Adolescents exhibit difficulties in behavioral regulation that become more evident when emotional contexts are involved, since these may hinder the development of socially-adaptive behaviors. The objectives of the present study were: to examine the influence of emotional contexts on adolescents’ ability to inhibit a prepotent response, evaluated by ERPs, and to determine whether sex differences in response inhibition are observed in adolescents in those contexts. Participants performed a prepotent response inhibition task (Go-NoGo) under 3 background context conditions: neutral, pleasant, and unpleasant. While no differences in accuracy were observed, the presence of emotional contexts did prolong reaction times compared to the neutral context. Also, the unpleasant context caused an enhancement of N2 amplitudes compared to the neutral and pleasant contexts. Also, N2 and P3 latencies were longer in emotional contexts than in the neutral condition during both correct responses and correct inhibitions. No sex differences were found in amplitude, but females showed longer N2 and P3 latencies than males. These results confirm the idea that, in adolescents, unpleasant pictures receive preferential attention over neutral images and so generate greater difficulty in response inhibition. Finally, results demonstrate that sex differences in inhibition control in adolescence were observed only in relation to time-processing. Difficulties in behavioral regulation in this population sector may become even more apparent when a romantic partner or peers are present, or when individuals are involved in emotionally-charged events. One biological factor that contributes to these difficulties is the incomplete maturation of the prefrontal lobes, together with over-activation of subcortical-limbic regions associated with affective responses and gratification9. This disequilibrium in the developmental processes of the limbic system and the prefrontal cortex underlies an imbalance between two systems: affective bottom-up regulation, and top-down impulse control10.Adolescence is a developmental period in which many physical, cognitive, emotional and physiological changes take place. It is widely-known that this transitional period between childhood and adulthood is often characterized by behavioral and emotional problems that can lead young people into risky situations and make them highly vulnerable to psychiatric illnesses and substance dependence11, behavioral regulation depends on the executive functions, but the inhibition processes that constitute a core component of those functions have not yet fully matured during adolescence. In fact, lower activity in the neural structures involved in response inhibition –including the right inferior frontal gyrus, the left dorsal and medial frontal areas and the cingulate gyrus, among others– has been described as being predictive of future involvement in such problem behaviors as consuming alcohol and other substances of abuse12.According to Barkley13.Managing daily life situations, meanwhile, demands that individuals perceive multiple environmental stimuli, and entails exerting attentional control and inhibiting prepotent responses in order to produce socially-adaptive behavior. Emotional stimuli in particular have a natural saliency that attracts attention and so modulates inhibitory control. One important factor to take into account in studies that seek to explore the effects of valence on cognition is the level of arousal that pictures with emotional content produce in an individual, as this could modulate the interference effect on the accuracy of both response and response inhibition16. In this regard, Brown et al.’s17 study with adults found higher activation of the inferior frontal gyrus (IFC) during inhibition trials compared to targets, and even higher levels when unpleasant pictures –instead of neutral ones– were presented as the background. Using a similar approach, Brown et al.18 subsequently conducted a study with adolescents. In the latter case, they found no differences in error inhibition rates, but determined that reaction times were shorter in neutral contexts than unpleasant ones. Their fMRI data revealed that the activation patterns of adolescents differed from those of adults, as the DLPFC and regions of the VLPFC showed greater activation in neutral than unpleasant contexts. These data suggest that differences between adolescents and adults may reflect not only contrasts in the activation level of those areas, but differences in certain neural structures involved in inhibition processing in the presence of emotional contexts.Some neuroimaging studies have addressed the effects of emotional stimuli on response inhibition in relation to brain activation. Two main interacting neural circuits are thought to be involved in emotional inhibition processing. The first is engaged-in-response inhibition, which involves the frontoparietal network; the other is a cortico-limbic circuit related to the top-down regulation of emotional stimuli19. The recording of brain electrical activity temporally-synchronized with specific stimuli (ERP) makes it possible to analyze precise temporal changes in cognitive and emotional processing. For this reason, the ERP technique has been used to examine the effects of emotional stimuli on inhibition processes. In this regard, two main ERP components have been identified in relation to response inhibition: N2 and P3. N2 occurs between 200 and 400 ms after stimulus onset and shows a maximum in frontal regions. The N2Go component is interpreted as reflecting responses to novelty and attentional control, whereas the N2NoGo would be more closely related to conflict resolution and inhibition processes associated with the functioning of the frontal and anterior cingulate cortices24. Some studies have demonstrated that N2NoGo amplitudes rise when task difficulty increases by reducing the reaction time deadline 25. Géczy, Czgler and Balázs26 proposed that increased N2 amplitudes in response to NoGo stimuli after Go cues could be related to more intense efforts to activate the response inhibition system and thus impede preparations for response execution. Another interesting assumption is that the N2NoGo reflects the closure of the inhibition process, or a response-monitoring process27. In summary, it is more likely that N2Go is associated with attentional control processes, while N2NoGo is related to cognitive control processes prior to response inhibition.The Go-NoGo paradigm is one of the most widely-used approaches to evaluate inhibition processing. Its aim is to generate a prepotent response by presenting a high rate of target stimuli (Go) to which the subject should respond, and a low rate of rare stimuli (NoGo) from which subjects should withhold their response28. Therefore, P3Go amplitude is enhanced at low probability targets, and has been related to such processes as stimulus evaluation, attention allocation, context updating, and subsequent memory storage30. In contrast, the P3NoGo component emerges 300–600 ms after stimulus onset, but with a frontocentral distribution. P3NoGo amplitude decreases and its latency delays with rising task difficulty as working memory requirements increase31. Also, P3NoGo amplitude is higher for motor compared to non-motor inhibition tasks32. This component appears to be linked to the later stage of the inhibition process that is closely related to the inhibition of motor responses34. It is generally accepted that the P3NoGo is related to the outcome of the inhibition process and reflects conflict inhibition processing,36 or an evaluation of the inhibitory process itself37.Turning to P3, this is a positive deflection which latency oscillates between 300 and 600 ms. It has been suggested that the parietal P3Go component is an analog of the target P3 or P3b component obtained in odd-ball paradigmsi.e., while larger P3 amplitudes were present in a positive context compared to a neutral one on NoGo trials, N2 showed higher responses in the negative context39. Another study evaluated distinct effects of two negative emotional context valences (threat and blood stimuli) on a Go/No-Go task40. Those authors found that the NoGo-N2 was larger in response to the threat than to the blood stimuli, whereas NoGo-P3 amplitude did not differ between the two conditions. Their interpretation of these results was that the threat stimuli facilitated performance of a prepotent response and enhanced conflict monitoring when action must be withheld. Killgore, Oki and Yurgelun-Todd41, meanwhile, studied the effects of two different unpleasant negative contexts on response inhibition, observing that the difference waves of P3 amplitude under disgusting contexts were smaller than under fearful ones, suggesting that disgusting distracters consume more attentional resources and, therefore, impair inhibitory control to a greater extent than fearful ones. Data from the aforementioned studies suggest that implicit emotional contexts may distract attention and, in so doing, affect inhibition processes. However, the first studies cited40 were conducted with mixed groups of men and women and did not test for sex differences, while the latter work42 involved only women. It is our contention that studying and comparing female and male adolescents in relation to the effects that emotional contexts exert on response inhibition could reveal additional data on whether sex differences exist during this important period, when top-down control is maturing. Eventually, it may be possible to relate such differences to behavioral and emotional regulation in social environments.Few ERP studies have addressed the disruptive effect of emotional information on attentional control. In this regard, researchers have identified differences that are a function of emotional context valence; 43 propose that the window of heightened vulnerability to risk-taking in this stage may be greater in magnitude and more protracted in males than females. This agrees with data from brain developmental studies that have revealed earlier brain development in females than males. Lenroot et al.44 observed that, for nearly all structures, the peak of gray matter volumes generally occurred approximately 1–2 years earlier in adolescent girls than adolescent boys, and that this difference was especially apparent in the frontal lobes. In addition, Killgore, Oki and Yurgelun-Todd41 found that at a higher age females show a progressive increase in left prefrontal cortex activation and a decrease in amygdala activation while processing fearful faces, but that males do not. This suggests a higher progressive modulation of the prefrontal cortex in the former compared to the latter. Unfortunately –and despite the importance of this topic– little attention has been paid to studying the effects of pleasant and unpleasant emotional contexts on inhibition processes in female and male adolescents, or to the neural mechanisms that underlie them.On the topic of sex differences in adolescents, Shulman, Harden, Chein and Steinberg48. Therefore, we could assume that unpleasant stimuli would cause greater inhibition interference in females, whereas males would find it more difficult to ignore pleasant stimuli while attempting to perform the inhibition task.We hypothesized that emotional contexts will have a deleterious effect on response inhibition processing, because their saliency draws attention and so will distract participants from the main task. Considering the higher modulation of the prefrontal cortex observed in female adolescents, it might be expected that they will show higher accuracy and lower neural activation than males during response inhibition when non-emotional stimuli are present. However, some studies have found that females are more sensitive than males to unpleasant/negative stimuli, while males are more reactive to pleasant/positive ones, especially images with erotic contentBased on this background, the objectives of the present study were twofold: (1) to examine the influence of emotional contexts on the ability to inhibit a prepotent response in adolescents, in relation to their behavioral performance and the time course of cognitive processing as measured by ERPs; and, (2) to elucidate whether sex differences in response inhibition under the emotional contexts are evident during adolescence.F = 0.07, p = 0.79, ŋp2 = 0.002; percentage