Purpose: Having a low level of physical fitness, especially cardiorespiratory fitness, appears to accelerate age-related aortic stiffening. Whereas, some studies have reported that trunk flexibility is a component of physical fitness, it is also negatively associated with arterial stiffening independent of cardiorespiratory fitness in cross-sectional studies. However, no long-term longitudinal study has determined whether poor trunk flexibility accelerates the progression of age-related aortic stiffening. We examined trunk flexibility and aortic stiffness progression in a 5-year longitudinal study.Methods and Results: A total of 305 apparently healthy men and women participated in this study (49.6 ± 9.5 years of age). Trunk flexibility was measured using a sit-and-reach test. Aortic stiffness was assessed using carotid-femoral pulse wave velocity (cfPWV) at baseline and after 5 years. Analysis of covariance (ANCOVA) was used to assess the association of the annual rate of cfPWV across flexibility levels . There were no significant differences in baseline cfPWV among the three groups . Annual ΔcfPWV was significantly higher in the low-flexibility group than in the high-flexibility group (P = 0.009). ANCOVA revealed an inverse relationship between flexibility level and annual ΔcfPWV . Multiple regression analysis revealed that baseline sit and reach  was independently correlated with ΔcfPWV following adjustment for baseline peak oxygen uptake, age, sex, body fat, heart rate, and cfPWV. The 5-year change in cfPWV was not significantly correlated with 5-year change in sit-and-reach performance (P = 0.859).Conclusion: Poor trunk flexibility is associated with greater progression of age-related aortic stiffening in healthy adults. However, we failed to confirm a significant association between 5-year change in aortic stiffness and 5-year change in trunk flexibility. The association between increased age-related increase in aortic stiffness and deterioration in flexibility due to age may require observation for more than 5 years. Aortic stiffness, as indexed based on pulse wave velocity (PWV), increases progressively with advancing age Lakatta,  and is aHaving a high level of physical fitness, especially higher cardiorespiratory fitness, is associated with the suppression of age-related arterial stiffening , which can be measured non-invasively, easily, quickly, and safely, may represent the phenotypic stiffness of various body parts. Flexibility is determined by connective tissue in the tendons, muscles, ligaments, and joint capsules Alter, . IncreasClinical Trials.gov, identifier: NCT00926744. For the purpose of this study, a total of 305 Japanese adults  were selected from among 1,125 participants. Participants underwent anthropometric measurements, physical fitness testing (cardiorespiratory fitness and flexibility), physical activity assessments, arterial stiffness measurement, and blood examinations (baseline measurement). They underwent the same tests  at 5-year follow-up (follow-up measurement). We excluded participants with a history of stroke, cardiac disease, or chronic renal failure, and those undergoing medical treatment for hypertension, dyslipidemia, or diabetes. We also excluded current smokers. Participants in this sample had an ankle-brachial pressure index between 0.9 and 1.3 at both baseline and the follow-up visit (during the observation period). All participants gave their written informed consent before participating in the study. The study was approved by the ethical committees of the National Institutes of Biomedical Innovation, Health and Nutrition, and Okayama Health Foundation, and the study was performed in accordance with the guidelines of the Declaration of Helsinki.The study population is part of the Nutrition and Exercise Intervention Study (NEXIS) cohort, registered at We measured carotid and femoral PWV (cfPWV) as indicators of aortic stiffness and blood pressure with a vascular test device  as described previously  as described previously  -specific sit and reach test results: lowest tertile, poor flexibility; middle tertile, mid-range flexibility; and highest tertile, high flexibility.We measured peak oxygen uptake as an indicator of cardiorespiratory fitness, which was measured according to the protocol of a graded exercise load using a cycle ergometer , as described previously , as described previously . We measured waist circumference using a tape measure. Body mass index (BMI) was calculated as measured weight in kilograms divided by the square of measured height in meters.Blood samples were obtained after at least 10 h of overnight fasting. Fasting plasma glucose and glycated hemoglobin (HbA1c) and serum levels of total cholesterol, high-density lipoprotein (HDL) cholesterol, and triglycerides were measured.t-test and McNemar's non-parametric test. In the ANOVA, Scheffe's method was used to identify significant differences among mean values.Means ± standard deviations were calculated for continuous variables. Analysis of variance (ANOVA) was used to assess the relationship of the continuous variables to categories of flexibility levels. The differences between baseline and follow-up measurements were assessed by paired Pearson's correlation coefficients were used to analyze the relationships between the 5-year changes in cfPWV and baseline values of factors known to influence vascular stiffness and 5-year changes in these variables.post-hoc comparisons were examined using a Bonferroni test. In ANCOVA, data were expressed as estimated marginal mean ± standard error.Analysis of covariance (ANCOVA) models were estimated to test differences in the annual rate of cfPWV [annual ΔcfPWV: (follow-up cfPWV—baseline cfPWV)/follow-up years] across flexibility levels. The annual ΔcfPWV was entered as a dependent variable; the tertile flexibility category was entered as a fixed factor; and baseline age, weight, body fat, systolic blood pressure (SBP), HR, cfPWV, peak oxygen uptake, moderate physical activity time, vigorous physical activity time, and sex were entered as covariates for adjustment. Pairwise A multiple regression analysis was used to determine the influences of baseline values of factors known to influence vascular stiffness and changes in these variables  on the annual ΔcfPWV.P values < 0.05 were considered statistically significant. Data were analyzed using SPSS software .P = 0.577). The paired t-test demonstrated that cfPWV increased significantly during the follow-up period in low and middle flexibility groups. The body fat, HbA1c, SBP increased significantly during the follow-up period in all flexibility groups.Table r = −0.16, P = 0.005); weight ; BMI ; waist ; cfPWV ; peak oxygen uptake ; and 5-year changes in SBP , diastolic blood pressure (DBP) , and HR . The 5-year change in cfPWV was not significantly correlated with the 5-year-change in sit-and-reach. Moreover, univariate regression analyses were used to assess the relationships between sit-and-reach values and the 5-year changes of cfPWV in different age categories . The Pearson's correlation coefficient was larger in older subjects than in young and middle-aged subjects . These results suggest that poor trunk flexibility is associated with greater progression of age-related aortic stiffening, especially in older adults.Pearson's correlation coefficients between the 5-year change in cfPWV and baseline and 5-year changes in other parameters were as follows: the 5-year change in cfPWV was correlated with baseline sit-and-reach (P for trend = 0.011). The annual ΔcfPWV was significantly higher in the low-flexibility group than in the high-flexibility group (P = 0.009). Moreover, we performed ANCOVA analyses by sex. In men, ANCOVA revealed an inverse relationship between flexibility level and the annual ΔcfPWV . In women, ANCOVA revealed an inverse relationship  between flexibility level and annual ΔcfPWV .Figure As shown in Table In this 5-year longitudinal study, the age-related increases in aortic stiffness were higher in the low-flexibility groups compared with those in the high-flexibility groups. Moreover, trunk flexibility was inversely associated with the annual rate of change in cfPWV. This association was independent of other confounders. These findings suggest that poor trunk flexibility is associated with greater progression of age-related aortic stiffening in healthy adults.The current study has several strengths that set it apart from previous studies. Previous cross-sectional studies indicated that poor trunk flexibility was associated with arterial stiffening  in the present study. Further research is necessary to determine the relationship between R577X genotype and age-related arterial stiffening. In general, there are gender differences in age-related arterial stiffening , and therefore may be a practical and achievable preventive strategy in older people. Studies have indicated that acute . Because the present study included subjects with a broad age range (27–66 years), our data may be less sensitive to cfPWV compared with previous reports. Our previous study suggested that trunk flexibility was correlated with arterial stiffness in older adults, but not in young adults  and −0.32 (older) and, in women, r = −0.02 (young) and −0.13 (older). These results seem to show greater sensitivity to cfPWV in older men. Therefore, there might be no significant differences in baseline cfPWV among the three groups. Fourth, we carried out this study as a sub-analysis of the NEXIS, which is an ongoing prospective study. Therefore, we used an observational study design. More research is needed to determine cause-and-effect relationships between flexibility and arterial stiffness.There are several limitations to the present study. First, the sit-and-reach test used as an indicator of flexibility is a multifactorial test, which may compromise the interpretation of information. We did not evaluate the flexibility of other body regions such as the elbow, shoulder, knee, or ankle. Further studies are required to refine our understanding of the link between flexibility and arterial stiffness. Second, arterial stiffness is influenced by the phases of the menstrual cycle. We could not control for the menstrual cycle in this study. However, the number of premenopausal women was relatively low (32%), and there were no significant differences among the three groups. Therefore, we think that this factor had a small effect on aortic stiffening. Third, there were no significant differences in baseline cfPWV among the three groups (The present longitudinal study suggests that poor flexibility is associated with greater progression of age-related aortic stiffening. This association was independent of known confounders including cardiorespiratory fitness. Therefore, trunk flexibility may be an effective measure for preventing age-related aortic stiffening.YG designed the work, acquired, analyzed and interpreted the data, and wrote the first draft of manuscript. HM planned, supervised the study, and acquired the data. KY conceived the idea for the study, wrote the first draft of the manuscript, and acquired and interpreted the data. RK, HO, and NM acquired and interpreted the data. SS interpreted the data. MM designed the work, interpreted the data, wrote the first draft of the article, and supervised the study. All authors read and approved the final manuscript.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.


