Music and language are human cognitive and neural functions that share many structural similarities. Past theories posit a sharing of neural resources between syntax processing in music and language Patel, , and a d Music and language are both universal human cognitive functions, but the degree to which they share cognitive resources is a long-standing debate in cognition. Theorists have argued for a shared evolutionary origin Mithen, , as wellOne theory that has influenced research in the structures of music and language is the Shared Syntactic Integration Resource Hypothesis (SSIRH), which postulates an “overlap in the neural areas and operations which provide the resources for syntactic integration” Patel, . The hypOne piece of evidence in support of the shared processing of musical and linguistic syntax comes from a reading time study in which musical and linguistic syntax were manipulated simultaneously . Twenty five (52%) participants identified as female, and 23 as male. Thirty eight (79.1%) reported that their first language was English, three were native speakers of English and one other language, and seven had a language other than English as their first language. Other than English, participants' first languages included Chinese (Mandarin), Arabic, Thai, Japanese, Spanish, French, German, Vietnamese, and Bengali. Sixteen participants (33.3%) spoke more than one language. All participants had normal or corrected-to-normal vision and reported being free of psychiatric or neurological disorders. Informed consent was obtained from all subjects as approved by the Ethics Board of Psychology at Wesleyan University.Fifty six undergraduate students from Wesleyan University participated in this study in return for course credit. A recording error resulted in the loss of data for 8 out of the 56 total students, and so 48 participants' data were used in the final analysis. Of the remaining participants, all reported normal hearing. Twenty eight participants (58.3%) reported having prior music training, averaging 6.8 years . Reading time (RT) comparisons between different conditions were controlled for length of segment because the critical regions are always the same number of words , 3 musical syntax violation trials, 1 linguistic syntax violation trial, 5 musical syntax plus rhythmic violation trials, 1 linguistic plus musical syntax violation trial, 2 linguistic syntax plus rhythmic violation trial, 2 trials with all 3 violations, and 30 sentences with no violations. Therefore, in a given module only 37.5% of trials contained any violation. Half of the sentences in a given module were assigned a “yes” question, the other half were assigned a “no.” The order of the trials was randomized for each subject.Before beginning the experiment, the participants gave informed consent and completed a short background survey. The participants were then instructed to pay close attention to the sentences being read, rather than the chord progressions that were heard over the headphones. Then, the participants ran through a set of practice trials. After the practice trials, in the actual experiment the experimenter selected one of the 12 possible experimental modules at random. Participants were instructed to press the spacebar on the keyboard as soon as they had read the sentence segment, and then wait for the next segment to be presented. Pressing the spacebar caused the current sentence segment to disappear and an indicator button labeled “I read it” to light up. The following segment appeared at a fixed IOI regardless of when the current segment disappeared. After the end of each sentence, a yes-or-no comprehension question was displayed, at which point participants answered the question by pressing Y or N on the keyboard. Answering the comprehension question cued a new trial. The experiment lasted ~20 min. Examples of different types of trials are shown in a video demo in the Supplementary Materials accompanying this manuscript.RT and response data were saved as text files from MaxMSP, and imported into Microsoft Excel and SPSS for statistical analysis. RTs were log-transformed to normal distribution for statistical testing. Only RTs pre-critical, critical, and post-critical regions for each trial were used for analysis. Filler trials were, therefore, excluded from analysis . Of the remaining trials, trials with RTs that were two or more standard deviations from the mean of log-transformed critical region RTs were excluded as outliers, resulting in a range of 102.76–816.74 ms. These criteria led to the exclusion of 92 (7.20%) of observations from critical regions in Experiment 1.n = 41) and non-native English speakers . Similarly, no significant differences were observed between participants who reported musical training (n = 29) and those who reported no musical training . To check for interactions between linguistic syntax and native English speaker experience, an ANOVA was run on the dependent variable of log-transformed RT with the fixed factor of linguistic syntax (congruent vs. incongruent) and the random factor of native English speaker status (native vs. non-native English speaker). No significant interaction between native English speaker status and linguistic syntax was observed . Similarly, to check for interactions between musical syntax and musical training, an ANOVA with the fixed factor of musical syntax (congruent vs. incongruent) and the random factor of musical training  showed no interaction between musical syntax and musical training . As we observed no main effects or interactions that were explainable by native English speaking experience or musical training, results were pooled between native and non-native English speakers, and between musically trained and untrained subjects.No significant differences were observed in log-transformed RTs between native English speakers  = 16.38, p < 0.0001].On comprehension questions, participants performed significantly above chance in all conditions , as well as a significant main effect of language  and a significant main effect of rhythm  and a marginally significant effect of music . Means and SDs of RTs are given in Table A Three-way ANOVA on the dependent variable of log-transformed RT during the critical region (log_RT_CR) was run with fixed factors of language (two levels: congruent and incongruent), music (two levels: congruent vs. incongruent), and rhythm , with subject number as a random factor. Results showed a significant three-way interaction among the factors of linguistic, musical and rhythmic expectancies . In contrast, the interaction between language and music was not significant at the rhythmically early condition  or the rhythmically late condition  . Results showed that for the rhythmically on-time condition, there was an interaction between language and music . In the rhythmically late condition only, musical syntax violations produced larger difference scores at the critical region; however this difference was not significant. In the rhythmically early condition and on-time conditions, musically in-key trials yielded larger difference scores than musically out-of-key trials at the critical regions, although these differences were not significant . Twenty (57.1%) participants identified as female, and 15 (42.3%) as male. Twenty-eight (80%) reported that their first language was English, and seven had a language other than English as their first language. Other than English, participants' first languages included Spanish, Chinese, and Thai. Twenty-four participants (68.6%) spoke more than one language. Informed consent was obtained from all subjects as approved by the Ethics Board of Psychology at Wesleyan University.A new group of 35 undergraduate students from Wesleyan University participated in Experiment 2 in return for course credit. From these participants, all reported normal hearing, normal or corrected-to-normal vision, and no psychiatric or neurological disorders. Twenty-five participants (71.4%) reported having prior music training, averaging 5.9 years  Lab Suite in Judd Hall at Wesleyan University. An Apple iMac was used for the experiment, with MaxMSP software for all stimulus presentation and response collection.The same experimental patch on MaxMSP and 12 experimental modules with the 48 sentences borrowed from Slevc et al.  were useSimilar to Experiment 1, participants were instructed to read sentences carefully, and hit the spacebar as soon as they had read a sentence segment. After running through a practice set, the participants began the actual experiment. The experimenter selected one of the twelve possible experimental modules at random. At the end of each trial, participants answered the “yes” or “no” comprehension question, queuing the next trial.RTs and comprehension question responses were saved as text files from MaxMSP, and imported into Microsoft Excel, and SPSS for statistical analysis. Only RTs at the pre-critical, critical, and post-critical regions for each trial were used for analysis. Filler trials were, again, excluded from analysis . The same parameters and methods of outlier exclusion were used from the previous experiment, resulting in a RT range of 123.63–1121.40 ms. These criteria led to the exclusion of 19 (1.97%) of observations in Experiment 2. RTs were also log-transformed to normal distribution for statistical tests.t(34) = 0.96, n.s.]. Similarly, an ANOVA with the fixed factor of linguistic syntax and the random factor of native English experience showed no significant interaction . As we observed no differences that were explainable by native English speaking experience, results were pooled between native and non-native English speakers.Results between musically trained and non-musically trained subjects were pooled because music was not a factor in this experiment. No significant differences were observed in log-transformed RTs between native English speakers and non-native English speakers . This suggests that the added variable of musical expectancy further taxed participants' attention from the task-relevant comprehension questions in Experiment 1.Participants performed significantly above chance  = 7.69, MSE = 0.001. p = 0.009], a significant effect of rhythm , and no significant two-way interaction . Mean and SD RTs are shown for each condition in Table A Two-way ANOVA on the dependent variable of log-transformed RT during the critical region was run with the factors of language and rhythm. Results showed a significant main effect of language [F1, 4 = 7.69,Results from Experiment 2 showed main effects of language and rhythm, validating the use of this novel task. There was also a higher comprehension accuracy compared to Experiment 1, but no interactions between the two factors of linguistic syntax and rhythmic expectancy .Taken together, Experiment 2 helps to validate the paradigm used in Experiment 1. By simplifying the experiment to remove the factor of music, some attentional resources may have been restored, resulting in higher comprehension accuracy overall, as well as main effects of language and rhythm with no interaction between the two.The goal of the current study is to examine how rhythmic expectancy affects the processing of musical and linguistic syntax. Experiment 1 shows main effects of language, music, and rhythm, and specificity of the interaction between musical and linguistic syntax in the rhythmically expected condition only. These data patterns confirm that rhythm affects the sharing of cognitive resources for music and language, and is largely consistent with SSIRH Patel,  and DAT As it is possible that results were due to the complexity of our design, Experiment 2 simplifies the design by eliminating the factor of music altogether. Results of Experiment 2 show superior comprehension accuracy compared to Experiment 1, and main effects of language and rhythm without an interaction between the two factors. The main effects help to validate our adaptation of the original self-paced reading time paradigm  vs. involuntary (exogenous) attention might shed light on this distinction (Hafter et al., In both experiments, effects of rhythm were observed in response to visually-presented sentence segments. While the rhythmic aspect of language might generally manifest itself more readily in the auditory than the visual modality, this effect observed from the visual manipulations suggests that rhythmic expectation for language is not limited to auditory processing, but may instead pervade the cognitive system in a modality-general manner, affecting even the visual modality. As visual detection and discrimination are both modulated by rhythm Grahn,  and musiResults appear to be independent of musical training and native English speaker experience. The link between linguistic and musical grammar processing could have been expected to vary by musical and linguistic expertise: children who perform well on phonemic or phonological tasks also outperform their counterparts in rhythmic discrimination as well as pitch awareness (Loui et al., In sum, the current study demonstrates that rhythmic expectancy plays an important role in the shared processing of musical and linguistic structure. The subject of shared processing of musical and language structure has been central to music cognition, as is the question of how rhythm affects attentional entrainment. While providing support for an overlap in processing resources for musical and linguistic syntax, the current results also suggest that perturbations in rhythmicity of stimuli presentation tax these attentional resources. By offering a window into how perturbations of rhythmic and temporal expectancy affect musical and linguistic processing, results may be translatable toward better understanding and possibly designing interventions for populations with speech and language difficulties, such as children with atypical language development (Przybylski et al., The 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.


