased risk of ADHD symptoms in young adulthood. The findings of this study are discussed within the existing literature and suggestions for future research are outlined in order to replicate these findings in other adult populations.Child maltreatment is associated with higher levels of ADHD symptoms in a nationally representative sample of young Danish adults.Co-occurring (multiple) types of maltreatment display stronger associations with ADHD symptoms with attenuated effects for sexual abuse.Males confer stronger associations with ADHD symptoms than females.Findings underscore the importance of exploring the role of childhood trauma in young adults with ADHD symptoms. The effects of abuse on a child's development and behavior are complex and multifaceted. Several factors can potentially harm and disrupt the natural development of children, and child maltreatment covers a broad spectrum of factors that can affect a child's emotional and psychological wellbeing. According to developmental theory and research Perry, , child mMany individuals, who were previously defined as having a combination of attentional and behavioral problems, are now receiving a clinical diagnosis as ADHD, and being treated with prescription drugs. The prevalence of ADHD in Denmark among children 0–5 years old has increased by more than 600% in the last 10 years  in order to find distinguishing and overlapping symptoms . A child protection case was objectively defined as a case where the council had provided support for the child and the family or placed the child with a foster family due to concerns about the wellbeing and development of the child. This assessment took place according to files of the local social workers.A stratified random probability survey was conducted in Denmark between 2008 and 2009. Statistics in Denmark randomly selected 4,718 participants born in 1984. The Danish Research Council founded the study that was approved by the Danish Data Protection Agency. To increase the number of participants who had experienced childhood abuse and neglect, children who had been assigned child protection service status by the Danish authorities was oversampled using a 1:2 ratio , being unreachable (13%), or illness or disability (2%). The sample consisted of 1,579 males (52%) and 1,401 females. The majority of the sample either owned or rented their own private accommodation (93.7%), and almost half was married or cohabiting (46.0%).Participants were asked 20 questions across four domains of childhood maltreatment: physical abuse ; psychological abuse ; neglect ; and sexual abuse . Four latent classes of abuse emerged: non-abused, emotionally abuse, sexual abuse, and abused overall. Retrospective reports on abuse or neglect by the parents or guardians were obtained on all four of these latent classes, utilizing items that described different experiences of physical neglect, emotional abuse, neglect, or serious sexual abuse =40.07, p<0.001). The overall abuse group reported higher levels of ADHD symptoms  with minimal differences to the emotional abuse group  and sexual abuse group . Post hoc analysis indicated that only the non-abused group differed significantly from the three abuse groups with much lower levels of ADHD .A one-way analysis of variance (ANOVA) was conducted on the abuse typologies and overall ADHD score. The results indicated significant differences between the abuse groups on ADHD scores 2(3) =88.44, p<0.001]. Cross tabulations revealed that the overall abuse latent class was especially associated with ADHD symptoms, with a total of 35.9% scoring higher than the cut off score on the ASRS  participants were above the cut off, scoring symptoms compatible with a probable ADHD diagnosis. A Chi-square analysis indicated significant association between childhood trauma groups and ADHD symptoms . Males were significantly more likely than females to report probable ADHD (OR=1.56). The three abuse classes showed a significantly increased risk of developing ADHD symptoms relative to the non-abused class. The strongest effect was evident in the overall abuse class with over a fivefold increase in probable ADHD. Participants in the emotional abuse class were over three times more likely to develop ADHD symptoms and in the sexually abused class were twice as likely to report probable ADHD.The aim of this study was to explore the association between different types of childhood maltreatment and ADHD symptoms. Similar to previous research, the results revealed a strong relationship between survivors of different kinds of childhood abuse and ADHD indicators. Overall, individuals whom had self-reported history of abuse also reported higher ADHD symptoms.The findings indicate that different forms of child maltreatment conferred a 2- to 5-fold increase in risk for adult ADHD symptoms. A possible explanation for these findings is that the potential symptom domains  are comparable