Individuals with schizophrenia have heterogeneous impairments of the auditory processing system that likely mediate differences in the cognitive gains induced by auditory training (AT). Mismatch negativity (MMN) is an event-related potential component reflecting auditory echoic memory, and its amplitude reduction in schizophrenia has been linked to cognitive deficits. Therefore, MMN may predict response to AT and identify individuals with schizophrenia who have the most to gain from AT. Furthermore, to the extent that AT strengthens auditory deviance processing, MMN may also serve as a readout of the underlying changes in the auditory system induced by AT.n = 105) were compared to baseline MMN data from the ESZ group.Fifty-six individuals early in the course of a schizophrenia-spectrum illness (ESZ) were randomly assigned to 40 h of AT or Computer Games (CG). Cognitive assessments and EEG recordings during a multi-deviant MMN paradigm were obtained before and after AT and CG. Changes in these measures were compared between the treatment groups. Baseline and trait-like MMN data were evaluated as predictors of treatment response. MMN data collected with the same paradigm from a sample of Healthy Controls . Reduced Double-Deviant MMN was associated with greater general cognitive impairment in ESZ individuals (p = .020). Neither ESZ intervention group showed significant change in MMN. We found high correlations in all MMN deviant types  between baseline and post-intervention amplitudes irrespective of treatment group, suggesting trait-like stability of the MMN signal. Greater deficits in trait-like Double-Deviant MMN predicted greater cognitive improvements in the AT group (p = .02), but not in the CG group.Compared to HC, ESZ individuals showed significant MMN reductions at baseline (In this sample of ESZ individuals, AT had no effect on auditory deviance processing as assessed by MMN. In ESZ individuals, baseline MMN was significantly reduced relative to HCs, and associated with global cognitive impairment. MMN did not show changes after AT and exhibited trait-like stability. Greater deficits in the trait aspects of Double-Deviant MMN predicted greater gains in global cognition in response to AT, suggesting that MMN may identify individuals who stand to gain the most from AT.NCT00694889. Registered 1 August 2007. Dysfunction in the coordination of neural activity during auditory processing is well-documented in individuals with schizophrenia , 2. ImpaTraining of auditory processing has evolved as a target in the development of interventions for the cognitive impairments characteristic of schizophrenia. The computerized auditory training (AT) program we have studied simultaneously targets feedback and feed-forward operations in the auditory system . The traTwo randomized clinical trials (RCT) that investigated the effectiveness of AT in schizophrenia reported improvements in general cognitive performance, with moderate to large effect sizes observed in individuals with recent-onset (d = 0.56) and chronic (d = .86) schizophrenia-related disorders , 17. DesWhy do some individuals with schizophrenia show evidence of robust cognitive improvement, while others do not? The observed variability in cognitive gains could be due to the heterogeneity of impairment of the prefrontal-temporal neural systems that underlie auditory processing. Investigating the neurophysiological bases of auditory system dysfunction in schizophrenia could therefore be key to identify predictors of response to AT and guide mechanistically informed, personalized treatments. Among the various indexes of early auditory processing, mismatch negativity (MMN), an event-related potential (ERP) that is elicited pre-attentively when an infrequent deviant sound violates an established pattern of repeated standard sounds , has beeN-methyl-D-aspartate receptor (NMDAR) function [Echoic sensory memory can be modulated by means of pharmacological agents, and these effects can be reliably assessed by changes in MMN. Because MMN depends critically on function , the modfunction , 33. Furfunction . In addifunction , 36.The reduced MMN amplitude, well documented in chronic schizophrenia, first episode psychosis, and even individuals at risk for psychosis –40 is thBesides being consistently reduced across the course of illness, MMN has other important characteristics that support its role as a potential vulnerability marker for schizophrenia and its use as a repeated measure in intervention studies, including substantial heritability , lack ofDespite a general convergence of research findings showing reduced MMN amplitude in response to a Frequency-Deviant  or a DurAnother hypothesis for inconsistent reports of MMN amplitude reduction is based on the high degree of heterogeneity in the fundamental mechanisms of short term synaptic plasticity found in schizophrenia –68. AlthFrom a methodological point of view, a third possible explanation for inconsistent reports of reduced MMN amplitude in individuals with schizophrenia may lie in the magnitude of the psychophysical deviation and in the different types of deviant stimuli used to elicit MMN, since it appears that distinct neural populations process different dimensions of auditory deviance –72. GiveAbove and beyond its role as a predictor of treatment response, and because of its trait-like stability, MMN may serve as an indicator of neurophysiologic changes in the central auditory system resulting from AT. Studies conducted with healthy participants support this hypothesis, showing persistent MMN improvements after auditory discrimination training –75. SimiIn this study, we assessed cognitive performance and auditory MMN in individuals with schizophrenia randomly assigned to intensive auditory training (AT) or computer games (CG) before and after the intervention. We enrolled participants with a schizophrenia-spectrum illness within 5 years of treatment initiation or illness onset. We hypothesized that individuals in AT but not in CG would have a non-zero relationship between baseline MMN and cognitive gains following treatment. Regarding the directionality of this relationship, our leading hypothesis was that individuals with greater auditory processing deficits, as indexed by reduced MMN amplitude, would show greater cognitive improvements after AT, but not after CG. The competing hypothesis was that normal MMN amplitude would reflect intact short term plasticity and therefore help identify the individuals who would show the greatest cognitive gains after AT, but not after CG. Finally, we hypothesized that individuals with schizophrenia would show increased MMN amplitude after AT but not after CG, and that MMN changes would correlate with cognitive changes in the AT group but not in the CG group.Fifty-six participants early in the illness course of schizophrenia, schizophreniform, or schizoaffective disorder (ESZ) completed an auditory cognitive training treatment study protocol at the University of California, San Francisco  and completed MMN measures. This sample comprises the subset of participants from the total trial sample (which has been reported upon previously ) who agrn = 52) were on a stable dose for at least one month prior to study participation. Four participants were not taking antipsychotic medications during the study. The study was approved by the Institutional Review Board of University of California, San Francisco. Adult participants and parents of minors provided written informed consent, and minors provided written assent.All ESZ participants had achieved outpatient status for at least 3 months, and participants taking antipsychotic medications (n = 27) or computer games (n = 29). ESZ participants were loaned laptop computers and participated in the intervention at home, except for 1 training subject who preferred to participate in the laboratory. ESZ participants were asked to participate for 40 h , followed by post-training assessments.Eligible ESZ participants completed a battery of clinical, neuropsychological and EEG assessments. Baseline assessments were conducted prior to randomization. ESZ participants were randomly assigned to auditory training (The auditory training (AT) program, provided by Posit Science Corporation, has been described in detail previously . It consThe computer games (CG) control condition controls for the effects of computer exposure, contact with research staff, monetary compensation, and non-specific engagement of attention, executive functions, and motivation. ESZ participants in the CG condition rotated through a series of 16 different commercially available games for the same number of hours as participants in the AT group, playing 4–5 games on any given day .All ESZ participants received compensation for study participation, and payment was contingent on study participation and not performance . During the intervention, ESZ participants were free to receive treatments by clinicians who were not involved in the study .z-scores using age-appropriate normative data provided in testing manuals and age-appropriate, published normative data. We used the Global Cognition score  as our primary cognitive outcome.All assessment staff were blind to group assignment. Eligibility diagnoses were determined using the Structured Clinical Interview for DSM-IV TR . SymptomWe assessed three types of MMN using a two-Deviant paradigm and a Single-Deviant paradigm. The two-Deviant paradigm assessed Frequency-Deviant MMN and Duration-Deviant MMN. In this paradigm, 80% of the stimuli were standard tones , 10% were duration deviants , and 10% were frequency deviants . The Single-Deviant paradigm assessed the MMN elicited by a Frequency + Duration Double-Deviant (DBL) stimulus. In this paradigm, 90% of the stimuli were standard tones , and 10% were double-deviants . Across paradigms, all tones had 5 millisecond rise/fall times and were presented with a 500 msec stimulus onset asynchrony at 78 dB sound pressure level via Etymotic ER3-A insert earphones. The paradigms were administered in four separate blocks  lasting approximately 5 min each, with each block comprising a fixed pseudorandom sequence of 615 tones. The order of the four MMN blocks was randomized. In order to reduce the influence of attention on the MMN measurements, participants were instructed to ignore auditory stimuli while performing simultaneously a computerized picture-word matching task presented on a video display that required a button press response on each trial  .EEG activity during the MMN paradigms was recorded in the ESZ participants before and after the AT or CG intervention. To confirm the presence of MMN amplitude reduction in the ESZ participants at baseline, we used EEG data from age-matched healthy control (HC) participants who had completed the same MMN paradigm in the context of other research studies. HC participants were recruited by advertisements and word-of-mouth. Those with a past or current DSM-IV Axis I disorder  or a first-degree relative with a psychotic disorder, and or with a history of substance dependence or abuse within the past year, a history of a significant medical or neurological illness, or a history of head injury resulting in loss of consciousness were excluded.EEG data were acquired using a high-impedance BioSemi Active Two recording system and a 64-channel electrode cap . Continuous EEG