Identifying and characterizing anti-CRISPR proteins opens an avenue to explore and control CRISPR–Cas machineries for the development of new CRISPR–Cas based biotechnological and therapeutic tools. Past studies have identified anti-CRISPRs in several model phage genomes, but a challenge exists to comprehensively screen for anti-CRISPRs accurately and efficiently from genome and metagenome sequence data. Here, we have developed an ensemble learning based predictor, PaCRISPR, to accurately identify anti-CRISPRs from protein datasets derived from genome and metagenome sequencing projects. PaCRISPR employs different types of feature recognition united within an ensemble framework. Extensive cross-validation and independent tests show that PaCRISPR achieves a significantly more accurate performance compared with homology-based baseline predictors and an existing toolkit. The performance of PaCRISPR was further validated in discovering anti-CRISPRs that were not part of the training for PaCRISPR, but which were recently demonstrated to function as anti-CRISPRs for phage infections. Data visualization on anti-CRISPR relationships, highlighting sequence similarity and phylogenetic considerations, is part of the output from the PaCRISPR toolkit, which is freely available at  Bacteria protect themselves from bacteriophage (phage) infections through a variety of different mechanisms, including the CRISPR–Cas adaptive immune system and restriction modification systems. To counteract different CRISPR–Cas systems, phages have evolved protein inhibitors known as anti-CRISPRs . IdentifSeveral strategies have been used to identify anti-CRISPRs ,5, incluet al. developed a random forest based model, which was fed with features, including protein length, whether it was annotated, and its mean hydrophobicity (doi: https://doi.org/10.1101/2020.01.23.916767). Using this model, a diverse array of anti-CRISPRs were predicted and made publicly accessible. While this warehouse stores many potential anti-CRISPRs for later experimental confirmation, it does not allow researchers to perform their own anti-CRISPR predictions. Eitzinger et al. developed an eXtreme Gradient Boosting based predictor AcRanker, and fed their model with features, including amino acid composition (AAC) and grouped dimer- and trimer-frequency counts based on the physicochemical properties of these amino acids   ,32. To md model) .F-value and Matthews correlation coefficient (MCC)  curve, with its AUC (area under the curve) value calculated, was used to visualize the prediction performance of a predictor.The proposed method was rigorously and extensively validated based on the 5-fold cross-validation test, an additional independent test, and prediction capability was further investigated using case studies. Performance measurements include Sensitivity (SN), Specificity (SP), Accuracy (ACC), nt (MCC) , which aThe architecture of the PaCRISPR server consists of two components: a client web interface and a server backend Figure .https://jquery.com/), Bootstrap (https://bootstrapdocs.com/) and their extension packages. Specifically, the sequence similarity was visualized by BlasterJS (https://www.java.com/) server development suite, including Struts 2 (https://struts.apache.org/) and Hibernate (https://hibernate.org/).The client web interface is responsible for interacting with users through the input and output displays, and to process the service logic including the illegal character detection, sequence validation and format. The former was implemented by JSP, CSS, jQuery  dependent on the e1071 package for SVM modelling (https://CRAN.R-project.org/package=e1071). The BLAST program (version 2.8.1+) (https://metacpan.org/pod/CGI) program was written to string together these steps within a single thread.The server backend is responsible for executing the whole prediction process, including encoding features, making predictions, and generating visualize-ready data. The prediction program was written in R language ( 2.8.1+)  was used 2.8.1+)  was usedhttp://gearman.org/). The client web interface simply puts the user's submissions (each of them as a job) into the queueing system, where the Perl idle threads, maintained in a daemon thread pool with customizable size, pull and execute the jobs. During the whole process, the MySQL database (https://www.mysql.com/) is used to store intermediate and final results, as well as synchronize messages between the client web interface and the server backend. In this way, the architecture brings better user experience by decoupling the client web interface that requires prompt response speed and the server backend that handles time-consuming jobs. This also makes the architecture amenable for expansions to add new computational facilities to meet the increasing demand in predicting ever accumulating genome-scale data.The client web interface interacts with the server backend through a fast and lightweight queueing system, implemented using the Gearman framework (N = 10 in this work), and subsequently averaged as the final performance results. Similarly, all independent tests were conducted 10 times, each of which was executed based on a balanced independent dataset comprised of the 26 independent positive samples and 26 randomly chosen negative samples. All models adopted a default cut-off threshold of 0.5 to keep the balance of sensitivity and specificity. The final ensemble model of the PaCRISPR toolkit was obtained through integrating the four single evolutionary feature-based models. The PaCRISPR toolkit was additionally benchmarked with two homology-based baseline predictors and the AcRanker toolkit (http://acranker.pythonanywhere.com/) -based predictor was implemented based on the HMMER toolkit   using thre.com/) . The BLAre.com/) . Each of toolkit . The cutF-value, MCC and AUC values reflect its comprehensive predictive power, while a balance of SN and SP  demonstrated its capability in identifying both positive and negative samples.In both test cases, similar observations were obtained Figure : (i) ModAll machine learning based models, including ours and the AcRanker, obviously outperformed the homology-based baseline predictors on the independent test, with a remarkably better recognition capability of anti-CRISPRs Figure . The homWhen predicting long non-anti-CRISPR proteins from phages and bacterial MGEs, both PaCRISPR and AcRanker achieved high true negative prediction accuracy . SpecifiTowards the end of this study, 5 new anti-CRISPR proteins were discovered, providing perfect case studies to compare the proposed method with its peers in terms of their generalized prediction capabilities. These proteins are significantly different from all anti-CRISPRs in the training dataset with less than 10% sequence similarity Figure .Four anti-CRISPRs from AcrIIA16 to AcrIIA19 were found to inhibit the class II-A CRISPR–Cas systems across multiple host species . No singAcrIII-1  inhibits type III CRISPR systems (including both type III-A and III-B subtypes) and was identified in a range of viruses and plasmids from archaeal species . This maIn summary, PaCRISPR (and its single models trained with PSSM-AC and RPSSM) successfully identified 4 out of 5 case study anti-CRISPRs. In contrast, both BLAST-based and HMM-based predictors failed to predict any of the newly identified proteins as anti-CRISPRs (data not shown). Both models rely on sequence similarity-based comparisons between the query proteins and previously known anti-CRISPRs, and in the case of each of the newly identified anti-CRISPR proteins above, the candidates show very low similarities with previously known anti-CRISPRs. Using the same threshold of –5, as used for the independent test, AcRanker picked out only AcrIIA17 and AcrIII-1 as anti-CRISPRs. However, the low prediction scores it gives to these new proteins would lead to a low rank in the context of genome-scale prediction.Streptococcus agalactiae strain GB00548. This contig encodes 93 proteins, only one of which is an anti-CRISPR, the novel AcrIIA21 discovered by AcRanker  from the genome sequence of AcRanker . We thenCompared to AcRanker, PaCRISPR generally ranked known anti-CRISPRs in higher ranking positions . For eacPaCRISPR is a publicly available and user-friendly web server for predicting anti-CRISPRs and analysing their relationships with known anti-CRISPRs Figure . PaCRISPTo distinguish between predicted and known anti-CRISPRs, each of the proteins will be marked with either an ‘Exp.’ if it has been experimentally validated or ‘Pred.’ with a prediction score. For validated anti-CRISPRs, users will be provided with a link to the relevant page in either the Anti-CRISPRdb  or the uDiscovery of new anti-CRISPRs opens an avenue for manipulating CRISPR–Cas machineries as a tool in gene editing or gene therapy, and provides new insights into phage interactions with their bacterial hosts. In this work, we have presented PaCRISPR, an ensemble and universal predictor to accurately and efficiently identify anti-CRISPRs from genome- and metagenome-derived sequence data. PaCRISPR aims at mining and extracting distinctive patterns and characteristics from known anti-CRISPRs by incorporating multiple evolutionary features within an ensemble framework. Compared with sequence-based features, the evolutionary features were demonstrated more suitable to predict this type of highly evolutionary proteins. Having been extensively and rigorously benchmarked in terms of prediction accuracy and robustness, PaCRISPR can identify novel anti-CRISPRs and outperforms other toolkits with a significant performance improvement. PaCRISPR differentiates itself with an extended interactive environment where users can tentatively annotate the putative anti-CRISPR based on the sequence similarity and phylogenetic analysis. It is anticipated that PaCRISPR could serve as a useful preliminary screening toolkit to identify potential anti-CRISPRs, and therefore expedite the discovery of novel anti-CRISPRs for their subsequent experimental validation. To ensure the PaCRISPR toolkit remains competitive and up-to-date, it will be periodically upgraded as new anti-CRISPRs are identified and experimentally validated.http://pacrispr.erc.monash.edu/. All datasets used in this study have been uploaded to the Supplementary Data at NAR online and can also be downloaded via http://pacrispr.erc.monash.edu/download.jsp.The PaCRISPR toolkit is freely available at gkaa432_Supplemental_FilesClick here for additional data file.