in both ADHD and PTSD  either ADHD youth or abused children.This study is not without limitation. First, several definitions of maltreatment and abuse have been presented in literature, presenting problems comparing the present study with previous findings in the child maltreatment field. The definitions of ADHD are even more diverse, and the diagnosis is used widely and across several dimensions of severity. This study also cannot determine specifically if the individual's responses lead to an ADHD diagnosis, as many of the examined symptoms can relate to a range of psychiatric disorders, for example, anxiety disorders, PTSD, or ODD (Ford et al., The present study utilized a latent class analysis to identify a meaningful set of latent subgroups characterized by different experiences of childhood maltreatment. This provided an opportunity to examine outcomes that may be associated with certain types of maltreatment Additionally, co-occurrence of two or more types of abuse often happens, hence resulting in a more severe and broad range of possible consequences. Finally, this study presented the opportunity to examine the association between childhood maltreatment and symptoms of ADHD in young adults using a large national representative sample.The work was no supported by any funding agency.


Barrett's esophagus (BE) is a commonly undiagnosed condition that predisposes to esophageal adenocarcinoma. Routine endoscopic screening for BE is not recommended because of the burden this would impose on the health care system. The objective of this study was to determine whether a novel approach using a minimally invasive cell sampling device, the Cytosponge, coupled with immunohistochemical staining for the biomarker Trefoil Factor 3 (TFF3), could be used to identify patients who warrant endoscopy to diagnose BE.A case–control study was performed across 11 UK hospitals between July 2011 and December 2013. In total, 1,110 individuals comprising 463 controls with dyspepsia and reflux symptoms and 647 BE cases swallowed a Cytosponge prior to endoscopy. The primary outcome measures were to evaluate the safety, acceptability, and accuracy of the Cytosponge-TFF3 test compared with endoscopy and biopsy.In all, 1,042 (93.9%) patients successfully swallowed the Cytosponge, and no serious adverse events were attributed to the device. The Cytosponge was rated favorably, using a visual analogue scale, compared with endoscopy (p < 0.001), and patients who were not sedated for endoscopy were more likely to rate the Cytosponge higher than endoscopy . The overall sensitivity of the test was 79.9% (95% CI 76.4%–83.0%), increasing to 87.2% (95% CI 83.0%–90.6%) for patients with ≥3 cm of circumferential BE, known to confer a higher cancer risk. The sensitivity increased to 89.7% (95% CI 82.3%–94.8%) in 107 patients who swallowed the device twice during the study course. There was no loss of sensitivity in patients with dysplasia. The specificity for diagnosing BE was 92.4% (95% CI 89.5%–94.7%). The case–control design of the study means that the results are not generalizable to a primary care population. Another limitation is that the acceptability data were limited to a single measure.The Cytosponge-TFF3 test is safe and acceptable, and has accuracy comparable to other screening tests. This test may be a simple and inexpensive approach to identify patients with reflux symptoms who warrant endoscopy to diagnose BE. Barrett's esophagus is a condition in which the cells lining the esophagus (the tube that transports food from the mouth to the stomach) change and begin to resemble the cells lining the intestines. Although some people with Barrett's esophagus complain of burning indigestion or acid reflux from the stomach into the esophagus, many people have no symptoms or do not seek medical advice, so the condition often remains undiagnosed. Long-term acid reflux , obesity, and being male are all risk factors for Barrett's esophagus, but the condition's exact cause is unclear. Importantly, people with Barrett's esophagus are more likely to develop esophageal cancer than people with a normal esophagus, especially if a long length (segment) of the esophagus is affected or if the esophagus contains abnormally growing “dysplastic” cells. Although esophageal cancer is rare in the general population, 1%–5% of people with Barrett's esophagus develop this type of cancer; about half of people diagnosed with esophageal cancer die within a year of diagnosis.Early detection and treatment of esophageal cancer increases an affected individual's chances of survival. Thus, experts recommend that people with multiple risk factors for Barrett's esophagus undergo endoscopic screening—a procedure that uses a small camera attached to a long flexible tube to look for esophageal abnormalities. Once diagnosed, patients with Barrett's esophagus generally enter an endoscopic surveillance program so that