data were digitized at a rate of 1024 Hz, referenced offline to averaged earlobe electrodes, high-pass filtered at 1 Hz, and segmented into 1000 ms epochs time-locked to the onsets of the various types of auditory stimuli (−500 to 500 ms). Vertical and horizontal electro-oculograms, recorded from electrodes above and below the left eye and at the outer canthi of both eyes, respectively, were used to correct EEG for eye movement and blink artifacts using a regression-based algorithm . Additioz- scores. These z-scores represent, in standard units, the degree to which a subject’s (ESZ or HC) MMN amplitude deviates from the normal value expected for their age. Accordingly, more positive z-scores indicate smaller MMN amplitude relative to HC norms . For statistical comparisons of age-corrected MMN z-scores between the ESZ and HC groups, we only included the HC participants (n = 105) who were younger than the oldest ESZ patient (age = 36.17). Because z-scoring effectively sets the HC mean equal to zero, within-subjects effects were assessed in a separate model using raw MMN scores.Given the broad age range of our ESZ sample , additional steps were taken to control for normal brain development and aging effects. To this purpose, we used EEG data from 121 healthy control (HC) participants who had previously completed the same MMN paradigm. Normal aging effects on the MMN were modeled in the HC (age range 12–43 years) by regressing the MMN amplitudes on age separately for each Deviant type and electrode. Next, resulting regression models were used to derive predicted normal MMN amplitudes for each subject (ESZ and HC) based on their specific age. Finally, we divided the differences between observed and age-specific predicted MMN amplitudes by the standard error of regression (from the HC regression model), yielding age-corrected MMN MMN amplitudes averaged over the six fronto-central electrodes  were used in the analyses. To examine the contribution of Single-deviant MMNs to the Double-Deviant MMN, we performed regression analyses in which we regressed DBL MMN onto DUR and FREQ MMN, for ESZ and HC participants separately.z-scores were assessed using a 4-way repeated measures Analysis of Variance (ANOVA) with Group  as the between-subjects factor and Deviant Type , Fronto-Central Lead , and Lateral Lead  as within-subjects factors. Significant effects were parsed using follow-up F-tests of simple main effects. Greenhouse-Geisser non-sphericity correction was applied to within-subjects effects with more than two levels.Group differences between HC and ESZ in baseline MMN age-corrected z-scores and baseline MATRICS Global Cognition scores. Alpha was set to p = 0.05, two-tailed, for all statistical tests.All subsequent analyses were exclusively conducted on ESZ participants who completed baseline and post-training EEG and cognitive assessments. Cognitive variables were screened and normally distributed after winsorising of outlying values . Pearsond) were computed using the mean change scores of the AT and CG groups (post-treatment minus baseline) and the change score SDs of each subgroup.AT and CG groups were compared on change in Global Cognition and MMN age-corrected z-scores using repeated measures ANOVA. Effect sizes  as the between-subjects factor and Deviant Type , Fronto-Central Lead , and Lateral Lead , and Time  as within-subjects factors. Significant effects were parsed using post-hoc Tukey-Kramer tests. Greenhouse-Geisser non-sphericity correction was applied to within-subjects effects with more than two levels. Pearson correlations were used to examine associations between Global Cognition change scores and MMN z-change scores changes in AT participants.Change in MMN z-scores predicted changes in Global Cognition scores, we performed a separate regression analysis for each MMN Deviant type in which we regressed the Global Cognition change score (post-treatment minus baseline) on Treatment Group (AT vs CG), baseline MMN z-score, and their interaction (baseline MMN z-score* Treatment Group). The interaction effect tests whether the slope of the relationship between Global Cognition change and MMN significantly differs between the groups.To test whether baseline MMN z-scores across Treatment groups for each MMN type separately. The stability of the trait aspects of MMN was operationalized as the average of baseline and post-training MMN z-scores. Twenty eight of the HC participants completed a second EEG session approximately 6 months (6.7 ± 1.8) after their baseline EEG session, allowing for ICC calculation in this HC sub-group as well.The stability of the MMN signal was examined by calculating Intra-class correlations (ICCs) between baseline and post-training MMN raw and z-score, and their interaction. For all regression models, in case of statistically significant slope differences, we ran follow-up within-group Pearson correlations to estimate the strength of the correlation between the predictor and the dependent variable.Finally, we regressed Global Cognition change scores on Treatment Group (AT vs CG), trait-like MMN z-scores were moderately correlated . DBL MMN similarly correlated with FREQ MMN  and DUR MMN . In the ESZ group, baseline DUR and FREQ MMN z-scores were moderately correlated . DBL MMN similarly correlated with FREQ MMN  and DUR MMN . When the DBL MMN was regressed on both frequency and duration MMN in a multiple regression model in the healthy controls, both accounted for independent aspects of the variance of the DBL MMN , leaving 54% of the DBL MMN variance unaccounted for. A similar pattern of results was observed in the ESZ group . Thus, while in ESZ the DBL MMN shared more variance with the DUR MMN than with the FREQ MMN, it contained information over and above that provided by the Single-Deviant frequency and duration MMNs.In the HC group, baseline DUR and FREQ MMN Group x Deviant Type x Fronto-Central Lead x Lateral Lead repeated measures ANOVA of MMN z-scores are presented in p = .029) indicating an overall reduction of MMN amplitude (more positive z-score) in the ESZ relative to the HC, there was also a significant Group x Fronto-Central Lead x Deviant Type interaction (p = .005). This three-way interaction was parsed by examining the Group x Fronto-Central Lead effect for each deviant type. The Group x Fronto-Central Lead effect was significant for the DBL MMN (p = .006), but not for the DUR or FREQ MMN. Further parsing of this Group x Fronto-Central Lead interaction for DBL MMN indicated a significant Group effect at Frontal leads (p = .003), with ESZ showing MMN deficits compared to HC, and a non-significant Group effect at Central leads. We also parsed the three-way interaction by examining the Group x Deviant Type interaction separately at frontal and central leads, but in neither case was this effect significant. Taken together, these analyses indicate the presence of a DBL MMN amplitude deficit over frontal leads in the ESZ group compared to the HC group. Analysis of raw scores indicated a main effect of deviant type  that was driven significantly larger DBL MMN amplitude than DUR (p = .002) but not FREQ (p = .469) MMN. Ear-referenced ERP difference waveforms and scalp voltage topography maps of MMN amplitudes for HC and ESZ are presented in Results of the z-scores), for both the DBL MMN  and the FREQ MMN , but not for DUR MMN . Demographic data and mean MMN z-scores averaged across the six fronto-central leads for ESZ participants undergoing 40 h of AT vs. CG are presented in z-scores. Demographic data for HC are presented in Within the ESZ group, greater impairment in baseline Global Cognition was associated with greater deficits in MMN amplitude (more positive z-scores are presented in p = .059) and a trend-level Treatment Group x Time interaction  with small effect size (d = 0.25). Mean Global Cognition z-score changes and SDs were 0.25 ± 0.62 for the AT group and 0.01 ± 0.41 for CG group. Eighteen (67%) participants in the AT group and 8 (27%) participants in the CG group showed Global Cognition improvements beyond expected practice effects (0.2 SD) [X square = 9.54, p = .002).Mean Global Cognition pre- and post-treatment (0.2 SD) , a diffez-scores are presented in z-scores between treatment groups  and a significant Treatment Group x Lateral Lead x Time interaction . However, the Treatment Group x Deviant Type x Lateral Lead x Time interaction was not significant . The Treatment Group x Lateral Lead x Time interaction were parsed in several ways. The Treatment Group x Time effect was examined for each each lateral scalp site separately, collapsing across fronto-central leads and deviant types. Here we found no significant Treatment Group x Time interactions for left, midline or right scalp sites. Additional parsing of the interaction effect is presented in z-scores in AT and CG participants.Mean MMN and pre- and post-treatment t groups , we founz-score interaction terms from the regression models, tests of the common slope predicting change in global cognition from baseline MMNs revealed significant effects for DBL MMN (p = .048) and DUR MMN (p = .013), and trend-level effects for FREQ MMN (p = .056), with greater MMN deficits predicting greater cognitive improvements across groups.Next, we tested a series of multiple regression models to evaluate whether baseline MMN predicted changes in Global Cognition, and whether such predictive relationships significantly differed between the two treatment groups. Models were run for each MMN deviant type. There were no significant slope differences between the groups for any of the MMN deviant types. After dropping the non-significant Group x MMN z-scores induced by the AT and CG interventions, we examined the extent to which individual differences in MMN amplitude z-scores among ESZ participants showed trait-like stability over the pre-post test interval , ignoring the small influence of treatment effects. This was done by simply calculating ICCs between baseline and post-training MMN raw and z-scores in the HC and ESZ samples. We found fair to good test-retest correlations for all MMN deviant types (ICCs are presented in ps < .001), suggesting trait-like stability and durable reliability of the MMN signal, despite the minor changes induced by the two treatments or by time in the HC sub-sample.Given the non-significant changes in MMN amplitude z-scores for each MMN deviant type and for the frontal and central leads separately. These trait-weighted MMN measures, reflecting more temporally stable aspects of MMN, were then entered into a series of regression models examining their relationship with pre-post changes in Global Cognition and possible differences in the slopes of these relationships between the two treatment groups. While there were no significant slope differences between the groups for any of the FREQ or DUR MMN deviant types, there was a significant Treatment Group x time-averaged MMN interaction (p = .05) for the DBL MMN (see z-scores) significantly predicted greater improvements in Global Cognition in the AT group , whereas this relationship in the CG group did not approach significance . For scatter plots, see To best capture the trait aspects of MMN, and to enhance the reliability of the MMN measurements, we averaged the baseline and post-training MMN  MMN see . Follow-d = 0.25), although the larger study from which the current sub-sample was drawn did demonstrate significant benefits of auditory training relative to the computer game control condition, with a relatively large effect size for global cognition (d = 0.73) [In this study, we directly compared the sensitivity of MMN elicited by three types of auditory deviance  in individuals with a schizophrenia-spectrum illness (ESZ) who were, on average, within 2 years of their illness onset, and healthy controls (HC). We found reduced MMN amplitude irrespective of deviant types in the ESZ group relative to the HC group, consistent with prior studies –40, 87.  = 0.73) . ImportaConsistent with prior studies , 87, incGiven the potential for heterogeneity among individuals with ESZ in terms of which type of MMN is most affected by their particular variant of the illness, we included a Double-Deviant paradigm to increase sensitivity to variations in MMN deficits between ESZ participants. While we did not find significantly greater sensitivity of Double-Deviant MMN to ESZ relative to the corresponding Single-Deviant MMNs, consistent with our prior studies in ESZ  and in iz-scores), and not more intact MMN, were associated with larger cognitive gains induced by auditory training in ESZ. This supports the idea that those with greater auditory processing deficits stand to benefit the most from auditory training, but does not support the alternative hypothesis that greater MMN deficits may index impairments in the mechanisms of short-term synaptic plasticity on which auditory training depends, thereby identifying those who are less likely to show training-induced cognitive gains. Interestingly, this result contrasts with a recent report by Perez and colleagues [We also found that greater deficits in MMN amplitude   has rareOur findings provide preliminary evidence for a future personalized medicine approach that uses reliably measured baseline MMN to identify which individuals with schizophrenia are mostly likely to benefit from auditory training. Moreover, more longitudinal research is needed using optimally reliable MMN measurements to assess whether MMN progressively changes over time during the transition from psychosis-risk states to full-blown psychosis, particularly in light of recent evidence that greater MMN amplitude deficits are present in individuals at clinical high risk (CHR) who subsequently converted to psychosis , 87 and In conclusion, this study demonstrates that Double-Deviant MMN amplitude is a stable trait that is reduced in ESZ compared to HC. Moreover, the trait aspects of Double-Deviant MMN seem to predict response to auditory training over and above the Single-Deviant MMNs, with greater deficits  associated with larger cognitive gains. Given that the neuroanatomically distinct MMN generators associated with processing different dimensions of auditory deviance seem heterogeneously compromised in schizophrenia  and poss


The prognostic value of repeat transurethral resection of bladder tumor (TURBT) in patients with diagnosed high-risk, non-muscle-invasive bladder cancer (NMIBC) was investigated. We retrospectively reviewed the medical records of patients treated from October 2004 to December 2013 at Seoul National University who underwent repeated TURBT within 2–6 weeks after an initial resection. The study enrolled patients who had been diagnosed with NMIBC at both the initial and repeat TURBT; patients with muscle-invasive tumors on repeat TURBT were excluded. We used stepwise multivariate Cox regression models stratified by study to assess the independent effects of the predictive factors and estimated hazard ratios (HRs) from the Cox models. We investigated a total of 198 patients who were diagnosed with high-risk NMIBC. In logistic regression analyses, number of bladder tumors , initially high tumor grade , and presence of carcinoma in situ lesion  correlated with residual tumor in the repeated-TURBT specimen. T1 stage in repeated-TURBT significantly correlated with recurrence  and progression  in multivariate analysis. The high grades of tumors in repeated-TURBT also significantly correlated with progression but not recurrence in the multivariate analysis . In this study, the pathologic findings in repeated-TURBT correlated with recurrence and progression in high-risk NMIBC. Repeated-TURBT is valuable because it can predict the recurrence and progression of high-risk NMIBC in addition to obtaining accurate pathologic findings. Approximately 80% of all newly diagnosed bladder cancers are non-muscle-invasive bladder cancer (NMIBCs) . TransurThe accurate diagnosis, staging, and subsequent prognostication of bladder cancer are achieved by TURBT, and repeated TURBT (re-TURBT) may change treatment strategy in high-risk NMIBC; this includes visibly recurring or residual tumors and muscular tissues around the initial TURBT scar, which can require random biopsy. When a muscle-infiltrating tumor is detected and upstaged at the re-TURBT, cystectomy or one of the bladder preservation protocols should be considered .The benefit of re-TURBT is that it avoids staging errors as well as complete resection of residual tumor; in this sense, the re-TURBT may guide the surgeon toward a more appropriate therapeutic decision, which could potentially impact the prognosis and the curative rate of the technique . A numbeA number of studies have investigated the clinical value of re-TURBT, and most have focused on upstaging to muscle invasion bladder cancer or the benefits of restaging TURBT; however, few studies have investigated the meaning of re-TURBT pathology findings for patients with NMIBC. Thus, the objective of this study was to determine re-TURBT pathology as a prognosis risk factor in patients with a diagnosis of NMIBC.The Institutional Review Board of Seoul National University Hospital approved this study (H-1611-096-809). Because we retrospectively performed our investigation, the IRB waived the need for informed consent documents from our patients. Patient information was anonymized and de-identified before we carried out the study, and we carried out all study procedures in accordance with the Declaration of Helsinki guidelines.We retrospectively evaluated data collected from patients who underwent initial TURBT and re-TURBT between October 2004 and December 2013 at a single institution. All visual tumor findings were recorded on operation record sheets by the surgeon immediately after each operation, including tumor size, number, appearance, and location. Then, based on the EAU guideline, the finding of initial TURBT was indicated as re-TURBT, re-TURBT performed within 2–6 weeks after initial resection. At the re-TURBT, the bladder was reassessed to detect any residual tumors or missed lesions, and the previously resected sites and the peripheral areas around them were resected during the re-TURBT. All of the post-TURBT pathologic findings are reported using the 2004 World Health Organization (WHO) classifications. All histologic slides were reviewed by our pathologist, and we changed the previous recordings that used the 1973 WHO grade classifications (G1/G2/G3) to use the 2004 classifications (low grade/high grade).In this study, we only analyzed patients who had been diagnosed with NMIBC at both the initial TURBT and the re-TURBT; therefore, we did not include patients who showed muscle-invasive tumors in re-TURBT. We also did not include patients for whom the surgeon documented ‘‘incomplete resection” or felt that biopsy alone was adequate after re-TURBT.The point of our study was to evaluate the correlations between pathologic re-TURBT findings and recurrence or progression of high-risk NMIBC; we defined recurrence as detection and development of bladder cancer with any stage regardless of grade and progression as worsening of T stage to T2 during follow-up.We used multivariate logistic regression analysis to assess the different risk factors for presence of residual tumors. Specifically, we used multivariable Cox regression analyses to calculate the hazard ratios for disease recurrence and progression, and we calculated the recurrence-free survival (RFS) and progression-free survival (PFS) curves using the Kaplan-Meier method and compared them with the log rank test. We analyzed the data using Statistical Package for Social Sciences, version 21.0 , obtaining the analytic statistics using the chi-square test or Fisher’s exact test; the differences were significant if P < 0.05.A total of 206 patients were diagnosed with NMIBC after re-TURBT during the study period. We excluded eight patients we felt had had incomplete resections and/or biopsies only, leaving 198 patients who were suitable for analysis. Patient demographics and tumor characteristics are described in Of the 198 patients who had been treated with re-TURBT, we found residual tumors in 107 (54.0%) of them. There were significant differences in baseline characteristics between the no residual tumor group and the residual tumor group concerning tumor grade, multiplicity and concomitant CIS . Among tA total of 103 patients showed tumor recurrence during follow-up. Univariate analysis revealed tumor number, lymphovascular invasion (LVI), residual tumor in re-TURBT, high grade in re-TURBT, and T1 stage in re-TURBT as significant predictive determinants of recurrence ; in addiDuring follow-up, a total of 19 patients developed disease progression, all of whom had high-grade T1 tumors at the initial resection. Ten patients had high-grade T1 tumor after re-TURBT, and five had concomitant CIS after initial and re-TURBT. Eight patients received radical cystectomy, but 11 patients denied it or any further treatment. Among the patients who refused further treatment, six died during follow-up, and others did not visit our department; otherwise, all patients who received radical cystectomy in this analysis survived during our follow-up. Multivariate analysis revealed that stage T1 (HR 2.806 p = 0.029), high grade in re-TURBT (HR 2.152 p = 0.08), and age (HR 1.068 p = 0.014) were significant predictors of disease progression . For patFor patients who showed residual tumor on re-TURBT (107 patients), 47.9% patients were received intravesical instillation treatment. Overall 2-year recurrence-free survival (RFS) rates in the no treatment group were poor compared to those in the intravesical treatment group  . HoweverWhereas most re-TURBT studies have focused on upgrading to muscle-invasive cancer, few have analyzed the clinical meaning of the pathologic findings of re-TURBT in high-risk NMIBC patients. For this reason, we wanted to understand the correlations between re-TURBT pathology and recurrence and prognosis in these patients.The initial step in managing bladder cancer is complete resection of the transurethral bladder tumor. Some reports have found that the rates of residual bladder tumors perceived in re-TURBTs range from 33% to 76% , and ourHerr et al. published the largest repeat TURBT series, summarizing unpublished data on routine repeat TUR and including 1,312 patients with NMIBC . They foGrims et al. demonstrated that tumor stage and grade at initial TURBT predicted residual tumor after re-TUR in their univariate analysis , but ourIt was notable that the presence of residual tumors on re-TURBT was associated with higher risks of tumor recurrence compared with patients who had no tumors on re-TURBT. Similar results were reported in