of correct inhibitions: F = 0.27, p = 0.60, ŋp2 = 0.008; reaction times: F = 2.40, p = 0.12, ŋp2 = 0.06).Behavioral results are shown in Table F = 1.13, p = 0.32, ŋp2 = 0.03) or correct inhibitions among conditions  = 1.20, p = 0.30, ŋp2 = 0.03). However, the presence of emotional background contexts did prolong reaction times when compared to the neutral context  = 6.42, p = 0.003, ŋp2 = 0.15). Behavioral data is presented in Table There were no statistically-significant differences in the percentage of correct responses  = 1.74, p = 0.19, ŋp2 = 0.04) when comparing Go versus NoGo trials. However, N2 latency was longer during NoGo than Go trials  = 8.39, p = 0.007, ŋp2 = 0.19).P3: The significant interactions found – = 3.30, p = 0.04, ŋp2 = 0.08; and sex × trials × topographic distribution: F = 3.52, p = 0.03, ŋp2 = 0.09)– show that both females and males had higher P3 amplitudes during NoGo than Go trials in all contexts  and in all regions .F = 4.26, p = 0.02, ŋp2 = 0.11; and trials × topographic distribution, F = 3.51, p = 0.04, ŋp2 = 0.09. These results demonstrated prolonged P3 in NoGo trials in all contexts  and all the cerebral regions explored .The analysis of P3 latency showed two significant interactions: trials × context, N2NoGo: The amplitude of this component showed significant differences with respect to its scalp distribution. A significant interaction of trials × topographic distribution  = 8.68, p < 0.001, ŋp2 = 0.20) was found, and the post hoc analysis demonstrated that the N2NoGo amplitude was lower in the parietal than in the frontal (p < 0.001) and central areas (p = 0.002).p2 = 0.07).N2NoGo latency did not reach the level of significant differences regarding its topographic distribution  = 2.76, p = 0.07, ŋP3NoGo: The analysis of the P3NoGo amplitude showed a significant interaction of sex × trials × topographic distribution  = 3.52, p = 0.03, ŋp2 = 0.09). Post hoc comparisons revealed that P3NoGo showed higher voltages in the central areas than in the frontal and parietal regions ; however, these differences were only observable in male participants. In addition, the post hoc analyses subsequent to the interaction trials × laterality  = 3.51, p = 0.04, ŋp2 = 0.09) showed that P3NoGo had higher voltages at midline than in the left and right hemispheres in the central  and parietal regions .F = 3.51, p = 0.04, ŋp2 = 0.09). Post hoc comparisons demonstrated that P3NoGo latency was longer in the frontal than the central (p = 0.003) and parietal regions (p < 0.001).When analyzing the P3NoGo latency, an interaction between trials and topographic distribution emerged  = 8.68, p < 0.001, ŋp2 = 0.20). Post hoc analyses suggested that N2Go amplitude was significantly lower in the parietal areas than in the frontal (p < 0.001) and central regions (p = 0.002).F = 49, p = 0.54, ŋp2 = 0.01).The analysis of the N2Go latency did not reveal any significant differences between the conditions examined  = 3.52, p = 0.03, ŋp2 = 0.09) was found when analyzing the P3Go amplitude. Post hoc comparisons demonstrated that P3Go showed higher voltages in the frontal areas than in the central (p < 0.001), and parietal regions (p = 0.004) in both females and males. In addition, another significant interaction of trials × topographic distribution × laterality was found  = 41.86, p < 0.001, ŋp2 = 0.55). Post hoc comparisons indicated that the P3Go voltage magnitude was higher at midline than in the right parietal area (p = 0.002).F = 3.51, p = 0.04, ŋp2 = 0.09), which revealed that it was more prolonged in the frontal than the central (p = 0.003) and parietal regions (p < 0.001).The latency of the P3Go component also showed an interaction effect of trials × topographic distribution  = 12.70, p < 0.001, ŋp2 = 0.27). As it can be observed in Fig. p = 0.001) or pleasant ones (p = 0.006). Also, a significant sex × topographic distribution × context interaction was found  = 2.93, p = 0.03, ŋp2 = 0.08). Here, the post hoc analysis suggested that the differences between the contexts were restricted to females in the frontal and central areas .F = 18.88, p < 0.001, ŋp2 = 0.35), as latency was longer in the pleasant and unpleasant contexts than in the neutral one .The analyses of the N2 latency demonstrated a significant effect of contexts  = 2.26, p = 0.11, ŋp2 = 0.06).F = 15.34, p < 0.001, ŋp2 = 0.31), as post hoc comparisons indicated that it was longer for both emotional contexts than for the neutral one .However, the P3 latency analysis was significant  = 0.005, p = 0.94, ŋp2 = 0.001). However, the same analysis for N2 latency demonstrated a laterality × sex interaction  = 3.48, p = 0.04, ŋp2 = 0.09). Post hoc comparisons depicted a significantly longer latency for N2 in females than males that was spread widely across the scalp .P3. Similarly, analyses of the P3 amplitude  = 0.06, p = 0.80, ŋp2 = 0.002) revealed no significant sex-related differences.F = 4.38, p = 0.02, ŋp2 = 0.11), and subsequent post hoc comparisons suggested that females had longer P3Go latencies than males (p < 0.001).In contrast, the analysis of the P3Go latency did show a significant sex × trials interaction , when their results were compared to those of adults who had performed a similar task in a previous study51, they clearly showed lower inhibition abilities. While the adults obtained an overall (sex and condition) average of 84% for correct inhibitions, the adolescents reached only 64%. This poor inhibition accuracy was accompanied by faster responses in adolescents. These data suggest a higher susceptibility to interference from irrelevant stimuli in adolescents compared to adults; an effect that could be attributed to the incomplete maturation of the prefrontal cortex, together with enhanced activation of sub-cortical regions in the presence of affective stimuli53.No effect of context conditions was found in relation to behavioral accuracy, a result that might be explained as a top-down compensatory mechanism for the emotional interference. However, the presence of pleasant and unpleasant background contexts did prolong reaction times in relation to the neutral context condition. In this regard, Brown 55. The unpleasant background context caused an enhancement of N2 amplitudes compared to the neutral and pleasant contexts, though P3 amplitude did not change as a function of context. In addition, N2 and P3 latencies were longer in both emotional contexts than in the neutral one. It is noteworthy that the context effects on ERPs were present during both correct responses (Go) and correct inhibitions (NoGo), with no differences between them regarding conditions. However, our results did replicate those described in the literature in relation to the higher P3NoGo amplitudes and longer latencies compared to the Go trials, a finding that has been interpreted as showing that inhibitory processes are more demanding than executive motor functions57.Turning to brain activity, the effects of emotional contexts observed in our work, agree with results from other studies in which emotional stimuli interfered with cognitive tasks39. It has been reported that higher NoGoN2 amplitudes are associated with more efficient inhibitory control58. In contrast, P3 amplitudes showed no significant differences among conditions in either the Go or NoGo trials. Albert et al.39 did observe larger P3 amplitudes in the positive context in NoGo trials and suggested that stopping responses to positive stimuli are more difficult than to negative ones. As well, Buodos et al.40 found that the NoGo-N2 was larger in response to the threatening stimuli than the blood stimuli, whereas NoGo-P3 amplitude did not differ between the two conditions. Their interpretation of these results was that the threatening stimuli enhanced conflict-monitoring related to response inhibition. However, while some authors have also associated P3NoGo with motor inhibition59, others indicate that it may be related to the evaluation of the inhibitory process60. Therefore, it appears that these processes were less affected by the presence of emotional contexts in adolescents.The increased N2 amplitudes in the unpleasant context may suggest that higher top-down attentional control was required to accomplish both response and response inhibition. These results concur with those found by other authors60. In the present study, the difficulty that adolescents experienced in achieving inhibition was precisely due to the need to inhibit, first, the interference from the background distracter and, second, from the prepotent response itself. This may be reflected in the increase in the N2 amplitude associated with attentional top-down control requirements. It may be that due to the saliency of the emotional stimuli, their distracter effect increased task difficulty, provoking the increase in N2 amplitude and the prolonged latencies of both components.The longer N2 and P3 latencies observed for both emotional backgrounds compared to the neutral one indicate the need for additional processing time due to their saliency, which concurred with the longer reaction times observed in the emotional contexts. It has been reported that N2NoGo amplitude and latency also vary with task difficulty, since they increase as this parameter increases, while the NoGoP3 amplitude was not affected. In addition, the latencies of both components increased with the degree of task difficulty51, adolescents showed an overall effect in both Go and NoGoERPs. Therefore, the distracting effect of the emotional stimuli observed in adolescents indicates that they experienced difficulties in exercising interference control over the background emotional stimuli at different levels, including attentional control, conflict resolution and inhibition processes associated with the functioning of the frontal and anterior cingulate cortices22. The discrepancy in inhibition accuracy between adolescents and adults might be explained by an incomplete maturation of the top-down frontal mechanisms, which is even more visible when emotional backgrounds are present, suggesting that the activation of the limbic structures (bottom-up regulation) could add to adolescents’ difficulties with those mechanisms, as other authors have proposed10.It is important to mention that while adults suffered a context effect only on the NoGo trials in a similar task, reflected in the enhancement of N2 amplitudes in the unpleasant compared to the neutral and pleasant contexts63, where a greater disrupting effect on attentional control of the unpleasant compared to the pleasant stimuli has been found. Those results support the assumption that an attentional bias toward negatively-valenced stimuli is due to their saliency for survival64. Alternatively, it is argued that the higher effect of negative stimuli depends on the level of arousal that emotional stimuli provoke in individual subjects60, which may explain the absence of valence differences, since in the present experiment the pleasant and unpleasant stimuli used were balanced in their levels of arousal.No significant effect of valence was observed in N2 amplitude. These data disagree with previous studies44. While this would predict better inhibition processing, there are also reports that contextual stimuli –especially negative ones– exert greater attraction in women than men48. If this is true, then it could counteract women’s potentially better inhibition abilities. However, the data from the present study suggest that the deleterious effect of emotional contexts was similar in both sexes, though the female adolescents seemed to spend more time on stimuli processing, as shown by the differences in N2 and P3 latencies.On the other hand, no sex-based differences were found in the amplitudes of the brain responses of these adolescents. There is evidence that some areas of the brain, including the frontal lobes, develop earlier in girls than boys40 or only with women43, or by means of fMRI that considered sex differences in adults65 or male adolescents18, it is not possible to directly compare our results with those of other studies. The most similar study is the previous one we performed in adults, since it involved a similar task and addressed sex differences51. Therefore, our results provide new clues to the effects of emotional stimuli on brain electrical activity during response and response inhibition when sex is considered as an important variable.Since previous research