dysplastic cells can be identified as soon as they appear and removed using endoscopic surgery or “radiofrequency ablation” to prevent cancer development. However, although endoscopic screening of everyone with reflux symptoms for Barrett's esophagus could potentially reduce deaths from esophageal cancer, such screening is not affordable for most health care systems. In this case–control study, the researchers investigate whether a cell sampling device called the Cytosponge coupled with immunohistochemical staining for Trefoil Factor 3  can be used to identify individuals who warrant endoscopic investigation. A case–control study compares the characteristics of patients with and without a specific disease. The Cytosponge is a small capsule-encased sponge that is attached to a string. The capsule rapidly dissolves in the stomach after being swallowed, and the sponge collects esophageal cells for TFF3 staining when it is retrieved by pulling on the string.The researchers enrolled 463 individuals attending 11 UK hospitals for investigational endoscopy for dyspepsia and reflux symptoms as controls, and 647 patients with Barrett's esophagus who were attending hospital for monitoring endoscopy. Before undergoing endoscopy, the study participants swallowed a Cytosponge so that the researchers could evaluate the safety, acceptability, and accuracy of the Cytosponge-TFF3 test for the diagnosis of Barrett's esophagus compared with endoscopy. Nearly 94% of the participants swallowed the Cytosponge successfully, there were no adverse effects attributed to the device, and those participants that swallowed the device generally rated the experience as acceptable. The overall sensitivity of the Cytosponge-TFF3 test (its ability to detect true positives) was 79.9%. That is, 79.9% of the individuals with endoscopically diagnosed Barrett's esophagus were identified as having the condition using the new test. The sensitivity of the test was greater among patients who had a longer length of affected esophagus and importantly was not reduced in patients with dysplasia. Compared to endoscopy, the specificity of the Cytosponge-TFF3 test (its ability to detect true negatives) was 92.4%. That is, 92.4% of people unaffected by Barrett's esophagus were correctly identified as being unaffected.The case–control design of this study means that its results are not generalizable to a primary care population. Also, the study used only a single measure of the acceptability of the Cytosponge-TFF3 test, Nevertheless, these findings indicate that this minimally invasive test for Barrett's esophagus is safe and acceptable, and that its accuracy is similar to that of colorectal cancer and cervical cancer screening tests. The Cytosponge-TFF3 test might, therefore, provide a simple, inexpensive way to identify those patients with reflux symptoms who warrant endoscopy to diagnose Barrett's esophagus, although randomized controlled trials of the test are needed before its routine clinical implementation. Moreover, because most people with Barrett's esophagus never develop esophageal cancer, additional biomarkers ideally need to be added to the test before its routine implementation to identify those individuals who have the greatest risk of esophageal cancer, and thereby avoid overtreatment of Barrett's esophagus.http://dx.doi.org/10.1371/journal.pmed.1001780.Please access these websites via the online version of this summary at Barrett's esophagus and gastroesophageal reflux diseaseThe US National Institute of Diabetes and Digestive and Kidney Diseases provides detailed information about esophageal cancer (in English and Spanish)The US National Cancer Institute provides information for patients and health professionals about Barrett's esophagus  and about esophageal cancerCancer Research UK (a non-profit organization) provides detailed information about gastroesophageal reflux and on esophageal cancer The UK National Health Service Choices website has pages on the complications of Heartburn Cancer Awareness Support is a non-profit organization that aims to improve public awareness and provides support for people affected by Barrett's esophagus; the organization's website explains the range of initiatives to promote education and awareness as well as highlighting personal stories of those affected by Barrett's esophagus and esophageal cancerdiagnosis and management of Barrett's esophagusThe British Society of Gastroenterology has published guidelines on the guidelines for gastroesophageal refluxThe UK National Institute for Health and Care Excellence has published Barrett's Esophagus Campaign is a UK-based non-profit organization that supports research into the condition and provides support for people affected by Barrett's esophagus; its website includes personal stories about the conditionThe  In a multi-center case-control study, Rebecca Fitzgerald and colleagues examine whether a minimally invasive