Introduction: Autonomic dysfunction is a common and disabling non-motor symptom of Parkinson's disease (PD). We aimed to understand autonomic dysfunction in PD motor subtypes, the pattern of sympathetic skin response (SSR) to motor asymmetry, and the association of SSR with autonomic and motor dysfunctions.Methods: A total of 101 PD patients of Han Chinese were included. Unified PD rating scale (UPDRS), scales for outcomes in PD-autonomic symptoms (SCOPA-AUT), orthostatic hypotension, and SSR were evaluated.Results: SCOPA-AUT and incidences of orthostatic hypotension and absent SSR were worse in the subtype of postural instability gait disorder (PIGD) than the subtypes of tremor dominant and intermediate. SSR latency and amplitude were asymmetrical corresponding to the accentuation of motor severity. Patients with absent SSR had worse UPDRS and SCOPA-AUT scores. SSR parameters of the severe side in patients with SSR showed no independent association with the scores.Conclusion: Our results support that autonomic dysfunction is more severe in the PIGD than other subtypes and demonstrate an asymmetry of SSR in PD patients. Absent SSR may indicate worse autonomic and motor symptoms, but SSR parameters are not sufficient to evaluate the severity of the dysfunctions. Parkinson's disease (PD) is a common neurodegenerative disease caused by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta. Patients not only suffer from motor inconveniences and non-motor symptoms such as cognitive impairment, psychosis, and autonomic dysfunction but also impaired quality of life . SimilarAutonomic dysfunction is one of the most common non-motor symptoms in PD and is considered to be heterogeneous as well . All regSympathetic skin response (SSR) is a simple and non-invasive method used to evaluate skin sympathetic sudomotor function based on conductance changes in response to stimuli , 13. ThiA total of 101 PD patients of Han Chinese ethnicity consisting of 49 men and 52 women were recruited in this study. The mean age of the patients was 65.2 ± 8.3 years. All patients were diagnosed by two movement disorder neurologists according to the UK Parkinson's Disease Society Brain Bank Criteria. Excluded participants were those with a family history of PD, secondary and atypical parkinsonism, or confounding factors potentially involving autonomic symptoms , on medications potentially influencing autonomic symptoms , and being diagnosed with cognitive impairment which would otherwise compromise our assessment. All included patients signed written informed consents. This study was approved by the ethics committee of the Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University.Clinical information including age at onset, disease duration, L-dopa equivalent daily dosage (LEDD), scale for outcomes in PD for autonomic symptoms (SCOPA-AUT), and non-motor symptom assessment scale (NMSS) was collected through face-to-face interviews. Unified PD rating scale (UPDRS), Hoehn–Yahr stage, orthostatic hypotension, and SSR were evaluated 12 h after stopping antiparkinsonian medication, a practically defined OFF medication state . The sevEach of the patients was classified into subtypes of tremor-dominant (TD), postural instability and gait disturbance (PIGD), or intermediate as described previously , 21. In P < 0.05 was considered statistically significant.Data were analyzed using the statistical package of Predictive Analytics Software 19.0  for Windows. Distribution normality was evaluated by the Kolmogorov–Smirnov test. Differences in gender, orthostatic hypotension frequency, and absent SSR frequency between PD subtypes were assessed using the Chi square test. Differences in age, age at onset, UPDRS-Total score, UPDRS-III score, and SCOPA-AUT score were analyzed by one-way analysis of variance. Differences in disease duration, Hoehn–Yahr stage, LEDD, and NMSS score were analyzed using the Kruskal–Wallis test or Mann–Whitney test. Multivariate analysis was performed by multiple linear regression model or binary logistic regression model using the stepwise forward method with subtypes, gender, age, age at onset, duration, LEDD and UPDRS-III score as covariates. Differences in SSR parameters between the severe and less-severe sides were analyzed by the Wilcoxon signed-rank test. Spearman rank correlation was used to test the association between SSR parameters and scores of SCOPA-AUT or UPDRS-III. A two-tailed P > 0.05) among the three subtype groups (P < 0.001), UPDRS-III (P = 0.013), Hoehn–Yahr stage (P < 0.001), NMSS (P < 0.001), SCOPA-AUT (P < 0.001), orthostatic hypotension frequency (P = 0.011), and absent SSR frequency (P = 0.001). Numeric results of all these measurements were higher in the PIGD subtype compared to the other two subtypes. However, for the ones with SSR latency and amplitude detected, SSRs of the severe side in both upper and lower limbs showed no significant difference among the three subtypes  and orthostatic hypotension , urinary (P = 0.002), cardiovascular (P = 0.001) and thermoregulatory regions  and lower limbs , but not associated with SSR latencies of both limbs , but not associated with other ones (The patients with absent SSR had worse UPDRS-Total, UPDRS-III, and SCOPA-AUT scores compared with those with SSR . The latth limbs . In contth limbs . Analysiher ones . We alsoher ones . To clarher ones .Autonomic dysfunction is a frequent and disabling complication of PD. In this study, we analyzed autonomic symptoms in a currently largest group of Han Chinese PD patients. The results disclose a heterogeneity of dysautonomia in PD subtypes, an asymmetry of SSR, and the role of SSR absence and parameters in autonomic and motor symptoms.PIGD and TD are two typical motor subtypes of PD. A number of studies have demonstrated that certain non-motor symptoms differ among these two subtypes as well as the subtype in between , 22–24. While motor manifestations often develop and persist asymmetrically in PD , 9, we oOrthostatic hypotension is a key manifestation of cardiovascular dysautonomia in PD and is believed to be caused by degeneration of postganglionic sympathetic neurons which resulted from alpha-synuclein deposition. Such incidence in PD is around 30–40% . HoweverIn conclusion, our results support that autonomic dysfunction is more severe in patients of the PIGD subtype and demonstrate an asymmetry of SSR in PD. As evidenced by the analyses with SCOPA-AUT and UPDRS-III scores, absent SSR may indicate worse autonomic and motor symptoms, but SSR parameters of the severe side are not sufficient to evaluate the progression of the dysfunctions.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Ethics Committee of the Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University. The patients/participants provided their written informed consent to participate in this study.J-YW, M-YW, R-PL, YL, and XZ examined the patient and acquired and analyzed all clinical data. J-YW, M-YW, W-YZ, and BO assessed the orthostatic hypotension and sympathetic skin response. J-HZ and J-YW reviewed literatures and drafted the manuscript. XZ and J-HZ supervised the study. All authors read, revised, and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Teachers’ healthy and effective functioning at work is impacted by the demands they face and the resources they can access. In this study, person-centered analysis was adopted to identify distinct teacher profiles of demands and resources. We investigated teachers’ experiences of two job demands , two job resources (teacher collaboration and input in decision-making), and one personal resource (self-efficacy for teaching). Using data from the Teaching and Learning International Survey (TALIS) 2013, the study involved 6,411 teachers from 369 schools in Australia and 2,400 teachers from 154 schools in England. In phase one, latent profile analysis revealed five teacher profiles that were similar across the two countries: the Low-Demand-Flourisher (12%), Mixed-Demand-Flourisher (17%), Job-Resourced-Average (34%), Balanced-Average (15%), and Struggler (21%). The profiles were differently associated with two background characteristics (teacher gender and teaching experience) and two work-related well-being outcomes . In phase two, we extended our analysis to the school-level to identify school profiles based on the relative prevalence of the five teacher profiles within a school. Indeed, a yield of large scale datasets such as TALIS is that there are sufficient units at the school-level to enable institutional insights, beyond insights garnered at the individual teacher-level. Two school profiles that were similar in both countries were revealed: the Unsupportive school profile (58%) and the Supportive school profile (42%). The Supportive school profile was associated with higher school-average teacher job satisfaction and occupational commitment than the Unsupportive school profile. Taken together, the findings yield knowledge about salient teacher and school profiles, and provide guidance for possible interventions at the teacher- and school level. Teaching work is complex. The extent to which teachers thrive at work involves a delicate balance between the demands placed upon them and the resources they can access to support them in their work . A growiA small, but growing body of research has conducted person-centered examinations of teachers’ experiences at work e.g., . HoweverThe aims of the current study, therefore, were to identify profiles of demands and resources experienced by teachers and then to explore the extent to which distinct profiles are predicted by teachers’ background characteristics and associated with meaningful differences in workplace well-being outcomes. We also investigated school-level profiles by identifying the proportion of the teacher-level profiles evident within different subpopulations of schools , along with links to school-average well-being outcomes. Data from the Teaching and Learning International Survey (TALIS) 2013 were used in our study. Key strengths of using large scale datasets like TALIS is that sampling is considered to be nationally representative, and sufficient numbers of schools are sampled to allow insights at the institutional level, beyond insights garnered at the individual teacher-level. Our study was conducted with teachers from Australia and England in order to provide evidence of the generalizability of these profiles, and to help guide national and international research, practice, and policy. Taken together, the findings have important implications for understanding demand-resource profiles among teachers and whether particular combinations are better aligned with well-being outcomes. The findings also have the potential to yield knowledge about salient school profiles and provide guidance for the development of appropriate intervention at the teacher- and school-level.We rely on the job demands-resources JD-R;  theory aThere are many job demands , job resources  and personal resources  that impact teachers’ work . In the barriers to professional development, which reflects teachers’ experience of factors preventing them from accessing the training necessary for their ongoing learning and development as a teacher. Barriers to professional development may include financial constraints, lack of appropriate opportunities, limited support from leadership, or limited time to complete such activities . Disruptive behavior has consistently been identified in research as challenging for teachers , two job resources (teacher collaboration and input in decision-making), and one personal resource (self-efficacy for teaching). In phase one of analysis, we identified demand-resource profiles among teachers in Australia and England. We then compared the solutions across the two countries to ascertain the generalizability of our results, as well as generalizability of links between the profiles and teachers’ background characteristics  and two well-being outcomes . In phase two of analysis, we extended our examination to the school-level where we sought to identify school profiles based on the relative frequency of teacher profiles. We then compared these profiles across the two countries to verify the generalizability of these results, and tested whether the school profiles displayed different levels of school-average teacher job satisfaction and occupational commitment. The sample comprised 6,411 teachers from 369 schools in Australia and 2,400 teachers from 154 schools in England who participated in the Teaching and Learning International Survey (TALIS) 2013. TALIS