on the effect of emotional contexts on inhibition processes was conducted using either ERPs with mixed groups of adults of both sexesMore research is needed to achieve a broader understanding of the brain mechanisms involved in the regulation of emotional stimuli in adolescents and adults, and of the factors that foster individual differences. Furthermore, studies considering adolescent groups with special conditions in which poor impulse control is a core characteristic –eg. impulsivity, ADHD, borderline personality, and substance abuse– would be very valuable in designing effective behavioral training programs and therapies designed to generate greater adaptive behavior in social environments.In summary, these results confirm the assumption that implicit, unpleasant pictures receive preferential attention over neutral images and so generate more difficulties in inhibition processing in adolescents. However, our findings only demonstrated sex differences related to the time processing of control inhibition in this population.A total of 36 right-handed teenagers participated in the study; 18 females  and 18 males . All were attending pre-college classes and showed above normal IQ according to the vocabulary and cubes WAIS sub-tests. The exclusion criteria were neurological or psychiatric antecedents, DSM-IV criteria for ADHD, and ongoing medical treatment. Though participation was voluntary, subjects did receive a modest financial compensation. All parents signed an informed consent form. The study was approved by the Ethical Board of the Institute of Neuroscience of the University of Guadalajara. All experiments in the present study were performed in accordance with approved guidelines and regulations.Subjects were seated comfortably in a dimly-lit, sound-attenuated, electrically-shielded room. A Go/NoGo response inhibition task was performed under 3 context conditions: neutral, pleasant and unpleasant emotional contexts. The stimuli consisted of colored arrows presented in the middle of a computer screen with bars on the left or right edge of the screen that appeared against a black background at a viewing distance of 60 cm. Subjects were instructed to press a key when the arrow coincided in both direction and color  with the bar (Go), but to withhold their response when it did not match (NoGo). The experimental design is shown in Fig. 66 were used. Thirty-five images with pleasant content and 30 others with unpleasant content were selected for the two contexts. The pleasant and unpleasant images were chosen on the basis of scores for valence and arousal using a 1-9-point analog scale from similar age groups. Images were assessed by 27 women and 17 men. The values of valence and arousal are shown in Table A total of 65 images from the International Affective Picture System (IAPS)Stimuli duration was 300 ms and the inter-trial interval was 1500 ms. Each condition consisted of 240 trials  and lasted approximately 6 min. The order of presentation of the conditions was randomized across subjects. The parameters measured were: percentage of correct responses, percentage of correct inhibitions, and reaction times. At the beginning of the experimental session, all participants received training to ensure that they clearly understood the task.67.EEG were recorded continuously at Fp1, Fp2, F3, F4, F7, F8, C3, C4, T3, T4, T5, T6, P3, P4, O1, O2, Fz, Cz and Pz, according to the 10/20 International System, with linked earlobe reference attached to an elastic cap. A Medicid 5 amplifier from Neuronic S.A. was used. The band pass was set at 0.05–50 Hz, and the sample rate at 500 Hz. Electrode impedance was kept below 5 KOhms. Electrooculograms (EOG) were recorded through electrodes placed on the supraorbital and infraorbital eye regions to eliminate segments contaminated by eye-movement artifacts. Twenty EEG artifact-free epochs from correct responses and correct inhibitions were independently averaged for each emotional condition of interest. Equal numbers of Go and NoGo trials were obtained. Epochs were defined as 100 ms before stimulus onset (baseline) and 800 ms post-stimulus. Electrode sites and scoring windows were selected by examining grand-average and individual waveforms in order to evaluate the typical recording sites analyzed in the literature for the Go/NoGo paradigm. N2 was identified as the most negative peak in the interval between 200 and 350 ms, while P3 peak amplitude was measured individually in the window between 350 and 600 ms after stimulus presentation. N2 and P3 amplitudes and latencies were analyzed for the frontal, central and parietal sites at midline  and in the left  and right hemispheres  because previous studies had reported higher amplitudes for N2 and P3 at these locations while performing a Go/NoGo paradigmp-values (p < 0.05) when appropriate, together with Bonferroni correction for pairwise comparisons.A mixed factorial analysis (2 × 3) was used to evaluate the behavioral measures, including factors –sex: male, female– and context: neutral, pleasant and unpleasant. Another mixed factorial analysis was used to analyze amplitude and latency data based on the following factors: sex , trials , contexts , topographic distribution  and laterality . Greenhouse-Geisser correction was used to adjust degrees of freedom and Tables 3 and 4


Keloid is a benign fibrous growth, which presents in scar tissue of predisposed individuals. It is a result of irregular wound healing, but the exact mechanism is unknown. However, several factors may play a role in keloid formation. To date, there are no studies of keloids in Syria, and limited studies on Caucasians, so we have investigated the risk factors of keloids in Syrians (Caucasians), and this is the main objective of this study..Diagnosis of keloids was clinically made after an interview and physical examination. We did a histopathological study in case the physical examination was unclear, anatomical sites, age of onset, number of injured sites (single\multiple) and family history.The following information was taken for each patient; sex, Blood groups (ABO\Rh), cause of scarring. There were 209 (80.7 %) patients with keloids in a single anatomical site compared to 50 (19.3 %) patients with 130 keloids in multiple anatomical sites, 253 (97.68 %) patients with keloids caused by a single cause for each patient compared to 6 (2.32 %) patients with keloids caused by two different causes for each patient.We have studied the clinical characteristics of 259 patients with keloids,130 (50.2 %) females and 129 (49.8 %) malesKeloids could follow any form of skin injury, but burn was the most common (28.68 %). Also, keloids could develop at any anatomical sites, but upper limb (20 %) followed by sternum (19.17 %) was the most common. Over half of the patients developed keloids in the 11–30 age range. 19.3 % (50/259) of patients had family history, 76 % (38/50) of them had keloids located in the same anatomical sites of relative, also, 66 % (33\50) of them had keloids caused by the same cause.p = 0.01) compared with other blood groups, spontaneous keloids in patients with blood group A (p = 0.01), acne in males (p = 0.0008) compared to females, acne in someone who has a previous acne keloid (p = 0.0002), burn in someone who has a previous burn keloid (p = 0.029), family history, especially for spontaneous (p = 0.005), presternal (p = 0.039) and shoulder (p = 0.008) keloids, people in second and third decades (p = 0.02) (p = 0.01) respectively.The following information was found to be statistically significant; people with blood group A  were significant in keloid formation in Syrians. Keloid is a benign fibrous growth, presents in scar tissue of predisposed individuals, extends beyond the borders of the original wound, doesn’t usually regress spontaneously, and tends to recur after excision . It is aThere is a clear genetic component given the correlation with family history, high occurrence in identical twins , 5, highTo date, there are no studies of keloids in Syria, and limited studies on Caucasians, so we investigated the risk factors of keloids in Syrians (Caucasians), and this is the main objective of this study.This study was conducted between March/2013 and August/2015 in the departments of dermatology, at Tishreen and Alassad Hospitals, Lattakia, Syria.The diagnosis of keloids was clinically made after an interview and physical examination. We did a histopathologic study if the physical examination was unclear, especially for genitalia, buttock, palm and sole keloids, and for other uncertain scars.All patients with keloids were included into this study, while patients with hypertrophic scare were excluded. Hypertrophic scars are defined as raised scars that remained within the boundaries of the original lesion, often regressing spontaneously after the initial injury and rarely recurring after surgical excision. In contrast, a keloid scar is defined as a dermal lesion that spreads beyond the margin of the original wound, continues to grow over time, does not regress spontaneously and commonly recurring after excision .The following information was taken for each patient: Sex , blood groups (ABO and Rh), cause of scarring or form of skin injury  and other), the age of onset , anatomical sites , the number of injured anatomical sites (single/multiple), family history, the anatomical site and the cause of relative when family history is positive.Note: Multiple sites refer to keloids found in a multiple number of anatomical sites as opposed to multiple keloids found in the same anatomical site. Therefore, the presence of more than one scar in the same anatomical site was not considered to be multiple keloids. Also, a single site keloid refers to a keloid or a number of keloids found in only one anatomical site.We have to compare some of our data  with similar society data, so we got the relative frequency of blood groups from National Blood Transfusion Center (NBTC) of Lattakia for 2015, and the relative frequency of sex and age groups from the Syrian Census Center (SCC) for 2011, which was the last data of SCC because of war conditions. We needed chi-square test to compare that data, so we created a miniature group containing 259 people reflecting society data . We considered it as a control group.http://www.quantpsy.org/chisq/chisq.htm was used for comparing data. A P-value of less than 0.05 was considered statistically significant.Microsoft Excel 2012 and Microsoft Word 2012 were used for tables and figures. Calculation for the chi-square test on We studied the clinical characteristics of 259 patients with keloids,130 (50.2 %) females and 129 (49.8 %) males. There were 209 patients with keloids in single anatomical site compared to 50 patients with 130 keloids in multiple anatomical sites, and 253 patients with keloids caused by a single cause for each patient compared to 6 patients with keloids caused by two different causes for each patient. According to that, we had 259 patients with 339 keloids spread over 15 anatomical sites, and 265 causes of scarring as a final result to 7 different forms of skin injury (Table p = 0.79) (Table p = 0.036)  and males 129 (49.8 %), with statistical significance for developing keloids neither male nor female compared to control ( % and map = 0.0008)  (28.68 %), and trauma (20\265) (7.55 %) was the least Table . Causes 8) Table . Also, k8) Table . Distrib8) Table  (Table 3p = 0.01) (Table P = 0.01) (Table p = 0.7)  found statistical significance for developing keloids in people with blood group A (1) Table  Fig. 2)p = 0.01)1) Table . Howeverp = 0.029) (p = 0.0002) respectively  of patients had 130 keloids in multiple anatomical sites as opposed to 80.7 % (209/259), who had a keloid in a single site  and not with number of causes (265) or number of sites (339)). As at (Table atients 29 and not causes 25 or numbp = 0.002) (p = 0.047) (p = 0.006) respectively (Table 19.3 % 50/259) of patients had family history, 76 % (38/50) of them had keloids located in the same anatomical sites of relative. Also, 66 % (33\50) of them had keloids caused by the same cause Table  0/259 of Over half of the patients developed keloids in the 11–30 age range Table .Fig. 6ReP = 0.02) (P = 0.01) respectively, as opposed to age group A (first decade), that had statistical significance for not developing keloids compared with control (p = 0.04). (Table Both age groups B and C (second and third decades) had