Working memory (WM) is essential for normal cognitive function, but shows marked decline in aging. The importance of selective attention in guiding WM performance is increasingly recognized. Studies so far are inconclusive about the ability to use selective attention during WM in aging. To investigate the neural mechanisms supporting selective attention in WM in aging, we tested a large group of older adults using functional magnetic resonance imaging whilst they performed a category-based (faces/houses) selective-WM task. Older adults were able to use attention to encode targets and suppress distractors to reach high levels of task performance. A subsequent, surprise recognition-memory task showed strong consequences of selective attention. Attended items in the relevant category were recognized significantly better than items in the ignored category. Neural measures also showed reliable markers of selective attention during WM. Purported control regions including the dorsolateral and inferior prefrontal and anterior cingulate cortex were reliably recruited for attention to both categories. Activation levels in category-sensitive visual cortex showed reliable modulation according to attentional demands, and positively correlated with subsequent memory measures of attention and WM span. Psychophysiological interaction analyses showed that activity in category-sensitive areas were coupled with non-sensory cortex known to be involved in cognitive control and memory processing, including regions in the prefrontal cortex and hippocampus. In summary, we found that older adults were able to recruit a network of brain regions involved in top-down attention during selective WM, and individual differences in attentional control corresponded to the degree of attention-related modulation in the brain. •Effective selection of targets amongst distractors for working memory (WM) in aging.•Category-selective attention leads to improved recognition memory in aging.•Reliable top-down attention modulation in category-sensitive cortex in older adults.•Modulation of category-sensitive cortex correlated with attention and WM span.•Category activity coupled with control and memory regions for effective selection. In addition, we administered a surprize subsequent recognition memory test to investigate the functional consequences of selective attention on memory for attended and ignored items. We recruited a relatively large sample of older adults for greater statistical power due to higher behavioral variability in older groups , and to We found that older adults were able to use top-down attention to modulate category-sensitive cortex to support performance on a selective WM task. Selective attention to individual exemplars of face and house stimuli significantly affected subsequent recognition memory for these items, demonstrating that attention can have durable functional consequences in older adults. Category-sensitive cortex and the PFC were modulated according to task demands, and attention-related modulation was predictive of our behavioral measure of attention (effect of attention on subsequent memory) and with individual WM capacity within an older group.22.1Eighty-one healthy older adults (aged 60–87) were recruited from the community via local media and public advertisements. Of these, 75 participants completed the current experiment. Five participants were excluded from the analysis due to cortical abnormalities observed in their T1-structural scans. The remaining 70 participants  were 60–87 years old (68.5 ± 0.85 years), had 16.0 ± 0.44 years of education, scored ≥26 on the Mini-Mental State Examination MMSE, , and had2.2Stimuli consisted of digital color photographs of faces and houses with 350 × 350 resolution . The task was presented using a display with spatial resolution of 1024 × 768 pixels and refresh rate of 60 Hz, projected onto a screen and reflected onto a mirror inside the scanner .The task was programmed and run in Presentation2.32.3.1A selective-WM task with images of faces and houses tested the ability to focus on items from one category to guide WM performance A. In eacAt the start of each block, an instruction cue (2000 ms) indicated if the upcoming block required attention to faces (“REPEATED FACES”) or to houses (“REPEATED HOUSES”). Prior to each stimulus, a reminder (“F” or “H”) was presented (2000 ms) to ensure participants were aware of the current task block, followed by the face or house stimulus (1000 ms). Each block consisted of ten trials. For each stimulus, its associated target (stimulus repeat) could be presented after one to three intervening stimuli, or not presented at all. Target (repeated) images, if presented, were only shown once. This was the case for both the attended and ignored stimulus categories, even though no response was required for the ignored category. Intervening stimuli were face or house images with equal probability. Stimuli from the attended blocks were not presented in the ignore blocks (e.g. faces in attend-face ignore-house blocks were not presented in the ignore-face attend-house blocks), and vice versa. Participants were not explicitly informed of these details but were simply asked to respond when they saw an image of the attended category presented a second time at any point during the block.There were 18 trials for each stimulus-attention condition  with novel images , and 12 trials for each condition with a repeated image , giving 120 trials in total.2.3.2Participants were given a surprise subsequent-memory test after the main experiment, once they were outside of the scanner. On each trial, participants were presented with a face or house image which had either been shown in the main selective WM task (old) or an image they had not seen before (new). They judged whether they remembered seeing this image (old) or not (new) by making a forced-choice response on a keyboard. Participants were presented with 72 ‘old’ images  and 73 ‘new’ images  in a pseudo-randomized order. Half of the ‘old’ images were from the ‘attend’ blocks and the other half from the ‘ignore’ blocks. Images were the same across participants. One participant did not complete the memory test.2.4At the beginning of the session, the experimenter provided a demonstration of the task accompanied with verbal instructions from a prewritten script. Participants proceeded to perform the task in a practice session, which consisted of two attend-faces and two attend-houses blocks. The experimenter provided verbal feedback on their performance. Once the experimenter was satisfied that the participant understood the task, the participant proceeded with the task in the scanner.For the main selective-WM experiment, participants completed six attend-faces blocks and six attend-houses blocks in the scanner in alternating order. In each block, there were ten trials, lasting a total of 30 s, followed by 14 s of rest with central fixation. Eye movements were not monitored. After the scan session, participants were given a subsequent memory test.2.52.5.1To characterize performance on the task in each block type, we computed the hit rate and mean correct reaction times for the face and house blocks separately. Hit rate was the number of responses when a target was presented (repeated stimulus in the attended category) divided by the total number of targets. To characterize the effect of category-based selective attention for WM on subsequent memory, we computed the proportion of correctly responded ‘old’ trials for stimuli that were presented in the fMRI session for attended faces, ignored faces, attended houses, and ignored houses separately. To examine subsequent-memory performance in general, we computed the proportion of correct responses for all stimuli in the memory test, separated into ‘old’ and ‘new’ stimuli. Participants correctly judged images as ‘old’ if they had been presented in the main experiment, and correctly judged images as ‘new’ if they were not presented previously.To test the effect of category-based attention during selective WM on subsequent memory, a repeated-measures ANCOVA was performed on proportion ‘old’ stimuli remembered (responded ‘old’), with factors Attention  and Stimulus Category , with covariates age, education, and gender. Follow up t-tests were used to test for differences between conditions, and Cohen's d was used to determine effect sizes. Statistical analyses for behavioral performance were performed using Matlab R2015a, Matlab's Statistics Toolbox, and R version 3.2.1  using the afex package .2.5.2Participants completed a neuropsychological battery and questionnaires separately to the current experiment. Individual differences in the ability to filter task-irrelevant distractors and enhance task-relevant items in WM tasks have been linked to differences in WM capacity e.g. . Therefo2.63 voxel size, repetition time (TR) = 2000 ms, and echo time (TE) = 30 ms. Flip angle was set to 78°. A fieldmap image was acquired with parameters as follows: 3.5-mm3 voxel size, TR = 488 ms, first TE = 5.19 ms, second TE = 7.65 ms. A high–resolution whole-brain T1-weighted structural image (MPRAGE) was acquired for registration purposes, with 1 mm3 voxel size, TR = 2040 ms, TE = 4.7 ms. A resting-state scan and a separate task scan were also acquired, which do not form part of this investigation.Functional and structural MRI data were acquired on a 3 T Siemens TIM Trio System  using a 32-channel head coil at the Oxford Centre for Clinical Magnetic Resonance Research (OCMR). An EPI-BOLD contrast image with 32 slices was acquired with 3-mm2.7http://www.fmrib.ox.ac.uk/fsl). Pre-processing consisted of head-motion correction  Version 6.00, part of FSL  with local autocorrelation correction . Data weIn the main analysis, four contrasts were used. To localize brain areas preferentially activated by face stimuli to identify region-of-interests (ROIs) in the fusiform gyri (FG), we tested for regions that evoked greater blood oxygen-level dependent (BOLD) activity for face compared house stimuli (FA & FI > HA & HI). To localize areas preferentially active for house stimuli to identify ROIs in the parahippocampal gyri (PHG), we tested for regions that evoked greater activity for house stimuli compared face stimuli . To test for areas that were modulated by attention during selective WM for faces, we contrasted Face Attend with Face Ignore (FA > FI). For selective WM for house stimuli, we contrasted House Attend with House Ignore (HA > HI). All contrasts used here did not have motor responses. Note that the contrasts used to define face-sensitive and house-sensitive areas were orthogonal to the attention contrasts and therefore do not suffer from selection bias .Group-level analyses were carried out using FMRIB's Local Analysis of Mixed Effects FLAME; . Z Gaus. Grey-ma2.8A conjunction masking analysis on the group-level z-statistic images (FWE cluster-corrected) was performed to localise attentional control areas involved in both face attention (FA > FI) and house attention (HA > HI). To compute the conjunction image, we multiplied the FA > FI contrast z-statistic image with the HA > HI contrast z-statistic image. Each contributing contrast was thresholded to include only significant clusters at the whole-brain level, so that voxels that do not overlap in the