is a survey run by the OECD every 5 years and was chosen for this study as it yields comprehensive and nationally representative data on demands and resources relevant to teachers’ work. Sample selection for TALIS 2013 involved a two-stage probability sampling design to ensure a representative sample of schools and of teachers within those schools (for full details see SD = 12) years, and had an average teaching experience of 16 (SD = 11) years. Most teachers (84%) were working full-time, and almost all (99%) had attained ISCED Level 5A (bachelor’s degree) or higher. The Australian teachers taught at ISCED Level 2 (lower secondary) and/or ISCED Level 3 (upper secondary). Just over half (55%) of the participating schools were publicly managed, and most (74%) had fewer than one-third students from low-SES backgrounds. Most schools (62%) had a male principal with an average age of 55 (SD = 7) years and an average experience as a principal of 9 (SD = 7) years. The schools were located in hamlets/villages , small towns , towns , cities , and large cities . There were on average 18 (SD = 5) teachers per school.Starting with the Australian sample, participating teachers were 57% female, had an average age of 43 (SD = 10) years, and had an average teaching experiences of 12 (SD = 9) years. Most teachers (87%) were working full-time, and almost all (97%) had attained ISCED Level 5A (bachelor’s degree) or higher. The entire English sample taught at ISCED Level 2 (lower secondary). Just over half (55%) of the participating schools were publicly managed, and most (76%) had fewer than one-third of their students from low-SES backgrounds. Most schools (64%) had a male principal with an average age of 50 (SD = 7) years and an average experience as a principal of 7 (SD = 6) years. The schools were located in hamlets/villages (4%), small towns (18%), towns (41%), cities (20%), and large cities (16%). There were on average 16 (SD = 4) teachers per school.Participating teachers from England were 63% female, had an average age of 39 . Items were scored on a scale from 1 (Strongly disagree) to 4 (Strongly agree). Reliability was assessed with coefficient omega1 and was adequate for the Australian (ω = 0.74) and English (ω = 0.74) samples. The scale displayed 7% variance (intraclass correlation [ICC] = 0.07) at the school-level. Although this is somewhat modest, it does warrant multilevel analyses . Items were scored on a scale from 1 (Strongly disagree) to 4 (Strongly agree). Reliability was satisfactory  and the scale demonstrated adequate variance at the school-level (ICC = 0.14).Teacher collaboration was assessed with items from the TALIS “Teaching in General” scale . Items were scored from 1 (Never) to 6 (Once a week or more). Reliability estimates were adequate  and the scale demonstrated modest, but adequate, variance at the school-level . Items were scored on a scale from 1 (Strongly disagree) to 4 (Strongly agree). Reliability was satisfactory  and the scale demonstrated adequate variance at the school-level (ICC = 0.14).Teacher self-efficacy was assessed with items from the TALIS “Teaching in General” scale that encompasses three types of self-efficacy: self-efficacy for classroom management , self-efficacy for instruction , and self-efficacy for student engagement . Items were scored on a scale from 1  to 4 (A lot). Reliability was satisfactory  and there was modest, but adequate variance at the school-level , self-efficacy was modeled as a higher-order factor.Job satisfaction and occupational commitment were assessed with items from the TALIS “About Your Job” scale. For job satisfaction, 3 items were used . For occupational commitment, 4 items were used . Items for both scales were scored on a scale from 1 (Strongly disagree) to 4 (Strongly agree). Because the outcomes were modeled at both the teacher- and school-level, we assessed reliability at both levels. Reliability was satisfactory at the teacher  and school  levels. The two scales also demonstrated adequate variance at the school-level  estimator was used in all models. Missing data were 5–8% for all variables . Missing data were handled with full information maximum likelihood procedures.All analyses were conducted using Mplus 8.4 . In our Confirmatory factor analyses (CFA) were run for each country separately to obtain estimates of correlations among the two background characteristics, the five demands and resources, and the two outcomes see the . Factor M = 0, SD = 1) for each country. All analyses relied on an assumption of conditional independence, meaning that any covariance between indicators is assumed to be entirely explained by the latent profile variable, given that we did not have any a priori theoretical or empirical reason for relaxing this assumption  allows comparison of a k-profile model with a k−1 profile model to see if the former model results in an improvement in fit relative to the latter. Finally, we created elbow plots of the AIC, CAIC, BIC, and SSA-BIC indices. In these plots, the profile at which point the slope noticeably flattens is another indicator of an appropriate solution , Consistent Akaike Information Criteria (CAIC), Bayesian Information Criteria (BIC), and sample-size-adjusted Bayesian Information Criteria (SSA-BIC) smaller values reflect better fit. The solution . We alsoAfter determining the optimal solution for each country, we next undertook tests of profile similarity to determine the extent to which the profile solutions could be considered to be comparable across Australia and England . These tplus MODEL CONSTRAINT option, which relies on the multivariate delta method for tests of statistical significance  and likelihood of profile membership (predictive similarity), as well as the equivalence of the associations between profile membership and the outcomes (explanatory similarity) across countries . For thece e.g., .In phase two, our aim was to extend the teacher-level  findings to consider the extent to which school-level  profiles could be identified. More precisely, these analyses sought to identify school profiles characterized by distinct proportions of teacher profiles. Thus, rather than estimating profiles  based on the means and variance of profile indicators, this second set of analyses identified school profiles based on the relative frequency of the various categories of the L1 latent profiles  reported low barriers to professional development, average disruptive behavior, high teacher collaboration, high teacher input, and high self-efficacy. This profile was thus labeled Mixed-Demand-Flourisher to reflect the mixed blend of low to average job demands, coupled with high job and personal resources. Teachers corresponding to profile 3 (21% of the sample) reported slightly below average barriers to professional development, average disruptive behavior, high teacher collaboration, high teacher input, and average self-efficacy. We labeled this profile Job-Resourced-Average to reflect the above average job resources, and average job demands and self-efficacy. Teachers corresponding to profile 4 (15% of the sample) reported average barriers to professional development, average disruptive behavior, average teacher collaboration, average teacher input, and average self-efficacy. We labeled this profile Balanced-Average to reflect the matching average levels observed across all demands and resources. Teachers corresponding to profile 5 (34% of the sample) reported high barriers to professional development, high disruptive behavior, low teacher collaboration, low teacher input, and low self-efficacy. We labeled this profile Struggler to reflect this blend of high job demands, and low job and personal resources.Teachers corresponding to profile 1 (12% of the sample) reported low barriers to professional development, very low disruptive behavior, high teacher collaboration, high teacher input, and high self-efficacy. This profile was thus labeled We next tested the predictive and explanatory similarity of this solution by including predictors  and outcomes to this final model of distributional similarity. In terms of predictive similarity, the CAIC, BIC and SSA-BIC decreased when equality constraints across countries were included for the predictive paths see , thus sup < 0.05). The Mixed-Demand-Flourisher profile displayed the highest levels of job satisfaction (M = 3.64) and commitment (M = 3.54), followed by the Low-Demand-Flourisher profile (M = 3.53 for job satisfaction and M = 3.46 for commitment), then by the Job-Resourced-Average profile (M = 3.36 and M = 3.23), followed by the Balanced-Average profile (M = 3.13 and M = 3.09), and finally by the Struggler profile (M = 2.88 and M = 2.89).In terms of explanatory similarity, the CAIC, BIC and SSA-BIC increased when profile-specific outcome levels were constrained to be equal across countries see , suggestp ≤ 0.05), with one exception. Starting with job satisfaction, the Mixed-Demand-Flourisher (M = 3.53) displayed the highest levels, followed by the Low-Demand-Flourisher profile (M = 3.41), then by the Job-Resourced-Average profile (M = 3.23), followed by the Balanced-Average profile (M = 3.06), and finally by the Struggler profile (M = 2.59). For occupational commitment, the Mixed-Demand-Flourisher displayed the highest levels (M = 3.56), followed by the Low-Demand-Flourisher profile (M = 3.44) and then equally by the Job-Resourced-Average (M = 3.18) and Balanced-Average (M = 3.13) profiles, which did not differ from one another, and finally by the Struggler profile (M = 2.75).For England, all mean comparisons were also statistically significant (p < 0.01). The exception was related to the Balanced-Average profile, which displayed similar levels of job satisfaction in both countries. For occupational commitment, there were no significant differences between countries, with the sole exception of the Struggler profile for which levels of occupational commitment were higher in Australia than in England (p <0.01).Finally, we also tested mean differences within matching profiles across the two countries. All profiles except one from the Australian sample displayed higher levels of job satisfaction when compared with the matching profiles from the English sample  school-average job satisfaction (M = 3.01) and occupational commitment (M = 3.04) than the Supportive school profile . The same was true for England, where the Unsupportive school profile also displayed significantly lower (p ≤ 0.01) school-average job satisfaction (M = 2.79) and occupational commitment (M = 2.92) than the Supportive school profile . Finally, the two L2-profiles from the Australian sample displayed higher levels of job satisfaction than the matching profiles estimated in the English sample (p < 0.01). In addition, the Unsupportive school profile from the Australian sample displayed higher levels of occupational commitment than the matching profile from the English sample (p ≤ 0.01), but no differences in occupational commitment were observed for the Supportive school profile across country.We next tested the explanatory similarity of this solution by including outcomes to this final model of distributional similarity. As for the single level models, the CAIC, BIC and SSA-BIC increased when profile-specific outcome levels were constrained to be equal across countries see , suggestWe used person-centered analyses to identify demand-resource profiles among teachers and schools across representative samples from two different countries. In phase one, five teacher-level demand-resources profiles were identified in both the Australian and English samples: the Low-Demand-Flourisher, Mixed-Demand-Flourisher, Job-Resourced-Average, Balanced-Average, and Struggler. Results showed that male teachers and less experienced teachers were more likely to be members of less adaptive profiles in both countries . The profiles were also associated with different levels on both well-being outcomes  in each country. More precise cross-country comparisons showed that profile-specific levels of job satisfaction were higher in the Australian sample than levels observed in the matching English profiles, whereas few differences in occupational commitment were evident.In phase two, we extended our analyses to the school-level. In both countries, we found evidence of two profiles of schools: a Supportive school profile comprising relatively similar levels of the Mixed-Demand-Flourisher, Job-Resourced-Average, Struggler, and Low-Demand-Flourisher profiles, and an Unsupportive school profile comprising much higher levels of the Struggler