statistical significance for developing keloids compared with control P = 0.0 (P = 0.0). Table .The distribution of causes and anatomical sites according to age groups were demonstrated at Table  Table 99 respectp value  to reduce false positive.This mean for each variable  we have 7 independent tests. Therefore, because of multiple tests, we lowered the p < 0.0001), and for no developing spontaneous keloids (p = 0.01) (Table In age group A 0-10Y), there was statistical significance for developing burn keloids  (0.008) (p = 0.002) respectively (Table In age group B 11–20Y), causes had almost coordinated distribution. The shoulder (17.76 %) followed by sternum (15.88 %) were the most common sites for developing keloids in this age group. There was statistical significance for developing sharp wound, acne and earlobe keloids , there was also a coordinated distribution of causes.p = 0.002) (Table The upper limb (26.7 %) followed by sternum (16.7 %) were the most common sites. We found statistical significance for developing scalp keloids in this group (2) Table .In age group D (31–40Y), causes had almost coordinated distribution considering the absence of acne keloids. The sternum (24.14 %) was the most common site for developing keloids in this age group without statistical significance.p = 0.00002) (p = 0.0002) (p = 0.003) respectively , there was statistical significance for developing surgical keloids in these groups (ly Table .p = 0.003) (p = 0.0002) respectively (Table The sternum was the most common site for developing keloids in these groups (28.6 %) (53.8 %) (77.8 %) respectively with statistical significance for developing presternal keloids only in F and G groups  to demonstrate some of the risk factors of keloids in Syrians (Caucasians).Incidence of keloids was equal in females and males. That agree with most previous studies , 11–13. p = 0.0008), because only males have acne keloidalis nuchae. Also, the severity of acne is higher in them [Burn was the most common cause of keloid formation in our study (28.68 %), but laceration was the most common one in a Jamaican study . We can  in them .The most common anatomical site for developing keloids differs according to race, traditions and conditions of study’s society, generally keloids tend to occur on highly mobile sites with high tension , 17. In p = 0.01), that may be partly explained by the association between the effect of red cell antigens A , which has not been previously reported. This result confirms the effect of red cell antigens A in development of keloids, as we discussed before.People with blood group A have high probability to develop keloids compared with other blood groups  and othal cells , and disal cells . Spontanal cells . Howeveral cells  (13.13 %p = 0.029) (p = 0.0002) respectively, because both acne and burn could affect multiple sites more than other causes, which is more located. This means there was high probability to develop acne or burn keloids in another anatomical site in a patient who had a previous acne or burn keloid respectively. In our study, the upper limb was the most common anatomical site for developing keloids in patient with multiple sites (46 %), and sternum was the most common one in patients with a single site (26.84 %), while the earlobe was the most common site in both multiple (24 %) and single (48 %) sites in the Jamaican study [Few studies discussed the development of keloids in single versus multiple anatomical sites , 19. Thele sites 6 %, and le sites 6 %, and le sites 6 %, and an study  found stOnly (6/259) (2.32 %) of our patients had keloids caused by two different causes, compared to 15.9 % of patients in another study . This rep = 0.002), which usually appears in the second decade ) (Table p = 0.047), (p = 0.006) respectively. These results reflect the importance of the cause and anatomical site in heredity of keloid. Some recent studies confirmed the hereditary of spontaneous keloids [Most keloids occur sporadically, but some cases are familial, 19.3 % of our patients had family history, which was lower than previous studies , 14. 76 )) Table , and for keloids . Other s keloids .p = 0.04), because people in this decade are not stimulated by sexual hormones (higher incidence of keloid formation during puberty) [P = 0.02) (P = 0.01) respectively, and tend to decrease in older , especially on upper and lower limbs. This is a logical result, because most of burn accidents exist in younger children especially on extremities. Occurrence of acne keloids was higher in age group B compared with other groups (p = 0.008), because the peak in prevalence and severity of acne occurs in second decade [p = 0.002) and earlobe (p = 0.002) keloids in this age group. Also, development of scalp keloids was higher in age group C compared with other groups (p = 0.002), because most cases of acne keloidalis nuchae occur in persons aged 14–25 years [p = 0.00002) (p = 0.0002) (p = 0.003) respectively, especially on sternum. These results reflect an increase of open heart surgeries in older people, especially for males who were older than forty compared to females in the same age (p = 0.036).Although keloids could occur at any age, they were rare in first decade (age group A) (puberty) , most lider Fig. , because) P = 0.0 respectid decade . High fr25 years   have an important role in keloid formation and consequentially in predicting a keloid’s behavior in response to treatment and prognosis. In this study we have demonstrated the significance of the previous factors in keloid formation in Syrian population. Therefore, we have to generalize these results, especially for dermatologists and surgeons. These observations could indicate a genetic basis in keloid formation, which justifies the need for genetic studies and more studies about families with keloids to define the type of heredity in our patients.  had keloid formation. That suggests kind of sex-linked heredity, which has not been previously reported).


NSCs) are self‐renewing, pluripotent and undifferentiated cells which have the potential to differentiate into neurons, oligodendrocytes and astrocytes. NSC therapy for tissue regeneration, thus, gains popularity. However, the low survivals rate of the transplanted cell impedes its utilities. In this study, we tested whether melatonin, a potent antioxidant, could promote the NSC proliferation and neuronal differentiation, especially, in the presence of the pro‐inflammatory cytokine interleukin‐18 (IL‐18). Our results showed that melatonin per se indeed exhibited beneficial effects on NSCs and IL‐18 inhibited NSC proliferation, neurosphere formation and their differentiation into neurons. All inhibitory effects of IL‐18 on NSCs were significantly reduced by melatonin treatment. Moreover, melatonin application increased the production of both brain‐derived and glial cell‐derived neurotrophic factors  in IL‐18‐stimulated NSCs. It was observed that inhibition of BDNF or GDNF hindered the protective effects of melatonin on NSCs. A potentially protective mechanism of melatonin on the inhibition of NSC's differentiation caused IL‐18 may attribute to the up‐regulation of these two major neurotrophic factors, BNDF and GNDF. The findings indicate that melatonin may play an important role promoting the survival of NSCs in neuroinflammatory diseases.Neural stem cells ( Spinal cord injury can be resulted from neural trauma, inflammation or degeneration, and it is a serious clinical disorder. The patients suffer from their immobility, and it also poses enormous economic burden to their family and also to the society Cytokine IL‐18 is a member of the IL‐1 family N‐acetyl‐5‐methoxytryptamine), synthesized from the pineal gland as well as in the peripheral tissues, plays the important roles in various physiological processes including the circadian rhythm, reproduction and the cerebrovascular and neuroimmuno‐endocrine functions Melatonin  in the presence of melatonin (10 ng/ml) and/or luzindole (5 μM). All mouse experiments are carried out in accordance with the relevant institutional and national guidelines and regulations, approved by the Animal Care and Use Committee of Peking Union Medical College Hospital and conform to the relevant regulatory standards.All experimental protocols had been approved by the Clinical Research Ethics Committee of the Peking Union Medical College Hospital. NSCs were obtained from 13.5‐day‐old embryos of Wistar rats using an established method Cells were fixed with 3.5% formaldehyde, permeabilized with 0.1% Triton X‐100, blocked with 3% BSA and 0.05% Tween‐20 in PBS and then incubated with primary antibody(dilutions 1:5000) ; Sigma‐Aldrich, Oakville, ON, Canada) overnight at 4°C. Nuclei were stained with 4, 6‐diamidino‐2‐phenylindole (DAPI) (Sigma‐Aldrich). Fluorescence was measured by a confocal microscope .Total protein was extracted from cells and separated using 10% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis gel. Proteins were transferred onto a nitrocellulose membrane , and the membrane was blocked with non‐fat milk. Then, the membrane was probed with primary antibodies (dilutions 1:5000) (Sigma‐Aldrich) and bound with horseradish peroxidase‐conjugated secondary antibodies. Chemiluminescent signal was developed using the ECL kit .Enzyme‐linked immunosorbent assay (ELISA) was performed to measure brain‐derived neurotrophic factor (BDNF) and glial cell‐derived neurotrophic factor (GDNF) levels in the supernatant of cell culture according to the manufacturer's instructions . The absorbance was measured at a 450‐nm wavelength.Cell proliferation was measured using a Cell Counting Kit‐8  according to the manufacturer's instructions. The cells were cultured for 0, 24, 48 or 72 hrs, and the absorbance was measured at a 450‐nm wavelength.Total RNA was extracted using Trizol Reagent (Invitrogen) from cells following to the manufacturer's instructions. PCR was performed with specific primers in 20 μl PCR mixtures for 35 cycles. The levels of mRNA were measured by SYBR Green quantitative PCR performed on the iQ5 Real‐Time PCR Detection System . The primers used for PCR amplification were shown in Table t‐test was performed for comparison between two groups, and an analysis of variance (anova) was used to compare multiple groups and followed by Student's t‐test for comparison between two groups. P < 0.05 were considered statistically significant.Data were expressed as means ± standard deviation (SD). Student's The cells cultured in neurobasal medium supplemented with bFGF and B27 proliferated and formed neurospheres on the second day after culture Fig. A and B. CCK‐8 assay showed that IL‐18 (1–100 ng/ml) significantly repressed NSC proliferation in a dose‐dependent manner Fig. A. In addper se had a profound effect to promote NSCs proliferation. When melatonin co‐incubated with IL‐18, it partially reversed the inhibitory effect of IL‐18 on NSCs  were expressed in NSCs Fig. A and B. BDNF and GDNF are two important growth factors involved in NSC survival and proliferation. In this regard, ELISA results showed that melatonin drastically increased contents of both BDNF and GDNF and these were also blocked by luzindole Fig. A and B. It was observed that both BDNF inhibitor (TrkB‐Fc) and GDNF inhibitor (MAB212) also partially reduced the beneficial effects of melatonin on the NSC proliferation and neurosphere formation under the IL‐18 challenging Figs A,B. The via its induction on antioxidant and anti‐inflammatory enzymes et al. et al. et al. NSCs are immature, self‐renewing and undifferentiated precursor cells with an potential to differentiate to astrocytes, neurons or oligodendrocytes The results identified the molecular mechanisms as to how melatonin would increase the survival rate and differentiation of NSCs and provided novel evidence for clinical application of melatonin as a neuroprotective agent in neuroinflammatory diseases, especially, considering the low or non‐toxicity of this molecule.The authors declare no conflict of interest.Z. L, X.Y L, X. Y, J.X. S set up the idea for writing the manuscript; Z. L, X. Y, J.X. S. collected the data regarding the manuscript; Z. L, X. Y, J.X. S analysed the data; Z. L, X. Y, J.X. S. wrote the original manuscript in English; and M.T.V. C, W. K. K. W., D. X. T revised the manuscript, worked on the English and made the final version of the manuscript. All authors reviewed the final version of manuscript.