two images would have a value of zero. The resulting image was binarized. Only voxels that were significant at Z > 3.09, or p < 0.001 in both contrasts were selected.2.93 cubes) were transformed into single-subject space. We ensured no spatial overlap between the masks in all participants, and that each participant's ROIs laid within the vicinity of the face and place-sensitive areas (3) were constructed and centred on the single-participant defined FGs and PHGs from each hemisphere to extract mean beta estimates in the FA, FI, HA, and HI conditions. Since we were mainly interested the effect of attention on BOLD activity in category-sensitive cortex, and not hemispheric differences, beta estimates were averaged across hemispheres in the FG and the PHG in each condition.To test for attentional modulation in category-sensitive areas, we focused on the posterior fusiform FG;  within tve areas . Within To examine top-down modulation during category-based attention in face-sensitive and house-sensitive regions, repeated-measures ANCOVAs were performed on BOLD activity (beta estimates) for each bilateral ROI  with factors Attention  and Stimulus Category ; with age, education, and gender as covariates. Follow-up pairwise t-tests were used to assess condition differences. We tested for a relation between attentional modulation  and the effect of attention on subsequent memory (memory accuracy difference between attended images versus ignored images) using Spearman's partial correlation, controlling for age, education, and gender. We also tested for a relation between attentional modulation and digit span, controlling for age, education, and gender. Finally, we also tested if attentional modulation was correlated with age, controlling for education and gender. Correlation coefficients were compared by using Fisher's r-to-z transformation and tested for significant differences . Partici2.10To test for individual differences in attention-related modulation of BOLD for faces (FA > FI) and houses (HA > HI) that corresponded to i) the effect of attention on subsequent memory, ii) WM capacity, and iii) age; subsequent-memory difference scores for faces (proportion remembered attended face images – proportion remembered ignored face images) and houses (proportion remembered attended house images – proportion remembered ignored house images), mean digit span, and age were included as regressors in the GLM. Education and gender were included as regressors of no interest. To test for relations between these measures and activity in attention-modulated regions, we ran a group analysis with only education and gender as covariates of no interest and constructed masks from two contrasts  to localise the areas that showed significant attentional modulation. Two group-level analyses for each contrast were performed with age as a regressor and education and gender as covariates of no interest, using the activation masks as pre-threshold masks to restrict the correlations to areas that exhibited attentional modulation in each respective contrast. To test the relation between areas that showed significant attentional modulation with digit span and subsequent memory, a group analysis was conducted with age, education, and gender as covariates to obtain the activation masks for the two main contrasts. Group-level analyses were then performed to test for a relation between the attentional modulation for faces (FA > FI) and digit span using the activation mask from the face attention contrast, and attentional modulation for houses (HA > HI) and digit span using the activation mask from the house attention contrast. The activation mask from the face-attention contrast was used to test for a relation between the attentional modulation for faces and the subsequent memory difference score for faces, and the activation mask from the house-attention contrast was used to test for a relation between modulation for houses and the subsequent memory difference score for houses. In both analyses, age, education, and gender were included as covariates of no interest. A group-level analysis without pre-threshold masking produced very similar results.Information on significant clusters including peak locations, cluster size, statistical values, and peak MNI coordinates is reported in the Appendix (fMRI cluster tables). The first local maximum value reported for each cluster is the peak activation within that cluster. The Harvard-Oxford cortical and subcortical structural atlases (in FSL) were used as a guide for labelling anatomical locations of the local maxima.2.11We used psychophysiological interaction (PPI) analyses  to test We tested for areas that showed greater coupling with bilateral FG (as PPI seed) when encoding faces that were attended versus ignored (FA > FI). We performed the same analysis for bilateral PHG for attended versus ignored house stimuli (HA > HI). For each PPI, we included four regressors: 1) the contrast of interest (e.g. FA – FI); 2) the time course of the seed ROI ; 3) the element-wise product of the first two regressors, representing the PPI; and 4) both conditions (e.g. FA + FI). The fourth regressor was included to model the shared variance of the two regressors not modelled by the contrast itself. In each analysis, all other task regressors noted in the main analysis were also included . Group-l2.12https://neurovault.org/collections/4742/(Unthreshholded whole-brain maps are available at ons/4742/.33.13.1.1Performance accuracy was near ceiling for both attend-face (hit rate: 0.97 ± 0.01) and attend-house (hit rate: 0.95 ± 0.01) blocks (hit-rate difference: t (69) = 1.80, p = 0.08, d = 0.25), and participants were slightly faster during attend-face (628 ± 8.93 ms) compared to attend-house blocks  = -2.05, p = 0.045, d = −0.28). Performance was not significantly correlated with age in the attend-face blocks , attend-house blocks , or the difference between attend-face and attend-house blocks .3.1.2p2 = 0.47; p2 = 0.01) and no difference in the attention effect between stimulus categories  = 0.62, p = 0.43, ηp2 = 0.009).Older adults performed relatively well on the subsequent memory test for faces (proportion correct for attended and ignored old images and new images: 0.78 ± 0.01) and houses (0.71 ± 0.01). They were much more likely to remember images that were attended (0.74 ± 0.02) in the main experiment compared to images that were ignored (0.60 ± 0.02)  = 57.78, p = 1.49e-10, η3.23.2.1Older adults recruited a common set of brain regions during selective WM for faces and during selective WM for houses, including PFC, temporal-occipital cortex including IT gyrus, fusiform gyrus, and lateral occipital cortex (LOC) . Areas a3.2.2p2 = 0.83) and houses in the PHG  = 461.87, p = 5.68e-31, ηp2 = 0.88). Both regions were strongly modulated by attention to stimulus category. There was a significant effect of category-based attention in the FG  = 34.99, p = 1.48e-07, ηp2 = 0.36), with greater BOLD for attended versus ignored stimuli, but no significant interaction with Stimulus Category  = 2.74, p = 0.1, ηp2 = 0.04). There was a significant effect of attention for houses in the PHG  = 20.16, p = 3.03e-05, ηp2 = 0.24) and an interaction between Attention and Stimulus Category  = 19.60, p = 3.80e-05, ηp2 = 0.23) where BOLD activity was greater for the attended-houses compared to ignore-houses conditions (t (69) = 6.37, p = 1.91e-08, d = 0.52) but not in the attend-faces versus ignore-faces conditions (t (69) = 1.37, p = 0.18, d = 0.13; difference between conditions: t (67) = 4.48, p = 2.88e-05, d = 0.62).Older adults were able to use top-down attention to modulate category-sensitive visual cortex during selective WM A. ReplicThe degree of top-down modulation in bilateral FG positively correlated with effect of attention on subsequent memory for faces (memory difference between attended versus ignore face images; rho = 0.37, p = 0.003; Among brain regions that showed attention-related modulation during selective WM for faces, we found that activity in right posterior IT including the LOC and posterior FG positively correlated with the effect of attention on subsequent memory for faces (p = 0.001, cluster-corrected; 3.2.3To identify control-related brain regions in which activity co-varied with activations in category-sensitive cortex depending on category-based selective attention, we used a PPI analysis. Activity in bilateral FG was coupled with bilateral precuneus and cuneal cortex, left superior parietal lobe, and right postcentral gyrus when participants were ignoring faces compared to attending faces (FI > FA; p's < 0.02, cluster-corrected; 4We tested a large group of older adults and found that they were able to use top-down control to modulate category-sensitive cortex in a selective WM task. Older adults used category-based attention to encode target items into WM and suppress distractor items. Their effective use of selective attention was evident from their high degree of accuracy during task performance. Better subsequent memory of attended images relative to ignored images further showed that selective attention to individual items had functional consequences for subsequent recognition memory. Category-sensitive cortex showed greater activation to the attended versus ignored category during WM encoding, and the degree of modulation was associated with behavioral consequences of selective attention and WM span. Purported control regions, including dorsolateral PFC, IFC, and ACC, were recruited in both types of category attention in concert with category-sensitive cortex, and the degree of coupling between FG and the right hippocampus, STS, and OFC was correlated with the effect of attention on subsequent recognition memory for faces.As a group, older adults recruited greater activity for attended compared to ignored faces in the FG and attended compared to ignored houses in the PHG, as reported in other studies in younger  and oldeThe PFC plays a key role in top-down attention , and infSome researchers have attributed general age-related cognitive deficits specifically to a decline in inhibitory control , and impIn the current study, older adults showed strong recruitment of non-sensory brain regions implicated in cognitive control and memory, along with appropriate modulation in category-sensitive cortex which positively correlated with behavioral performance but not with age. Some studies have reported older adults show strong or over-recruitment of control areas including frontal and parietal cortex e.g. , whereasOur study shows that older adults are able to use selective attention to control the contents of WM and could be used to support other cognitive processes that undergo age-related decline. Despite effective selective WM in older adults as a group, there were individual differences in the degree of top-down modulation in relevant brain regions, and this variability was predictive of WM function. Our study suggests that studying the variability in behavior and brain patterns in older adults is a promising way to characterise the neural systems that support cognitive abilities in old age, and future studies should use multiple tasks with varying levels of cognitive demand to reveal how the brain markers adapt or break down in different cognitive scenarios in normal aging.None.