profile. For both countries, the Supportive school profile was associated with significantly higher levels of school-average job satisfaction and occupational commitment. Cross-country differences in outcomes levels were also apparent, and are discussed below.As noted, five demand-resource profiles were evident at the teacher-level for both Australia and England. Two of these profiles were named flourishers, the Low-Demand-Flourisher and Mixed-Demand-Flourisher, and in combination made up almost one-third of the sample. Given the nature of these two profiles , this is a positive finding. The third profile, the Job-Resourced-Average, represented around one-fifth of the sample. Thus, around 1 in 5 teachers in the sample experienced average job demands and self-efficacy, but simultaneously felt well-supported (above average job resources). The fourth profile, the Balanced-Average, represented 15% of the sample. These teachers appear to experience relatively similar (and average) levels of demands and resources. Finally, the Struggler represented around one-third of each sample. Thus, 1 in 3 teachers in the sample experienced high job demands, low job resources, and low self-efficacy.Taken together, the findings are important as they provide insight into the groups of teachers that work in schools. The findings are also significant because of the close overlap in the profiles demonstrated across the two countries—which may have occurred given the historical and socio-cultural similarities of the two contexts e.g., . Importar = 0.22). Conversely, our findings showed that while personal resources and job resources appeared to be at similar levels for most profiles , they were less aligned for the Job-Resourced-Average profile. Taken together, these findings highlight the merits of person-centered research given that it is able to access the experiences of different subpopulations of teachers.The findings provide a nuanced understanding that complements prior variable-centered results showing that job and personal resources are typically positively correlated. For instance, In both countries, the background characteristics were associated with the profiles in similar ways, showing that male teachers were more likely to correspond to the Job-Resourced-Average, Balanced-Average, or Struggler profiles than to either of the two Flourisher profiles. It is possible this finding occurred because, relative to male teachers, female teachers have been shown to report stronger perceptions of job resources generally , greaterTurning to the outcomes, the Mixed-Demand-Flourisher profile displayed the highest levels of job satisfaction and occupational commitment in both countries. This was followed by the Low-Demand-Flourisher profile. These results are understandable given the nature of the two Flourisher profiles, and given that low demands and high resources have been associated with well-being in prior variable-centered research e.g., . The sigMoving along to the other profiles, the Job-Resourced-Average profile typically displayed the third highest levels of the well-being outcomes, followed by the Balanced-Average profile. In comparing these two profiles, the major differences occurred in job resources, which underscores prior research on the importance of contextual supports for teachers e.g., . The JobFinally, the Struggler profile displayed the lowest outcomes. Alongside the mismatch between (high) demands and (low) resources that this profile experienced, low collaboration and input in decision-making may mean that teachers in this profile experience a reduced sense of autonomy e.g.,  and loweTaken together, the findings involving the outcomes complement knowledge gained from prior variable-centered research by providing a more nuanced understanding of the associations that demands and resources have with outcomes related to well-being. Prior research has clearly documented that job demands are typically negatively associated with well-being, whereas the reverse is true for resources e.g., . In a coIn terms of cross-country comparisons, significantly higher levels of job satisfaction were evident in most of the profiles from the Australian sample when compared with the matching profiles from the English sample. For occupational commitment, only the Struggler profile displayed higher levels in the Australian sample than in the English sample. Additional research is needed to understand precisely why these findings occurred, but it may be related to increases in compliance and reductions in professional autonomy that have been documented in England over the past decade e.g., —such worAs noted earlier, large scale datasets from surveys like TALIS enable insights into phenomena at a school-level that are often inaccessible with smaller datasets. In our study, two profiles were evident at the school-level in both countries. The first we called the Unsupportive school profile and comprised 58% of participating teachers. The second was the Supportive school profile (comprising around 42% of participating teachers), and included a substantially greater proportion of the two Flourisher profiles, and a smaller proportion of the Struggler profile than the Unsupportive school profile. The higher proportion of schools corresponding to the Unsupportive school profile is in accord with the growing attention toward the escalating demands faced by many teachers and schools in Australia and England. Indeed, there is growing attention to teachers’ workload, burnout, and attrition from government working groups  and not-Notably, our results are some of the first to examine demand-resource profiles at the school-level among teachers, and the first to test profile similarity at the school-level across country. Our findings are important because limited research has examined multilevel associations using JD-R theory . MoreoveTurning to the outcomes, the Supportive school profile was associated with significantly higher levels of school-average job satisfaction and occupational commitment. This finding contributes to the literature by highlighting that the particular combination of teacher types within a school is associated with school-average teacher well-being. It is possible that this finding occurred because the Supportive school profile was characterized by a higher proportion of the two Flourisher profiles, which had more positive outcome levels. When teachers’ resources outweigh their demands, then teachers are better equipped to undertake their work and are likely more satisfied with their jobs and committed to their profession e.g., . It is aBecause the current study is one of the first to consider demand-resource profiles among teachers, we emphasize that more research is needed to ascertain the generalizability of our findings. Nonetheless, we do provide some tentative suggestions for practice. Notably, a key contribution of person-centered research is that it allows implications for practice that are targeted more closely to the needs of particular subpopulations of teachers and schools. For example, teachers in the Flourisher profiles would likely benefit from efforts to further boost (or at least maintain) their access to resources. For the Struggler profile, efforts may want to focus on reducing demands and increasing resources. Turning to the school-level, efforts targeted at the Unsupportive school profile may focus on school-wide efforts to boost resources and reduce job demands, whereas efforts focused on the Supportive school profile may focus on further boosting job and personal resources.In terms of practices that can help boost resources and reduce demands, reducing barriers to professional development is likely important. As noted earlier, barriers include budget constraints, availability, support from leadership, and time e.g., . EffortsIn terms of resources, engagement in professional learning communities and instructional rounds (observing other teachers) are two effective methods for encouraging teacher collaboration e.g.,  and boosConsideration of several limitations is important in interpreting findings from the current study. First, the use of TALIS 2013 provides significant strengths . Nonetheless, this type of data does come with some limitations. In particular, our study was cross-sectional in nature, which means that we are unable to ascertain causal ordering between the profiles and the outcomes, nor whether teachers’ profile membership fluctuates over time. Longitudinal modeling  will be an important avenue to explore in future. Second, we focused on five demands and resources. As noted earlier, our selection of factors was firmly based in JD-R theory, conceptual reasoning, and prior empirical research. Nonetheless, in future it will be important to consider a different range of factors to see what else is salient in teachers’ experiences. Third, our study employed data from teachers. Of course, teachers’ perceptions are essential given that it is their interpretations that may influence their well-being outcomes. Nonetheless, in future it will be interesting to consider other markers of demands and resources to see how perceptions of demands and resources align with measures taken from other informants . Fourth, our study was conducted among teachers from Australia and England. Examining the extent to which similar profiles can identified, or not, in other countries (including non-English speaking countries) should be an important upcoming research focus. As noted earlier, teachers’ experiences of demands and resources in Australia and England are mirrored in many other countries e.g., . NonetheThe aim of the current study was to establish whether different profiles of teachers could be identified based on their experiences of demands and resources, and, if so, to ascertain which profiles are more aligned with well-being. We conducted our examination at the teacher- and school-level among teachers from Australia and England. Findings revealed five teacher profiles that were similar across the two countries: the Low-Demand-Flourisher, Mixed-Demand-Flourisher, Job-Resourced-Average, Balanced-Average, and Struggler. Notably, the profiles differed in relation to two well-being outcomes, with the Mixed-Demand-Flourisher typically evincing the highest levels of job satisfaction and occupational commitment. Two school-level profiles that were similar in both countries were identified based on the prevalence of the five teacher profiles: the Unsupportive and Supportive school profiles. Of note, the Supportive school profile was associated with higher school-average teacher job satisfaction and occupational commitment. Taken together, the findings yield novel understanding about different subgroups of teachers and schools, and hold implications for practice at the teacher- and school-level.The datasets used in our research are available from the OECD.