To improve its air quality, Beijing, the capital of China, has implemented high-cost pollution control measures mainly focused on shifting its energy mix. However, the effectiveness of these measures has long been questioned, especially given the recent problem of severe haze. The main study objectives are to achieve independent, although indirect, information on Beijing’s air pollution by measuring the level of polycyclic aromatic hydrocarbon (PAH) contamination in topsoil and to examine how soil contamination reflects energy consumption. Soil sampling data from two years, 2004 and 2013, were used. The key findings are as follows: 1) although the total PAH content in the topsoil did not significantly decrease from 2004 to 2013, the composition changed considerably; 2) as of 2013, vehicle emissions replaced coal combustion as the leading source of soil PAHs, which validates the existing policy measures regarding vehicle purchasing and traffic volume; 3) the regional transport of atmospheric pollutants, as indicated by the contribution of coking sources in 2013, is not negligible; and 4) appropriate policy measures are needed to control the growing practice of burning biomass. Overall, this study demonstrates that the PAH contamination in topsoil represents an informative indicator of Beijing’s energy consumption and overall environmental quality. Based on long-term monitoring data of SO2, NO2, CO and PM10, this review found that the control measures are improving Beijing’s air quality. Citywide monitoring of PM2.5 started in Beijing after 2012, but the trend of PM2.5 during the study period remains unclear. Overall, more studies are needed to evaluate the cost and benefits of Beijing’s air pollution control.Significant air pollution in Beijing has long been a topic of concern10This study used a unique variable, polycyclic aromatic hydrocarbon (PAH) contamination in topsoil, to interpret the evolution of energy consumption and air quality in Beijing. PAHs are a group of toxic and carcinogenic pollutants ubiquitous in urban environments13141517182021231325262829This study investigated PAH contamination in the topsoil of Beijing’s downtown and suburban areas based on a unique dataset from two years. Two large field campaigns, one organized in 2004 and the other in 2013, generated two snapshots of the contamination. As most PAH compounds are fairly persistent in natural conditions, the snapshots reflect the average contamination situation prior to the campaigns. By comparing the two snapshots, the evolution of PAH contamination during the past decade was assessed. Source apportionment analyses were performed using Positive Matrix Factorization (PMF) to examine the change of PAH sources from 2004 to 2013. PMF is a receptor model approach that allows the optimal use of available data because it can retain missing data points and/or values that are below the detection limit in its calculation. PMF also considers data uncertainty by assessing the quality and reliability of each data pointOur study focused on the downtown and suburban districts of Beijing. All 16 PAH compounds were detected in the soil samples (45 samples in each campaign) collected within the study area. In the downtown districts, the median concentrations of ∑16 PAHs in 2004 and 2013 were 699.44 ng/g and 500.27 ng/g, respectively. In the suburban districts, the median concentrations were 183.20 ng/g and 177.11 ng/g, respectively, indicating a much lower level of PAH contamination than in the downtown districts. The source profiles derived for both 2004 and 2013 are given in 32333435353438eq) as a risk indicator. The BaPeq concentration mainly reflects the magnitude of coal combustion and vehicle emission sources. The average BaPeq values of all samples were 139.48 ng/g and 310.42 ng/g for 2013 and 2004, respectively, indicating a significant reduction of the potential risk associated with soil contamination. eq values. The interpolation was finished in ArcGIS for Desktop (version 10.2.1) using the inverse distance weighting method. As shown in the figure, the risk level substantially decreased from 2004 to 2013 for most of the downtown area, and its spatial pattern changed significantly. In 2004, two contamination hotspots were in the southeastern and northern regions , the total annual energy consumption of Beijing increased from 46.482 million tons of SCE  in 2003 to 71.777 million tons of SCE in 2012. The energy mix also changed significantly. The annual coal consumption in Beijing has been decreasing since 2005 , and the40422.5 in Beijing, respectively. All the above results are consistent with our findings on soil PAHs : naphthalene (NAP), acenaphthene (ACE), acenaphthylene (ACY), fluorene (FLO), phenanthrene (PHE), anthracene (ANT), fluoranthene (FLA), pyrene (PYR), benz(a)anthracene (BaA), chrysene (CHR), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), indenopyrene (IcdP), dibenzanthracene (DahA), and benzoperylene (BghiP).−1. The oven temperature was programmed to increase from 60 °C to 280 °C at a rate of 5 °C min−1 and maintained at 280 °C for 20 min. All solvents, hexane and dichloromethane, were HPLC- grade . Silica gel  was heated at 450 °C for 4 hours, kept in a sealed desiccator, and reactivated at 130 °C for 16 hours prior to use. Sodium sulfate was pre-baked at 650 °C for 4 hours and then kept in a sealed desiccator. The working standard solution was prepared by diluting a stock standard from a commercial mixed standard  with hexane.The laboratory sample analysis and quality control used in 2004 were described in a previous paper8, acenaphthene-d10, phenanthrene-d10, chrysene-d12, and perylene-d12), and the surrogate recoveries were 65% to 127%. For each batch of samples, two procedural blanks were processed, and all data were blank-corrected. The relative percent difference between individual PAH compounds identified in method duplicate samples was <15%. The detection limits were 0.30 ng/g to 0.60 ng/g based on a 10 g soil sample.All data were subject to strict quality control procedures. Quantification was performed using an internal calibration method. The samples were spiked with five surrogate standards  is a principal component analysis (PCA)-based receptor model with non-negative constraints:Xn×m is a matrix of sample concentrations that can be decomposed into Gn×p, a matrix of source contributions, and Fp×m, a matrix of source profiles, plus a residual matrix, En×m. n and m denote the number of samples and the number of pollutant species, respectively, and p is the number of influential sources. The physical nature of the influential pollution sources can be interpreted from Fp×m based on prior information such as measured profiles of specific sources and emission inventories. Each row in Fp×m represents an influential source , and the sum of the row elements  quantifies the overall contribution of the source. PMF minimizes the following objective functionwhere Sij represents the “uncertainty” in the jth species for the ith sample and Eij  represents the error corresponding to Sij.where sij based on the method detection limit (MDL) of targeted pollutant species as:In this study, the EPA PMF 5.0 programxij represents elements of the concentration matrix Xn×m and EF is a measurement error fraction. EF was set to 0.2 in this study. In PMF analyses, the value of p has to be carefully determined to ensure physically meaningful results while keeping the matrix dimensionality lowp value. The theoretical Q value should be approximately equal to the number of degrees of freedomwhere p was varied between 3 and 6 and eventually set to 4, for which the Q values calculated by Because NAP is a relatively volatile and unstable compound, it was excluded from the source apportionment, which was aimed at determining the long-term characteristics of PAH sources. Two PMF models were established for the sample sets of 2004 and 2013. eq) as a risk indicatoreq can be calculated as follows:We used the carcinogenic equivalent concentration of BaP . The TEF values are 0.001 for NAP, ACY, ACE, FLO, PHE, FLA and PYR; 0.01 for ANT, CHR and BghiP; 0.1 for BaA, BbF, BkF and IcdP; and 1 for BaP and DahAeq values to these limits and defined five levels of risk: I) BaPeq ≤ 10 ng/g (no risk); II) 10 ng/g < BaPeq ≤ 100 ng/g (low risk); III) 100 ng/g < BaPeq ≤ 500 ng/g (medium risk); IV) 500 ng/g < BaPeq ≤ 1000 ng/g (high risk); and V) BaPeq > 1000 ng/g (extremely high risk).where TEFHow to cite this article: Li, J. et al. PAH contamination in Beijing’s topsoil: A unique indicator of the megacity’s evolving energy consumption and overall environmental quality. Sci. Rep.6, 33245; doi: 10.1038/srep33245 (2016).