Non-adherence to medication remains a major challenge in the long-term management of patients with schizophrenia. Next to lack of insight into the illness, adverse effects of antipsychotic drugs, cognitive deficits, poor therapeutic alliance, reduced quality of life, missing social support, and negative attitudes toward medication are predictors of non-adherence. This study examined potential correlations between attitudes toward antipsychotic drug therapy, subjective well-being, and symptom change in patients with chronic schizophrenia.30 patients with schizophrenia starting monotherapy with a new-generation antipsychotic were included into the study. The Drug Attitude Inventory (DAI) and the Subjective Well-being under Neuroleptic Treatment Scale, short form (SWN-K), were administered after 2, 4, and 12 weeks of treatment. At the same points in time and at baseline, psychopathological symptoms were rated by means of the Positive and Negative Syndrome Scale (PANSS), and functioning was assessed by means of the Global Assessment of Functioning Scale (GAF). Antipsychotic induced side effects were evaluated by using the Udvalg for Kliniske Undersogelser (UKU) Side Effect Rating Scale.p = .003) and 12 (p = .001), respectively. Neither the DAI total score nor the SWN-K total score changed significantly over the course of time. The severity of symptoms was not correlated with drug attitude at any time point but was negatively correlated with wellbeing at weeks 2  and 4 . There was no significant correlation between DAI and SWN-K total scores at any time point.Study participants had a mean age of 37.5 ± 9.7 years, baseline symptoms were mild. The PANSS total score improved significantly from baseline to weeks 4 (Next to showing that the DAI and the SWN-K measure different aspects of subjective experiences during antipsychotic treatment these findings emphasize the use of both instruments to optimize adherence to medication. Despite advances in the pharmacotherapy of schizophrenia poor-adherence to antipsychotic medication remains a major challenge in the long-term treatment of patients. Most studies reported on high frequencies of partial and non-adherence in this patient group with different factors being relevant in this context : next toMost patients with schizophrenia, if not acutely psychotic or experiencing severe cognitive impairments, are able to complete self-rating scales in a meaningful way, and it has been shown that patients’ and psychiatrists’ perspectives on antipsychotic treatment differ considerably –13. SeveAlthough some authors claimed that both instruments examine quality of life , 23, merThe aim of this study was to assess the associations between subjective well-being and attitudes towards antipsychotic medication as well as the interrelations between these issues and both symptoms and functioning in patients with chronic schizophrenia starting monotherapy with a new-generation antipsychotic drug.Patients aged between 18 and 65 years and starting treatment with a new-generation antipsychotic drug were included into a prospective longitudinal study to build a drug monitoring register. Subjects were recruited via a specialized outpatient service for patients with psychotic disorders or they were inpatients. They met the diagnostic criteria of schizophrenia, schizotypal and delusional disorders according to ICD-10 and signed informed consent forms as approved by the local ethics committee. They did not suffer from any other axis I disorder, including substance abuse. Diagnoses were confirmed using chart information and reports from clinicians who had treated these patients. Patients who had previously been receiving antipsychotic medication underwent a washout period of 3–5 days. Antipsychotics were chosen by the psychiatrists treating the patients, dosing followed clinical needs within the recommended labeled dose ranges.At baseline as well as after 2, 4, and 12 weeks of treatment, psychopathology was rated by means of the Positive and Negative Syndrome Scale (PANSS) . At the After 2, 4, and 12 weeks of treatment, the patients’ subjective well-being as well as their subjective response to and their attitudes towards medication were assessed by means of the short form of the SWN (SWN-K) and the DAI, respectively.The SWN-K represents a self-rating Likert scale with 6 response categories  . It consThe DAI, on the other hand, is a 30-item self-report questionnaire consisting of statements about perceived effects and benefits of antipsychotics with which the patient can agree or disagree. It is divided into 7 factors: (1) subjective positive feelings related to antipsychotics , (2) subjective negative feelings attributed to the drugs , (3) health/illness-dependent drug intake: patients’ model of health , (4) patients’ confidence in the physician , (5) control: patients’ attitudes toward the locus of control in taking medication , (6) prevention: patients’ belief in the effect of antipsychotics in forestalling relapse , and (7) harm: patients’ concerns with potential toxic effects . Each item oft the DAI is scored 1 or 2, depending on whether the answer selected by the patient indicates a negative or positive view of medication. Raw scores of the DAI  were linearly transformed to a common range from 0 to 100 to facilitate interpretation . For the1. Values of G1 above 0.5 or below − 0.5 were regarded as sizable deviation from normality indicating the need for non-parametric testing. Changes of PANSS, GAF, SWN-K and DAI over the course of time were analyzed by repeated-measures ANOVA or by Friedman test and subsequent Wilcoxon matched-pairs tests, depending on the variable type . Missing observations were omitted listwise in the repeated-measures analyses . However, missing values were very rare as all patients completed the SWN-K and 29 of 30 (96.7%) completed the DAI at all planned assessment times . Associations of SWN-K and DAI scores with patient characteristics, symptomatology (PANSS), side effects (UKU) and GAF were investigated by means of the Spearman rank correlation coefficient, as part of the variables involved were non-normally distributed.Prior to the analysis all metric variables were checked for deviations from normality by investigating their skewness, Gn = 30 patients provides sufficient power (1-β = 0.8) to detect in a repeated-measures ANOVA under standard assumptions  effect sizes of f = 0.236 for the comparison of 4 assessments and f = 0.258 for comparing 3 assessments. These are medium effect sizes according to Cohen’s classification [n = 30 is sufficient to detect with a power of 1-β=0.8 and a two-tailed type-one-error of 0.05, Spearman rank correlation coefficients r ≥ 0.49 a in a non-parametric correlation analysis. This is a medium to large correlation according to Cohen’s classification (close to the threshold of 0.5 indicating large correlations).The sample size of n = 7, aripiprazole: n = 7, clozapine: n = 6, olanzapine: n = 3, quetiapine: n = 3, ziprasidone: n = 2, sertindole: n = 2). The mean doses and ranges of antipsychotic medication were as follows: amisulpride 428.6 ± 138 mg (200–600 mg), aripiprazole 19.3 ± 15.5 mg (15–30 mg), clozapine 366.7 ± 75.3 mg (300–500 mg), olanzapine 16.7 ± 5.8 mg (10–20 mg), quetiapine 700 ± 173.2 mg (600-900 mg), ziprasidone 140 ± 28.3 mg (120–160 mg), and sertindole 16 ± 0 mg.Demographic and clinical characteristics of the study sample are summarized in Table The time course of disease severity as indicated by PANSS scores, subjective well-being , and attitudes toward medication  is depicted in Table Correlations between SWN-K and DAI and the interrelations between these scales and both symptom level (PANSS) and functioning (GAF) are depicted in Table r = −.362, p = .05) and 12 , and with the DAI total score at week 2 . Additionally, the side effect failing memory was negatively correlated with the SWN-K total score at week 12 . No further correlation between side effects and both subjective well-being and drug attitude was found.Regarding correlations of patient characteristics, i.e. age, gender, duration of illness, and inpatient vs. outpatient, with SWN-K and DAI scores, no significant correlation was found. Side effects that were most frequently reported within the observation period of 12 weeks (found in ≥10% of patients) were as follows: increased salivation (18.3%), concentration difficulties (15.8%), asthenia (15.8%), weight gain (15%), tension/ inner unrest (15%), hypokinesia (12.5%), failing memory (11.7%), akathisia (10%), and diminished sexual desire (10%). The item tension/ inner unrest was negatively correlated with the SWN-K total score at weeks 2 (The aim of the current study was to assess a potential association between subjective well-being and attitudes toward antipsychotic medication as well as the interrelations between these issues and both symptom severity and functioning in chronic schizophrenia patients starting treatment with a new-generation antipsychotic drug. As expected, both symptoms and global functioning improved over the course of antipsychotic treatment. Most previous studies reported on an improvement of patients‘ subjective well-being over the course of treatment –39. In oThe negative correlation between the SWN-K total score and both the PANSS total score and the general psychopathology subscore is in line with the literature , 43, andOur finding of a significant correlation of global functioning with patients’ subjective well-being corroborates the results of previous studies , 45. AccSimilar to the SWN-K total score and in line with a previous report , the DAINotably and in contrast to our finding of a consistent association between patients’ subjective well-being and functioning, the DAI total score did not correlate with the GAF score at any time point, which corroborates previous findings . HoweverRegarding the impact of antipsychotic-induced side effects on subjective well-being and drug attitude, inconsistent correlations have been found for tension/ inner unrest and failing memory. This finding is supported by previous research that found that the psychiatrists’ beliefs about the tolerance of a particular antipsychotic drug do not necessarily reflect patients’ well being and attitude toward medication . \AdditiWhen interpreting our data, one has to consider a number of limitations. First of all, the sample size was rather small and early noncompliers, who usually stop attending mental health services, have not been included. Furthermore, the fact that only patients with a generally good attitude toward medication took part in the study limits the generalizability of our findings. Lastly, we did not thoroughly assess the reason for switching antipsychotic medication in patients who had previously been receiving antipsychotic drugs.In summary, our findings emphasize that, particularly in chronic schizophrenia patients experiencing mild symptoms, the differences between the SWN and DAI may give good reason for the use of both instruments during antipsychotic treatment in clinical practice and/ or research to systematically monitor and adjust treatment with antipsychotic medication. In so doing it may help finding the optimal dosage and type of antipsychotic medication for an individual patient and therefore enhance adherence.