Multiple studies have tried to establish the distinctive profile of individuals with Asperger's syndrome (AS). However, recent reports suggest that adults with AS feature heterogeneous cognitive profiles. The present study explores inter-individual variability in children with AS through group comparison and multiple case series analysis. All participants completed an extended battery including measures of fluid and crystallized intelligence, executive functions, theory of mind, and classical neuropsychological tests. Significant group differences were found in theory of mind and other domains related to global information processing. However, the AS group showed high inter-individual variability  on most cognitive tasks. Furthermore, high fluid intelligence correlated with less general cognitive impairment, high cognitive flexibility, and speed of motor processing. In light of these findings, we propose that children with AS are characterized by a distinct, uneven pattern of cognitive strengths and weaknesses. Autism spectrum disorders (ASD) are a group of neuro-developmental conditions that compromises social interaction , and presents restricted, repetitive interests and activities  and were assessed by a psychiatrist. Their diagnosis was based on the criteria established by Diagnostic and Statistical Manual of Mental Disorders (DSM-IV)  was employed as a measure of effect size for the significant effects. In addition, we included an inferential test used to assess variance equality between two groups (only significant differences were reported). To further assess inter-individual differences, we conducted MCSA and compared each participant with the mean of the control group on every measure. We followed the method of Towgood et al. .Group differences were analyzed through an ANCOVA test using age as a covariate. Eta squared  tests to assess whether the number of measures in which AS individuals obtained sub- and supra-normal performance was associated with individual differences in FI. Finally, we used Spearman's rank correlations to examine the association between FI and neuropsychological measures. The significance of all correlations was corrected for multiple comparisons using the Sidak method. The adjusted α level after correction was set at 0.002. The α value for all other statistical tests (not related to correlation) was set at 0.05.F = 5.72, p = 0.024, n2 = 0.175], visuo-spatial constructional ability , and ToM   = 17.34, p = 0.000, n2 = 0.344], but not to RAVLT acquisition  or RMET .Table p = 0.000), RCFT copy (p = 0.016), and story memory immediate recall (p = 0.001). Consistent with previous reports , working memory (digit span and arithmetic from WISC III), motor processing speed (coding and symbol search from WISC III), cognitive flexibility (TMT-B), response inhibition (Stroop task), and logical memory (story memory from WRAML), no significant differences were observed in these measures. However, the SDs of some measures were higher in the AS group , followed by SM immediate recall (37%), coding from WISC III (32%), and arithmetic (27%). There were no participants who performed only supra-normally.H = 8.37, p = 0.015). After that, we used Mann-Whitney tests for pair-wise comparisons. Participants with superior FI scores displayed a smaller number of measures with sub-normal performance than children with inferior  and average  FI scores. No significant differences between AS children with average and inferior FI  were observed. These results suggest that AS children with higher FI have a lower probability of showing deficiencies in other domains.Therefore, we used the Kruskal-Wallis test to compare the number of measures with sub- and supra-normal performance according to the patients' FI score. Following Towgood et al. , we sepaFinally, to further explore the influence of FI on other domains, we conducted a correlation analysis between FI (RPMT) and neuropsychological measures in each group . A significant correlation was also found between FI and motor processing speed   across cognitive tasks. At the individual level, higher FI was associated with less cognitive difficulties and high cognitive flexibility and motor processing speed. To our knowledge, this is the first application of MCSA in children with AS and the first report of partial influence of FI on the cognitive profile of these patients.The results demonstrated significant group differences between AS and control children in RMET. This suggests that AS children have difficulties in inferring the mental states of others. However, the MCSA showed that only a few children of the AS sample performed sub-normally, confirming that group differences are not always a reliable index of deficits in the AS group. Furthermore, as in the case of AS adults , verbal communication (receptive vocabulary), and repetitive interests and activities (cognitive flexibility). Moreover, the group exhibited both sub- and supra-normal performance in domains excluded from diagnostic criteria, such as information processing styles , processing speed (coding from WISC III), and working memory (arithmetic from WISC III).In support of our first hypothesis, subtle differences across cognitive domains were not revealed by the group-type analysis. Thus, in domains associated with the AS diagnosis, this group included a high proportion of individuals with sub-normal performance, while in other domains, AS children obtained either sub- or supra-normal performance.For instance, although AS and control groups were similar regarding FI, the former showed high inter-individual variability on the RPMT. Indeed, this group displayed the greatest variability in FI. Most previous reports described FI as either an intact or superior ability in individuals with AS  in neurological patients  studies. The lateral prefrontal cortex and posterior parietal regions are the neural substrates subserving the relation between abstract reasoning (FI) and performance on EF tasks among neurotypical adults , which require an extended number of participants, could be robust models to predict the role of FI in cognitive profiles.noise in the data, which was attributed to limitations of the AS diagnosis, leading to multiple subgroups of patients (Towgood et al., Furthermore, the observed heterogeneity may allow for alternative explanations. For instance, clinical measures of autistic symptomatology have been associated with cognitive heterogeneity in adults with AS (Hill and Bird, We suggest that heterogeneity is a defining feature of the cognitive profile of children with AS. These findings have important implications for the treatment, identification, and assessment of these individuals. The DSM-5 has included AS within ASD, which incorporates even more variability in symptoms and behavior among these individuals. The challenge for clinical practice is to work with extensive and flexible neuropsychological assessments that allow for the identification of both deficits and strengths in individuals with AS, so as to hone individual treatment.The data showed that AS children, as a group, present common difficulties in ToM and global information processing. At the individual level, they demonstrated a wide range of variation in most of the cognitive functions evaluated. Thus, despite extensive research seeking a typical cognitive profile of individuals with AS, the evidence suggests that this syndrome is characterized by an uneven pattern of cognitive strengths and weaknesses. The present report of inter-individual cognitive variability in children with AS aligns with similar findings in autism and adults with AS.Moreover, a detailed MCSA revealed that individual differences in FI may be associated with this heterogeneous profile. The data showed that high FI was related to fewer cognitive impairments. In addition, FI was associated with cognitive flexibility and motor processing speed only in AS children. The current report is the first to highlight the possible influence of FI on AS cognitive profiles. Indeed, superior abilities in abstract reasoning could compensate or reduce AS children' vulnerability to develop other deficits in cognitive abilities. Further research is needed to elucidate the relationship between FI and the cognitive functioning of these individuals.The 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.