Fluorine is a bone‐seeking element ubiquitously present in the environment and widely used in many oral hygiene products. In humans, excessive intake of fluoride may cause dental and skeletal fluorosis. However, endemic fluorosis does not appear to develop in a proportion of individuals exposed to the same levels of fluoride. The mechanisms by which mammalian cells resist fluoride are still unclear. In this study, we developed strains of mouse L‐929 cells resistant to different levels of fluoride. High‐throughput RNA‐sequencing analyses of the fluoride‐resistant L‐929 cells indicated that massive changes in global gene expression occurred, compared with the wild‐type L‐929 cells. The main biological processes and functions changed were associated with the extracellular region and matrix, response to stress, receptor binding, and signal transduction. This indicated that high doses of fluoride not only exerted stress on L‐929 cells but also induced functional pathways that helped them adapt to the presence of fluoride or to expel it. These data should prove useful in identifying cellular processes or transporters/channels that play central roles in adaptation to or expulsion of fluoride in humans. ECMextracellular matrixFoxOforkhead box OFRfluoride‐resistantKEGGKyoto encyclopedia of genes and genomesMAPKmitogen‐activated protein kinaseMTTmethyl thiazolyl tetrazoliumNF‐kappa Bnuclear factor kappa BODoptical densitiesPI3Kphosphoinositide 3‐kinaseWTwild‐typem) fluoride. On the other hand, accumulated data suggest that fluoride is toxic to bacteria, fungi, plants, animals, and humans at high concentrations or doses Fluorine is a bone‐seeking element and is ubiquitous in the environment. Compared with chloride and iodide, two halides that have been well studied for their effects on living organisms, much less is known about the biological importance of fluoride and the mechanisms by which cells respond to this anion − or other inorganic anions across cell membranes F is identified to function as fluoride‐specific F−/H+ antiporters in bacteria and lower eukaryotes Because of its ubiquitous presence in the environment and its toxic effects, organisms most likely have evolved mechanisms of resistance to fluoride. Previous studies suggested that a carrier protein could have already existed within the cellular membrane acting as a barrier to fluoride or as a transporter of fluoride, protecting the activity of intracellular enzymes sensitive to fluoride In the present study, we developed strains of mouse L‐929 cells resistant to different levels of fluoride. High‐throughput RNA‐sequencing analyses of the fluoride‐resistant (FR) L‐929 cells indicated that, compared with the wild‐type (WT) L‐929 cells, massive changes in global gene expressions occurred within the induced FR L‐929 cells. The main biological processes and functions changed were associated with extracellular region and matrix, response to stress, receptor binding, and signal transductions etc., indicating high dose fluoride not only exerted stress on L‐929 cells but also induced functional pathways that helped them adapt to or expel the fluoride. Our data may be valuable in identifying cellular processes and transporters/channels that play central roles in adapting to/expelling fluoride in mammalian cells in the future.−1 penicillin, and 100 μg·mL−1 streptomycin. The 2000 p.p.m. (105.2 mm) fluoride stock solution was prepared by solving 2.2 g sodium fluoride (Sigma) powders in 500 mL MilliQ water . Different concentrations of fluoride media (work solution) were prepared by mixing different volume of fluoride stock solution with the culture medium.The culture medium was Eagle's Minimum Essential Medium  supplemented with 10% FBS , 100 U·mL2 atmosphere. During subculture, cells were treated with 0.25% (wt/vol) trypsin/0.02% (wt/vol) EDTA and then were resuspended and seeded with culture media. Countess Automated Cell Counter  were used to count cells and measure average cell size.Wild‐type L‐929 cells  were maintained in culture media at 37 °C under a 5% COin vitro by sequential exposure of WT L‐929 cells to 10 p.p.m. (0.526 mm) fluoride concentration gradient ascending media. The induction was initiated by subcultivating L‐929 cells in fluoride medium of low fluoride concentration. Cell proliferation rate was determined by the methyl thiazolyl tetrazolium (MTT) assay. The induction cycles continued till the proliferation rate of the WT L‐929 cells in the fluoride media became normal and were repeated at a higher fluoride concentration till the desired FR L‐929 cell strain was established.Fluoride‐resistant L‐929 cells, which are capable of proliferating in media of fluoride concentrations that would otherwise be cytotoxic to WT L‐929 cells, were induced Poly(A) mRNA was isolated using beads containing oligo (dT) from total RNA. Purified mRNA was then fragmented and used as templates. The first‐strand cDNA was synthesized with random hexamer primers and the second‐strand cDNA was synthesized using buffer, dNTPs, RNase H, and DNA polymerase I. Short double‐stranded cDNA fragments were purified with a QIAquick PCR purification kit  and ligated to Illumina sequencing adaptors. The amplified library was sequenced on an Illumina HiSeq™ 2000  sequencing machine by paired‐end sequencing.Differentially expressed genes (fold change ≥ 2) between WT and 30 p.p.m. FR L‐929 cells were identified using the significance of digital gene expression profiles.Total RNA was extracted using the Trizol reagent  according to the manufacturer's instructions. RNA was reverse‐transcribed into cDNAs by the AMV Reverse Transcription System . Quantitative RT‐RCR was performed with the SYBR Green QPCR system  with GAPDH as an internal control.t‐test was used. When P value is less than 0.05, significant differences were determined.Experimental data were reported as mean ± SEM. To compare between the different of two groups, two‐tailed Student's m) fluoride caused slight to moderate reduction in cell growth. Cells tended to adapt to present fluoride concentration around 3 days after the treatment. Inhibition was pronounced with 30 p.p.m. (1.579 mm) fluoride and showed no obvious debilitation during 2‐week observation period. In the presence of 60 p.p.m. (3.158 mm) or higher concentrations of fluoride, extended cell death occurred quickly after the addition of fluoride and almost all cells died within 3 days.Wild‐type L‐929 cells were shocked with different concentrations of fluoride by exposing cells to prepared fluoride media. MTT assay was performed according to scheduled time duration and the optical densities (OD) at the wavelength of 570 nm were obtained. As shown in Fig. m) fluoride media till cell proliferation rate was back to normal, which we considered that the strain was adapted to 5 p.p.m. (0.263 mm) fluoride. Next, cells were subcultured in media with 10 p.p.m. (0.526 mm) fluoride. Four weeks later, the growth rate was normal again and 10 p.p.m. FR strain was obtained. Such cells were then stepwise exposed to 10 p.p.m. (0.526 mm) gradient ascending fluoride media from 20 p.p.m. (1.052 mm) to 90 p.p.m. (4.737 mm). Fluoride sensitivity was determined by MTT test when cell confluence reached 90% to confirm the adaptation of cells to fluoride. Cells would not be exposed to higher concentration of fluoride until the growth rate became normal and the adaptation was obtained. The proliferation curves of WT and FR L‐929 cells were showed in Fig. Initially the WT L‐929 strain was exposed to 5 p.p.m.  and the highest 90 p.p.m. (4.737 mm) fluoride concentration so that the gene expression changes may be significant but not biased. We used three batches of independently cultured WT and 30 p.p.m. FR L‐929 cells in the study. Cluster analysis of the global gene expression patterns showed that the three independent 30 p.p.m. FR L‐929 cells naturally grouped together, indicating the 30 p.p.m. (1.579 mm) fluoride exerted significant changes on gene expressions within L‐929 cells , 6 of them were related to responses to external stimuli  fluoride was first a strong external stress for the survival of the L‐929 cells. Level 4 analyses of the biological process showed that the top three biological processes (with the highest percent of genes) were ‘cell communication’, ‘multicellular organismal development’, and ‘system development’  fluoride stress.We next performed gene ontology analyses of the 385 genes whose expressions were significantly changed in the 30 p.p.m. FR L‐929 cells. Analyses of the biological processes that the 385 genes were involved showed that many genes possibly belong to stress response  fluoride stress elicited massive rearrangements of extracellular molecules as well as their cytoplasmic counterparts. The above information suggested that the L‐929 cells made a lot of changes across the cellular membrane to adapt the toxic stress of the 30 p.p.m. (1.579 mm) fluoride.Analyses of the cellular components that the 385 genes belong to showed that most of them were closely associated with the extracellular compartment, including extracellular matrix (ECM), organelles, and vesicles etc. Fig. A,B. Deepm) fluoride elicited many signaling transduction activities in the L‐929 cells, which could also play very important roles in helping the cells in adaptation to the toxic fluoride rich environment.Analyses of the molecular functions the 385 genes were associated with showed that a majority of them were involved in protein and receptor binding  pathway analyses showed that, of the six major functional areas, three of them were involved, which are ‘environmental information processing’, ‘organismal systems’, and ‘Human diseases’ Fig. A. The toPlin2, Clec11a, Ereg, Cxcl1, Inhba, Loxl3, Col1a2, Fmod, Il1a, Ccl20, Slit1, and Ctgf were consistent with the expression fold change data from RNA‐seq studies. These data indicated that our RNA‐seq studies of the global gene expression changes in the 30 p.p.m. FR L‐929 cells were reliable.To validate the gene expression changes in the 30 p.p.m. FR L‐929 cells from the RNA‐seq data, we further performed real‐time RT‐PCR experiments on a list of genes that participated in multiple biological process and pathways concluded from our gene ontology and KEGG pathway studies Fig. A. These Toxic effects of excessive fluoride on organisms including mammals have been well documented crcB RNA motif) that binds to fluoride and regulates the expression of genes in response to this anion in a wide variety of bacteria and archaea FF proteins have been shown to function as fluoride‐specific F−/H+ antiporters Recently, some new clues emerged with the discovery of the fluoride riboswitch  in culture media m) and could recover to normal within 2 weeks, while 30 p.p.m. (1.579 mm) and higher concentrations of fluoride caused more pronounced inhibition of cell growth. Fluoride above 60 p.p.m. (3.158 mm) killed almost all the cells within 1 day. Early studies in 1970s reported that by stepwise increase in fluoride concentration, cells adapted to grow at fluoride concentrations that would kill them Developing mammalian cells that can adapt to high level of fluoride is believed to be one of the best ways in vitro by sequential exposure of WT L‐929 cells to gradient ascending fluoride concentration in culture media. By using high‐throughput RNA‐sequencing technology, we, for the first time, systemically analyzed the global gene expression changes after fluoride adaptation of mammalian cells. We identified 385 genes that exhibited at least twofold changes in 30 p.p.m. FR mouse L‐929 cells compared with WT ones. Of the 385 genes, 265 genes were up‐regulated and 120 genes down‐regulated. Gene ontology analyses for these 385 genes showed that, response to stress, multiorganismal and system development, extracellular region of the cells, as well as protein and receptor binding are the main biological processes and molecular functions the FR L‐929 cells changed to adapt to the fluoride toxin. These data suggested that the high concentration of fluoride is firstly a strong stress for the mammalian cells to respond. The cells also tend to form a unity to adapt to the toxic stress and then stimulate downstream signaling pathways to up‐regulate the inner system of expelling the fluoride toxin. KEGG pathway analyses indicated that these downstream pathways that may play key roles in helping expelling fluoride out of cells are ‘ECM‐receptor interaction’, ‘MAPK signaling pathway’, ‘PI3K‐Akt signaling pathway’, ‘P53 signaling pathway’, ‘NF‐kappa B signaling pathway’, ‘FoxO signaling pathway’, and ‘Focal adhesion’ etc. These results further indicated that, even as simple as the addition of one toxin as an anion of fluoride, the cells respond to it in a very complex way to systemically change the functional units expressed within cells for adaptation or expelling the fluoride toxin.In the present study, FR strains of L‐929 cells were successfully induced Although we did not identify the single key gene/protein that determine fluoride resistance of mammalian cells in the current study, our approach using next‐generation RNA‐seq in FR mammalian cells should facilitate future studies in dissection of molecular pathways and the development of more appropriate FR mammalian cell model systems. We believe our data can be used as an important tool for novel genes and gene feature discovery in FR mammalian cells in later more detailed studies.In summary, under the limitation of this study, we developed FR strains of mouse L‐929 cells and systemically analyzed the global gene expression changes after fluoride adaptation of these mammalian cells. To our knowledge, our study showed, for the first time, that main biological processes and functions changed within the induced FR strains of L‐929 cells were associated with extracellular region and matrix, response to stress, receptor binding, and signal transductions etc., which indicate that high‐dose fluoride not only exerted stress on L‐929 cells but also induced functional pathways that helped them adapt to or expel the fluoride.BL and YL conceived and designed the project; SR, WZ, LW, and JW performed the experiments; NS, YL, and BL analyzed data; SR and NS wrote the paper; BL and NS made manuscript revisions. All authors read and approved the final manuscript.Table S1 Transporters/Channels among the 385 changed genes. [Correction added after online publication on 21 June 2017: Table S1 revised].Click here for additional data file.