cell sampling device could be used to identify patients who warrant endoscopy to diagnose Barrett's esophagus. It is estimated that 5% to 15% of adults in the Western world suffer from reflux symptoms . On the The importance of BE lies in its potential to progress to esophageal adenocarcinoma through intermediate dysplastic stages. Although the absolute risk of malignant progression is low ,10, thisThresholds for endoscopy referral have been the subject of vigorous debate. The major US medical societies and British Society of Gastroenterology recommend endoscopic screening for patients with multiple risk factors for esophageal adenocarcinoma, i.e., reflux symptoms, male, age above 50, high body mass index (BMI), and high waist-to-hip ratio ,13. OnceHence, there is a renewed interest in determining whether a more systematic diagnosis of BE coupled with endoscopic therapy could lead to lower mortality from esophageal adenocarcinoma. Endoscopic screening for all patients with reflux symptoms would not be affordable or justifiable for most health care systems. There is therefore a need for a more systematic, cost-effective, and patient-friendly diagnostic test for BE. We have developed a minimally invasive non-endoscopic cell collection device, called the Cytosponge, coupled with immunohistochemical staining for a single biomarker, Trefoil Factor 3 (TFF3), that could provide such a test. The device was piloted previously and consists of a 30-mm polyurethane sponge, contained within a capsule, which is attached to a string . The caphttp://www.stard-statement.org/; see We conducted a case–control study to determine the sensitivity and specificity of the Cytosponge-TFF3 test for the detection of BE compared with endoscopy and biopsy as the reference standard see . The stun = 13) were crossed over to the case arm. Participants were recruited from across 11 centers that were geographically dispersed across the UK , with documented histopathological evidence of intestinal metaplasia (IM) on at least one biopsy in the course of their endoscopic history. Controls were individuals referred to endoscopy because of dyspepsia and/or reflux symptoms. Participants who were initially enrolled as control patients but were then diagnosed with BE at endoscopy , and symptoms; administered the Cytosponge; and then performed an endoscopy procedure. Following each procedure, acceptability measures were collected. The Cytosponge was administered by a research nurse, and the whole process, including instructing the patient, took less than 10 min. Twenty-seven nurses were involved in administering the device over the course of the study across the different sites. After retrieval, the Cytosponge was placed in SurePath Preservative Fluid  and kept at 4°C until transportation to the laboratory.BE patients who happened to undergo a second surveillance endoscopy for clinical purposes during the study period were invited to swallow the Cytosponge again.The primary outcomes were sensitivity and specificity of the Cytosponge-TFF3 assay for detecting BE compared with endoscopy, as well as the safety and acceptability of the device. Secondary endpoints were the sensitivity of the TFF3 screen in patients with dysplasia and in patients with BE who swallowed the Cytosponge on more than one occasion.Anonymized samples were processed to paraffin blocks and cut into consecutively numbered sections. Immunostaining was carried out on slides 2 and 15 for TFF3  using standard protocols on a BOND-MAX autostainer  according to GCP/GLP standards in the Tissue Bank laboratory housed within Addenbrooke's Hospital, Cambridge. One of two independent researchers (E. W. or C. S. R.) and a gastrointestinal histo-cytopathologist (M. O.) scored the slides in a binary fashion as either positive (≥1 TFF3-positive goblet cell) or negative. The expert histo-cytopathologist had the final say on TFF3 status. The scorers were unaware of the clinical diagnosis of the patient.The gastroscopies were carried out by a local study endoscopist within an hour of the Cytosponge procedure, and patients had the choice of sedation or local anesthetic spray. A strict protocol was followed. Biopsy samples were taken from the cardia as well as 2 cm above the squamocolumnar junction in all participants, and in BE cases diagnostic biopsies were collected following the recommended Seattle surveillance protocol . DiagnosAcceptability for both the Cytosponge and endoscopy was recorded by the participant using a visual analogue scale in which 0 represented the “worst experience ever,” 10 represented the “best experience ever,” and 5 indicated “neither pleasant nor unpleasant” . This acp-values were two-sided. Statistical analyses were carried out using Prism V5.01, SPSS version 19, or R version 3.Statistical analyses were conducted by the Cancer Prevention Trials Unit (B. V. N. and P. D. S.). Assuming a sensitivity of 80% and a specificity