The behaviors of and mechanisms acting in Ti–6Al–4V alloy during low-temperature superplastic deformation were systematically studied by using a Gleeble-3800 thermocompression simulation machine. Focusing on the mechanical behaviors and microstructure evolution laws during low-temperature superplastic compression tests, we clarified the changing laws of the strain rate sensitivity index, activation energy of deformation, and grain index at varying strain rates and temperatures. Hot working images based on the dynamic material model and the deformation mechanism maps involving dislocation quantity were plotted on the basis of PRASAD instability criteria. The low-temperature superplastic compression-forming technique zone and the rheological instability zone of Ti–6Al–4V were analyzed by using hot processing theories. The dislocation evolution laws and deformation mechanisms of the grain size with Burgers vector compensation and the rheological stress with modulus compensation during the low-temperature superplastic compression of Ti–6Al–4V were predicted by using deformation mechanism maps. Ti–6Al–4V, which is characterized by formability, biocompatibility, toughness, low density, high strength, anti-corrosion, and excellent high-temperature properties, is the most widely used Ti alloy both commercially and industrially . Due to Titanium alloys, especially the rudder surface of air-to-underwater missiles that are widely used in aerospace, hardly deform at room temperature. The superplastic temperature of materials is generally above half of the melting point, and the strain rate must be within an appropriate range. The largest elongation of superplastic alloys is at least 100% and is reportedly more than 2000% in some alloys . HoweverSuperplastic deformation mechanisms are classified by three major processes. These include dominant grain boundary sliding, compatible diffusional creep, and dislocation creep ,9. SuperIn this work, the low-temperature deformation of Ti–6Al–4V was investigated by using the flow curves detected from compression trials at varying tensile temperatures and strain rates. A new deformation mechanism map and a processing map of variable rheological instability concerning dislocation quantity were drawn for the two-phase Ti alloy. These were used to forecast the deformation mechanism and to quantify the dislocation quantity. This work can theoretically underlie the utilization of low-temperature deformation maps of Ti–6Al–4V .−4–0.05 s−1. Three samples per condition and each specimen were deformed to a real strain of 0.6 and then immediately water-cooled to ambient temperature [Multidirectional forging Ti–6Al–4V sheets in bar diameter of 50 mm were selected. The gauge part of each specimen was 12 mm long and 8 mm in diameter . The expperature ,26.2O containing 20 mL of HF and 40 mL of HNO3, and then observed using a LEICA Q550IW optical microscope . The average particle size and volume fraction of the β phase were detected on an OLYMPUS M3 image-meter . The microstructures were characterized on a Tecnai G2 20 transmission electron microscope  after mechanical grinding, punching, and twin-jet chemical polishing in an electrolyte of 5% HCLO4, 35% C4H9OH, and 60% CH3OH at a voltage of 20 V at 25 °C.In the metallographic tests, each specimen was polished mechanically, etched in 100 mL of HThe microstructure of the initial multidirectional forging of Ti–6Al–4V consisted of the scattered α phase (dark contrast) and the band-structured β phase (bright contrast) . The vol−1 (−1), the upper yield effect occurred.The deformation is nonuniform, and the deformed specimen is barrel-shaped due to the inevitable interfacial friction between the tested specimen and dies. Essentially, the computed flow stress–strain plots overestimate the real ones. For these reasons, the impact of friction on flow stress must be taken into account, which makes the stress–strain curves closer to the real ones. The details of flow stress correction have previously been reported ,28. The −1 a–d. The −1 a–d. At t−1 c,d. This−1) was selected for analysis , and when the strain ε was close to 0.6, it started to reached the stable state (zone III). The softening effect of this alloy during high-temperature deformation fell within the strain of ε = 0.14–0.57. Based on the above method, the strain ranges of the softening effect at different strain rates to reach the alloy temperature of 860 °C have been summarized in The rheological strain curve under the typical thermal deformation conditions σ=kεnQ  kJ/mol  kJ/mol . Q elevaThe activation energy of thermal deformation of Ti–6Al–4V is far larger than the activation energy for self-diffusion cited in the literature . This wad is average grain size; L is the linear intercept of metallopraphical microstructures. −4, 10−3, 5 × 10−3, and 5 × 10−2 s−1. Based on Equation (4) and The microstructure evolution rules of metal materials during high-temperature deformation considerably affect the hot forming properties. The grain size was computed as follows : (4)d=1.d is the grain size, σ is the stress, p is the grain index, 0D is the diffusion coefficient, b is the Burgers vector, G is the shear module, k is the Boltzmann constant, and R is the ideal gas constant. The Arrhenius constitutive relation universal model  was exprp, which reflects the effect of grain size on the thermoforming performance. The grain index at 860 °C is 2.31. With the above method, the p of this alloy at other temperatures can be determined  and dissipation assisting energy (J). The consumed energy or dissipation energy (G) is mostly converted to thermal energy and slightly stored as crystal defect energy in the material. The dissipated energy, resulting from structural changes during deformation, is called the assisting dissipation energy (J).According to the dynamic materialogic model, the plastic history of this material can be regarded as an energy dissipative system, and the energy dissipation is decided by the material processing rheological behaviors and obeys the power-law Equation : (6)σ=k×J and G is decided by the material rheological properties, and the J to G ratio is called the strain rate sensitivity index : The allocation between T) and strain (ε): At given temperature  and (11), the expression of power dissipation rate  and (14), we plotted the power dissipation rate and rheological instability of Ti–6Al–4V during thermal compression and thereby acquired the thermal processing maps . The cha−1, and the thermal deformation falls within the rheological instability zone , which does not fall within the instability zone. Thus, Ti–6Al–4V has very high thermal forming performance at low-temperature and low-strain-rate areas and at medium-temperature and medium-strain-rate areas. The strain rate of this alloy within 800–840 °C is 0.034–0.05 sity zone . The strSevere dislocation motion occurred during the thermal deformation of Ti–6Al–4V. Therefore, based on the Ruano–Wadsworth–Sherby (RWS) deformation mechanism, we introduced the dislocation model via a constitutive equation and plotting deformation mechanism maps that involved the dislocation quantity, grain size, strain rate, and rheological stress. m, and p are material constants, E is Young’s modulus, b is Burgers vector, D is the diffusion index, including the lattice index LD and the crystal boundary index gbD. The high-temperature deformation of metals can be described by a constitutive equation : (15)ε˙iThe internal dislocation root count of single crystal grains ( follows : (16)ni=d/b) × 10−7) and the normalized flow stress with modulus normalization ((σ/E) × 104) of Ti–6Al–4V at the above temperatures were calculated (The RWS deformation mechanism maps involving dislocation count at 830 or 890 °C were drawn in lculated . 7) (6.85 × 108) (52) (71) (368) dislocation polygon zone. The mechanism was the dislocation glide controlled by lattice diffusion, with a stress index of 7 (7)  (5425) (5816) (368) dislocation polygon zone, and the mechanism was the dislocation glide controlled by lattice diffusion with a stress index of 5. At 830 °C and a very high strain rate, Ti–6Al–4V fell within the (3.95 × 10dex of 7 . As the 6)  (6488) (4258) (415) dislocation polygon zone  The m of Ti–6Al–4V at 800–890 °C with a strain rate of 5 × 10−1; 860–875 °C and strain rate at 0.001–0.005 s−1; the rheological instability zone was at 800–840 °C and strain rate at 0.034–0.05 s−1. (2) The dynamic deformation mechanism map (DMM) thermal processing showed the machine-shaping of Ti–6Al–4V as follows: 800–840 °C and strain rate at 0.0005–0.001 s(3) The deformation mechanism of RWS Ti–6Al–4V involving dislocation count was clarified. The dislocation quantitative relationship of this alloy based on the modulus compensation stress and Burgers vector compensation was determined, and the low-temperature superplastic compression deformation mechanism was predicted.