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Drug addiction is a chronic relapsing disease leading to allostatic alterations in the brain’s reward, stress, and executive function systems . The etiBidirectional interactions between the nervous system and immune system, collectively known as the “neuroimmune system”, regulate a wide range of physiological and pathological processes . While sGranulocyte-colony stimulating factor (G-CSF) is a ~20-kDa protein produced by bone marrow stromal cells, endothelial cells, macrophages, and fibroblasts, as a peripheral hematopoietic growth factor . Both G-In a series of rodent experiments, Kutlu et al.  investigex vivo fast-scan cyclic voltammetry to assess dopaminergic function in the nucleus accumbens (NAc)—a region heavily implicated in reward processing. Repeated systemic administration of G-CSF increased electrically-evoked dopamine release in the NAc, without altering dopamine uptake or transport. Acute application of G-CSF onto the brain slice failed to alter NAc dopamine transmission, indicating that G-CSF does not act directly at dopaminergic terminals in the NAc, but perhaps on their cell bodies in the ventral tegmental area (VTA) or other converging circuits . Finally, quantitative polymerase chain reaction revealed that repeated G-CSF treatment suppressed pro-inflammatory cascades in the NAc, as demonstrated by reduced TNF-α gene expression, while acute G-CSF increased the expression of several dopaminergic genes   . CollectA previous study by the same group delineated the link between G-CSF and the biobehavioral response to cocaine. Specifically, Calipari et al.  found thFuture work is warranted to determine the extent to which G-CSF influences behavioral correlates of drug and natural rewards, as well as its regulatory effects in other addiction-related brain regions. As a first pass, Mervosh et al.  used an As a whole, the aforementioned data elucidate novel mechanisms whereby peripheral administration of an anti-inflammatory cytokine (G-CSF) enhances the activity of a central reward circuit (mesolimbic dopamine system) and modulates both motivational and cognitive aspects of reward processing. The sucrose threshold task used in this study should be highlighted as this behavioral economics self-administration protocol allows for the dissociation of A) the animal’s preferred level of consumption when effort is low (i.e. preferred intake level) and B) the maximal effort the animal will expend to maintain its preferred intake level (i.e. motivation to consume). The researchers determined that G-CSF increased both sucrose intake and motivation. As demonstrated by the reversal learning task, cognitive flexibility was also enhanced by repeated, but not acute, G-CSF injections. This is an important finding given that cognitive deficits play substantial roles in many neuropsychiatric diseases, including addiction, and most of the currently available pharmacotherapies do not address these impairments . A singlMedication development for neuropsychiatric disorders, especially addiction, is a complex, lengthy, and expensive process with low success rates. Therefore, the notion that G-CSF may represent an effective therapeutic target is of particular relevance and interest. Recombinant G-CSF is already approved by the FDA and has been tested in clinical trials for other indications—an advantage which may facilitate translation of the present preclinical findings into human research. As an initial bench-to-bedside step, proof-of-concept human laboratory studies may provide valuable information on the effects of G-CSF administration in patients with SUDs. As demonstrated by Calipari et al. , G-CSF dMF and FV were responsible for the conceptualization of this paper. MF, AB, HK, LH, and FV developed the rationale, wrote the first draft, critically reviewed the content, and approved the final manuscript for publication.This work was supported by NIH intramural funding ZIA-AA000218 , jointly supported by the NIDA Intramural Research Program and NIAAA Division of Intramural Clinical and Biological Research (MF); NIH Center on Compulsive Behaviors, funded by the NIH Deputy Director for Intramural Research (DDIR) Innovation Award (MF); NIAAA K99 AA027576 grant (LH); NIAAA K99 AA025408 grant (FV); and Developmental Exposure Alcohol Research Center (DEARC) at Binghamton University, P50 grant AA017823 (FV). The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the funders, which had no role in the development of this article.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To compare immediate postoperative results in patients receiving heparin-albumin-coated and non-coated circuits.A total of 241 patients undergoing on-pump cardiac surgery were divided into two groups: those receiving heparin-coated circuits  and those receiving non-coated circuits .vs. 560.38±90.20, respectively, P=0.001). In-hospital mortality and postoperative stroke rates and lengths of intensive care unit stay were similar between the groups; in contrast, in the heparin-albumin-coated group, patients had significantly better outcomes for hospital stay, drainage, and need for erythrocyte transfusion.Activated clotting times (ACT) during cardiopulmonary bypass (CPB) were significantly shorter in the heparin-albumin-coated group than in the non-coated group (355.64±34.12 Heparin-coated circuits and reduced level of systemic heparinization with 300 seconds of target ACT level in cardiac surgery under CPB are safe and result in a very satisfactory clinical course. Several CPB circuits with heparin-coated or surface-modifying agents are available. These systems have been shown to reduce inflammatory response and to provide better hemocompatibility.Activation of the complement cascade, oxidative stress, and coagulation pathways induced by cardiopulmonary bypass (CPB) resulting in systemic inflammatory response syndrome after open heart surgery may cause several complications, like bleeding or organ dysfunctions. Heparin is the most known anticoagulant used in CPB because of its rapid onset, effectiveness, ease of reversal by protamine, and low cost. While heparin is most known for its impact on the coagulation pathway, there is some evidence that heparin also affects fibrinolysis and platelet function independent of CPB. The effect on fibrinolysis and platelets may result in postoperative bleeding or increasing the transfusions.To avoid circuitry blood clotting and thromboembolic complications, systemic heparin is administered to both the patient and the circuits. Heparin coating may decrease the appropriate ACT levels, which are resulting in less transfusion and bleeding.Heparin dosage for anticoagulation during CPB is calculated with an empiric formula based on the patient’s body weight and preoperative activated clotting time (ACT). Commonly for initiating CPB, ACT length must be > 480 seconds with non-heparin-coated circuits. Some authors have reported that this full-dose anticoagulation approach unnecessarily exposes the patient to excessive blood losse.g., drainage, erythrocyte transfusion, postoperative stroke, and mortality) in patients receiving heparin-albumin-coated and non-coated circuits.In this study, we aimed to compare immediate postoperative results  before 2016 routinely, and by 2016 we started to use heparin-albumin-coated circuits . All perfusion tubing systems were completely coated, except for the cannulas.Hemochron® Jr. Signature plus Whole Blood Microcoagulation System  was used to measure ACT. This system uses silica, kaolin, and phospholipid as activators and measures the elapsed time between the start of the test and clot formation, and the ACT is automatically converted to a reference celite-based ACT value.Before CPB, 300 IU/kg of heparin  was administered intravenously to patients receiving non-coated circuits. Readministration of 5000 IU heparin boluses took place if the ACT was < 480 seconds. A reduced dose of heparin (150 IU/kg) was administered for systemic anticoagulation to patients receiving heparin-albumin-coated circuits. Readministration of 2500 IU heparin boluses took place if the ACT was < 300 seconds. ACT was repeatedly determined during CPB, after protamine administration, and two hours postoperatively. Myocardial protection consisted of intermittent antegrade administration of cold blood cardioplegic solution. After completion of CPB, heparin was antagonized with protamine in a ratio of 1:1.The amount of postoperative bleeding from the time of sternal closure until the drains were removed was recorded. Postoperative 24-hour drainage was used for analysis. Normovolemic anemia was accepted to a hematocrit level of 0.25 postoperatively; a level less than this was considered an indication for allogeneic red blood cell transfusion.2/min. Systemic moderate hypothermia (30 ˚C) was used in coronary and valve surgery; in aortic procedures, deep hypothermia (24 ˚C) was used. Standard adult sizes of the circuits were used and primed with 1000 mL of lactated ringer.CPB was performed with a Terumo® Advanced Perfusion System 1  with a non-pulsatile flow control and at a flow rate of 2.4 L/mAntiplatelet therapy was not stopped before surgery. Vitamin K antagonist was stopped and changed to low-molecular-weight heparin before surgery.t-test for continuous variables and the c2 or Fisher’s exact test for categorical variables. A P-value < 0.05 was statistically significant.Data were expressed as mean ± standard deviation for quantitative variables and as number and percentage for categorical variables. The groups were compared by Student’s vs. 560.38±90.20, respectively, P=0.001) In-hospital mortality rates, postoperative cerebrovascular event rates, and lengths of intensive care unit (ICU) stay were similar between the groups; in contrast, patients in the heparin-albumin-coated group had significantly better outcomes for hospital stay, drainage, and need for erythrocyte transfusion.The patients’ baseline demographic characteristics are summarized in This study revealed similar clinical outcomes of non-coated and albumin-heparin-coated circuits in terms of in-hospital mortality rates, postoperative cerebrovascular event rates, and lengths of ICU stay. Furthermore, better outcomes for hospital stay, amount of mediastinal drainage, and need for erythrocyte transfusion were achieved with coated circuits. In fact, it is obvious that the proposal of maintaining CPB using heparin in a lesser amount with a shorter ACT and, at the same time, using bioartificial surfaces will have positive results. In this study, the rational basis of this hypothesis was investigated with literature examples.. A heparin titration method used in a study to achieve conventional ACT level resulted in using low doses of heparin, which is associated with lower blood loss. In this recent study, we used low-dose heparin with titration model with a significantly lower ACT level.Conventionally, an empirical dose of heparin has been used to inhibit coagulation for initiating and maintaining CPB to achieve an ACT level > 480 seconds. Achieving this target ACT level by giving high doses of heparin is associated with significantly higher postoperative blood loss. Although the endpoint of our study is not to assay inflammatory responses, using more biocompatible surfaces is related with better outcomes. Tayama E. et al. showed reduced inflammatory response with heparin-coated circuits, but no benefits in clinical outcomes. In that study, heparin administration and target ACT levels were the same in both heparin-coated and non-coated groups. Similarly, there were no significant clinical outcomes in the study by Reser D. et al., in which three different biocompatible surface circuits were compared and heparin administration and target ACT levels were conventionally set. These consequences support our hypothesis.Heparin-coated surfaces do not only reduce the systemic heparinization, they also reduce the systemic inflammatory process and oxidative stress showed that a median ACT level  was safe and resulted in a very satisfactory clinical course in 5,954 patients undergoing on-pump CABG with heparin-coated circuits. In this recent study, mean ACT level in heparin-coated circuits was 355 seconds during the CPB and it was not associated with adverse clinical outcome.Another important debate is that if low target ACT levels are safe or not. Ovrum E. et al.. Therefore, the advantages of this system, like reduction in blood loss and reduction of ventilator dependence and length of hospital stay, make heparin-coated circuits more reasonable to use in terms of overall costs.Heparin coating the circuits increases cost and this technology is not routinely used for short-time devices due to higher initial costs. In our assay, we found no significant difference between the groups for 30-day hospital mortality and postoperative stroke rates, similarly to the literature.Limited use of anticoagulation during CPB and risk for stroke is another concern which was investigated previously, and stroke and mortality rates were comparable in heparin-coated circuits vs. conventional ones. A review by Mahmood S. et al. about heparin-bonded CPB circuit showed that perfusion with heparin-coated and heparin-polymer-coated bypass does not increase the risk of adverse effects but reduces blood loss, re-operation rates, ventilation time, length of ICU and hospital stays, and is also associated with improved biocompatibility.A meta-analysis demonstrates parallel results, that biocompatible circuits have a limited effect  on morbidity, leading to shorter ICU and hospital staysThe limitations of this study are its single-center nature, small sample size, and nonrandomized design. This study focused on immediate outcomes, so long-term follow-up data from randomized clinical trials will be needed to evaluate clinical outcomes.Comparable postoperative stroke and mortality rates were found in contrast with less blood transfusion, lower drainage, short periods of postoperative ventilator support, and reduced hospital stay seen in the study group. In conclusion, heparin-coated circuits and reduced level of systemic heparinization with a 300 seconds target ACT level in cardiac surgery under CPB are safe and result in a very satisfactory clinical course.