Dipyrone  is a non-steroidal anti-inflammatory drug commonly used both in human and in veterinary medicine. After oral administration, is broken down rapidly to metabolites which largely retain the activity of the parent drug. Its metabolites have analgesic, antipyretic and anti-inflammatory effects.p < 0.05) was proven in case: activated partial thromboplastin time; ratio of activated partial thromboplastin time; hemoglobin; hematocrit; mean corpuscular hemoglobin; mean corpuscular volume; red blood cells volume; white blood cells volume; prothrombin time index.The subjects were eight healthy male Large White post-suckling piglets, weighing between 5.0 to 7.4 kg, of ages 35 ± 10 days. The animals were administered MET (100 mg/kg) by an intramuscular (I.M.) injection. The study calculated the value of several hemorheological parameters. Significant impact of MET treatment  contains supplementary material, which is available to authorized users. The validation parameters that were determined through our work according to the LOO 4MAA↔ PT + [(Hb + Hct)/(RBCATT Pr atio)] generated the following values: Q2asym-Q2 = 0,063, Q2 = 0.8274, R2 = 0.8905, Q2-R2 = 0.0631, SS = 1573.36, PRESS = 271.57. At the same time, our work did not confirm the existence of the same dependency relation to 4AA↔ PT + [(Hb + Hct)/(RBCATT Pr atio)] (p > 0.05).As presented in the form of an arithmetic expression o)] Fig. , there wThe aim of the current study was a verification of piglets as a model reflecting changes in hematological parameters after a single I.M. injection of MET. Thrombocytopenia and pancytopenia after MET administration is related to bone marrow suppression. Mechanism covers drug-dependent suppression of the growth of myeloid progenitors, primitive multipotential progenitors and erythroid progenitors , 32. As Rhamdia sp. based studies . This relationship predictive value with respect to 4MAA, confirms the performed validation procedure. This means that hemorheological parameters after administration of MET may reflect the concentration of 4MAA in piglet blood plasma. Of note, the mechanism of the effect of the 4MAA on hemorheological parameters is significantly different from that of 4AA.In the course of the analysis, we have confirmed the existence of a 4MAA dependence↔ In our study, all the piglets showed tolerance to the dosages given out, and such high doses allowed to analyze precisely the impact of MET on hemorheological parameters. In our study, nine parameters showed significant differences to baseline. Most deleterious effect on blood parameters observed in current study were deep and prolonged decrease of PT and Hb concentration connected with decreased number of RBC. As in humans, the piglet model shown that MET and its metabolites contribute to significant increase of number of neutrophils. The resulting image changes converge to the observations that were noticed in humans. To sum up, our observations suggest that a piglet model is useful for studying MET and its metabolites impact on hemorheological parameters.


Carotid‐femoral pulse‐wave velocity (cf‐PWV) and brachial‐ankle PWV (ba‐PWV) are the 2 most frequently applied PWV measurements. However, little is known about the comparison of hypertensive target organ damage (TOD) with cf‐PWV and ba‐PWV.P<0.05). When cf‐PWV and ba‐PWV were both or separately put into the same full‐mode model after adjustment for confounders, only cf‐PWV, but not ba‐PWV, showed significant association with carotid intima‐media thickness and creatinine clearance rate (P<0.05). Similar results were observed in logistic regression analysis.A total of 1599 community‐dwelling elderly subjects (age >65 years) in northern Shanghai were recruited from June 2014 to August 2015. Both cf‐PWV and ba‐PWV were measured using SphygmoCor and VP1000 systems, respectively. Within the framework of comprehensive cardiovascular examinations, risk factors were assessed, and asymptomatic TOD, including left ventricular mass index, peak transmitral pulsed Doppler velocity/early diastolic tissue Doppler velocity (E/Ea), carotid intima‐media thickness, arterial plaque, creatinine clearance rate, and urinary albumin‐creatinine ratio were all evaluated. Both PWVs were significantly associated with male sex, age, waist/hip circumference, fasting plasma glucose, and systolic blood pressure, and ba‐PWV was also significantly related to body mass index. Both PWVs were significantly correlated with most TOD. When cf‐PWV and ba‐PWV were both or separately put into the stepwise linear regression model together with cardiovascular risk factors and treatment, only cf‐PWV, but not ba‐PWV, was significantly associated with carotid intima‐media thickness and creatinine clearance rate (Taken together, in the community‐dwelling elderly Chinese, cf‐PWV seems to be more closely associated with hypertensive TOD, especially vascular and renal TOD, as compared with ba‐PWV.http://www.clinicaltrials.gov. Unique identifier: NCT02368938.URL:  Pulse‐wave velocity (PWV) has been widely accepted in clinical practice as a gold standard of arterial stiffness,As a velocity, PWV can be conveniently measured as the distance divided by the time interval between 2 selected points of the arterial system.The Northern Shanghai Study is a prospective, ongoing, and multistage cohort study with the objective of investigating the cardiovascular risk assessment system in elderly Chinese. From June 2014 to August 2015, subjects were recruited if they were (1) aged 65 years or more, (2) local residents from urban communities in the north of Shanghai, and (3) available for long‐term follow‐up. Exclusion criteria included (1) severe cardiac disease (NewYork Heart Association Class IV) or end‐stage renal disease (chronic kidney disease >4), (2) malignant tumor with life expectancy less than 5 years, and (3) stroke history within 3 months. Finally, 1721 subjects were invited, of whom 1599 (92.9%) were enrolled. The Northern Shanghai Study was authorized and financially supported by the Shanghai municipal government (Grant ID 2013ZYJB0902) and was approved by the Ethics Committee of Shanghai Tenth People's Hospital. Written informed consent was obtained from all subjects after relevant information had been provided to them and their relatives.As previously described,2])=175×(plasma creatinine)−1.234×age−0.179 (women×0.79).Venous blood samples and urine samples were obtained in subjects after an overnight fast. Biological markers, including plasma low‐density lipoprotein (LDL) cholesterol, high‐density lipoprotein (HDL) cholesterol, plasma creatinine, and urinary microalbumin and creatinine were assayed by standard methods in the Department of Laboratory Medicine of Shanghai Tenth People's Hospital. Fasting plasma glucose was measured with the glucose‐oxidase method. Creatinine clearance rate (CCR) was calculated by the modified Modification of Diet in Renal Disease formula for Chinese as follows: CCR (mL/[min·1.73 mBlood pressure (BP) of each subject was measured in the morning with the electronic device by specialized physicians 3 times after at least 10 minutes of rest in the sitting position. The average of the 3 BP readings was calculated and used in the subsequent statistical analysis. Pulse pressure was defined as the difference between systolic (SBP) and diastolic BP.Bilateral brachial and ankle BPs were automatically and simultaneously measured to calculate the ankle‐brachial index  using the VP1000 system .The order of cf‐PWV and ba‐PWV measurements was randomly chosen by the coin‐tossing method. The cf‐PWV was measured using applanation tonometry , according to the European Expert Consensus on Arterial Stiffness.The ba‐PWV was measured using the VP1000 system (Omron) as previously described.The common carotid artery intima‐media thickness (CIMT) was measured using the MyLab 30 Gold cardiovascular system  with a 7.5‐MHz probe. As previously described,M‐mode and 2‐dimensional echocardiography were performed using the same device with a 3.5‐MHz probe, according to the guidelines of the American Society of Echocardiography.2  or LVMI ≥95 g/m2 . LV diastolic dysfunction was assessed by E/Ea and other evidence of abnormal LV relaxing and filling, such as enlarged left atrial volume and increased LVM.2 or LVMI>149 g/m2  or LVMI≥122 g/m2 .2), and microalbuminuria (UACR >30).Asymptomatic hypertensive TOD include cardiac, arterial, and renal TOD. Left ventricular hypertrophy was defined as LVMI ≥115 g/mP<0.05 was considered statistically significant.Quantitative and qualitative parameters were presented as means±standard deviation and numbers with the percentage in parentheses, compared between men and women by Student t test and chi‐squared test, respectively. Multivariate linear regressions were conducted to detect the association of conventional cardiovascular risk factors with cf‐PWV and ba‐PWV, such as sex, age, smoking, family history of premature CVD, waist/hip circumference, body mass index (BMI), fasting plasma glucose, LDL cholesterol, HDL cholesterol, and SBP. Pearson correlation analysis was applied to investigate the correlation of asymptomatic hypertensive TOD with cf‐PWV and ba‐PWV, respectively. Further, stepwise multivariate linear and logistic regressions were conducted to investigate the association of hypertensive TOD with cf‐PWV and ba‐PWV. Only variables staying in the final model were presented. In addition, cf‐PWV and ba‐PWV were both or separately put into the same full‐mode regression models after adjustments for confounders to detect the regression coefficients and odds ratios (ORs) of PWVs in hypertensive TOD. It is noteworthy that age and SBP were forced into all models of regressions because PWVs were strongly dependent on the 2 variables. Statistical analysis was performed using SAS software, version 9.3 . P<0.001), higher waist circumference , lower incidence of family history of premature CVD , lower HDL cholesterol , lower LDL cholesterol , lower pulse pressure , higher CIMT , lower CCR , and lower ba‐PWV .Characteristics of participants by sex are presented in Table P<0.04), whereas ba‐PWV was also significantly related to BMI (P=0.007). The total R2 values of the full models with cf‐PWV and ba‐PWV were 0.252 and 0.301, respectively.Conventional cardiovascular risk factors, including sex, age, smoking, family history of premature CVD, waist/hip circumference, BMI, fasting plasma glucose, HDL cholesterol, LDL cholesterol, and SBP, were put into a multivariate linear regression model to investigate their association with cf‐PWV and ba‐PWV. As shown in Table P<0.01), and ba‐PWV was also significantly correlated with all parameters (P<0.05), except CCR (P=0.077)  Table .P=0.001, incremental R2=0.005) and CCR . Consistently, when they were separately put into the same model, only cf‐PWV showed significant association with CIMT , and CCR   were forced into all models in the following linear and logistic regression analysis to investigate the association of cf‐PWV and ba‐PWV with hypertensive TOD. As shown in Table P=0.037) and microalbuminuria . Similarly, when they were separately put into the stepwise linear regression model, cf‐PWV showed significant association with arterial plaque  and microalbuminuria , whereas ba‐PWV was significantly associated with increased CIMT  and microalbuminuria  (Table P=0.045) (P=0.005), chronic kidney disease , and microalbuminuria , whereas ba‐PWV was significantly associated only with microalbuminuria  in Moreover, when both cf‐PWV and ba‐PWV were put into the stepwise logistic regression model in Table P=0.045) , panel