Interestingly, a development-specific pattern was observed during early postnatal life: whereas the increase in Bdnf expression, mainly driven by the modulation of Bdnf isoform IV was clearly visible after weaning at postnatal day (pnd) 30 in both sexes of TPH2−/− in comparison to TPH2+/+ rats, at early stages (pnd1 and pnd10) Bdnf expression levels did not differ between the genotypes, or even were downregulated in male TPH2−/− animals at pnd10. Moreover, to establish if hyposerotonergia may influence the response to a challenging situation, we exposed adult rats to an acute stress. Although the pattern of corticosterone release was similar between the genotypes, neuronal activation in response to stress, quantified by the expression of the immediate early genes activity regulated cytoskeleton associated protein (Arc) and Fos Proto-Oncogene (cFos), was blunted in both sexes of animals lacking brain serotonin. Interestingly, although upregulation of Bdnf mRNA levels after stress was observed in both genotypes, it was less pronounced in TPH2−/− in comparison to TPH2+/+ rats. In summary, our results demonstrated that serotonin deficiency affects neuroplastic mechanisms following a specific temporal pattern and influences the response to an acute stress.Dysregulations of the central serotoninergic system have been implicated in several psychopathologies, characterized by different susceptibility between males and females. We took advantage of tryptophan hydroxylase 2 (TPH2) deficient rats, lacking serotonin specifically in the brain, to investigate whether a vulnerable genotype can be associated with alterations of neuronal plasticity from the early stage of maturation of the brain until adulthood. We found a significant increase, in both gene and protein expression, of the neurotrophin brain-derived neurotrophic factor (BDNF), in the prefrontal cortex (PFC) of adult TPH2-deficient (TPH2 Serotonin 5-hydroxytryptamine (5-HT) was originally discovered as cardiovascular hormone but has later gained prominence as neurotransmitter or neuromodulator widely distributed in the central nervous system (CNS) and peripheral nervous systems. Nevertheless, by far the highest amounts of the monoamine are produced in the intestine and released into the blood stream, where it is taken up by platelets which transport it through the circulation and release it at sites of their activation.5-HT biosynthesis occurs from the essential aminoacid tryptophan in two steps, and tryptophan hydroxylase 2 (TPH2) is the rate-limiting enzyme in this process. As it has been discovered in 2003  rats  from early life until adulthood. Indeed, it is well known that there is a correlation between 5-HT and BDNF  rats to an acute stress focusing on BDNF modulations, since it is known that there is a strong relationship between acute stress response and neuroplasticity  and wild type (TPH2+/+) rats on the dark agouti background  was used for the basal measurements. A second cohort of adult male and female TPH2+/+ and TPH2−/− rats  was used for the stress experiment. For the developmental analysis male and female rats were sacrificed at pnd 1, 10 and 30 . All procedures were approved by the ethical committee of the local government .Animals were kept under standardized conditions with an artificial 12/12-h light/dark cycle (lights on at 6 a.m.), room temperature of 22°C, and approximately 80% humidity with access to food  and water +/+ and TPH2−/− rats were exposed to acute restraint stress: rats were placed in an air-accessible apparatus for 1 h. The size of the container was similar to the size of the animal, which made the animal almost immobile in it. Half of the stressed animals were sacrificed immediately at the end of the stress session (stress0’), while the other groups were sacrificed 60 min after the end of the stress (stress60’). Unstressed animals served as control group . Animals were sacrificed by decapitation and trunk blood was collected for the analysis of plasma corticosterone levels.Adult male and female TPH2Animals were sacrificed by decapitation and brain regions of interest were rapidly dissected, frozen on dry ice and stored at −80°C for the molecular analyses. Dissections were performed according to the atlas of Paxinos and Watson . In detaBdnf, long 3′-UTR Bdnf and the exons IV and VI, activity regulated cytoskeleton associated protein (Arc) and Fos Proto-Oncogene (cFOS) mRNA levels. An aliquot of each sample was treated with DNAse  to avoid DNA contamination. RNA was analyzed by TaqMan qRT-PCR instrument  using the iScript™ one-step RT-PCR kit for probes . Samples were run in 384 well formats in triplicate as multiplexed reactions with a normalizing internal control (36B4). Primers and probes  according to the manufacturer’s instructions and quantified by spectrophotometric analysis. Following total RNA extraction, the samples were processed for real-time polymerase chain reaction (RT-PCR) to assess total s Tables ,B were ps Tables .g for 15 min at 4°C to obtain the pellet corresponding to the crude synaptosomal fraction which was re-suspended in a buffer  with protease and phosphatase inhibitors. Total protein content was measured according to the Bradford Protein Assay procedure (Bio-Rad Laboratories), using bovine serum albumin as a calibration standard. Equal amounts of protein were run under reducing conditions on the criterion TGX precast gels (Bio-Rad Laboratories) and then electrophoretically transferred onto nitrocellulose membranes (Bio-Rad Laboratories). The blots were blocked with 10% nonfat dry milk and then incubated with the primary antibodies , 4° O/N; β-actin 1:10,000 (Sigma), 4°C, O/N). Membranes were then incubated for 1 h at room temperature with the appropriate secondary antibody . Immunocomplexes were visualized by chemiluminescence using the Western Lightning Clarity ECL (Bio-Rad Laboratories) and the Chemidoc MP imaging system (Bio-Rad Laboratories). BDNF expression was quantified by the evaluation band densities, normalized to the β-actin .Western blot analysis was used to investigate mature BDNF in the crude synaptosomal fraction. Tissues were manually homogenized using a glass-glass potter in a pH 7.4 cold buffer containing 0.32 M sucrose, 0.1 mM EGTA, 1 mM HEPES solution in the presence of a complete set of protease (Roche) and phosphatase (Sigma-Aldrich) inhibitors. The total homogenate was centrifuged at 2,500 rpm for 10 min at 4°C. The supernatant obtained was further centrifuged at 10,000 g, 4° and corticosterone was determined by an enzyme-linked immunosorbent assay (ELISA) using a commercial kit  according to the manufacturers’ instructions.Samples of blood from each rat were collected in MiniCollect K3EDTA  tubes. Plasma was separated by centrifugation for 10 min at 1,300 p < 0.05. Data are presented as means ± standard error mean (SEM). For graphic clarity, all qPCR and western blot results are normalized to male TPH2+/+ values that are taken as 100%.All the statistical analyses were conducted by using “IBM SPSS Statistics, version 24.” The effects of genotype and sex were analyzed with the two-way analysis of variance (ANOVA) with genotype and sex as independent factors. When appropriate, differences between the individual groups were analyzed by Fisher’s protected least significant difference (PLSD). The effects of stress, genotype and sex in the acute stress experiments were analyzed with three-way ANOVA with Fisher’s PLSD. Significance for all tests was assumed for Bdnf was modulated by the lack of serotonin, in the PF of both male and female adult rats. As shown in Figure F(1–52) = 10.232, p < 0.01) but not of the sex and interaction genotype × sex. Post hoc analysis revealed an increase of total Bdnf mRNA levels both in TPH2−/− male rats  and in TPH2−/− female rats . However, no difference was found between the sexes for both genotypes.We first investigated whether the mRNA expression of total Bdnf mRNA were paralleled by alterations in BDNF protein, we investigated the levels of the mature form of BDNF (mBDNF) in crude synaptosomal fraction  = 17.673, p < 0.001) with no genotype × sex interaction. As revealed by post hoc analysis, the levels of mBDNF were significantly increased in both male  and female  TPH2−/− rats in comparison to the TPH2+/+ counterparts  = 4.703, p < 0.05), with an up-regulation of Bdnf long 3’UTR mRNA levels specifically in male TPH2−/− in comparison to TPH2+/+ rats . Moreover, the significant effect of the sex (F(1–53) = 5.884, p < 0.05) reflected the increased expression of Bdnf long 3’UTR in female TPH2+/+ in comparison to male counterparts  = 14.017, p < 0.05) for Bdnf isoform IV .Similarly to the results obtained for the total form of the neurotrophin, we observed a significant effect of the genotype  = 4.580, p < 0.05) for Bdnf isoform VI. Indeed, Bdnf VI increased only in male TPH2−/− rats .On the contrary, as shown in Figure Bdnf expression, we investigated its mRNA levels in TPH2+/+ and TPH2−/− rats at different ages of life, namely pnd 1, 10 and 30  = 107.444, p < 0.001) that is reflected by a significant increase of total Bdnf transcripts in both genotypes, at pnd10 compared to pnd1 rats  but also at pnd30 compared to pnd10 animals  = 7.115, p < 0.01). Indeed, the developmental profiles of the Bdnf expression differed between the genotypes, with an equal expression at pnd1, decreased total Bdnf mRNA levels in TPH2−/− at pnd10 , and a significant increase at pnd30 .In male rats, we found a significant effect of the age (F(2–45) = 297.514, p < 0.001) for Bdnf long 3’UTR transcripts with an upregulation from pnd10 to pnd30 in both TPH2+/+  and TPH2−/− rats  = 287.421, p < 0.001) for Bdnf isoform IV, with an increase in both genotypes from pnd10 to pnd30 . Furthermore, there was a significant effect of the genotype (F(1–46) = 8.667, p < 0.01) and a significant genotype × age interaction (F(2–46) = 10.602, p < 0.001) with an upregulation of Bdnf isoform IV at pnd30 in TPH2−/− rats  = 823.232, p < 0.001) with an increase both in TPH2+/+ and TPH2−/− from pnd1 to pnd10  and also from pnd10 to pnd30 . Moreover, we found a significant effect of the genotype (F(1–46) = 10.912, p < 0.01) and a significant genotype × age interaction (F(2–46) = 19.039, p < 0.001) with an upregulation of Bdnf isoform VI only at pnd30 in TPH2−/− in comparison to TPH2+/+ rats of the same age  = 105.889, p < 0.001) for total Bdnf mRNA levels with a significant increase among the three ages. Actually, we observed an enhancement from pnd1 to pnd10 only in TPH2−/− rats , while starting from pnd10, an upregulation of total Bdnf mRNA levels was seen in both genotypes . Moreover, we found a significant increase of total Bdnf in TPH2−/− rats compared to TPH2+/+  only at pnd30, as supported by