Directiveness, Scope and Gradation, Set of Reference and Sign. On this basis, we then build seven alternative generic labelling systems that we test in a framed-field experiment enabling us to measure the effect of each component on food purchases in isolation. Our results show that an effective front-of-pack labelling system should be Food-Directive (instead of Diet-Directive) and be displayed on both healthy and unhealthy food. The reference set, which is across categories or within categories, produces the same average nutrition score but generates contrasting behavioural responses.The relative impacts on food purchases of many alternative front-of-pack nutritional labelling systems were tested, with various methods—from opinion pool to nationwide experiments. Clearly, some systems induce better purchasing responses, having better nutritional impacts on food baskets. Nonetheless, we still ignore what the ingredients of an efficient label are. Here, we propose guidance for label designers. To do so, we first propose a typology that breaks down established labelling systems into four elementary components:  Many front-of-packs labelling systems (FoPLs) have become established around the world. The best-known systems include the British Traffic Lights, the Scandinavian KeyHole, the New Zealand and Australian Health Star Rating, the Chilean Warning Labels and the French Nutri-Score among many others  and the ranking method . Our design components could thus be transposed to non-nutritional labelling, such as environmental labelling. We also do not consider the positioning issue of the FoPLs such as size or positioning on the pack.basic forms that the component may take, knowing that established or possible FoPLs may mix several of these basic forms; (iii) we give some examples from existing FoPLs, whose pictures and descriptions are presented in We identify four components of a typical FoPLs. In this section, we present each of these four components in turn as follows: (i) we define the component, (ii) we list Directiveness is defined as the extent to which the label conveys interpretative information. Directiveness may take three basic forms: Non-Directive, Diet-Directive and Food-Directive . Non-DirIn contrast, Directive FoPLs issue normative assessments. On the basis of criteria set by nutritionists and epidemiologists, they provide a judgement either on the total product or by nutrient. Nutrient-based schemes provide an analytical assessment. That is, they break down the assessment of food into several ratings. As with the UK Multiple Traffic Lights , these rSome FoPLs may mix several forms of directiveness. For instance, the UK Multiple Traffic Lights complement their Diet-Directive colour-coded recommendations with Non-Directive Nutritional Information and Reference Intakes. Similarly, the Health Star Rating may supplement their Food-Directive summary score with Non-Directive Nutritional Information. They are Hybrid FoPLs ,21,23.Directiveness may affect behavioural responses . Nutriti, Warned food and Both Recommended and Warned. The first form covers only the recommended foods. Examples include all the endorsement logos like the Keyhole . In stated shopping experiments, Food-Directive Nutri-Score led to a slightly healthier food basket than the Green Tick label (Note: The Green Tick label was created for the purposes of the study and was derived from the Keyhole system. The Tick label was attributed to products assigned to the dark and light green categories of the Nutri-Score)  but no dThe Set of Reference component describes the set with which the comparison of food or nutrient is made. Known also as Segmentation , it may Behavioural issues are the nature and the extent of induced substitutions. While it remains unclear whether food is evaluated relative to a particular category or in absolute terms , Within-The Sign component describes the type of semiology used for the labelling scheme. It may take four basic forms : writtenColour codes and ideograms, such as those borrowed from the highway code, are quickly and easily recognisable. They essentially serve the objective of facilitating the perception and the assimilation of labels by the consumers. Word and number processing is effortful and requires potentially more cognitive resources ,39. HoweFor our experimental purpose, among all the possible combinations of the four key components proposed in our typology, we narrow down the number FoPLs prototypes to seven. Our first prototype is a Non-Directive FoPLs similar to the Reference Intake system that displays the percentage of daily-recommended intake values per serving for each nutrient (RI). All other prototypes are either Diet-Directive (D) or Food-Directive (F) with colour-coded signs, each sign being a simple dot, ignoring systems that feature ideograms and texts. Each of the two last components takes two forms: Within-Categories (W) vs. Across-Categories (A) for the Reference Set component and Recommended (R) vs. Both Recommended and Warned (R&W) for the Scope component. As for the Gradation component, we use the Binary form for R and the 3-class form for R&W.With option D, each nutrient of a given product is graded as such. We only consider the density (g per 100 g) of the following three nutrients: salt, free sugar and saturated fatty acid. These nutrients are consensually recognized among nutritionists as the nutrients to be limited. Therefore, the Diet-Directive systems display three coloured signs, one for each of our three nutrients. Alternatively, only one coloured sign is displayed with option F. With Option W (respectively A), each food or nutrient is graded according to its rank within its food category . With option R&W, the following Scope and Gradation rule is used: the healthiest 1/3 is assigned a green sign, the unhealthiest 1/3 a red sign and the remaining 1/3 a colourless sign. Only the best third is rewarded with a green dot with option R. The sorting among nutrients is directly determined the nutrient content per 100 g  and the sorting among food is determined through an aggregated nutrition score that averages our three-nutrient content per 100 g weighted by the daily-recommended intake values Among the stem see : D-W-R, https://www.oqali.fr/oqali_eng/). Each category includes six, nine or twelve food items. With the help of renowned French nutritionists and consumption data, food items were chosen among the most frequently bought products in France, so as to model the existing range of nutritional quality in each food category. Products were proposed at current outside market prices. Posted prices had been observed in a local supermarket at the time of the sessions. Participants were aware of that.We use an e-shopping mock-up that includes a total of 273 food products in 35 familiar food categories corresponding to the usual classification used in self-service grocery stores in France and fitting the standard classification proposed by OQALI . On this basis, one can measure the relative effectiveness of the seven logos by comparing the extent of changes between logo formats (between-subject method). This is possible because each treatment differs only with respect to the labelling system. We consider not only average distances but also individual dispersion. In particular, individuals who improve, do not alter and reduce the nutritional quality of their baskets are distinguished. Non-parametric tests are used. With matched data  we use the Wilcoxon matched-pairs signed-ranks (WSR). For unmatched data , we use the Mann–Whitney test (MW). Finally, we use the Fisher Exact test (FE) when proportions are compared.This study aims at measuring the nutritional impact of different FoPLs on food shopping baskets. We first measure the relative distance (in %) between the reference basket and the label basket for each subject, and thus the changes, LIM score, a standard index that estimates the mean percentage of the maximal recommended values for our three nutrients of interest in this study, namely free sugar, salt and SFA .p-value < 0.001). With Diet-Directive schemes, significantly more participants do not change their basket , or more participants decrease the nutritional quality of their baskets .Food-Directive FoPLs induce greater nutritional impact compared to Diet-Directive FoPLs. When the entire food is graded, the impact is twice as large as when each nutrient is graded separately: the nutrition score decreases by −10.8% and −9.7% for F-A-R and F-W-R respectively and by −5.2% and −4.4% for D-A-R and D-W-R. Food-Directive FoPLs also induce improvement for a greater proportion of participants: 81.1% against 53.1% for the Diet-Directive FoPLs  due to the small sample size of F-A-R&W. RI does better than the two Diet-Directive schemes, although it should be considered that these two schemes only cover recommended nutrient contents with green dots. When comparing RI with D-A-R and D-W-R pooled together, its overall impact is significantly higher .Non-Directive RI average performance ranks fourth among our seven formats tested, with an average nutrition score decrease of −10.6%, with 76.9% of participants improving their nutritional performance. While RI seems less efficient than the corresponding Food-Directive schemes , there are no statistical differences  or both Recommended and Warned food/nutrients displayed . We also remind the reader that each grade (green or red), whatever the system, always contains one-third of the unit graded.t-test, p-value = 0.070; Mann–Whitney, p-value = 0.424). This is because R&W FoPLs generate more extreme effects both ways. On the one hand, significantly more consumers achieve large nutrition improvement  and, on the other hand, more consumers degrade the nutritional quality of their baskets .Overall, R&W FoPLs induce a 40% greater nutrition impact with an average Nutrition Score decrease of −14.2% for F-A-R&W and F-W-R&W compared to a decrease of −10.2% for F-A-R and F-W-R. Note that this difference is significant according to average, but not according to rank  is not significantly different from the one induced by Across-Categories FoPLs . However, this global effect is generated by contrasted changes in purchasing behaviours see . On averThis article aims to provide guidance for the design of labelling systems and thus to better understand the interplay between the