a previous study in which tumor-free status at re-TURBT was associated with fewer tumor recurrences and longer times to recurrence . A retroFew studies have analyzed re-TURBT pathology findings as prognosis risk factors; as mentioned above, most studies on re-TURBT in bladder cancer have focused on the stage migration and presence of the residual tumors. In this study, both the tumor pathology and the residual tumor status after re-TURBT were also associated with recurrence; in univariate analysis, stage T1 and high tumor grade in re-TURBT were significantly associated with residual tumor recurrence, and stage T1 in re-TURBT was still statistically significant in multivariate analysis.Herr et al. reported in their retrospective study that among 710 patients with bladder cancer, including 352 with pT1 tumors, 76% with residual pT1 tumors on restaging TUR progressed to muscle invasion, versus 14% who had no tumor or noninvasive  tumors . MoreoveOur patient population, however, had relatively few patients diagnosed with initial to residual T1 tumors compared with other studies, possibly because in this study, we excluded all incomplete resection patients at the initial TURBT and patients with visible residual tumors on re-TURBT; thus, our proportions of residual T1 may have been low compared with other studies. Nevertheless, in our multivariate analyses, residual stage T1 was a significant factor in disease recurrence and progression. These results agree with Herr et al.’s suggestion of immediate radical cystectomy for T1 patients after re-TURBT.LVI, the presence of tumor cells in lymphatic vessels and vascular walls, is emerging as a prognostic factor for bladder cancer . AlthougThe fact that local tumor control and accurate tumor staging depend on a complete TUR and reevaluation of the tumor base suggests that a restaging TURBT may be of value in assessing patients with bladder tumors , 22. Re-There were some limitations to our study. First, we did not include stage T2 among the re-TUR patients in this study, and it could have been meaningful to learn the prognoses of patients who had been diagnosed with NMIBC even after re-TURBT. However, excluding stage T2 patients may have affected our analysis of disease progression risk factors. Second, our study was limited by its retrospective design. Nevertheless, it is meaningful in that we had the largest proportion of patients with NMIBC after re-TURBT compared with other studies excluding Herr. However, there were few prospective studies about re-TURBT findings, and thus, a prospective study design with a larger sample is needed to confirm our presented results.In this study, tumor multiplicity, high tumor grade, and concomitant CIS were revealed to be independent risk factors for residual tumor after TURBT. In re-TURBT pathology, T stage is an independent predictor of NMIBC recurrence and progression, and high tumor grade in re-TURBT pathology was also an independent predictor of progression. From the results of previous reports as well as our own experience, re-TURBT is recommended to reduce the chance of residual tumor and to more accurately predict the prognosis in high-risk NMIBC.


A wide array of central nervous system complications, neurological deficits, and cognitive impairments occur and persist as a result of systemic cancer and cancer treatments. This condition is known as chemo brain and it affects over half of cancer survivors. Recent studies reported that cognitive impairments manifest before chemotherapy and are much broader than chemo brain alone, thereby adding in tumor brain as a component. The molecular mechanisms of chemo brain are under-investigated, and the mechanisms of tumor brain have not been analyzed at all. The frequency and timing, as well as the long-term persistence, of chemo brain and tumor brain suggest they may be epigenetic in nature. MicroRNAs, small, single-stranded non-coding RNAs, constitute an important part of the cellular epigenome and are potent regulators of gene expression. miRNAs are crucial for brain development and function, and are affected by a variety of different stresses, diseases and conditions. However, nothing is known about the effects of extracranial tumor growth or chemotherapy agents on the brain microRNAome.We used the well-established TumorGraft ™ mouse models of triple negative (TNBC) and progesterone receptor positive (PR+BC) breast cancer, and profiled global microRNAome changes in tumor-bearing mice upon chemotherapy, as compared to untreated tumor-bearing mice and intact mice. Our analysis focused on the prefrontal cortex (PFC), based on its roles in memory, learning, and executive functions, and on published data showing the PFC is a target in chemo brain.This is the first study showing that tumor presence alone significantly impacted the small RNAome of PFC tissues. Both tumor growth and chemotherapy treatment affected the small RNAome and altered levels of miRNAs, piRNAs, tRNAs, tRNA fragments and other molecules involved in post-transcriptional regulation of gene expression. Amongst those, miRNA changes were the most pronounced, involving several miRNA families, such as the miR-200 family and miR-183/96/182 cluster; both were deregulated in tumor-bearing and chemotherapy-treated animals. We saw that miRNA deregulation was associated with altered levels of brain-derived neurotrophic factor (BDNF), which plays an important role in cognition and memory and is one of the known miRNA targets. BDNF downregulation has been associated with an array of neurological conditions and could be one of the mechanisms underlying tumor brain and chemo brain. In the future our study could serve as a roadmap for further analysis of cancer and chemotherapy’s neural side effects, and differentially expressed miRNAs should be explored as potential tumor brain and chemo brain biomarkers. Initial reports on the cognitive changes associated with cancer chemotherapies appeared in the late 1970s to the mid-1980s, but received scientific attention starting only in the mid-1990s. Several studies reported significant cognitive changes in cancer survivors . These eThe clinical analysis of chemo brain’s molecular mechanisms is complicated due to varying treatment protocols, significant inter-patient variability (as a result of co-morbidities), and various other factors -12. HencRecent studies reported that cognitive impairments manifest before chemotherapy in 20%-30% of breast cancer patients and that these phenomena might be much broader than chemo brain alone. They may instead constitute both cancer and cancer treatment-associated cognitive changes reviewed in , therebyThe frequency and timing, as well as the long-term persistence, of chemo brain and tumor brain suggest they are epigenetic in nature. Epigenetic changes encompass meiotically heritable and mitotically stable alterations that regulate gene expression and genome stability; these include DNA methylation, histone modification, and non-coding RNA regulation . Among sAnalyzing these phenomena in tumor-bearing animals is needed to gain a full mechanistic understanding of both chemo brain and tumor brain. In a previous study, we analyzed gene expression and DNA methylation changes in the PFC tissues of triple negative breast cancer (TNBC) and progesterone positive breast cancer (PR+BC) mice. We noted that tumor growth caused changes in gene expression Aging, in press (2017). We used mouse TumorGraft ™ models of untreated TNBC and PR+BC, as well as TNBC TumorGraft animals treated with Doxorubicin/ Cyclophosphamide/ Paclitaxel (TNBC/DCP) and PR+BC animals treated with Topotecan (PR+BC/TOP) or Crizotinib (PR+BC/CRIZ) to analyze the roles of miRNAs in tumor brain and chemo brain.Here we report that extracranial malignant tumor growth had a profound effect on the microRNAome of the prefrontal cortex of experimental animals and caused changes in the levels of the miRNA processing machinery protein Ago2. Chemo and tumor brain-induced miRNA changes involved several miRNA families, such as the miR-200 family and miR-183/96/182 cluster, which were deregulated in PR+BC tumor-bearing and chemotherapy-treated animals. MiRNA deregulation was associated with altered levels of BDNF (brain-derived neurotrophic factor), a miRNA target that plays a key role in cognition and memory. Furthermore, deregulated miRNAs may serve as biomarkers of tumor brain and chemo brain.The NGS approach offers excellent technological opportunities to capture the entire repertoire of small RNAs and conduct a comprehensive analysis of the small RNAome .A total We identified all classes of differentially expressed (DE) small RNAs with a fold change > 2.0 and a false discovery rate cut-off of 0.05. Initial unsupervised hierarchical clustering was performed using all DE small RNAs. We noted a distinct separation between intact control PFC tissues and PFC tissues of tumor-bearing animals Figure . Next, wWhile there was an apparent separation between the small RNA and miRNAome profiles of the PFC tissues of the intact control and TNBC- and PR+BC-bearing animals, profiles of the PFC samples of tumor-bearing treated animals differed only from those of intact controls and not from the tumor-bearing untreated animals. Amongst samples, TNBC samples clustered better than the PR+BC samples. These differences may depend upon tumour transplantation, positioning and growth in each individual animal and host responses.Furthermore, sample clustering revealed they are primarily distinguished by the relative expression of a common set of miRNAs Figure . To gainNGS data revealed that the presence of a malignant extracranial tumor alone had a major effect on the miRNAome. We found that 5 miRNAs were up-regulated in the PFC tissues of TNBC bearing mice, as compared to intact controls. In the PFC tissues of PR+BC bearing mice, 33 miRNAs were up-regulated, and 4 miRNAs were down-regulated. Among those, miR-191-5p was up-regulated in both TNBC and PR+BC tumor-bearing groups Figure .Treatment of TNBC animals with the DCP regimen led to the upregulation of 18 and downregulation of 2 miRNAs in the PFC tissues of tumor-bearing treated animals. Two miRNAs (miR-191-5p and miR-100-5p) were up-regulated in TNBC and TNBC/DCP animals, as compared to intact controls Figure . InteresThe growth of PR+BC and subsequent chemotherapy treatments also affected the miRNAome profiles of PFC tissues of TumorGraft mice. There, treatment of PR+BC animals with crizotinib led to the upregulation of 13 microRNAs in the PFC tissues of experimental animals, as compared to intact controls. Topotecan treatment led to the upregulation of 37 and downregulation of 7 miRNAs Figure .We noted a commonality between all three groups : miR 200 family , miR-183/96/182 cluster, miR-30d-5p, and miR-191-5p were up-regulated, as compared to intact controls. Moreover, miRNAs of the let-7 family were up-regulated in the PFC tissues of PR+BC untreated and crizotinib-treated animals as compared to intact controls. MiR-409-3p and miR-22-3p were down-regulated in the PFC tissues of the PR+BC untreated and crizotinib-treated PR+BC animals. Similar to the TNBC TumorGraft mice, no differential miRNA expression was identified between PR+BC tumor-bearing treated and PR+BC tumor-bearing untreated mice.MiR-191-5p was up-regulated in all animal groups. Interestingly, miR-191 was recently reported to be a