the significant effect of the genotype (F(1–41) = 8,400, p < 0.01) and of the interaction age × genotype (F(2–41) = 9.843, p < 0.001).As shown in Figure Bdnf long 3’UTR  = 292.426, p < 0.001) with a robust upregulation at pnd30 compared to pnd10 in both TPH2+/+  and TPH2−/− , but we didn’t find any significant interaction of age × genotype.With respect to R Figure , we obseBdnf isoform IV was significantly modulated by the age (F(2–43) = 217.038, p < 0.001), but not by the genotype in young female rats. Accordingly, we observed an increase in both genotypes from pnd10 to pnd30 . Furthermore, the lack of serotonin at pnd30 induced an upregulation of Bdnf isoform IV in comparison to TPH2+/+/pnd30  = 535.298, p < 0.001) with the increase, both in TPH2+/+ and TPH2−/−, from pnd10 to pnd30  or 1 h later (stress60’).F(1–27) = 60.729, p < 0.001). Indeed, we found a strong increase in corticosterone levels in rats of both genotypes at stress0’ time point in comparison to BL conditions , while a significant decrease was observed in the group of rats killed at stress60’ time point compared to BL groups of the same genotype  as well as to stress0’ groups  = 5.239, p < 0.05), with an upregulation of corticosterone levels in stress0’ group compared to BL  and a decrease at stress60’ time point with respect to TPH2−/−/stress0’ .As shown in Figure Arc and cFos, in order to evaluate if the lack of serotonin in the CNS influenced the response to the restraint stress.Since we previously demonstrated the implication of the immediate early genes in the response to acute stress  = 1.668, p > 0.05) on the expression of Arc. Nevertheless, we found a significant effect of the stress (F(2–101) = 5.692, p < 0.01) with a genotype × stress interaction (F(2–101) = 7.181, p < 0.01). Accordingly, Arc mRNA levels were increased by stress exposure only in the TPH2+/+ rats of both sexes . Interestingly, this stress-mediated upregulation of Arc expression was completely blunted in TPH2−/− rats.As shown in Figure cFos expression was not significantly modulated by genotype × stress × sex interaction (F(2–90) = 2.170, p > 0.05). However, a significant effect of the stress (F(2–90) = 20.596, p < 0.001) with a genotype × stress interaction (F(2–90) = 19.110, p < 0.001) was observed. Post hoc analysis revealed a strong increase in cFos mRNA levels in TPH2+/+ male rats at both time points after stress in comparison to BL . The upregulation of the cFos mRNA levels peaked at stress0’ was markedly attenuated 1 h later , confirming previously published data  and stress60’ group . This characteristic pattern was again not observed in TPH2−/− female rats; moreover, 1 h after the stress exposure we found a significant downregulation of cFos mRNA levels in TPH2−/− rats  = 1.754, p > 0.05) on total Bdnf gene expression  = 112.032, p < 0.001). Accordingly, total Bdnf was increased in both TPH2+/+ and TPH2−/− male rats exposed to the acute stress and sacrificed at time point stress0’  and at time 60’ .The three-way ANOVA revealed no effect of the interaction genotype × stress × sex  as well as in TPH2−/− . Moreover, TPH2+/+stressed rats sacrificed 1 h after the stress exposure had higher mRNA levels compared to the stress0’ groups .In the female rats, the exposure to stress increased the expression of total F(2–98) = 6.490, p < 0.01) indicates that the upregulation found in TPH2+/+ was higher than the one observed in TPH2−/− rats.Furthermore, the significant genotype × stress interaction  = 3.203, p < 0.05). In particular, we found a significant genotype × sex interaction (F(2–101) = 7.205, p < 0.01) reflecting an upregulation of Bdnf long 3’UTR expression in female TPH2+/+/BL compared to the male counterpart . Moreover, the significant effect of stress (F(2–101) = 109.926, p < 0.001) indicated that Bdnf long 3’UTR was increased after the acute challenge in male  and in female , independently from the genotype.s Figure  were sig+/+/stress60’/male: +23%, p < 0.001 vs. TPH2+/+/stress0’/male; TPH2−/−/stress60’/male: 81%, p < 0.001 vs. TPH2−/−/stress0’/male).Moreover, in male rats the up-regulation observed 1 h after the stress exposure was significantly higher than the effect found at time 0’ in both genotype (TPH2F(2–101) = 4,368, p < 0.05) indicates that the increase in expression of Bdnf long 3’UTR due to stress exposure was more robust in TPH2+/+ than in TPH2−/− rats.Finally, the significant genotype × stress interaction  = 0.094, p > 0.05), but a significant effect of stress (F(2–101) = 66.292, p < 0.001). In male, Bdnf isoform IV expression was upregulated by stress in TPH2+/+ rats  with a further increase from stress point 0’ to 60’  and in TPH2−/− . Moreover, Bdnf IV was increased in unstressed TPH2−/− in comparison to TPH2+/+ rats .Analysis of s Figure  revealedBdnf isoform IV mRNA levels in both genotypes . Similar to total Bdnf and Bdnf long 3’UTR, the significant genotype × stress interaction (F(2–101) = 11.065, p < 0.001) was also observed for the Bdnf isoform IV, suggesting a different response to the challenging situation in the two genotypes in terms of magnitude of the effect.In female, exposure to the acute stress increased the Bdnf isoform VI mRNA levels  = 1.741, p > 0.05). However, the significant effect of the genotype (F(2–100) = 10.338, p < 0.01) supports the slightly higher mRNA levels found in unstressed male TPH2−/− compared to unstressed TPH2+/+ male rats .Similarly, s Figure  were not+/+ rats , while its levels were increased in TPH2−/− stressed rats sacrificed 1 h after the stress exposure in comparison to the animals of the same genotype killed immediately after the acute stress .Moreover, in females, isoform VI expression was downregulated specifically in TPH2Tph2 profoundly affects neuroplastic mechanisms from the first stage of life until adulthood and influences the response to an acute stress.The results of the present study highlight that the deletion of Bdnf mRNA levels in both adult TPH2−/− male and female rats. Interestingly, the contribution of different Bdnf isoforms to this increase is sex-specific: in male TPH2−/− rats, it is due to enhancement of all Bdnf transcripts investigated, while in females only the isoform IV is upregulated.We found a significant increase of total Bdnf, serotonin depletion in the adult CNS has an opposite effect, confirming the strict dependence of Bdnf expression on serotonergic status.These results are in line with our previous data obtained in a rat model with increased extracellular 5-HT levels, SERT-deficient rats , or even downregulation in Bdnf expression levels in TPH2−/− in comparison to TPH2+/+ pnd10 male rats were observed, suggesting that peripheral sources may compensate for the lack of central 5-HT synthesis during the early postnatal periods of life. Indeed, the blood brain barrier is not fully functional before pnd12 (Ribatti et al., Furthermore, it’s well known that ontogenesis is a critical period for brain adaptability and plasticity. Several lines of evidence support the fundamental role of 5-HT during neurodevelopment Lauder,  as well Bdnf isoforms we found in females in comparison to males may be due to the fact that different transcripts may have different subcellular localization (Chiaruttini et al., Moreover, the different regulation of Bdnf expression by activating their receptors and may modify the neurotrophin activity through methylation of the Bdnf promoter IV and V in the hippocampus (Moreno-Piovano et al., Bdnf transcript, it is not feasible to draw clear-cut conclusions on the consequences exerted by differences in isoform expression between male and female.Among these, sex hormones are known to contribute to BDNF modulations (Chan and Ye, +/+ male rats and a significant decrease 1 h later, while in females this pattern was not so pronounced and was statistically significant only in TPH2−/−, but not in TPH2+/+ rats. Corticosterone levels in females are highly influenced by the estrus cycle (Atkinson and Waddell, Seeing that, in basal condition, 5-HT deficiency was in some way compensated by neuroplastic mechanisms and considering the well-established relationship between the serotonergic system and the stress reactivity (Chen and Miller, Arc and cFos. As expected, in both males and female wild type rats we observed a clear stress response, with the slow increase in Arc expression during and 1 h after stress and quick upregulation at stress0 and downregulation 1 h later in cFos expression, confirming previously published data (Durchdewald et al., +/+ rats. Seen that Arc mediates the translation between neuronal activation-induced changes into sustained structural and functional modification at synaptic levels (Alberi et al., −/− rats may reflect an incorrect translation of the stimulus into a more stable outcome.Furthermore, the implication of neuronal activation in response to acute stress can be evaluated by the expression of immediate early genes (Ons et al., Arc in SERT-deficient rats in comparison to wild types (Molteni et al., 2 receptor antagonist ketanserin also blocks Arc induction in the PFC in response to a stress paradigm (Benekareddy et al., Moreover, we have previously shown that hyperserotonergia, evoked by the lack of SERT, has an opposite effect on the stress-induced neuronal activation, leading to a more pronounced upregulation of −/−, rats, this compensatory mechanism was not enough to restore the normal ability to react to the acute stress challenge, probably because of the complexity of the system and circuit involved that may be BDNF-independent. Even if Bdnf transcripts were upregulated in both genotypes in response to stress, the effects found in TPH2+/+ were stronger than those induced in the TPH2−/− rats. The fact that BDNF is upregulated by stress in the PFC even in the absence of serotonin, while the immediate early genes are not, suggests distinct stress-modulated neural circuits whereby one is serotonergic, and another is 5-HT independent.Finally, we found that, despite the increased levels of BDNF observed in TPH2Taken together our findings suggest that 5-HT deficiency in the brain modulates neuroplasticity probably to compensate for the lack of trophic support provided by serotonin starting after weaning until adulthood. Nevertheless, in response to a stressful condition the system is not able to properly respond by setting in motion the strategies to cope with an acute challenge.FC and NA: conception and study design. PB, PP, MT and GS performed experiments. PB, PP, FC and NA performed data analysis and interpretation of the data. PB drafted and FC, NA and MB critically revised the manuscript. All authors critically reviewed the content and approved the final version for publication.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.