defining features of labels and consumers’ purchasing behaviours. Real-life research on the effects of FoPLs on actual shopping behaviours is much needed . NeverthOur first result concerns the Directiveness component. Food-Directed systems are more efficient than Diet-Directed ones in improving food purchases nutritional score. Simplicity pays ,30. AggrOur second result concerns the Scope and Gradation component. Signalling both recommended and warned food is globally more effective than signalling only recommended food. First, systems that cover all foods are more informative. In this experiment, FoPLs with Recommended and Warned scope display one-third more coloured signs than FoPLs with Recommended scope only. As a result, they provide consumers with more opportunities for change. Second, this additional third of food is marked with a red sign. A red indicator, with its off-putting effect, may make a strong emotional impression on shoppers, transitory but yet significant . At the Our third result concerns the Set of Reference component. Across-Category and Within-Category schemes produce the same average impact on the nutrition score but very contrasted behavioural responses. The designer trade-off here is to choose between favouring intra-category substitutions and inter-category substitutions. Within-Category FoPLs enhance intra-category substitutions, to the detriment of inter-category substitutions. By requiring less effort and by appearing less costly for consumers (substitution of crisps by light crisps instead of radishes), they increase the number of substitutions. On the other hand, substitutions between categories generate greater nutritional gains per unit of substitution, which explains that even with a lower number of substitutions, Across-Category FoPLs achieve similar global nutritional improvement. Moreover, the Within-Category Set of Reference may induce potential perverse effects: dropping a bad product in a good category for a good product in a bad category with a negative effect on the diet . Note thOur fourth component, very poorly explored in our experiment, is the Sign used. For our six Directive systems, we used symbolic colours and, because we had only three grades, we used the classic red and green and a neutral white circle for the intermediate third. Red and green are universally recognized symbols for stopping and going because of traffic-lights. As soon as we used more than two grades, the behavioural meaning of other colours was not that clear . Beyond four colours, the question becomes more pressing.What makes a front-of-pack labelling system effective in improving the nutrition quality of shopping baskets? An initial trade-off is between Food-Directive and Diet-Directive options. Our results suggest that a simple aggregated coloured sign is welcomed for fast, simple and clear choices. Partial schemes, i.e., covering only a subset of the food supply , should be reserved to cases where lobbies threaten the very existence of an exhaustive display system. While the Set of Reference does not seem to matter on consumer’s average nutrition score, the choice between Within-Category and Across-Category schemes has important behavioural consequences and may trigger different responses from the supply side.Future research in the field may cover the following subjects. First, many existing labels mix several forms. For instance, Multiple Traffic Lights and Health Star Rating supplement their Directive forms (respectively coloured codes and summary score) with Non-Directive Nutritional Information or Reference Intake. Therefore, they speak to both fast-thinking and health-motivated consumers. Does the association of opposing forms produce additive effects? More generally, which forms are complements, and which are substitutes with negative composition effects? Second, we did not address the issue of the optimal number of classes for the Gradation component. A smaller number of classes may be more effective in behavioural responses. On the other hand, a higher number of classes allows for finer comparisons within categories and provides greater incentives for firms to change their recipes. This brings us to the last point: here, we evaluated only the effectiveness of labelling schemes in changing demand. It is, however, also important to know the effect on supply.


Antibiotic resistance is an escalating, worldwide problem. Due to excessive use of antibiotics, multidrug-resistant bacteria have become a serious threat and a major global healthcare problem of the 21st century. This fact creates an urgent need for new and effective antimicrobials. The common strategies for antibiotic discovery are based on either modifying existing antibiotics or screening compound libraries, but these strategies have not been successful in recent decades. An alternative approach could be to use gene-specific oligonucleotides, such as peptide nucleic acid (PNA) oligomers, that can specifically target any single pathogen. This approach broadens the range of potential targets to any gene with a known sequence in any bacterium, and could significantly reduce the time required to discover new antimicrobials or their redesign, if resistance arises. We review the potential of PNA as an antibacterial molecule. First, we describe the physicochemical properties of PNA and modifications of the PNA backbone and nucleobases. Second, we review the carriers used to transport PNA to bacterial cells. Furthermore, we discuss the PNA targets in antibacterial studies focusing on antisense PNA targeting bacterial mRNA and rRNA. Excessive use of antibiotics has led to an alarming situation when many bacterial strains developed resistance to these antibiotics. According to the World Health Organization, resistance to existing antibiotics, and slow rate of developing their new classes are currently among the greatest threats for human health ,2. BacteAntisense oligonucleotides, used to inhibit the synthesis of proteins essential for bacteria to sustain life, may be helpful in the fight against bacterial infections. One such oligonucleotide is the peptide nucleic acid (PNA) molecule that combines the properties of both peptides and nucleic acids. PNA were designed as synthetic analogues of DNA , which cInside bacteria, antisense PNA oligomers inhibit the translation process by binding to mRNA or the ribosome. The antisense effect of PNA is based on the formation of hydrogen bonds between the complementary PNA sequence and selected nucleic acid target. An important advantage of PNA is its selectivity and high-affinity binding. Thanks to that, it is possible to design PNA-based antimicrobials specific for particular genes in selected bacteria. In principle, PNA show huge potential to control the spread of resistant microorganisms. Unfortunately, the use of PNA in antibacterial applications encountered several crucial obstacles. The hydrophobicity of the PNA backbone causes problems with PNA solubility in aqueous solutions, which leads to PNA adopting compact structures susceptible to aggregation . One of To point-out the antibacterial potential of PNA and challenges facing any future therapeutic applications of these molecules, it is necessary to understand the structural and physicochemical properties of PNA. In this section, we present the most relevant PNA properties and structural fundaments of PNA complexes with nucleic acids.Besides the higher enzymatic stability, PNA has another important advantage: it hybridizes with complementary sequences of natural nucleic acids creating either duplexes or triplexes. So far, nearly 20 structures containing PNA oligomers have been solved by X-ray crystallography or nuclear magnetic resonance (NMR) including single-stranded PNA, PNA-PNA, PNA-DNA and PNA-RNA duplexes, and a triplex of double-stranded PNA with DNA . In most crystallized duplexes, the PNA terminus is extended with a lysine.The simplest duplexes observing the Watson–Crick base-pairing scheme are formed by single-stranded PNA with complementary strands of DNA ,32,33, RHowever, in general, classical PNA duplexes can be formed both in a parallel and antiparallel manner. Also, such PNA duplexes can form right- and left-handed P-type helices, characterized by a deeper and wider major groove, smaller angle, and larger displacement as compared to typical DNA and RNA helices. The P-type helix is 28 Å wide and, for comparison, classical helices composed of natural oligonucleotides are 23 Å (in the case of an A-helix) and 20 Å (B-helix) wide. The P-type helix has 18 base pairs per turn (as compared to A-helix – 11 and B-helix – 10). The PNA-DNA or PNA-RNA hybrids tend to be organized as B- or A-like helices, respectively .In addition to forming duplexes, single-stranded PNA can also bind to double-stranded DNA or RNA. Homopyrimidine PNA has the ability to bind a homopurine strand of a DNA duplex, opening the DNA helix and displacing the non-complementary DNA strand that forms the so-called P-loop . As a reApart from NMR and crystallography, the complexes with PNA have been investigated also by other experimental methods, e.g., isothermal titration calorimetry ,43, diffPNA are achiral molecules, but chiral centers can be introduced by adding amino acids into the PNA oligomer or at its terminus . As a result, CD can be observed confirming the helicity of PNA-PNA, PNA-DNA, and PNA-RNA duplexes .m) profiles of PNA-PNA and PNA-RNA 10-mer mixed-sequence duplexes were determined  prThe types of complexes presented in 1, see N-(2-aminoethyl)glycine backbone in the α-, β- or γ-position -one 33 turned out to be an effective thymine substitute in the PNA oligomers and increased PNA affinity in both duplex and triplex systems . C. C105].  