Many water quality models have been successfully used worldwide to predict nutrient losses from anthropogenically impacted catchments, but hydrological and nutrient simulations with limited data are difficult considering the transfer of model parameters and complication of model calibration and validation. This study aims: (i) to assess the performance capabilities of a new and relatively more advantageous model, namely, Hydrological Predictions for the Environment (HYPE), that simulates stream flow and nutrient load in agricultural areas by using a multi-site and multi-objective parameter calibration method and (ii) to investigate the temporal and spatial variations of total nitrogen (TN) and total phosphorous (TP) concentrations and loads with crop rotation by using the model for the first time. A parameter estimation tool (PEST) was used to calibrate parameters. Results show that the parameters related to the effective soil porosity were highly sensitive to hydrological modeling. N balance was largely controlled by soil denitrification processes. P balance was influenced by the sedimentation rate and production/decay of P in rivers and lakes. The model reproduced the temporal and spatial variations of discharge and TN/TP relatively well in both calibration (2006–2008) and validation (2009–2010) periods. Among the obtained data, the lowest Nash-Suttclife efficiency of discharge, daily TN load, and daily TP load were 0.74, 0.51, and 0.54, respectively. The seasonal variations of daily TN concentrations in the entire simulation period were insufficient, indicated that crop rotation changed the timing and amount of N output. Monthly TN and TP simulation yields revealed that nutrient outputs were abundant in summer in terms of the corresponding discharge. The area-weighted TN and TP load annual yields in five years showed that nutrient loads were extremely high along Hong and Ru rivers, especially in agricultural lands. Eutrophication is caused by excessive inputs of nitrogen (N) and phosphorus (P) in rivers and lakes; as such, this phenomenon has been extensively investigated ,2,3,4,5.The Hong–Ru River basin located in the mid-eastern China is representative of agricultural areas. However, the water quality of Hong–Ru River Basin does not satisfy the expected category III criteria, which are 1.0 mg/L and 0.2 mg/L for total nitrogen (TN) and total phosphorous (TP), respectively, as recommended by the Chinese Environmental Quality Standards for Surface Water . TherefoHydrological processes are strongly dependent on climate change, topography, soil properties, and catchment scale ,11. Hydr2 [A few well-known models, such as SWAT , SWRRB , and ANS2  and in n2 . HoweverIn this study, a parameter estimation tool (PEST) was used to calibrate hydrological and water quality parameters simultaneously; this multi-objective calibration is more appropriate than other calibration methods to increase constraints on hydrological and water quality processes because both processes are related to each other ,14,21,35The HYPE model is developed by SMHI to simulate stream flow and water quality on their way from precipitation through soil, river, and lakes to river outlets over time . The spaThe HYPE model simulates snow accumulation and snowmelt, evapotranspiration, surface runoff, infiltration, macropore flow, percolation, interflow, tile drain flow, quick base flow, slow base flow, snow depth, frost depth, river delay, and damping in a hydrological process ,32. The 2, which is a combination of Hong and Ru rivers, as they share the same outlet at the Bantai gauging station. A digital elevation model (DEM) was used to present the basin topography. The four discharge gauging stations  with daily time step data from 2006 to 2010 were chosen in the Hong-Ru River Basin  simulation was conducted for a preliminary model evaluation, which was ignored in the model evaluation. Model calibration (2006–2008) and validation (2009–2010) were performed as the input files were improved after one year of model warm up. The spatial and time series data prepared for the model setup are shown in In this study, land use and soil were respectively reclassified into 11 and eight types on the basis of the original sources by using ArcGIS. The whole Hong-Ru River Basin was divided into 92 sub-basins, and 66 SLCs were defined on the basis of the descriptions of HYPE. The number of SLCs was greatly reduced by aggregating similar classes into larger groups that might exhibit similar properties. The driving data of daily precipitation and daily mean temperature of the discharge simulation in each sub-basin were prepared in accordance with the required format of HYPE; missing values were replaced with data from Zhumadian station, which represents the whole basin. The atmospheric depositions containing dry and wet depositions of N and P were filled with values from neighboring cities and provinces because of limited studies and monitoring data. The agricultural nutrient inputs were obtained from a field survey. Diffuse and point sources were prepared after the statistics from the Environmental Protection Agency of Zhumadian was analyzed.Many parameters are related to hydrological and nutrient processes in the HYPE model. Some parameters are general, and other parameters are dependent on land use and soil type. Most of these parameters were derived from literature review and from previous modeling experiences, although only the most sensitive parameters were selected for calibration. Some parameters were held constant ,30. In tm is the total number of the observation groups of the observed discharge and nutrient concentrations from the gauging stations, n is the total number of the measured discharge/nutrient concentrations at each gauging station, and ω is the weight of the related objective function.2), NSE, PBIAS, and root mean squared error (RMSE) observation standard deviation ratio (RSR), were used to evaluate the agreement between simulated (Sim) and observed (Obs) values of daily discharge and daily nutrient loads. The four criteria were determined on the basis of the following equations:Cubic spline interpolations were used in this study ,24, and According to the watershed simulation evaluation guidelines described in a previous study , model sThe parameter calibration results and the related physical interpretations are presented in The most sensitive parameter is denitrlu (plain dry land–land use dependent), which corresponds to denitrification in soil. This result is consistent with that described in previous studies , which rThe discharge simulation results are separated into two sets. One set involves the comparison of two relatively low-discharge gauging stations, namely, Dingwan and Miaowan. The other set includes the comparison of two relatively high-discharge gauging stations, namely, Shakou and Bantai. The highest observed discharge values at Dingwan and Miaowan stations were 858 and 496 mm, respectively. The highest observed discharge values at Shakou and Bantai stations were 1620 and 2310 mm, respectively. The HYPE model captured peak flow relatively well during both calibration and validation periods at the four stations, especially at Miaowan, Dingwan, and Bantai stations in July 2007. Some mismatches between simulated peak flow and observed peak flow is mainly due to the continuous model setup of a daily time step ,49. The At the Mianwan and Dingwan stations, the residuals were intensely distributed around 0 as time passes. In summer in 2007, the residuals slightly fluctuated because of extreme climatological events, which changed groundwater base flow. This finding also occurred because the model did not well depict the complicated relationships between soil water and land use, especially in this case, which considered plain dry land ,51.In the validation period from 2009 to 2010 when precipitations were more than 40 mm, the peaks of the observed and simulated discharge were not presented well. This result is probably attributed to low soil water storage where agriculture lands are largely distributed . With thThe observed and simulated TN and TP concentrations and daily TN and TP load at the three gauging stations are illustrated in As the outlet of Suyahu reservoir, Shakou station yielded weaker simulations of TN loads than Dingwan station. The performance of the model  at Shako2 in July, August, and September; among these values, the highest load was simulated in July. This result corresponds well to the values in the months with a relatively high precipitation. The average total monthly precipitation values were 287 mm, 119 mm, and 66 mm in July, August, and September, respectively. The average total monthly loads in December, January, February, and March were relatively higher than those in April, May, and June. This difference can be contributed by the basic fertilizer inputs prepared for winter wheat in October in most areas of the agricultural land and additional fertilizer inputs at the end of February, although the corresponding precipitation was maintained at a relatively low level [ow level . Anotherow level . The low2, 189.4 kg/km2, and 52.4 kg/km2 were detected in July, August, and September, respectively. By contrast, the lowest yields of 8.2 kg/km2 and 10.5 kg/km2 were found in May and June, respectively. The yield in winter in December was 28.4 kg/km2, and this value was higher than those in February (19.2 kg/km2) and March (19.0 kg/km2). This difference is probably attributed to the reduced consumption of P and the decreased microbial population. In the first six months, the TP loads were lower than 19.2 kg/km2 in February, which could be the month when a low amount of P was used in winter wheat planting and less inputs were contributed by other nonpoint sources because point sources in the area were not the main contributors [The trend of TP load yield fluctuations was similar to that of TN load yield fluctuations. The highest yields of 437.2 kg/kmributors .The area-weighted TN and TP annual yields from 2006 to 2010 were determined  to inves2 to 58.07 tons/km2 for the five studied years. The initial TN load in 2006 ranged from 0.168 tons/km2 to 39.447 tons/km2. The sub-basins 33, 35, 44, 52, 55, and 88, which are located in the upstream mountain area, showed relatively low TN loads. This mountain area is dominated by forests and grasses (2 to 58.07 tons/km2 were found on the downstream section of the Ru River. The manure from livestock and poultry is responsible for the high TN loads, as indicated by high fertilizer inputs.The annual yields of the TN load varied from 0.066 tons/km grasses . The oth2 to 13.210 tons/km2 during 2006–2010. The differences in the spatial distributions of TP loads for five years were relatively more normal than the distributions of TN loads. The highest amount of TP loads occurred along the Ru River. In 2006 and 2007, TP loads were relatively more than those in 2008, 2009, 2010. This finding indicates that high rainfall conditions were more significant; thus, this phenomenon indirectly describes temporal variation in the annual loads of TP through surface flow with sediments [The annual yields of TP loads varied from 0.003 tons/kmediments . TP loadThe HYPE model has been successfully used in agricultural lands with crop rotation to assess the temporal and spatial variations of water quality in the Hong Ru River Basin in China. However, the model should be further evaluated with appropriate tools to check its uncertainties because numerous data were obtained through literature review and many historical monitoring data were insufficiently described. The simplified descriptions of the model simulations, such as transport and transformation of P and processes during extreme climatological events of agricultural land, can also be further developed. Crop rotation with more crop plantings under different circumstances in one year is also a challenging work. The simplified structures of the HYPE model should be considered in practical applications because the model can be modified easily on the basis of specific characteristics.HYPE was applied to the Hong–Ru River Basin with a daily time step. Discharge in this agricultural area from 2006 to 2010 was simulated; TN and TP concentrations and loads from 2006 to 2010 were also simulated. A multi-site and multi-objective calibration method was utilized with PEST to identify the relevant parameters. The most sensitive general parameters and land use/soil type parameters during hydrological simulations were wcep, rivvel, and cevp. The most sensitive general parameters and land use/soil type parameters during nutrient simulations were denitrlu, rivvel2, wprodn, wprodp, and sedpp. The optimized values were significantly satisfactory.The HYPE model captured the temporal and spatial variations of discharge very well in both the calibration and validation periods at the four gauging stations containing high flow and low flow circumstances. Among the NSEs, the lowest was 0.74. However, the model underestimated the water balances at Shakou station and the catchment outlet Bantai by 27.3% and 8.7%, respectively. Shakou is very close to the outlet of the regulated reservoir Suyahu, and this station is also located downstream of the two other upstream reservoirs, namely, Bantiao and Boshan, which influence the flow through Shakou and Bantai stations to some extent. The underestimation was also observed because HYPE simplified the descriptions of the lakes within the catchment and disregarded the effects of reservoir management on the main rivers and land use; as a consequence, the model performance was reduced. The headwater contributes most of the total runoff in this studied catchment.The HYPE model captured the temporal and spatial variations of TN and TP relatively well during both calibration and validation periods at the three gauging stations. The lowest NSE and PBIAS for TN are 0.51% and −48.6%, respectively. By contrast, the lowest NSE and PBIAS for TP are 0.54% and −38.6%. The seasonal variations of daily TN concentrations in the entire simulation period is not sufficiently significant, and these variations are different from the low and high concentrations in winter and summer reported in a previous study . This reThe average monthly TN and TP simulation yields revealed that nutrient outputs were mainly detected in summer by considering the corresponding discharge. The area-weighted TN and TP load annual yields showed that nutrient loads were extremely high along the Hong and Ru rivers, especially in the agricultural land. Annual area-weighted precipitation indirectly affected the transport and transformation of N to a greater extent than P.Developed on the basis of Swedish characteristics, the HYPE model can be used in agricultural areas with crop rotation in China; this model can also be widely used worldwide. This model can be considered as a good decision-making tool of environmental protection agencies to predict discharge and enhance agricultural management with limited related data.