CThere were 2 major findings in the present study. First, both cf‐PWV and ba‐PWV exhibited similar associations with various cardiovascular risk factors in the elderly, including male sex, age, waist/hip circumference, fasting plasma glucose, and SBP. Second, hypertensive TOD, especially vascular and renal TOD, may be more closely associated with cf‐PWV than with ba‐PWV in the study cohort of community‐dwelling elderly.For 2 decades, arterial stiffness was recognized as an important predictor of CVD and mortality. In theory, increased arterial stiffness is related to elasticity loss and reduced compliance in the arteries, which can further increase blood pressure.Both cf‐PWV and ba‐PWV, as the most common indexes of arterial stiffness, have been widely used in clinical practice, with greater usage of cf‐PWV in the Western countries and greater usage of ba‐PWV in Asia.2 values. The relatively small correlation coefficients and R2 values of the present study may be attributable to the complicated confounders and relatively weak association among the intermediate outcomes in the real‐world “healthy” elderly subjects. Nevertheless, it is also noteworthy that our findings were reliable because they were double checked by the multiple linear and logistic regression models and with many variables adjusted. As to the discrepancies existing between the cardiac and vascular/renal TOD in association with those 2 PWVs, the main reason responsible for them may be that cardiac structure and function indicated by LVMI and E/Ea were less affected by arterial stiffness as compared with vascular and renal abnormalities.In literature,Our results were consistent with previous studies and current guidelines,The findings of the present study need to be interpreted within the context of its limitations. First, as a cross‐sectional study, we focused only on the comparative analysis of cf‐PWV and ba‐PWV in association with cardiovascular risk factors and hypertensive TOD in the elderly but without any intervention or causality interpretation. With ongoing follow‐up studies, we will be able to provide more prospective data in the future. Second, we could not fully adjust for the influence of different medications on PWVs, such as various antihypertensive, antidiabetic, and antihyperlipidemic drugs, which may have differing effects on central and peripheral arterial stiffness.In light of the high burden of cardiovascular diseases on the aging society, an accurate assessment of arterial stiffness is of great importance and should be strongly recommended in the elderly. In the present study cf‐PWV was more closely associated with vascular and renal TOD in comparison with ba‐PWV in the elderly. However, the mechanisms remain incompletely understood. Therefore, further laboratory research is warranted to investigate the underlying mechanisms.Cf‐PWV seems to be superior to ba‐PWV in association with asymptomatic hypertensive TOD, especially vascular and renal TOD, in the community‐dwelling elderly Chinese population.This study was authorized and financially supported by the Shanghai Municipal Government . Dr Yi Zhang was supported by the National Nature Science Foundation of China .None.


Previous research has reported a negative impact of olfactory dysfunction on quality of life (QoL) and depressive symptoms. As self‐esteem was identified as a contributing factor to depression, this study aimed to investigate QoL, depressive symptoms and self‐esteem in patients with smell loss.Prospective controlled study.Department of Biomedical Imaging and Image‐guided Therapy, Medical University of Vienna, in co‐operation with the Department of Ear, Nose and Throat Diseases, Medical University of Vienna, Austria.Twenty‐two anosmic patients  and 25 healthy controls  participated in this study.WHOQOL‐BREF), depressive symptoms (BDI‐II) and self‐esteem (MSWS) were conducted.Olfactory performance was assessed using the Sniffin’ Sticks battery. In addition, psychological questionnaires that covered the topics quality of life (The results of this study revealed a decrease in QoL and reduced body‐related self‐esteem in anosmic patients. Furthermore, QoL and self‐esteem were correlated with depressive symptoms.As self‐esteem, QoL and depressive symptoms in anosmia interact with each other, we suggest that self‐esteem should be considered in the medical history, in order to provide a personalised intervention, adapted to the patient's needs. Many affected patients suffer from reduced QoL, and they have a higher risk to develop depressive symptoms.It has been established that olfactory dysfunction and depressive symptoms are associated in two ways. First, patients diagnosed with major depressive disorder often exhibit decreased olfactory function , and the study protocol was approved by the Ethics Committee of the Medical University of Vienna. All subjects were informed about the aim of the study and gave written, informed consent prior to inclusion.2.2Twenty‐two anosmic patients  and 25 healthy subjects  participated in this study. All subjects had no history of neurologic or psychiatric diseases, and no history of severe head trauma. Parts of this cohort had already participated in other studies of our research group.Self‐esteem is an important factor in anosmic patients.Self‐esteem, quality of life and depressive systems act as highly interactive factors in anosmic patients.Self‐esteem should therefore be assessed in medical history.Self‐esteem may be important to provide a treatment adapted to the patient's needs.2.3Olfactory performance was assessed using the Sniffin’ Sticks test battery , comprising the three subtests: the odour detection threshold test; the odour discrimination test; and the odour identification test. A detailed description of the testing procedure is presented in Hummel et al.2.4Quality of life depicts the general well‐being of a subject. Subjective QoL was assessed using the German version of the WHOQOL‐BREF,Depressive symptoms were evaluated using the German version of the Beck Depression Inventory II BDI‐II. This muSelf‐esteem reflects a person's subjective evaluation of his or her own worth. It includes beliefs and emotional states about oneself. Self‐esteem was investigated using the Multidimensionale Selbstwertskala MSWS, a Germa2.5t‐test was performed. The alpha level for statistical testing was set to α=0.05. In addition to traditional statistical parameters, the Bayes factor (BF)https://jasp-stats.org/). The BF is a relative likelihood ratio for the null hypothesis and the alternative hypothesis. High values represent a likelihood favouring the null hypothesis, low or negative values represent a likelihood favouring the alternative hypothesis.Statistical analysis was performed using the Statistical Package for the Social Sciences , version 20.0. For all test scores, mean and standard deviation were calculated. As all variables fulfilled the requirements for parametric testing, data were analysed using a one‐way multivariate analysis of variance (MANOVA). For group comparison of depressive symptoms, a two‐sample 33.1r=.054, P=.720, BF01=5.166; psychological: r=−.246, P=.096, BF01=1.428; social relationships: r=−.085; P=.571, BF01=4.708; environment: r=−.043, P=.774, BF01=5.283) or gender .The QoL questionnaire (WHOQOL‐BREF) scores were not significantly correlated with age . In a next step, all dependent variables were analysed separately. A significant effect of olfactory dysfunction was observed in the psychological domain =4.823, P=.033, ωp²=0.075; see Figure The four domains of the WHOQOL‐BREF were compared between anosmic patients and healthy controls using a one‐way MANOVA with the dependent variables physical health, psychological, social relationships and environment. Statistical analysis revealed a significant main effect of olfactory dysfunction on the combined variables =2.782, 3.2t(41)=0.898, P=.375, BF01=2.389). However, a significant negative correlation was obtained between the psychological domain of the WHOQOL‐BREF and the BDI scores , as well as the body‐related self‐esteem and the BDI scores  in the total study sample.No statistically significant difference in depressive symptoms was observed between anosmic patients and healthy controls  or gender . General self‐esteem was significantly correlated with the psychological domain , with social relationships , and with environment  of the WHOQOL‐BREF. Body‐related self‐esteem was associated only with the psychological domain .Self‐esteem (MSWS) scores were not significantly correlated with age  on the combined dependent variables. Detailed analysis, corrected for multiple testing using Bonferroni correction, revealed that olfactory dysfunction had a statistically significant effect on only one of the three subscales of the MSWS: body‐related self‐esteem =4.391, P=.046, ωp²=0.110), with decreased scores for patients with olfactory dysfunction =1.665,44.1p²=0.110) suggests that the reduced body‐related self‐esteem is not only statistically significant, but also clinically relevant. Although no statistically significant differences in depressive symptoms were observed between anosmic patients and healthy controls, the scores in the psychological domain were highly correlated with depressive symptoms.The main aim of the study was to investigate self‐esteem and QoL in anosmic patients compared to healthy controls. The results of this study revealed that anosmic patients experience a decreased QoL in the psychological domain, and significantly reduced body‐related self‐esteem. The medium effect size , resulting in a power (1−β)=0.81 with a total sample size of n=47 and medium effect sizes of |ρ|=0.35. For the analysis of self‐esteem, the statistical power decreased to 0.60 because of the reduced sample size. However, despite this reduced statistical power, a statistically significant difference was obtained. The study sample was recruited for another investigation conducted by our study group.This is the first study investigating the impact of self‐esteem on QoL and depressive symptoms in anosmic patients. Data of a homogeneous study sample of anosmic patients were acquired. However, there are still some limitations on this study. A potential limitation of this study is that healthy controls were significantly younger than patients with olfactory dysfunction. However, no significant correlations between age and the behavioural measures  were observed. Furthermore, data from the MSWS were available for only a part of the complete study sample. Another important issue is the application of correlation analyses in smaller samples. We therefore conducted a post‐hoc power analysis using the statistical program G*Power (5The present study is the first investigating self‐esteem in patients with anosmia. The results have shown that self‐esteem plays a crucial role in these patients. We assume that self‐esteem and QoL are important factors in the risk for developing depressive symptoms in patients with smell disorder. We therefore suggest collecting data regarding self‐esteem and depressive symptoms in patients with olfactory dysfunction, in order to grain a deeper insight into the complex interaction of these parameters and develop individually adapted therapeutic interventions according to the needs of the patient.K. Kollndorfer and V. Schöpf designed the study. J.L. Reichert, B. Brückler and V. Hinterleitner acquired the data and supported data analysis. K. Kollndorfer analysed the data. K. Kollndorfer and V. Schöpf wrote the manuscript. All authors revised the article critically.None declared.