Physical activity is well recognized for its bone health benefit. We examined the benefit of walk/run/jog on bone health using broadband ultrasound attenuation (BUA) of the calcaneus.Caucasian and African American males (n=593) and females  had their calcaneal BUA measured two years later after enrollment into the AHS-2. The association between calcaneal BUA (dB/Mhz) and the distance of walk/run/ jog level per week (miles) was assessed using multiple linear regression.P < .001), total calcium intake (P =0.31), total protein intake (P =0.38) and inversely associated with age (P < .001) and smoking (P < .05). Compared to women who did not walk/ run/ jog, women walking 10 or more miles per week had an increase in BUA by 4.08 (dB/Mhz) (Ptrend=0.03). Similarly, compared to men who did not walk/ run/ jog, men walking 10 or more miles per week had an increase in BUA by 5.97 (dB/Mhz) (Ptrend=0.01).In a multivariable model adjusted for important covariates, BUA was positively associated with BMI (We concluded that BUA is positively associated with walk/ run/jog after accounting for age, BMI, smoking status, calcium intake, protein intake and estrogen usage. Attenuation of ultrasound waves depends upon the bone’s microstructure, elasticity, anisotropy and mineral density . BUA is ry (DXA) –5. It isry (DXA) –11.Previous animal studies have shown some evidence that the mechanical loading stimulus occurring during weight-bearing exercise stimulates the bone remodeling process . SeveralIn this study, we ascertained the effect of walking/ running/ jogging on bone health using calcaneal BUA among Caucasian and African American males and females adjusting for demographic and lifestyle factors .2.2.1Subjects were enrollees in the Adventist Health Study 2 (AHS-2), a large National Cancer Institute (NCI) funded cohort study investigating the relationship between lifestyle factors and several disease outcomes. The study has been described in detail elsewhere  and consOne thousand and eleven subjects were randomly selected through a 2 stage sampling process to participate in a large calibration study. In addition, a sample of 1,119 subjects from the AHS-2 attended the pilot church clinics that were conducted in preparation for a nationwide clinic study. Of these study subjects, there were 2,037 Caucasian and African American males and females, who attended clinics arranged in conjunction with the AHS-2. We excluded those with a history of minor accidental fracture (n=260), those with missing data on physical activities (n=47) and BUA measurement (n=31), leaving a total of 1,699 Caucasian and African American males and females for the analysis.2.2e.g., subject who walked 2.5 miles would receive a score of 2 since it reflected a walking distance between 2 to 3 miles). An algorithm was developed based on the multiplication of the frequency score and the distance score and further categorized into six physical activity distance levels  per week.Physical activity was assessed using two questionnaire items focusing on subject’s physical activity during the last twelve months. Questions were part of a large self-reported baseline lifestyle questionnaire completed at enrollment into the AHS-2. The questions captured the distance and the frequency per week that the subject engaged in weight-bearing exercises such as walking, running or jogging. The question on the frequency of exercise was “How many of these “walk” or “run” or “jog” workouts do you usually do per week?” The response choices were: “less than once/week”, “1 time per week”, “2 times per week”, “3 times per week”, “4 times per week”, “5 times per week”, “6 or more times per week”. We assigned a frequency score of 0 to 6, respectively, corresponding to the choices given. The question on the distance of exercise was “How many miles do you average per “walk” or “run” or “jog” workout?” The responded choices were: “1/4 mile or less”, “1/2 mile”, “1 mile”, “1 ½ miles”, “2 miles”, “3 miles” and “4 or more miles”. We assigned a distance score of 0.25, 0.5, 1, 1.5, 2, 3 and 4 miles, respectively, corresponding to the choices given  reporting on the subject’s dietary intake during the last twelve months. Subjects were asked to report how frequently they consumed a food: “never”, “1–3 times per month”, “1 time per week”, “2–4 times per week”, “5–6 times per week”, “1 per day”, and “2 or more times per day”. In addition, subjects were given the size of an average serving of that particular food and asked to mark if they consumed the average size, half this size or one-and-a-half or larger of the average size. Based on this information from the FFQ, several nutrient indices  total calcium (mg)) were developed.2.4At enrollment, in addition to the FFQ and exercise information, participants completed questions on medical history, smoking and anthropometrics. There was also a female section which included menopausal status and estrogen usage. The question on fractures was asked “How many of your fractures (since the age of 35) were due to minor accidents ?” Those who reported one or more fractures were excluded from our study population (n=254).2.5Approximately two years after enrollment into the parent study, subjects were invited to attend a clinic either as part of the calibration study  or as pa2.6Chi-Square tests were used to determine the association between physical activity level per week and selected predictor variables . Multipl3.3.1P=0.08), however the significant association was observed in the internal comparison between those whose walk/ run/ jog was greater than 10 miles to those with lower level of physical activity (P < .05). There was no significant difference in the mean age of participants or BUA by the level of walking or running. However, there was a significant difference in BMI by the level of physical activity. Those who reported engaging in longer distance of walking or running had lower BMI compared to those whose weekly walk was 2.5 miles or less (Approximately 27% (n=465) of our participants reported the lowest level of physical activity (0 mile/wk) whereas 12% (n=203) reported walking or running more than 10 miles per week. No significant association was observed between physical activity level and BUA  and level 6 (>= 10 miles/ wk) significantly increased BUA by 4.26 dB/Mhz and 5.83 dB/Mhz, respectively, compared to the lowest level (0 mile/wk) . BUA was3.2P <0.001). BUA was negatively associated with a positive smoking history and positively associated with total daily calcium intake and protein intake, although these associations were not statistically significant.When further adjusting for smoking status, total calcium intake and total protein intake, the effect estimates for the variables in the basic model remained virtually unchanged and the associations of walk/run/jog with BUA was stable with an increase of 3.9 dB/Mhz and 5.64 dB/Mhz, respectively, for walk/ run/ jog at level 5 (7.6 – 10 miles/wk) and level 6 (> 10 miles/ wk) . A trend2), BUA significantly increased by 0.35 dB/Mhz and 0.57 dB/Mhz among males and females, respectively  Tables  and 4. Tectively  & 4.BUA was significantly higher among Blacks compared to Whites overall, and for both sexes separately, though much more so among females   & 4. Amo4.P < 0.001). The multiple regression model of the quantitative ultrasound index confirmed that the type of academic program students attended was the single most significant predictor variable in both genders [In our study, walking/ jogging more than 10 miles per week significantly increased calcaneus BUA by 4.08 dB/Mhz and 5.97 dB/Mhz among women and men, respectively. Our findings are in agreement with previous studies examining the effect of physical activity on calcaneal BUA in similar adult populations. Among postmenopausal women, a brisk walking of approximately 20 minutes per day for a period of 12-months can significantly improve calcaneal BUA by 3 dB/Mhz compared to control subjects . In the  genders .P <.05) in men belonging to the highest quartiles of the exercise index as compared to those in the lowest [Most previous studies among males consisted mainly examined younger men –29. Dalye lowest .It is well established that osteogenic stimulus requires mechanical loading on bone and that the relationship between the loading force and the osteogenic response is strongly linear within stress tolerance limits. Thus, high impact weight bearing exercise such as jumping has been found to produce the strongest osteogenic response ,31. DespIn our study, the benefit of walking or jogging more than 10 miles per week on calcaneal BUA was greater among men compared to women. A gender difference in the effect of exercise on BUA has previously been reported for adolescent boys and girls.In a cross-sectional study of Japanese students, 15 – 20 years of age, calcaneal BUA was significantly higher in the exercise group compared to non-exercise group by 8.7 dB/Mhz and 4.6 dB/Mhz in males and females, respectively . In an 8We found that, being Black was associated with greater BUA , when compared to White subjects. Thus, the effect of race was more pronounced among females compared to males. The observed difference in the effect of race may have been due to a higher peak bone mass achieved in the young adult years among Blacks compared to Whites. In our study, younger Black females (<=40 years) and younger Black males (<=40 years) had a significantly higher calcaneal BUA compared to their White counterparts by 7.04 dB/MHz and 6.91 dB/MHz, respectively (data not shown). Aloia et al found that among white and black women of the same height and weight, Black women have both a higher skeletal mass and lean mass . Many auOur findings of the effects of age and BMI are similar to that reported by others ,48,49. E5.Our findings suggest that walking, running or jogging can in addition improve calcaneal BUA in both males and females.5.1The strength of our study is the employment of our validated physical activity questionnaire ,56. A li