Alien species can severely disrupt the structure, function and stability of native communities. We evaluated the structure of pollination networks in the three main habitats and in the two seasons on the two most disturbed Galápagos Islands, and investigated how such structure is influenced by invasive plants. Alien plants integrate easily into the communities, but show low impact on overall network structure, except for an increase in network selectiveness. The highly invaded and low diversity humid zone showed the highest nestedness and the lowest modularity. Both pollinators and plants were more generalized during the hot season, when most plants were flowering and networks became more nested. Oceanic archipelagos are threatened by the introduction of alien species which can severely disrupt the structure, function and stability of native communities. Here we investigated the pollination interactions in the two most disturbed Galápagos Islands, comparing the three main habitats and the two seasons, and assessing the impacts of alien plant invasions on network structure. We found that the pollination network structure was rather consistent between the two islands, but differed across habitats and seasons. Overall, the arid zone had the largest networks and highest species generalization levels whereas either the transition between habitats or the humid habitat showed lower values. Our data suggest that alien plants integrate easily into the communities, but with low impact on overall network structure, except for an increase in network selectiveness. The humid zone showed the highest nestedness and the lowest modularity, which might be explained by the low species diversity and the higher incidence of alien plants in this habitat. Both pollinators and plants were also more generalized in the hot season, when networks showed to be more nested. Alien species (both plants and pollinators) represented a high fraction (∼56 %) of the total number of interactions in the networks. It is thus likely that, in spite of the overall weak effect we found of alien plant invasion on pollination network structure, these introduced species influence the reproductive success of native ones, and by doing so, they affect the functioning of the community. This certainly deserves further investigation. Sexual reproduction is an essential step for the life cycle of most plant species, and is chiefly limited by the quantity and quality of pollen grains arriving to their stigmas , and long-term impacts on the most pristine uninhabited islands. The main objective of this study was thus to assess the spatio-temporal variation of pollination interactions in the two most disturbed Galápagos islands and to determine whether and how alien plants may modify such interaction patterns. In addition, we investigated if alien species  differ from endemic and non-endemic natives in their integration into the pollination networks.2, which were formed by volcanic activity between 0.035 and 4.0 My ago   . These tThe Galápagos climate is characterized by two seasons. The hot/wet season, from January to May, is associated with frequent rain throughout the islands , and is 2), elevation (864 m vs. 735 m above sea level), and latitude . San Cristóbal is older (max. age = 4.0 My) than Santa Cruz (2.3 My) and is also more isolated from other islands , as follows: (i) all species with open flowers were identified, (ii) the number of individuals of each species was counted, (iii) for each species, the number of flowers on two individuals having an average flower display was counted and (iv) the number of open flowers for each species in the transect was extrapolated from the number of flowers per individual × number of individuals. For species with tightly clustered inflorescences (e.g. the capitula of Asteraceae) we scored each inflorescence as a flower, as this is the ecologically relevant unit visited by pollinators.At each site, regular focal flower censuses were performed to quantify the contact of flying animals, mostly insects, with the reproductive organs of open flowers. Each site was visited every other week, when climatic conditions allowed, from March to May (rainy season) and from July to September (dry season) in 2010. In 2011, the 12 sites were re-sampled but only during the flowering peak (hot season); this year, sampling took place between January and May. On each sampling day, flower diversity and abundance were measured along a 500 × 6 m transect (i.e. 3000 mFlower-visitors were censused during periods of 10 min in front of target plants (∼1 m away). On each census day, all species with open flowers (regardless of their anthesis stage and nectar production) were observed for at least two non-consecutive periods between 06:00 h (sunrise) and 22:00 h. Nocturnal censuses were made by means of red (low energy) l.e.d. headlights to avoid affecting insects. Species were arbitrarily selected for the different time periods to avoid censusing the same species always at similar times of the day. In each census period we recorded: (i) identity of the flowering plant species, (ii) number of open flowers observed on each individual plant (often only one branch was observed in the case of shrubs or trees), (iii) identity of each flower-visitor, (iv) number of individuals of each species visiting flowers and (v) number of flowers visited by each individual flower visitor. The sampling protocol resulted in 1145 h of flower visitation censuses  of 119 plant species of which 36 are introduced in the Galápagos. Plant identifications followed L), species specialization (d′) and species strength (st), for both pollinators and plants. Definitions of each of these parameters can be found in Appendix S1Supporting Information][see . In 2011, pL was higher in Santa Cruz than in San Cristóbal , and it was lower in the humid zone than in the other two habitats . In contrast, neither d′ nor st were significantly influenced by any of the predictor variables included in any of the models.Regarding z = 6.45, d.f. = 415 and z = 3.83, d.f. = 265, P < 0.001, respectively), although in 2011 aliens and non-endemic natives did not differ significantly  , despite the usual fluctuations in flower production and flower-visitors' presence/abundance. Thus, we focus on the temporal differences observed between seasons. All pollination networks were larger during the hot rainy season, when more plant species are blooming and more insects are flying, than in the cold dry season. Both pollinators and plants actually showed higher linkage levels in the hot than in the cold season, given the greater availability of partner species in the former. Moreover, networks were more nested in the cold than the hot season after controlling for network size, which influences this parameter. Such temporal difference in the degree of nestedness suggests that the interactions in the hot season tend to be more specific, with specialist species interacting more than expected with each other and less so with generalists.Alien pollinators were consistently found to visit fewer plant species than endemic pollinators and, at least one year, also visited fewer plant species than native pollinators, which suggests that these newly-arrived species are focusing flower visitation on species with particular traits. However, the fact that alien pollinators also showed lower levels of selectiveness than endemic pollinators implies that they tend to visit more abundant flower resources compared with endemic pollinators, which visit even rare flowers. Likewise, species strength was consistently lower for alien than endemic pollinators, indicating that the former are less important to plants. In a previous study focusing on the pollination networks of the arid zone in five Galápagos islands, we found that alien insects had more links than either endemics or non-endemic natives  and strength  were spatially and temporally consistent and not influenced by alien plants.Pollination networks are larger during the hot/rainy season, when most flowers are in bloom and more insects are flying, than in the cold/dry season. They are also more nested in the hot season, and thus probably more robust to disturbances. Pollinators visit more plant species, and plants are visited by more pollinator species, during the hot season. In the cold season, the number of insects is especially low in the humid zones and thus the number of pollinators visiting plants is also lower in that season and habitat. In contrast, both pollinator and plant selectiveness  which have displaced many native species in the invaded areas, mainly in the humid zones  and were less important to the plant community (i.e. showed lower species strength) than endemic and native pollinators. They, however, infiltrated the native communities of all habitats and in both seasons and currently represent over 40 % of all recorded pollination interactions. Alien plants, on the other hand, were visited by approximately the same number of pollinators as natives—but less than endemic plants—implying that they are also well integrated into the native communities. In this study, we found a rather feeble effect of alien plants on the structure of pollination networks. As previously mentioned in the methods, our study intentionally considered sites that are not completely disturbed by highly invasive species (e.g. BBVA Foundation and is also framed within project CGL-2013-44386-P financed by the Spanish Government. R.H. was supported by the Portuguese grant FCT-IF/00441/2013, and by the Marie Curie action FP7-2012-CIG-321794.This research is part of a larger project funded by A.T., J.M.O. and R.H. designed the study, S.C. collected the data and did some preliminary analyses, and A.T. and R.H. performed the final analyses. A.T. led the writing and all authors contributed to the text. All authors read and approved the final manuscript.None declared.