of 90% for the TFF3 screen, we needed to recruit at least 600 BE patients and 450 controls to ensure a 95% confidence interval of approximately ±3%. Statistics for continuous variables were expressed as medians and interquartile ranges (IQRs). The Mann-Whitney test and Kruskal-Wallis test were used to compare continuous and ordinal variables, respectively, between groups, and Fisher's exact test and the Cochran-Armitage trend test  were useTP53 mutations in Cytosponge samples can be used to diagnose high grade dysplasia (HGD) when compared with samples from patients without BE and patients with BE who had never developed dysplasia over a median follow-up of 6 y [TP53 mutations, we have modeled this in a hypothetical population of 10,000 individuals attending primary care for investigation of reflux symptoms. We used a sensitivity of 79.9% and a specificity of 92.4% for detection of any length of BE; for TP53 mutation detection we performed the calculations using a sensitivity of 86% (95% CI 65%–96%) and a specificity of 100% (95% CI 94.6%–100%) for detection of HGD.We recently demonstrated that analysis for p of 6 y . To undeIn all, 463 controls with dyspepsia and reflux symptoms and 647 BE cases were recruited; Three serious adverse events were reported under the Good Clinical Practice guidelines, none of which were attributed to the Cytosponge. One patient had onset of atrial fibrillation during radiofrequency ablation that was performed at the same time as the study endoscopy. One patient was admitted for a bleed from a biopsy site, which stopped spontaneously. One patient was found to have unsuspected esophageal varices during the endoscopy, but no evidence of bleeding or Cytosponge abrasions were seen. Three days later the patient had a hematemesis, and the varices were banded. It was not possible to determine the trigger for bleeding in this case.In terms of adverse events, 16.7% of patients had oozing of blood from a Cytosponge abrasion site (noted by the endoscopist); however, these abrasions did not require any intervention. The oozing was at worst similar to that seen at a biopsy collection site. There were anecdotal reports of a short-lived sore throat.p = 0.003). Using a fully adjusted logistic regression, increasing BMI was associated with more difficulty swallowing the Cytosponge; however, the age and gender of the patients had no impact. Of the 6%  of patients who were unable to swallow the device, 11.8% (8/68) were also unable to tolerate endoscopy.93.9% of patients  swallowed the Cytosponge successfully. The proportion of patients who were unable to swallow the device was significantly higher in the case arm than in the control arm . In all, 258/748 patients (34.5%) rated the procedures equally, and 276/732 patients (37.8%) preferred the Cytosponge. On the whole, men rated the Cytosponge better than endoscopy , and patients who were not sedated for endoscopy were more likely to rate the Cytosponge higher than endoscopy .When asked to rate the Cytosponge procedure using a visual analogue scale for acceptability with 0 representing the worst experience ever, 5 representing a neutral score, and 10 representing the best experience ever, 689/746 patients (92.4%) scored the experience as 4 or more, and 729/746 patients (97.7%) gave a score of 3 or more (mildly unpleasant or better), with a median value of 6 IQR 5–8; . OverallA diagnosis of BE using the Cytosponge relies on adequate sampling of the esophageal mucosa and the presence of TFF3 positivity. p < 0.001; p = 0.253; p < 0.001; p = 0.24, sensitivity range 91% to 98%).The sensitivity for diagnosing BE increased with the circumferential length of the BE segment up to 87.2% .The study design allowed for patients with BE to swallow a second Cytosponge if they attended for a further clinically indicated endoscopy during the time course of the study. As shown in TP53 mutations in Cytosponge samples can be used to diagnose HGD [TP53 testing, between zero and 40 patients would require endoscopy. Those testing negative for TP53 mutations would be scheduled for a repeat Cytosponge, expected to be between 2 and 5 y later, but the optimal interval is still to be determined. Similarly, for the true TFF3-positive patients with BE , between two and 14 cases would test positive for TP53 mutations, and approximately two of these would be confirmed dysplastic at endoscopy. Hence, following risk stratification, the endoscopy burden in the TFF3-positive arm would be significantly reduced from 974 to between two and 54 cases. On the other hand, one would expect 9,026 patients to test TFF3 negative, of which 63 would be false negatives. If sensitivity was felt to be paramount and these patients were given a second Cytosponge-TFF3 test, then one would find another 33 true positives at the expense of 681 false positives (based on our repeat Cytosponge-TFF3 test data). However, after risk stratification of these