regulator of brain-derived neurotrophic factor (BDNF) . BDNF isp = 0.00006) and in PR+BC/TOP and PR+BC/CRIZ , as compared to intact controls. Moreover, levels of BDNF in the PFC tissues of PR+BC/CRIZ were also significantly decreased, as compared to untreated PR+BC animals (p = 0.0011). In the untreated TNBC cohort, BDNF levels exhibited a trend to decrease (90% confidence level). No significant change was observed in the TNBC/DPC animals, albeit a down-regulation trend was noted (p < 0.10)  and in PR+BC/TOP and PR+BC/CRIZ  , there is still a lot to learn about their molecular mechanisms. As per the timing and long-term persistence of tumor brain and chemo brain, these phenomena might be epigenetic in nature and based on altered gene expression patterns affecting brain function.Epigenetic changes governing alterations in gene expression include DNA methylation, histone modification, and small RNA-associated silencing. Amongst the latter, miRNAs  are of special interest and significance. MiRNAs are important in brain development and in all CNS functions and processes. They regulate cellular proliferation, differentiation, repair and death, and take part in inflammatory responses. Aberrant expression of miRNAs underlies a wide array of neurological and psychiatric diseases, such as Alzheimer’s, Parkinson’s, and Huntington’s, stroke, traumatic brain injury, amyotrophic lateral sclerosis, autism, schizophrenia, and brain tumors .Even so, nothing is known about the role of miRNAs in the brain’s response to the combination of malignant non-CNS tumor growth and subsequent chemotherapy. This is the first study to analyze miRNA changes in tumor brain and chemo brain. We showed that TNBC and PR+BC tumor growth and presence significantly altered the miRNAome in the PFC tissues of TumorGraft animals. The PFC is a key regulatory region that receives input from all other cortical areas, and serves to coordinate executive function, motor, cognitive, and social behaviors, attention, and working memory , 56.While tumor growth-related miRNAome changes were observed in the PFC tissues of both breast cancer groups, they were more pronounced in the PFC tissues of PR+BC-bearing animals than in TNBC-bearing ones. The reasons behind these changes and their molecular mechanisms, along with potential cellular changes and behavioral repercussions need to be further studied and analyzed, and may depend upon the biology of tumors as well as chemotherapy responsiveness. Chemotherapy treatment of TNBC and PR+BC-bearing animals led to further alterations in the miRNAome, and amongst differentially expressed miRNAs, miR-191-5p was up-regulated in all experimental groups . While this miRNA has not been extensively researched, some studies have reported that miR-191-5p was up-regulated in major depressive disorders and down-regulated in Alzheimer’s disease (AD) . This miAlong with miR-191-5p, miR-100-5p was upregulated in TNBC animals and TNBC/DCP animals, as compared to intact controls. Recently, miR-100-5p was shown to be an upregulated circulating marker of Huntington’s disease . FurtherDCP treatment caused upregulation of the let-7 family miRNAs - let-7a, 7b, 7g, 7i - in the PFC tissues of TBNC mice, as compared to intact controls. These family members were also upregulated in the PFC tissues of PR+BC-untreated  and PR+BC/topotecan-treated mice . The let-7 family is highly conserved in both invertebrates and vertebrates, and was the second such miRNA family to be identified and characterized . Since tC. elegancs, several neurons have shown increased levels of let-7 as they age, whereby let-7 upregulation contributed to the decline of aging neurons’ regeneration potential [The let-7 family is involved in neural development and neuronal differentiation , 71. In otential . Let-7 motential . Severalotential , 78. Theotential . OverallThe most pronounced miRNAome changes were observed in the PR+BC group. PR+BC tumor growth and subsequent chemotherapy treatment with topotecan and crizotinib had profound effects on the miRNAome of TumorGraft animals’ PFC tissue. Both tumor growth and chemotherapy treatment led to the upregulation of the miR-200 family, as compared to intact animals. We observed the upregulation of eight miRNAs that make up a related miR-200 family  and a miR-183/96/182 cluster. These miRNAs are often co-transcribed and referred to as the miR-200/182 cluster. It was shown the entire miR-200/182 cluster is upregulated in acute herpes simplex virus 1 encephalitis .via the Zeb/miR-200 family feedback loop. Overall, much remains to be discovered about the miR-200 family’s roles in various diseases and conditions, including cancer and cancer treatment-associated CNS toxicity. The miR-183/96/182 cluster upregulated in the PR+BC tumor-bearing treated and untreated mice was implicated in hepatocellular carcinoma, breast cancer, and glioma [The miR-200 family is important for the proper balance between neuronal proliferation and differentiation during development . Its abed glioma -87. It hd glioma . BDNF isd glioma .The negative correlation between the levels of miRNAs that target BDNF and the levels of BDNF protein constitutes an interesting and important finding. We noted that BDNF levels were strongly downregulated in PR+BC tumor-bearing treated and untreated animals; however, TNBC treated and untreated animals exhibited only a trend. BDNF, along with the nerve growth factors neurotrophin-3 and neurotrophin-4/5, is a member of the neurotrophin family . BDNF is96. However, an animal model-based study of doxorubicin and cyclophosphamide chemotherapy-induced chemo brain showed that behavioral deficits  were paralleled by decreased neurogenesis and lowered serum BDNF levels without alterations to BDNF, mRNA, or protein levels in the brain [It was reported that depression, which might be a manifestation of tumor brain and chemo brain in cancer patients, is often associated with reduced BDNF serum levels , 97. Lowhe brain . In our While we did not see any changes in the Dicer levels, we noted a significant upregulation in the levels of Ago2 in the PFC tissues of PR+BC tumor-bearing treated and untreated animals. Elevated Ago2 levels may, in turn, contribute to the more profound changes in miRNA expression observed in the PR+BC groups. Upregulation of the Ago2, protein that partakes miRNA production and execution of miRNA-mediated gene silencing was implicated in the regulation of cocaine addiction and anorexia . MoreoveHere we focused on tumor brain and chemo brain as induced by the growth of TNBC and PR+BC tumors, both stage IV. In the future, it would be prudent to look at the miRNAome as a function of tumor stage and grade, as well as to analyse miRNAome deregulation caused by other tumor types. Additionally, we focused our attentions on the PFC; however, previous studies have suggested chemo brain also manifests in the hippocampus . TherefoIn sum, growth of malignant TNBC and PR+BC tumors altered the miRNAome of tumor bearing treated and untreated animals’ PFC. The observed changes may have opposing functional consequences - some positive and protective and some, negative and deleterious. Among altered miRNAs, miR-183/96/182 cluster and miR-191-5p both target and downregulate BDNF. Here their increased expression was paralleled by a decrease in BDNF. Low BDNF levels have been shown to decrease neuronal survival, growth, differentiation and plasticity, and were reported in a wide array of neurological diseases and conditions. Meanwhile, miR-22 is a well-studied neuroprotective molecule  and its Some of the observed miRNA changes could be associated with cellular senescence and aging. Amongst these, oxidative stress-induced upregulation of the miR-200 family caused senescence ., and stFor a bigger picture of tumor brain and chemo brain’s mechanisms, we would proceed with identifying molecular pathways that were aberrantly expressed and regulated by miRNAs As such, it would be prudent to analyse the mechanisms, timing, and potential feedback loops between miRNA and mRNA regulation and expression, and integrate transcriptome, small RNAome, as well as methylome. More studies are needed to dissect the effects of miRNAs on signalling pathways and to discern the interplay between the various signalling pathways underlying tumor brain and chemo brain in the context of the entire signalome and interactome, as well as their functional outcomes. We focused on miRNAs, molecules that constitute the largest proportion of differentially regulated small RNAs. In the future, we would seek to analyze the roles of other small RNA molecules, such as tRNAs, snoRNAs, snRNA and others. These could prove to be important tumor brain and chemo brain biomarkers. In summary, this study is the first to show miRNAome deregulation in tumor brain and chemo brain, and may serve as a roadmap for further analysis of cancer and chemotherapy’s neural side effects. In the future, in order to gain a full understanding of tumor brain and chemo brain, molecular and cellular data need to be put in context of neuronal structure analysis and behavioral changes.nu/nu athymic mice; Harlan Laboratories, Indianapolis, IND) and further propagated as earlier described. Animal experiments were approved by Institutional Animal Care and Use Committee protocols. Following tumor propagation, when TumorGrafts grew larger than 200 mm3, the animals were divided into groups of three to five. Drug dosage and drug combinations were applied consistent with individual physicians’ recommendations and in consultation with the patient. The starting volumes differed between individual TumorGraft models because of the varying doubling time involved. As per agreed protocol, TNBC PDX-bearing TumorGraft animals were treated with Doxorubicin/Paclitaxel/Cyclophosphamide (n = 4 treated and 4 untreated), and PR+BC animals with Topotecan (PR+BC/TOP) or Crizotinib (PR+BC/CRIZ) (n = 3 treated and untreated). Intact animals  served as baseline controls. The aforementioned chemotherapy agents were prepared according to the manufacturer’s specifications, and Champions Oncology ran all chemotherapy treatments. Tumor growth was strictly monitored; tumor dimensions were regularly measured and tumor volumes were calculated as previously described [We used the well-established TumorGraft technology developed by Champions Oncology, Inc. ; the frozen brain tissues of TumorGraft mice carrying TNBC and PR+BC patient-derived xenografts (PDX) were provided by Champions Oncology, Inc. Patients diagnosed with TNBC and PR+BC had their tumors engrafted into mice to generate personalized TumorGraft mouse models for precision oncology approaches. The patients provided their informed consent, which covered the use of tumor material for research purposes. TumorGrafts were generated, as previously described -110. In https://cutadapt.readthedocs.io/en/stable/ ). The quality of the sequenced reads after adapter trimming was assessed using FASTQC software (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/). The bowtie v.1.1.2 version [Small RNA sequencing was conducted using Illumina next generation sequencing technology as previously described . In brie version  was used version . MultiplWestern immunoblotting was conducted as previously described -117. In Statistical analysis (Student’s t-test) was performed using the Microsoft Excel software package.