Objectives. We aimed to test the hypothesis that metabolic syndrome (MetS) is significantly associated with cognitive decline (CoD) in elderly adults and further assess whether MetS and inflammation have a significant joint effect on CoD. Methods. Data (n = 2975) from the U.S. National Health and Nutrition Examination Survey (1999–2002) in participants aged ≥60 years who had Digit Symbol Substitution Tests (DSS: a standard measure of cognitive function) were studied. CoD was defined as those in the lowest quintile of DSS score. MetS was defined as having ≥3 of 5 MetS traits (large waist circumference (WC), high blood pressure (BP), elevated glucose, triglycerides, and decreased high density lipoprotein cholesterol). Results. Of 2975 participants, the prevalence of CoD (DSS score <25) was 12.1%. After adjusting covariates, individual large WC, high BP, elevated glucose level, and MetS were significantly associated with CoD in logistic regression models (P < 0.001). There was a significant dose-response relationship between an increased number of MetS traits and CoD (P < 0.001). A significant joint effect of MetS and CRP on the odds of CoD was observed. Conclusion. The study, using a nationally representative sample, extended previous studies by highlighting a significant MetS-CoD relationship and a joint effect of MetS and CRP on CoD. These novel findings add to our understanding of the association of neurometabolic disorders and cognition and have implications that may be relevant to primary care practice. Impaired cognitive function is emerging as one of the greatest health threats of the twenty-first century. As the life expectancy of the population has increased so too has the prevalence of cognitive decline (CoD) and dementia, largely in the form of Alzheimer's disease, which now affects almost 50% of adults over the age of 85 in the United States . This stThe NHANES, using a cross-sectional study design, has been conducted since 1971. This study has been redesigned to conduct and combine data once in a two-year cycle since 1999. In the NHAES, participants are selected using a multiple sampling approach to recruit a nationally representative sample of approximately 10,000 participants aged 2 months and older in each two-year cycle [The NHANES consists of interviews, physical examinations, and data from blood sample analysis. All measurements were processed per standard protocols. Household interviews were conducted prior to the examination to collect demographic, medical history, and health behavioral information. Race and ethnicity were determined by self reports. Examinations were carried out in mobile examination centers. Blood samples were obtained at the examination centers. Participants were asked to fast for 12 hours prior to the morning examination or 6 hours prior to the afternoon or evening examination. To ensure the quality of blood sample collection and to minimize hemolysis of blood samples, all phlebotomists were certified and trained in standardized laboratory procedures. Serum lipid profiles, metabolic indicators, and CRP concentrations were measured using standard methods. Chronic conditions, including hypertension, coronary heart disease, diabetes mellitus, and stroke were participants' self-reported medical history that were diagnosed by a physician or health professional. A detailed description of the survey and its sampling procedure is available at the NHANES website .In the 1999–2002 NHANES, cognitive function was tested using the WAIS III  Digit Symbol Substitution Test (DSS)—Coding module and was conducted during the household interview for those aged 60 years and older. In this coding exercise of DSS, participants copy symbols that are paired with numbers. Using the key provided at the top of the exercise form, the participant draws the symbol under the corresponding number. The score is the number of correct symbols drawn within a period of 120 seconds. One point is given for each correctly drawn symbol completed within the time limit. The maximum score is 133 with a lower score indicating poorer cognitive function. Sample items are provided for initial practice. Participants who are unable to complete any of the sample items do not continue with the remainder of the exercise .Several criteria for diagnosing MetS have been proposed by health organizations , 9. In tIn the analysis, we included the following variables as covariatesage (years), sex , race/ethnicity , marital status , and smoking status (ever-smoking versus never).t-test for continuous variables, and Chi-square for categorical variables. Serum TG, glucose, and CRP concentrations are log-transformed because their values have a skewed distribution. The relationship between age and DSS score is tested using linear regression analysis for White, Black, and Hispanic groups separately. In the second group analysis, we classified participants into 5 groups by quintiles of the distribution of DSS scores. Subjects who had DSS scores in the lowest quintile  were classified as having CoD. We then tested adjusted mean differences in each MetS trait and CRP across quintiles of DSS using general linear models, with adjustment for age, sex, and race/ethnicity. Trends of changes in means of MetS traits and CRP across DSS quintiles were tested using linear regression analysis. In the third group analysis, we estimated odds ratios of individual MetS traits and MetS for the prevalence of CoD using multivariate logistic regression modeling technique with adjustment for a set of covariates. Then, three models were conducted using the total combined sample of all racial/ethnic groups: (1) in Model 1, age, sex, and race/ethnicity were adjusted, (2) in Model 2, age, sex, race/ethnicity, marital status, and cigarette smoking status were adjusted, and (3) in Model 3, the covariates that were adjusted in Model 2 plus CRP were adjusted in order to test any potential mediate effect of CRP with MetS on CoD. In this analysis, serum CRP level was dichotomized. Subjects who had serum CRP values in or higher than 75% distribution in the total study participants were classified as having a high CRP value (>0.60 mg/L). In the final analysis, we examined the combined effects of MetS (yes versus no) and CRP (>0.60 mg/dL versus ≤0.60 mg/L) on CoD by three ages  and sex.A serial analysis was conducted using univariate and multivariate analysis approaches to adjust for potential confounders. In the first group analysis, we described the characteristics of the study participants by sex. Differences in the study variables between genders were tested using P value <0.05 was considered as having statistical significance.All data analyses were conducted using SAS software version 9.2 . SAS Statistics package for Surveymean, Surveyfreq, Surveyreg, and Surveylogistic procedures were used to take into consideration the complex multistage sampling data collection used in the NHANES. A two-sided P < 0.05, or <0.01), except for the differences in serum TG and the prevalence of stroke and diabetes (P > 0.05).P = 0.02). P < 0.001). Black and Hispanic had significantly lower mean DSS scores than White across age groups (P < 0.01).Of 2975 participants aged ≥60, the prevalence of CoD (DSS score <25) was 12.4% in males, and 11.79% in females (P < 0.01), except for nonsignificant changes in mean DBP (P = 0.158) and log-TG values (P = 0.144).P < 0.001, Models 1–3). The values of odds ratios in those with ≥3 MetS traits combination significantly increased after adjustment including CRP.The main findings from the study are that MetS significantly predicts the odds for the prevalence of CoD in subjects aged ≥60 years old. Of 5 MetS traits, large WC , high BP, and impaired glucose metabolism had significantly independent effects on the risk of the prevalence of CoD. Elevated CRP had potential meditative and combined effects with MetS on the risk of CoD. These associations were independent of a set of covariates. These findings add to growing evidence that aggressively controlling MetS and CRP may offer an important primary prevention strategy to prevent or attenuate CoD in the elderly.MetS, first described about 40 years ago, is a multifactorial disorder of substantial heterogeneity, represented by a concurrence to central obesity that also includes impaired glucose metabolism, dyslipidemia, and high BP and that depicts a risk status for both type 2 diabetes mellitus, and coronary artery disease . A possiMetS and 3 of 5 MetS traits, central obesity, high BP, and elevated fasting glucose had significant associations with CoD in our present study. There have been several reports of MetS link to deficits in memory, executive functioning, processing speed, and overall intellectual functioning, although the potential mechanisms by which MetS and its traits are associated with the risk of CoD still need to be studied using data from large-scale prospective studies and clinical trials –14, 18.To our knowledge, the present study is the first to test a dose-response association between an increased number of MetS traits and CoD using a nationally representative sample in elderly adults aged 60 and older. This finding highlights the magnitude of the number of MetS traits on CoD. Furthermore, a significant joint effect of MetS and CRP on CoD is observed. The degrees of this joint effect were different in men and women. It appeared that it had a similar effect in those with MetS and elevated CRP in men regardless ageing, but it varied in women. Although the present study is unable to test the mechanism of the associations of MetS and CRP with CoD, the findings of the study provide us with important evidence that aggressive treatment for MetS and elevated CRP could have immediate protect effects on improving cognitive function.n = 428) and Hispanic (n = 557) were much smaller than White (n = 1823). Further studies are needed to address in-depth racial/ethnic differences in MetS and cognitive function. Finally, the associations between MetS, CRP, and CoD may be underestimated due to possible selection and survival biases. For example, subjects who had serious disease status, including those who were hospitalized or died, were not included in the NHANES.Several limitations may exist in the present study. Firstly, this study used data from baseline 1999–2002 NHANES sample that had a cross-sectional study design. Therefore, it cannot be applied to interpret any causal-effect relationship of MetS and CRP with CoD. Secondly, although a significant difference in mean DSS scores was observed by racial/ethnic groups , we are There are also several advantages of the present study. Firstly, regarding studies in elderly populations, the present study is one of the few large-scale studies that had a nationally representative sample of those aged 60 years and older. Secondly, the study takes the advantages of the NHANES design, data collection, measurements, and lab tests that are validated and standardized under the support of the National Center for Disease Control and Prevention. Finally, using a standard and robust statistics analysis approach, the present study has demonstrated a strong and significant dose-response relationship between MetS traits and CoD that enhances the prevention of CoD through treatment of MetS.In conclusion, the study, using a nationally representative sample, extended previous studies by highlighting a significant MetS-CoD relationship and a joint effect of MetS and CRP on CoD. These novel findings add to our understanding of the association of neurometabolic disorders and cognition and have implications that may be relevant to primary care practice.