positive cases using TP53 sequencing, again the number of endoscopies would be dramatically reduced to between two and 39  next-generation sequencing approach, we recently demonstrated that screening for nose HGD . We modeThe Cytosponge-TFF3 test is safe and generally acceptable to patients with symptomatic reflux or dyspepsia undergoing investigation, and for those patients with BE undergoing surveillance. The test has a sensitivity of around 80% for diagnosing BE; sensitivity increases with BE segment length and is not compromised in the presence of dysplasia. The specificity of the test is 92%.In the previous pilot study in which the primary endpoints were feasibility, safety, and acceptability, 504 patients were recruited . AlthougThe Cytosponge-TFF3 test sensitivity for diagnosing BE increased to nearly 90% when the device was swallowed a second time. The commonest reason for a false-negative test (76%) was the absence of columnar cells on the specimen, rather than failure of the TFF3 antibody to recognize BE when columnar cells were present. Hence, in clinical practice, it would be possible to advise patients with a TFF3-negative test and no or very few columnar cells seen on the specimen to attend for a repeat Cytosponge-TFF3 test, similar to being recalled for a repeat cervical smear because of an inadequate specimen. Alternatively, the Cytosponge-TFF3 test could be performed twice in everyone, in a manner analogous to the serial fecal occult blood testing undertaken in some countries as part of a colorectal cancer screening program. It is also possible that the device could be engineered to maximize collection of cells at the gastroesophageal junction.p = 1.0). However, the correlation between false-positive TFF3 results and the presence of Helicobacter pylori nearly reached significance, with a p-value of 0.0825. H. pylori infection may lead to the presence of IM within the stomach, which may have been missed on the gastric cardia biopsy used to assess the presence of cardia IM, but which may nevertheless have been sampled by the Cytosponge. It is also known that up to 10% of patients presenting to endoscopy for dyspepsia have a diagnosis of gastric IM, a known precursor lesion to gastric cancer [False positives are also an important consideration for any screening or diagnostic test. Analysis of our Cytosponge-TFF3 test data in which there were 34/445 (7.6%) false positives suggests that these were not due to IM at the gastric cardia, nor were they due to very short tongues of BE. Patients with BE tongues of 1 cm or less with no IM present on biopsies were included in the control cohort as these patients are likely to have irregular Z lines, but there was no correlation between these types of patients with very short Barrett tongues and false-positive TFF3 results . In terms of sample logistics, the samples were stored at 4°C and could be kept for several weeks prior to processing. The sample processing and antibody staining were performed in a laboratory housed within the hospital pathology department, and as such followed clinical laboratory standards. Because of the binary nature of the TFF3 scoring, this test would be amenable to automated image scanning. Overall, this study was conducted using methods relevant to routine clinical practice, and nothing emerged that would seem to preclude the implementation of such a diagnostic test.The Cytosponge-TFF3 test could be used as a first-line diagnostic test in patients with multiple risk factors for BE  wWhile the case–control design was appropriate to obtain accuracy data with narrow confidence intervals, this design has known limitations in terms of the ability to generalize the data to the primary care population consulting for reflux symptoms. Care must be taken not to extrapolate the negative and positive predictive values directly. In addition, the acceptability data were limited to a single measure. In the future it will be important to collect more in-depth qualitative data on acceptability.The Cytosponge-TFF3 test can diagnose BE in a manner that is acceptable to patients and logistically feasible across multiple centers. This test may substantially lower the threshold for investigating patients with reflux, as part of a strategy to reduce population mortality from esophageal adenocarcinoma. Analysis of a full panel of risk-stratification biomarkers enriched further for dysplasia and applied to this cohort is ongoing, and a randomized controlled trial of the Cytosponge-TFF3 test compared with the current standard of care for patients with reflux and dyspepsia symptoms in primary care is in preparation. The data from such studies will be required prior to implementation of the Cytosponge-TFF3 test on a population level.S1 Checklist(DOC)Click here for additional data file.S1 Fig(PPTX)Click here for additional data file.S1 Protocol(PDF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.