This meta-analysis was updated with results from a new trial and final data to reassess the efficacy and safety of bevacizumab combined with chemotherapy in ovarian cancer (OC).clinicaltrial.gov databases. Outcomes included the progression-free survival (PFS), overall survival (OS), objective response rate (ORR) and common adverse events. The hazard ratio (HR), risk ratio (RR) and odds ratio (OR) were pooled when the meta-analysis was performed.Randomized controlled trials (RCTs) were searched in PubMed, EMBASE, Cochrane clinical trials, Web of Science and Five RCTs with 4994 patients were included. In overall newly diagnosed OC, bevacizumab combined with chemotherapy did not significantly improve PFS  or OS . In the high-risk progression subgroup, the addition of bevacizumab significantly improved PFS  and OS . In recurrent OC, the addition of bevacizumab to chemotherapy significantly extended PFS  and OS . The ORR was improved  in the overall population. Bevacizumab increased the incidence of hypertension , proteinuria , bleeding , GI perforations , arterial thrombosis events  and venous thrombosis events .Bevacizumab combined with chemotherapy significantly improved PFS and OS in both patients with high-risk of progression and patients with recurrent OC, with an increased incidence of common adverse events. However, no statistically significant survival benefit was identified in the front-line settings. Ovarian cancer (OC) carries one of the worst prognoses among gynecological tumors and is the fifth cause of cancer death among women . The staBevacizumab is an anti-VEGF antibody that has demonstrated activity in ovarian cancer . SeveralFurthermore, previously reported preliminary data could not accurately reflect the survival benefit. The most recently published meta-analysis indicated that OS was improved by bevacizumab combined with chemotherapy for newly diagnosed OC , whereasWe initially identified 147 articles from all searched databases, and 24 articles were retained after a full-text screening for inclusion in our review after excluding duplicates, reviews, and phase I and phase II trials. Finally, 5 randomized controlled trials with 4994 patients met our inclusion criteria  . Considering the large heterogeneity between newly diagnosed settings , the random-effects model was utilized for further subgroup analysis. The benefit on PFS in patients with a high risk of progression was significant   0.85, 95% confidence interval (CI) 0.70-1.02, 2 = 0%) and the patients with a high-risk of progression  .Bevacizumab had a significantly better OS in both the recurrent setting   have been reported, and the pooled odds ratio (OR) for ORR was 2.37 , proteinuria , wound healing disruption , bleeding , GI perforations , arterial thrombosis events , and venous thrombosis events   with large heterogeneity. According to our analysis, the heterogeneity was mainly derived from the difference FIGO stages of the recruited patients. Thus, we performed further analysis in patients with a high risk of progression who were predefined in the ICON7 trial and matched all the recruited patients in the GOG-218 trial. The results indicated that the PFS was significantly improved in this subgroup . Moreover, for the non-high-risk patients, bevacizumab did not improve OS  or PFS . This finding implies that the benefit from bevacizumab is associated with prognostic factors [Four trials  were designed to observe the PFS as the primary endpoint. For recurrent ovarian cancer, bevacizumab combined with chemotherapy reduced the HR of progression by 47% compared with chemotherapy alone. For patients with a high risk of progression, the addition of bevacizumab reduced the HR of progression by 24% compared with chemotherapy alone. Notably, the pooled HR of PFS for the newly diagnosed setting was 0.85  relative to placebo [Even if a pathological complete response is achieved, the recurrence rates are greater than 40%. Thus, almost all patients will eventually die from recurrence . The mos placebo .Several problems must be addressed for bevacizumab-containing therapy in ovarian cancer. Firstly, the optimum dose of bevacizumab in the treatment of ovarian cancer is undefined . The ICOThis updated meta-analysis included 5 RCTs with 4994 patients, whereas the previous publication contained 4 RCTs with 4246 patients. One additional trial, GOG-213, had final results published in abstract form from conference proceedings. Moreover, the final data from the OCEANS trial (OS) and the ICON7 trial  were published recently. Previously reported preliminary data could not accurately reflect the survival benefit. The conclusion of the most recent meta-analysis showed that bevacizumab combined with chemotherapy improved PFS and OS in the front-line setting in ovarian cancer instead of improving the OS in the recurrent setting . HoweverThe main limitation of this meta-analysis is the clinical heterogeneity across the studies, including the different tumor stages, the length of follow-up, the dose of bevacizumab, and the chemotherapy regimens. Secondly, we pooled HR for time-to-event data rather than assessing individual patient data. Thirdly, some negative trial results may not be published, thus excluding unpublished trials may introduce bias. Finally, this meta-analysis only included 5 RCTs, which was insufficient to analyze sensitivity.clinicaltrial.gov. We used the search terms “bevacizumab”, “Avastin”, “chemotherapy”, and “ovarian cancer” in various combinations. In addition, only phase III randomized trials were restricted in the search strategy. The language of an article published was not restricted.The literature search focused on randomized controlled trials published from database inception to May 2016. Studies comparing bevacizumab plus chemotherapy with chemotherapy alone were eligible for inclusion. We searched the PubMed, EMBASE, Web of Science and Central  databases, and we also searched In order to explore the general efficacy of bevacizumab and to avoid biased conclusions, we used the same criteria as the previous meta-analysis . SelectiThe risk of bias was assessed using the Cochrane Collaboration's tool. The risk of bias assessment was judged by three categories for each study: low risk (+), unclear risk (?) or high risk (-) of bias . Data fr2 > 75% indicated considerable heterogeneity [As a time-to-event outcome, PFS and OS were evaluated by using the HR. The HR and two-sided 95% CI were extracted directly from the trial reports. The Chi-squared test and Cochran Q-test were used to evaluate heterogeneity among trials, and Iogeneity . We poolThis updated meta-analysis indicates that bevacizumab combined with chemotherapy significantly improved PFS and OS in both patients with high-risk of progression and patients with recurrent OC, with an increased incidence of common adverse events. However, no statistically significant survival benefit was identified in the front-line settings. ORR is improved in overall population by the addition of bevacizumab.