The central nervous system seems to have an important role in fatigue and exercise tolerance. Novel noninvasive techniques of neuromodulation can provide insights on the relationship between brain function and exercise performance. The purpose of this study was to determine the effects of transcranial direct current stimulation (tDCS) on physical performance and physiological and perceptual variables with regard to fatigue and exercise tolerance. Eleven physically active subjects participated in an incremental test on a cycle simulator to define peak power output. During 3 visits, the subjects experienced 3 stimulation conditions  in a randomized, counterbalanced order to measure the effects of tDCS on time to exhaustion at 80% of peak power. Stimulation was administered before each test over 13 min at a current intensity of 2.0 mA. In each session, the Brunel Mood State questionnaire was given twice: after stimulation and after the time-to-exhaustion test. Further, during the tests, the electromyographic activity of the vastus lateralis and rectus femoris muscles, perceived exertion, and heart rate were recorded. RM-ANOVA showed that the subjects performed better during anodal primary motor cortex stimulation (491 ± 100 s) compared with cathodal stimulation (443 ± 11 s) and sham (407 ± 69 s). No significant difference was observed between the cathodal and sham conditions. The effect sizes confirmed the greater effect of anodal M1 tDCS . Magnitude-based inference suggested the anodal condition to be positive versus the cathodal and sham conditions. There were no differences among the three stimulation conditions in RPE (p = 0.07) or heart rate (p = 0.73). However, as hypothesized, RM- ANOVA revealed a main effect of time for the two variables (RPE and HR: p < 0.001). EMG activity also did not differ during the test accross the different conditions. We conclude that anodal tDCS increases exercise tolerance in a cycling-based, constant-load exercise test, performed at 80% of peak power. Performance was enhanced in the absence of changes in physiological and perceptual variables. Among the many factors that influence fatigue and exercise tolerance, central nervous system related factors seem to play a signfiicant role. As early as 1890, Angelo Mosso provided preliminary evidence on the effect of cognitive activity on fatigue . FurtherIn this context, the use of neuromodulation techniques that modulate the function of the cerebral cortex might increase our understanding of the factors that govern exercise performance. Transcranial direct current stimulation (tDCS) is a noninvasive neuromodulation technique that delivers continuous, low-intensity electrical current, causing significant changes of cortical excitability . The effInitial evidence has supported the implementation of tDCS in studies on fatigue, exercise tolerance, and recovery between training sessions ,9,10. CoNevertheless, most existing studies have used isometric exercise for a small muscle mass ,10,11. OThe objective of this study was to determine the effects of tDCS on time to exhaustion and physiological responses during high-intensity constant-load exercise and on mood before and after exercise. Based on earlier findings, we hypothesized that: a) anodal tDCS would increase exercise tolerance , irrespective of changes in heart rate and electromyographic (EMG) activity, whereas cathodal tDCS would fail to improve, or worsen, performance; and b) anodal tDCS conditions would moderate and delay the increase in RPE over time, explaining the improvement in performance.Participants were deemed eligible using the following criteria: (1) age between 18 and 30 years; (2) males (3) no diagnosis of neurological, or psychiatric disorders; (4) no drug or alcohol abuse; (5) not enrolled in another trial involving weight training; (6) being active . Fifteen physical active subjects met our inclusion criteria and agreed to participate initially in the study. Two subjects dropped out for personal reasons, one subject dropped out without providing any reason, and one because he was afraid to receive the stimulation. Thus, the sample consisted of 11 male subjects aged 21–31 years (M = 26 SD = 4), see http://www.randomization.com. All subjects received all stimulation conditions in a counterbalanced order.This study was a single blinded, randomized, placebo-controlled, crossover study with a repeated measures design. First, the subjects were seen at the laboratory for consenting and scheduling. The order of stimulation conditions were then randomized using the following site: The subjects visited the laboratory four times . In the pre-experimental session, the subjects performed an incremental test to determine peak power. In the subsequent visits, all procedures were the same, except for the stimulation conditions, which were previously randomized.First, the transmitter strap of the heart rate monitor was fixed to the subject’s chest and the subject was asked to sit comfortably in a chair. The electrodes for tDCS were maintained on the subject’s head with a rubber band. After the beginning of tDCS, the skin of the thigh was prepared and electrodes were placed for EMG recording. At the end of tDCS, the subjects answered a questionnaire for the assessment of mood state. Next, the subjects walked to a cycle simulator and started a standard warm-up, pedalling at 40% of peak power for 4 min, followed immediately by a test for EMG normalization. Finally, after 5 min of rest, the subjects performed a constant-load test (time to exhaustion). The subjects were asked to pedal at 60 to 90 rpm until voluntary exhaustion against a resistance corresponding to 80% of the peak power obtained in the pre-experimental test . Feedback about cycling cadence was provided and displayed on a screen positioned in front of the subject. The criterion adopted for interruption of the test was a decrease in the required pedalling cadence without recovery for more than 5 s. Immediately after the end of exercise, the subjects again filled the mood state questionnaire.For the incremental test, the subjects warmed up for 2 min at 100 W. Next, the workload was increased by 50 W at intervals of 2 min until voluntary exhaustion or inability to maintain the established minimum cadence (60 rpm) for more than 5 s. The power achieved in the last complete stage added to the product of the percent time spent in the stage of exhaustion by the standard increment (50 W) was defined as the peak power output. The position of the cycle ergometer was adjusted individually and the specifications were recorded so that it could be reproduced in the subsequent visits.During the following three visits, the subjects performed a constant-load test in which they were asked to pedal until exhaustion at 80% of the load achieved in the incremental test. The test was interrupted due to voluntary exhaustion or when the subjects could no longer maintain the pedalling cadence (60 to 90 rpm) for more than 5 s. The Velotron Dynafit Pro™ cycle simulator , with a capacity of up to 1500 W, was used for all physical tests.The electrical current was applied with a portable apparatus consisting of four main components: electrodes (anode and cathode with an area of 35 or 36 cm2), ammeter , potentiometer (component that permits manipulation of current intensity), and three batteries (9 V) to generate the current. The electrodes were wrapped in a saline (150 mM NaCl)-soaked sponge. The equipment used was built by an electronic engineer and has been previously approved by our Local Ethics Committee.The EEG 10–20 international system  was usedDespite reports of some side effects, tDCS is considered to be a safe technique . The mosA torque-velocity test was used to normalize EMG activity during the time-to-exhaustion test . After wEMG activity was measured continuously during the test in the three experimental sessions. Data were collected using an 8-channel EMG system . The sampling frequency of the electromyographic recordings was 2000 Hz. The common mode rejection ratio was ≤ 95 dB.The following muscles were analysed: vastus lateralis (VL), and rectus femoris (RF). Standards of the International Society of Electrophysiological Kinesiology (ISEK) were followed for the EMG data collection. The active electrodes , with an interelectrode (centre to centre) distance of 2 cm, were placed after shaving of the site, asepsis with alcohol and curettage to reduce impedance of the skin. The reference electrode was placed on the olecranon.The electrodes were located on the muscles analysed placed according to Surface Electromyography for the Non-Invasive Assessment of Muscles (SENIAM)  recommenRoot mean squares (RMS) and median frequencies (mF) obtained at intervals of 5 s were the indices used to evaluate the activity of the RF and VL muscles . The datAll subjects self-rated overall perceived exertion using the Borg scale 6–20 points) [ points [The volunteers filled out the Brazilian version of the Brunel Mood Scale (BRUMS) immediately before and after the experimental test session . The queHeart rate was measured continuously during all tests with an RS800 heart rate monitor , with beat-to-beat recordings using the Polar Precision Performance software  for subsequent analysis. Heart rate was analysed using the mean of 5 s obtained at the end of each minute of the test.2 = 0.06. The sample size was calculated using G*Power 3.1.7 considering power of 0.80, a correlation among the measurements of 0.85, nonsphericity correction ϵ = 0.75 and as aforementioned a partial η2 of 0.6. Using these parameters, sample size calculation resulted in a sample size of 11 subjects.For the sample size calculation, given this was an exploratory study we used a moderate effect size. If the results were not significant that would indicate that there would be no differences between the groups or the difference would be so small that would not be relevant. As we used ANOVA as the main method of analysis, a moderate effect size would be a partial ηhttp://www.sportsci.org/resource/stats/index.html were used for these analyses. In addition, the effect size (Cohen d) [http://www.sportsci.org/resource/stats). Our spreadsheet including raw data is available at the following link: http://datadryad.org/review?doi=doi:10.5061/dryad.30p8j.The assumptions of Normality and sphericity of the data were checked using the Shapiro-Wilk test and the Mauchly test and all the data showed normal distribution and sphericity. Repeated measures analysis of variance (RM-ANOVA) followed by the Bonferroni post hoc test were used for comparison of time to exhaustion in the constant-load test and of BRUMS scores between the