Intravitreal antivascular endothelial growth factor (anti-VEGF) therapy has been widely used for the treatment of neovascularization (NV) secondary to age-related macular degeneration (AMD). This study aimed to compare the efficacy among different subtypes of neovascular age-related macular degeneration (nAMD).  PubMed, Embase, and the Cochrane Library were searched for eligible studies. We performed meta-analysis using Review Manager 5.3 and Stata/SE 12.0. μm. At 12 months, the mean changes in Early Treatment of Diabetic Retinopathy Study (ETDRS) letters were 6.38, 8.12, and 9.37, while the MT decrease was 126.51, 126.52, and 139.85 μm, respectively. However, statistically significant difference was only found between type 1 and 3 in vision improvement, both in the short term (p=0.0002) and long term (p=0.01).  A total of 24 studies met our inclusion criteria and were included in the systematic review. At 3 months, the mean logarithm of the minimum angle of resolution (logMAR) improvements were −0.09, −0.18, and −0.23 for type 1, 2, and 3, respectively, while the mean macular thickness (MT) changes were −104.83, −130.76, and −196.29  The reactivity to VEGF inhibitors varied among different subtypes of nAMD. The efficacy of intravitreal anti-VEGF therapy in type 3 nAMD was statistically better than type 1 when considering vision improvement at 3 and 12 months. Thus, the lesion subtype is a predictor for the treatment outcome which can help guide prognosis. Age-related macular degeneration (AMD) is a progressive chronic disease of the central retina and a leading cause of vision loss worldwide  which baThe classification of nAMD was first developed in 1991 , which wIntravitreal antivascular endothelial growth factor (anti-VEGF) therapy has been identified to possess the potential to stabilize or even improve visual acuity in nAMD by clinical trials . SeveralTwo independent reviewers (J. Li and J. Xu) performed a systematic search in PubMed, Embase, and the Cochrane Library on March 16, 2018, for articles focusing on the efficacy of intravitreal anti-VEGF on different subtypes of nAMD. The search strategy was [(“nAMD” AND “subtype”) AND “anti-VEGF”] using the MeSH or Emtree terms as well as free words. Here, “nAMD,” “wet AMD,” and “exudative AMD” were searched for “nAMD”; “Type 1,” “PCV,” “occult,” “poorly-defined,” “sub-RPE,” “Type 2,” “classic,” “well-defined,” “subretinal,” “Type 3,” “RAP,” and “intraretinal” were for different subtypes of nAMD, while “pegaptanib,” “bevacizumab,” “ranibizumab,” “aflibercept” and “conbercept” were searched for “anti-VEGF.”The eligibility criteria were as follows: (1) any subtype of nAMD, (2) treatment-naive nAMD, (3) anti-VEGF monotherapy, (4) efficacy measured by vision improvement or macular thickness (MT) changes, and (5) efficacy measured at 3 months or 12 months. Those studies which were not in English with no full text or irrelevant data were excluded. Any disagreements about the inclusion of an article for full review were resolved by a third researcher (P. Lu).“Risk of bias” of each included article was assessed using “risk of bias table,” which was suitable for both randomized and nonrandomized studies, according to the Cochrane Handbook for Systematic Reviews of Interventions Version 5.1.The characteristics extracted from the eligible articles included the first author's name, publication year, country where the study was conducted, subtype of nAMD, sample size, gender, mean age of the sample, vision criteria for recruitment, anti-VEGF use, and study design. Besides, PCV was marked in the column of subtype of nAMD in The main outcome of this meta-analysis was efficacy comparison of intravitreal anti-VEGF therapy for three subtypes of nAMD. The efficacy was measured by vision improvement or macular thickness (MT) changes at 3 or 12 months; therefore, these data were extracted for further analysis. Since meta-analysis was a second source, the relevant data were extracted either directly from the article or by extrapolation. In this study, no authors were contacted for the raw data of each patient, thus we could not adjust some different factors such as visual acuity at baseline.I2 ≥ 50% and/or a Q-statistic of p<0.05 was evidence supporting the presence of heterogeneity, in which the random-effects modeling method was needed. Otherwise, the fixed-effects modeling method was applied. In addition, publication bias and sensitivity analysis were conducted to study the relevant bias as well as stability and reliability of the outcomes.All the statistical analyses in our study were completed using Review Manager (RevMan) version 5.3  and Stata version 12.0 . The effect value in this meta-analysis was the mean difference (MD). Before the analysis, the study heterogeneity was tested using both the I-squared and chi-squared test statistics. An A total of 1147 articles were identified, and their records were included in EndNote X8 . After removing 242 duplicates, the remaining 905 articles were screened based on the titles and abstracts by two reviewers according to our inclusion criteria. Any disagreements about the inclusion of an article for full review were resolved by the third researcher. Full-text assessment was conducted on the rest of the 116 articles. Finally, 24 articles –36 were “Risk of bias” of these studies was assessed by “risk of bias table,” and the outcome is summarized in Among the included studies, 4 were randomized controlled trials (RCT), 12 were retrospective interventional studies (RIS), 6 were clinical trials, and the left 2 were case reports. Altogether, 2594 patients were involved in this review. The demographic characteristics of the studies were summarized in μm, respectively. It was obvious that the efficacy varied among different subtypes of nAMD; however, the subtype difference was only statistically significant between type 1 and type 3 in vision improvement (p=0.0002).μm, respectively. Although the efficacy differed in the three types of nAMD, the statistically significant subtype difference still only existed between type 1 and type 3 in vision improvement (p<0.0001).The long-term (12 months) efficacy comparison was presented in Funnel plots for short-term and long-term vision improvement comparison are shown in The impact of individual data was examined for sensitivity analysis, which was conducted on the outcome of vision improvement at 3 and 12 months . When omp=0.0002) and long term (p=0.01). We could draw a conclusion that the reactivity to VEGF inhibitors varied among different subtypes of nAMD. Thus, the lesion subtype could be considered as a predictor for the treatment outcome.To our knowledge, this was the first meta-analysis quantitatively comparing treatment efficacy of intravitreal anti-VEGF therapy for three subtypes of nAMD. A total of 24 articles were included in our systematic review. In order to assess the efficacy, the relevant data about vision improvement and MT changes at 3 months or 12 months were extracted for further pooled analysis. We found that although the efficacy of type 2 nAMD was superior to type 1 and inferior to type 3, statistically significant difference was only found between type 1 and 3 in vision improvement, both during the short term  and optical coherence tomography (OCT). Nevertheless, OCTA (and a possible OCTA classification) may offer additional information on treatment-response and anti-VEGF resistance by providing morphological information and quantitative measures , but that the clinical relevance of such findings is still debated , 38, 39.Some limitations did exist in our systematic review. Firstly, this meta-analysis consisted mostly of nonrandomized studies which were more likely to be influenced by various kinds of biases. Hence, it was not surprising that almost all of them were at relatively high risk when we assessed the risk of bias as shown in In summary, we believe that this meta-analysis, comparing treatment efficacy among different types of nAMD, is of great significance to clinical practice. The results indicate that lesion subtype is a predictor for the treatment outcome which could help guide prognosis.