Purpose: This study is to exam self-esteem related brain morphometry on brain magnetic resonance (MR) images using multilevel-features-based classification method.Method: The multilevel region of interest (ROI) features consist of two types of features: (i) ROI features, which include gray matter volume, white matter volume, cerebrospinal fluid volume, cortical thickness, and cortical surface area, and (ii) similarity features, which are based on similarity calculation of cortical thickness between ROIs. For each feature type, a hybrid feature selection method, comprising of filter-based and wrapper-based algorithms, is used to select the most discriminating features. ROI features and similarity features are integrated by using multi-kernel support vector machines (SVMs) with appropriate weighting factor.Results: The classification performance is improved by using multilevel ROI features with an accuracy of 96.66%, a specificity of 96.62%, and a sensitivity of 95.67%. The most discriminating ROI features that are related to self-esteem spread over occipital lobe, frontal lobe, parietal lobe, limbic lobe, temporal lobe, and central region, mainly involving white matter and cortical thickness. The most discriminating similarity features are distributed in both the right and left hemisphere, including frontal lobe, occipital lobe, limbic lobe, parietal lobe, and central region, which conveys information of structural connections between different brain regions.Conclusion: By using ROI features and similarity features to exam self-esteem related brain morphometry, this paper provides a pilot evidence that self-esteem is linked to specific ROIs and structural connections between different brain regions. Self-esteem is defined as the degree that people evaluate and accept themselves  using multilevel-ROI-features-based classification method. Multilevel ROI features consist of ROI features and similarity features, which are extracted from T1-weighted structural brain MR images. These two types of features are complementary to each other in revealing neuroanatomical information about self-esteem. In order to reduce the dimension of features and select the most discriminating regions related to self-esteem, a mixed feature selection pattern is adopted in this study. A machine-learning-based multi-kernel support vector machine (SVM) is constructed to train the optimal classifier.3. Each participant receives a structured clinical interview by a psychiatrist to rule out any psychiatric or neurological diagnoses. None of them has received stimulant or hypnagogic medication previously. All subjects have normal or corrected to normal vision and are right-handed. After the assignment, all participants are rewarded by giving small gifts or financial payments. The study is approved by the Ethics Committee of the Third Affiliated Hospital of Soochow University. Written informed consents are obtained from all subjects.Characteristics of all subjects are shown in Table N = 34), while the low self-esteem group consists of the low 50 percent (N = 34) of all subjects.All participants have performed the Rosenberg Self-esteem Scale (RSES) test  and t(j) represent the cortical thickness values for the i-th and j-th ROIs. σ is defined as i and σj representing the standard deviation of cortical thickness for the i-th and j-th ROIs. Owing to the symmetrical property of the similarity map, only the upper triangular elements of the matrix are adopted to construct the feature vector. For each subject, 3003 elements in the upper triangular part are concatenated to compose a long feature vector.where The classification is performed using different feature types, including GM volume, WM volume, CSF volume, union of the above three volumes, cortical thickness, cortical surface area, similarity features, and the multilevel ROI features. The union of GM volume, WM volume, and CSF volume are constructed by a junction of the three volumes into a long vector.t-test is used to select the features that their p-values are smaller than the threshold (p < 0.05). Then, the dimension of the retained features is further reduced by the minimum redundancy and maximum relevance (mRMR) method  kernel function. We compare the classification performance using linear kernel function and RBF function (nonlinear), which shows that the RBF kernel can significantly improve the classification performance. Thus, we choose RBF kernel function to construct the kernel matrix for each feature type. Second, these two kernel matrixes are integrated into a multi-kernel matrix with appropriate weighting factor , NSFC grants (61501452), SSTP program (ZXY201426), Jiangsu Key Research and Development programs , postdoctoral research funding of Jiangsu province (1501089C), BKLN program, Suzhou Scientific and Technologic programs , and SND Medical Plan Project (2016Z010).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.


Recent neuroimaging research suggests sex-related brain differences in smoking addiction. In the present pilot study, we assessed gender-related differences in brain activation in response to cigarette-related video cues, investigating non-smokers, smokers, and ex-smokers.post hoc analysis of non-smokers , smokers , and ex-smokers . Participants performed a block-design functional magnetic resonance imaging paradigm contrasting smoking with control cue video exposures. Data analyses included task-related general linear model, voxel-based morphometry of gray matter (GM), and tract-based spatial statistics of white matter (WM).First, we compared 29 females (28.6 ± 5.3) vs. 23 males (31.5 ± 6.4), regardless of current smoking status to assess global gender-related effects. Second, we performed a First, the global effect regardless of current smoking status revealed higher activation in the bilateral superior frontal gyrus and anterior cingulate cortex (ACC) for females compared to males. Second, the analysis according to current smoking status demonstrated higher activation in female vs. male smokers vs. non-smokers in the superior frontal gyrus, anterior and posterior cingulate cortex, and precuneus, and higher activation in female vs. male ex-smokers vs. non-smokers in the right precentral gyrus, in the right insula and ACC. No structural differences were found in GM or WM.The current study identifies gender-related brain functional differences in smokers and ex-smokers compared to non-smokers. The current work can be considered as a starting point for future investigations into gender differences in brain responses to cigarette-related cues. On the basis of recent data from the WHO, it has been estimated that 250 million women worldwide smoke daily . BetweenFurthermore, neuroimaging techniques, such as positron emission tomography (PET) and functional magnetic resonance imaging (fMRI), allowed the detection of gender-related differences in brain functions associated with addiction, identifying in dependent women higher activation in frontal regions related to craving stimuli compared to men , 9.Recent fMRI studies focusing on heavy smokers established a link between blood oxygenated level-dependent (BOLD) signal in brain areas related to nicotine craving and the sex of participants , 11. In Although gender effects during nicotine addiction are suggested, none of the previous studies contrasted male and female smokers to a non-smoking group to control for the effects of stimulus administration. Moreover, the previous works investigated current smokers, while sex-related differences during long-term nicotine abstinence (in ex-smokers) have yet to be investigated.Given these previous findings, we hypothesized the presence of gender-related brain functional differences during tobacco dependence and long-term tobacco abstinence. fMRI in a task-based paradigm, contrasting smoking with control cue videos was used in three populations: non-smokers, active smokers, and ex-smokers (more than 1 year of smoking abstinence). In a first step, we analyzed global gender-related differences in smoking cue processing regardless of the current smoking status by contrasting 29 females vs. 23 males. In fact, as researches on passive smoking demonstrate, it is not an unusual situation that even non-smokers are exposed to smoking cues on a regular basis . In the The presented work is an extension of an fMRI dataset previously published by Zanchi et al. . DetailsThis study was approved by the local ethics committee . All the experimental procedures were carried out in accordance with the approved guidelines and with the principles of the Declaration of Helsinki. All the participants gave written informed consent prior to inclusion. Fifty-one subjects were recruited by word of mouth and local advertising. The inclusion criterion for non-smoking status was the absence of any regular lifetime use of tobacco. Inclusion criteria for smokers were: (a) 10 or more cigarettes smoked per day over at least 2 years, (b) no intention to quit smoking in the next month, and (c) ability to maintain 15 min of smoking abstinence prior to scanning. Inclusion criteria for male and female ex-smokers were: (a) abstinence from cigarettes for more than 12 months prior to the study and (b) previous smoking period of at least 2 years with 10 or more cigarettes per day. Non-smokers, smokers, and ex-smokers groups matched for gender and age, with smokers and ex-smokers matching also for years of smoking and number of cigarettes smoked per day. Within each group, males and females were also matched for age, years of smoking, and years of abstinence. Exclusion criteria for all participants were history of drug or excessive alcohol abuse/dependence, major medical disorders, and use of medications at the time of brain scanning that could affect brain function . The final sample included 9 female  and 8 male  non-smokers, 10 female  and 8 male smokers , and 10 female  and 7 male  ex-smokers  with a standard 32 channel head coil. Subjects underwent an fMRI examination including a functional scan, a 3D T1-weighted structural scan, and a diffusion tensor imaging (DTI) sequence. For technical details regarding the fMRI parameters, please see Ref. . During Statistical analyses were conducted using GraphPad Prism  and FSL .t-tests. Non-normally distributed variables, notably number of cigarettes a day and rating scores for the smoking cue videos, were tested using Mann–Whitney non-parametric tests.First, a D’Agostino–Pearson omnibus test was performed to check for normal distribution. Then, we investigated demographic and behavioral differences between males and females overall separately and for each group . Normally distributed variables, notably age and years of smoking, were analyzed using unpaired P values <0.05 were considered as significant.Processing and analysis of imaging data were performed using FSL FEAT  and white matter (WM) gender-related differences, a voxel-based morphometry (VBM) and a tract-based spatial statistic (TBSS) analysis were performed in FSL  and in the rating scale scores of smoking videos between males and females for each group Table .In the task-related GLM, we considered the contrast of smoking vs. control cue videos.First, we assessed the main effect of gender regardless of current smoking status. The comparison “females vs. males” revealed significantly greater activation in the bilateral superior frontal gyrus and anterior cingulate cortex (ACC) for female smokers  and non-smokers (lowest level). These results show that smokers indeed perceived more craving than non-smokers and ex-smokers during smoking cue videos. This finding was discussed already in our previous article , and it The current study identifies for the first time gender-related brain functional differences in smokers and ex-smokers when compared to non-smokers. The current work can be considered as a starting point for future investigations on this topic.This study was approved by the Ethics Committee of the University Hospital of Geneva. All the experimental procedures were carried out in accordance with the approved guidelines and with the principles of the Declaration of Helsinki. All the participants gave written informed consent prior to inclusion.The dataset supporting the conclusions of this article is available at the University Hospital of Geneva. For repository, please contact Prof. Sven Haller.Conceived and designed the experiments: DZ and SH. Performed the experiments: DZ and SH. Data analyses: DZ, AB, and SH. Writing the paper: DZ, AB, SB, and SH. All the authors read and approved the final manuscript.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.