different stimulation conditions. Two-way ANOVA for repeated measures was used for analysis of EMG, HR and RPE response, using the stimulation conditions and time as main factors. Alpha level was set at p<0.05. In addition, magnitude-based inference was applied to evaluate the time to exhaustion . For thiCohen d)  was calcThe intervention was well tolerated by all subjects. Only one subject reported weak headache after anodal stimulation.A one-way repeated measure ANOVA was conducted to compare the effect of stimulation type  on time to exhaustion. There was a significant effect of stimulation type, Wilks’ Lambda = 0.33, F  = 9.14, p = 0.007. The Bonferroni corrected post hoc tests  found the time to exhaustion was significantly larger in subjects receiving anodal stimulation as compared to those receiving cathodal  and sham  stimulation . However2 = 0.65, nonsphericity correction ϵ = 0.85) presented value equal to 1.Additionally, the post hoc analysis to compute achieved power on time to exhaustion results based on the parameters generated by ANOVA  = 3.730, p = 0.07)  or heartFurthermore, no significant differences were observed in any of the domains of the BRUMS scales. The peak power achieved in the torque-velocity test did not differ between conditions . EMG activity also did not differ during the test among the different conditions .The main finding of our study is the increase of exercise tolerance in the time-to-exhaustion test after anodal tDCS, irrespective of differences in physiological variables , consistent with our initial hypothesis. However, in contrast to our expectations, perceptual and affective variables  did not differ between experimental conditions.Specifically, our results are consistent with Cogiamanian et al. , who repIn this study, the configuration of the electrodes was selected to modulate motor cortex excitability and facilitation in both hemispheres , becauseAnother methodological strength of our study was our decision to fix the constant-load test at 80% of peak power in a previous incremental test. This method shows good sensitivity when the objective is to evaluate factors that alter endurance exercise performance and allows us to analyse physiological responses in a controlled manner . The heaSimilarly, to our study, in which tDCS increased the time to exhaustion, other groups demonstrated that tDCS improve performances in other tasks by enhancing muscle strength. For instance, Tanaka et al.  observedIn contrast to our initial hypothesis, rating of perceived exertion (RPE) was unaltered between experimental conditions. It is possible that the area of stimulation (Cz) does not interfere with perceptual responses. The perception of effort is related to activity in the insular cortex, anterior cingulate cortex, thalamus, and frontal operculum. Further, an increase in RPE correlates with an increase in central motor commands , as measured by electroencephalography at the Cz . This reA study on direct electric stimulation in patients who were undergoing surgery reported that when the electrical stimulus was applied to the parietal cortex, the subjects believed that they had performed movements. However, no EMG activity or movement was observed. Conversely, stimulation of the premotor cortex triggered mouth and contralateral limb movements, which the patients denied that they made . These dIn our study, no difference in EMG responses or heart rate was observed between the 3 experimental conditions. This finding was expected, because physiological variables are well controlled in time-to-exhaustion tests, as in this study , in whicWe suggest that the mechanisms of longer exercise tolerance that is mediated by anodal tDCS are related to an increase in intracortical facilitation and motor cortex excitability . Thus TDIn particular, Tanaka and Watanabe  developeThus, the output  from the primary motor cortex is regulated primarily by the balance between inhibition and facilitation, leading us to speculate that anodal tDCS at the Cz has a facilitatory effect for longer periods, increasing exercise tolerance.Nevertheless, we agree that the balance between inhibitory and excitatory information in the primary motor cortex determines the amount of motor output and, consequently, the end of exercise. In addition, tDCS alters corticospinal excitability, and thus, the effects might not be restricted to changes in intracortical excitability but might extend, for instance, to distant areas of the motor corticospinal tract.We cannot directly confirm that the changes in excitability and intracortical facilitation are responsible for altering the performance observed in our study. However, some authors showed that changes in excitability and intracortical facilitation are responsible for changing the performance of locomotor muscles , 34. In Therefore, intracortical facilitation probably occurs when time to exhaustion increases, but further studies are necessary to confirm this hypothesis. Furthermore, the tDCS montage can influence other brain areas related to performance, such as parietal cortex or occipital. A more specific stimulation should be applied to investigate each area separately.Our results suggest that anodal tDCS increases exercise tolerance in a cycling-based, constant-load exercise test, performed at 80% of peak power. Performance was enhanced in the absence of changes in physiological variables, such as heart rate and EMG, and perceptual variables, such as RPE, during the test. Moreover, anodal tDCS did not alter the profile of mood states before or after the time-to-exhaustion test versus cathodal and sham conditions. These results, if confirmed in time-trial protocols, will advance the theoretical and applied use of tDCS in improving athletic performance and its examination in future studies to better understand the neurophysiological mechanisms of exercise tolerance and fatigue.Additional studies should be conducted to confirm our results—for instance, experiments that determine whether tDCS alters performance in pacing exercises, such as time-trial exercise, and studies that measure the effects of tDCS as recovery technique between exercise sessions. In addition, whether tDCS can be used as a tool for recovery in athletes with signs and symptoms of non-functional overreaching and overtraining should be examined. tDCS might be applied not only to cycling activities but also to tasks that require the acquisition and retention of new motor skills and to high-precision sports, such as shooting and archery.


The aim of this study was to investigate potential differences in neural structure in individuals with Asperger syndrome (AS), high-functioning individuals with autism spectrum disorder (ASD). The main symptoms of AS are severe impairments in social interactions and restricted or repetitive patterns of behaviors, interests or activities.Diffusion weighted magnetic resonance imaging data were acquired for 14 adult males with AS and 19 age, sex and IQ-matched controls. Voxelwise group differences in fractional anisotropy (FA) were studied with tract-based spatial statistics (TBSS). Based on the results of TBSS, a tract-level comparison was performed with constrained spherical deconvolution (CSD)-based tractography, which is able to detect complex  fiber configurations. In addition, to investigate the relationship between the microstructural changes and the severity of symptoms, we looked for correlations between FA and the Autism Spectrum Quotient (AQ), Empathy Quotient and Systemizing Quotient.TBSS revealed widely distributed local increases in FA bilaterally in individuals with AS, most prominent in the temporal part of the superior longitudinal fasciculus, corticospinal tract, splenium of corpus callosum, anterior thalamic radiation, inferior fronto-occipital fasciculus (IFO), posterior thalamic radiation, uncinate fasciculus and inferior longitudinal fasciculus (ILF). CSD-based tractography also showed increases in the FA in multiple tracts. However, only the difference in the left ILF was significant after a Bonferroni correction. These results were not explained by the complexity of microstructural organization, measured using the planar diffusion coefficient. In addition, we found a correlation between AQ and FA in the right IFO in the whole group.Our results suggest that there are local and tract-level abnormalities in white matter (WM) microstructure in our homogenous and carefully characterized group of adults with AS, most prominent in the left ILF. Asperger syndrome (AS) is a neurodevelopmental autism spectrum disorder (ASD), which affects 0.6 to 0.7% of the population . The maiin vivo investigation of anatomical brain connectivity [Functional imaging studies have shown that there is reduced long-distance and increased short-distance connectivity in ASD –6. Recenectivity –10. In wectivity . Mean diectivity . In partectivity –15.The findings of diffusion MRI studies in ASD are not consistent. Both decreased and increased FA values have been reported in many WM tracts in ASD, compared to typically developing control subjects , 17. MosThe aim of our study was to investigate potential differences in FA and MD in adults with ASD. To narrow down the variation in the cognitive profile of the subjects, we only investigated high-functioning individuals with AS. Individuals with AS and autism share the same core symptoms, but according to DSM-IV, subjects with AS do not have a clinically significant delay in speech and cognitive development, although in DSM-5, autism and AS (among others) were placed on the same spectrum of autistic disorders.As different analytic approaches may also contribute to the varying results in DWI studies in ASD, we have chosen to use multiple approaches to confirm that the results support each other. In our previous histogram-based study, we used a dual approach, a skeletonized white matter histogram and a whole-brain tractography histogram, and found a global increase of FA values in adult males with AS compared to age-, sex- and IQ-matched controls, suggesting a more coherent neural tract organization in the individuals with AS . This rat-test was used to test for group differences. There were no significant differences in total IQ, verbal IQ, performance IQ, FRT, Eyes test or SQ, whereas individuals with AS had significantly higher AQ scores (P = 0.0000001) and significantly lower EQ scores than controls (P = 0.0017) . The patients were recruited from a private neuropsychiatric clinic  in Helsinki and from the neuropsychiatric clinic in Helsinki University Central Hospital. Only individuals fulfilling ICD-10  criteria, diagnosed by experienced clinicians specialized in developmental neuropsychiatry, were included in the study. Both individuals with AS and controls had a full psychiatric evaluation before inclusion in the study. Diagnostic process for the AS group included full developmental history, acquired using multiple sources of information . Benton Facial Recognition Test (FRT)  and Read2. Echo time (TE) was set to the minimum (approximately 98 ms). Repetition time (TR) was 10 s, and the number of excitations (NEX) was one. The imaging area covered the whole brain with 53 contiguous axial slices. The acquired in-plane resolution of the slices was 1.875 mm × 1.875 mm, and the thickness of the slices was 3.0 mm. The matrix size was 128 × 128.The MR images were acquired with a Signa VH/i 3.0 T scanner with HDxt upgrade . A quadrature receiving eight-channel high-resolution brain array coil was used . The maximum field gradient amplitude of the MRI system was 40 mT/m with a slew rate of 150 T/m/s. A high-order shimming with 24-cm field of view (FOV) was applied prior to DW imaging. A spin echo pulsed sequence of 60 unique gradient orientations arranged on the unit sphere was used. Eight nondiffusion weighted B0-images were acquired and all of the 60 orientations were imaged twice, resulting in 120 diffusion-weighted images in total. The b-value, which controls the diffusion weighting, was 1000 s/mmt-test. All of the parameters of the affine transform (12 degrees of freedom) were tested both for absolute and relative differences [P < 0.05) in any of the parameters.The DW acquisitions of the two groups were tested for possible differences in subject motion using a two-tailed ferences . No grouThe voxelwise statistical analysis of the diffusion data was carried out using TBSS , which ba priori information of tract location [CSD-based tractography was performed in ExploreDTI together with the preprocessing of the data , 45. Thelocation . The tralocation . The semlocation . The invlocation  and the A voxelwise comparison of FA, MD and CP values between individuals with AS and age, sex and IQ-matched controls was performed with TBSS, and the findings were confirmed by CSD-based tractography.P <0.05) in the following tracts  is minimized in terms of adverse contributions of GM or cerebrospinal fluid to WM voxels. Furthermore, no smoothing is needed. However, abnormalities in the WM might reduce the FA value of a tract locally, and it is possible that the voxel chosen to the skeleton is not actually in the center of the tract. Therefore, we transformed the TBSS results back to native space of each subject to confirm that a given point in the skeleton was derived from the correct anatomically corresponding region. A limitation of TBSS is that by studying only the voxels on the skeleton with the highest FA, a large part of WM remains uninvestigated. The tract-level analysis with the CSD-based tractography method complements the more local TBSS approach, as it is not limited to the voxels with the highest FA in the cross-section of the tract.P value = 0.02633). However, Bonferroni correction for multiple comparisons is very conservative and does not account for the correlation between the FA values of different tracts. Thus, in regard to the previous global finding and the results of TBSS, it is possible that true increases in FA may be present in also other tracts than left ILF. No significant changes were found in MD, CP or LI in any of the tracts between the two groups.CSD-based tractography showed an increased FA in the right IFO, left ILF, right and left SLF and left UNC in individuals with AS compared to controls, as shown in Table P = 0.06417). No correlation was found between SQ and FA in any of the WM tracts.To see if the findings in FA correlated with a specific area such as empathy or systemizing skills, we performed a correlation analysis of AQ, EQ and SQ with FA. Results are shown in Table Anisotropic diffusion is primarily caused by dense packing of axons and their cell membranes . Other tIncreased FA indicates that the coherence of the WM tracts in subjects with AS may be higher than in controls . HoweverAlthough most of the previous studies in adults with ASD have reported decreased FA values –23, incrCSD-based tractography has been previously used in only two studies in ASD , 33. McGThe inconsistent findings in the previous studies may be partly due to the use of different methods, but also to the heterogeneity of the samples, as in ASD, the diversity of symptoms is large and variation in the degree of their severity among individuals varies a lot. Differences in age and IQ between the different samples may also affect the results, as it has been shown that FA values correlate with IQ  and thatThe large amount of different analysis methods makes it more difficult to compare different studies with each other, as each of the methods has its limitations , 71, 72.Thus, TBSS and CSD-based tractography complement each other, and both indicate increased FA in individuals with AS. Furthermore, these results are supported by our earlier findings of globally increased FA in subjects with AS , but proLimitations of our study include a relatively small sample size. However, our results survived corrections for multiple comparisons and thus, can be considered robust. Finland is an isolated and genetically homogeneous country , which cWe investigated local microstructural differences in the WM of individuals with AS by comparing their FA, MD and CP values to those of age-, sex- and IQ-matched controls. We used a dual approach and performed first a more local voxelwise analysis, TBSS, and then confirmed the findings with a tract-level analysis approach, CSD-based tractography. Our results suggest that there are widely distributed abnormalities in the WM tracts of adults with AS, and that these are most pronounced in the left ILF. The changes were not explained by differences in the complexity of the microstructural organization of the WM. Furthermore, these results are in line with our earlier findings of globally increased FA in subjects with AS .Our study was restricted to adult males with AS. In the future, longitudinal studies beginning in the childhood and following the diagnosis and possible rehabilitation should be performed to more thoroughly understand the neural deficits and the effects of age and IQ in ASD. Furthermore, as ASD is a very broad and heterogeneous disorder, the study samples should be as homogeneous as possible, and different symptom domains should be investigated separately. Finally, the WM tracts could be segmented and thus, short parts of the tracts could be investigated separately in addition to looking at the mean FA of the whole tract.UR has two academic degrees, a Master of Science (M.Sc.) in Technology from Aalto University (2010) and medical degree from the University of Helsinki (2011), acquired simultaneously. In the beginning of her medical studies she was chosen to a graduate program for medical doctors, and she will soon finish her doctoral studies, during which she has attended multiple scientific conferences and courses and visited research groups abroad. She’s especially interested in neuropsychiatry, diffusion magnetic resonance imaging and applying the newest technology to clinical studies. JS is a researcher and a psychologist. He has a PhD in psychology. He is currently working as a clinical psychologist helping children and adults with neurodevelopmental disabilities and as a post-doctoral researcher in the field of cognitive neuroscience. TR received his M.Sc. (Tech.) in 2009 from Helsinki University of Technology , Finland. He did research on mineral processing technology in the Control Engineering group of Aalto University from 2005 to 2010. From 2010 to 2012 he worked as a consultant for healthcare management in Nordic Healthcare Group, Finland. In 2012 he started his PhD about diffusion MRI and constrained spherical deconvolution at the iMinds-Vision Lab, Department of Physics, University of Antwerp, Belgium under the supervision of Ben Jeurissen, Alexander Leemans and Jan Sijbers. TNvW is a pediatrician and a child neurologist. She did her PhD on Asperger syndrome, and she is a specialist in diagnostics, neuroimaging and molecular genetics of AS. TNvW has a wide experience in neuropsychiatric diagnostics, and she treats both children and adults with AS. She is the managing director of the Neuropsychiatric Rehabilitation and Medical Centre Neuromental in Helsinki, Finland. SL is an adjunct professor of psychiatry at the University of Helsinki. He has ten years of experience with developmental neuropsychiatric disorders and is especially interested in clinical manifestations of ADHD and ASD in adulthood. In addition, he has done research on mood disorders and chronobiology. PR is an MD and a PhD. He has two specialties regarding neuropsychiatric disorders: he is a pediatric neurologist and an adolescent psychiatrist. In addition, he has five years of brain imaging experience with neuropsychiatric disorders at UCLA PET-Center and Neuropsychiatric Center. In Finland, he has clinical work experience with neuropsychiatric disorders including Asperger syndrome and other ASD, and he works at the Helsinki University Central Hospital Adolescent Psychiatry Clinic. His main field is evaluating neuropsychiatric patients and their comorbid conditions in addition to medical treatment and rehabilitation. PT is an adjunct professor of psychiatry at the University of Helsinki. He has more than ten years of experience on developmental neuropsychiatric disorders and is especially interested in clinical manifestation of ASD in adulthood. In addition, he has done research in forensic psychiatry. AL is a physicist who received his PhD in 2006 at the University of Antwerp, Belgium. From 2007 to 2009, he worked as a postdoctoral researcher at the Cardiff University Brain Research Imaging Center (CUBRIC), Cardiff University, Wales, United Kingdom. In 2009, he joined the Image Sciences Institute (ISI), University Medical Center Utrecht, the Netherlands, where he currently holds a tenured faculty position as Associate Professor. His current research interests include modeling, processing, visualizing and analyzing diffusion MRI data for investigating microstructural and architectural tissue organization. He heads the PROVIDI Lab and is the developer of the software ExploreDTI. MS is a professor of cognitive neuroscience in Aalto University. He is very experienced in noninvasive study of neural basis of cognitive functions such as attention, speech perception, auditory perception and multisensory integration. Now his research is focused on studying brain activity in naturalistic conditions.