complex . Most CP3K, which was first synthesized by Vaara and Porro in 1996 [3K efficiently transports PNA in vitro, both to gram-negative and gram-positive cells [3K-PNA conjugates drastically decreases in the presence of blood serum [3K is not an ideal candidate for a PNA transporter and its future medical use is doubtful.The mechanism of cell penetration by CPP may be different for different bacteria. The most commonly used CPP that transports PNA into bacterial cells is the synthetic peptide (KFF) in 1996  based onve cells . Despiteod serum . Moreoveod serum ; for com4XB  [rpoA gene, which is the key gene for the survival of Listeria monocytogenes. The authors confirmed that (RXR)4XB, TAT and (RFR)4XB, are the most effective in introducing PNA into L. monocytogenes. The same peptides were tested by the Patenge group [gyrA gene in order to inhibit the growth of Streptococus pyogenes. Out of 18 different peptides, TAT, oligolysine (K8), and (RXR)4XB, effectively inhibited the growth of the tested strains.Several other CPP have been tested as PNA carriers in vitro, including (RXR)alanine) , the TATalanine) , and manalanine) ,113. Abualanine)  tested tge group  and conjThe first protein identified as involved in the transport of peptide-PNA conjugates is an inner membrane protein SbmA . Ghosal E. coli than the conjugate with the ethylene glycol linker. Many other linkers were tested, e.g., a stable triazole ring [The effectiveness of PNA delivery into bacteria using CPP can be modulated by the linker between the PNA and peptide (using either a degradable or non-degradable one) ,118. Theole ring  or degraole ring ,120. We ole ring .Note, that CPP as PNA carriers are not universal because the transport of CPP may be strain dependent. Additional obstacle in the use of CPP is that they may be cytotoxic to eukaryotic cells and cause hemolysis of erythrocytes ,122. The12  into methicillin-resistant Staphylococcus aureus.An interesting and innovative approach was proposed in the work Readman et al. . The selE. coli . The autonjugate . The traonjugate , TDNs ef12 and TDN have proven that we do not have to limit ourselves to cationic peptides and completely different (non-peptidic) types of transporters could be considered and tested.Despite these few other strategies to deliver PNA to bacteria, the covalent conjugation of PNA and CPP is still the most popular one, mainly due to well-developed and relatively easy synthesis protocols. These protocols allow quick changes of the peptide sequence, PNA attachment site, and the linker type. However, this way of delivering PNA to bacteria is not ideal. Peptide uptake depends on bacterial strain and, at high concentrations, CPP exhibit toxicity to both bacterial and eukaryotic cells. Importantly, the new carriers such as vitamin B3K-PNA conjugates aimed at various targets and bacterial strains. Note, that the summary in 3K carrier may work differently in different strains. Similarly, it is impossible to compare the activity of PNA with classical antibiotics. Nevertheless, the MIC values in In this section, we summarize the PNA sequences used as antibacterials. Since it is impossible to list all the PNA-targeted genes and strains, we overview the most promising reports presenting the lowest minimal inhibitory concentrations (MIC) necessary to inhibit bacterial growth. In the last two decades, many mRNA encoding essential genes in clinically pathogenic bacteria have been validated as possible targets for antisense PNA . The mosE. coli acpP gene that encodes the acyl carrier protein, a protein crucial in fatty acid biosynthesis [acpP gene is conserved among gram-negative bacteria and has become a frequent PNA target in various human pathogens such as: Brucella suis [Haemophilus influenza [Pseudomonas aeruginosa [fabI gene, also involved in fatty acid biosynthesis, was targeted in E. coli and S. aureus [E. coli growth was also inhibited by PNA directed at mRNA involved in the folate biosynthesis (folA and folP) [The first reported antibacterial PNA targeted the mRNA transcript of ynthesis . The acplla suis , Haemophnfluenza , Pseudomruginosa . In addi. aureus . The E. nd folP) .E. coli [To improve the antimicrobial activity, PNA have also been used in combination with antibiotics. Dryselius et al. and Castillo et al. analysed synergistic interactions between PNA targeting fatty acid and folate synthesis pathways and a series of conventional antibiotics used against E. coli ,132. AntfolP gene) in an early step of folate biosynthesis, and trimethoprim that inhibits dihydrofolate reductase (of the folA gene) in the later step of this pathway. They synergy was observed if the anti-folA PNA was combined with sulfamethoxazole, but no synergy was detected if anti-folP PNA was combined with trimethoprim [acpP gene (involved in biosynthesis of fatty acids) exhibited synergistic interaction with trimethoprim, whose target is unrelated [S. pyogenes for the combination of anty-gyrA PNA with levofloxacin and novobiocin, agents that share the same target, namely the gyrase enzyme [Dryselius et al. examined the effects of the combinations of PNA and drugs against folate biosynthesis: sulfonamides that target dihydropteroate synthase , E. coli (murA), Klebsiella pneumoniae (murA), and S. Typhimurium  in a mixed culture, selectively killed bacteria [Interestingly, PNA targeted to specific sites of selected genes in bacteria . These facpP gene significantly inhibited the growth of E. coli strains in mice [L. monocytogenes growth in vitro, in cell culture, and in the Caenorhabditis elegans infection model. They also demonstrated that the PNA sequence did not adversely affect mitochondrial protein synthesis [Importantly, the efficacy of the antimicrobial peptide-PNA targeting mRNA was also demonstrated in a mouse model of infection. Tan et al. showed that injection of the antisense peptide-PNA targeting the  in mice . Moreoveynthesis .E. coli cell growth. For example, Good et al., used PNA oligomers to strand-invade and disrupt peptidyl transferase center (PTC) and α-sarcin loop of the 23S rRNA in the 50S ribosome subunit [E. coli AS19 cells , not only an effective PNA carrier is needed but also a more PNA-susceptible target. Thus, future efforts should also focus on the search for novel PNA targets that go beyond mRNA encoding essential proteins. Despite these limitations, PNA shows promise in antibacterial studies because of its high binding affinity to RNA and strand-invasion capability. Studies highlighted in this review point to effective antibacterial PNA sequences. However, the majority of PNA sequences were tested in vitro and many questions on the PNA use in vivo still remain to be answered. Few reports have shown PNA efficacy in animal models of infection using clinically relevant doses, but most studies were performed in non-human models. Therefore, how PNA affects the interferon response and emergence of bacterial resistance remains to be seen.It is worth noting that PNA use has already been successful in detection of bacterial pathogens. The use of PNA in diagnosis of bacterial infections has gained a lot of attention because it is critical to quickly recognize a particular pathogen to administer proper medication. The new pathogen identification platform based on the interaction of γ-PNA with double-stranded DNA shows promise in diagnostics . Thus, PThe number of studies related to the use of PNA in antibacterial applications is constantly growing, with PNA as a diagnostic tool for detecting pathogens paving the way. Thus, in the future the development of PNA-based antibiotics could become an alternative approach in the fight against multi-drug bacterial resistance. Other possibilities of PNA are yet to be discovered.


Subjective measures of sedentary behaviour (SB) (i.e. questionnaires and diaries/logs) are widely implemented, and can be useful for capturing type and context of SBs. However, little is known about comparative validity and reliability. The aim of this systematic review and meta-analysis was to: 1) identify subjective methods to assess overall, domain- and behaviour-specific SB, and 2) examine the validity and reliability of these methods.The databases MEDLINE, EMBASE and SPORTDiscus were searched up to March 2020. Inclusion criteria were: 1) assessment of SB, 2) evaluation of subjective measurement tools, 3) being performed in healthy adults, 4) manuscript written in English, and 5) paper was peer-reviewed. Data of validity and/or reliability measurements was extracted from included studies and a meta-analysis using random effects was performed to assess the pooled correlation coefficients of the validity.R] range − 0.01- 0.90) with logs/diaries (R = 0.63 [95%CI 0.48–0.78]) showing higher criterion validity compared to questionnaires (R = 0.35 [95%CI 0.32–0.39]). Furthermore, correlation coefficients of single- and multiple-item questionnaires were comparable . The reliability of SB measures was moderate-to-good, with the quality of these studies being mostly fair-to-good.The systematic search resulted in 2423 hits. After excluding duplicates and screening on title and abstract, 82 studies were included with 75 self-reported measurement tools. There was wide variability in the measurement properties and quality of the studies. The criterion validity varied between poor-to-excellent (correlation coefficient [Logs and diaries are recommended to validly and reliably assess self-reported SB. However, due to time and resources constraints, 1-item questionnaires may be preferred to subjectively assess SB in large-scale observations when showing similar validity and reliability compared to longer questionnaires.CRD42018105994. BaseVarious devices  and quesSeveral self-reported tools  have been developed recently to measure SB. These tools vary from single-item questions to extensive questionnaires about SB considering various domains. Currently, some reviews compared the validity and reliability of these tools , 16. HowCRD42018105994) and the ‘Preferred Reporting Items for Systematic Reviews and Meta-Analyses’ (PRISMA)  = 0.63 [95% CI 0.48–0.78], I2: 95%) were substantially higher than the coefficients of the questionnaires . Furthermore, correlation coefficient estimates of the questionnaires with ≥10-item ) did not differ much from the questionnaires with fewer items  .The correlation coefficients of logs and diaries (correlation coefficient estimate [n = 27), fair (n = 16), and poor (n = 8) quality. Most studies with a lower quality of the study were limited by a small sample size and calculation of correlation coefficients instead of ICCs. The time interval between the first and second assessment ranged between 0.5 h and 15 months, but most studies had an interval of 1–2 weeks , Bland-Altman plots with mean difference and limits of agreement (N = 2), and kappa values (N = 2). The ICC of the test-retest reliability of the subjective measures of SB ranged between 0.44 and 0.91 , in which the quality of most studies (i.e. level of evidence) was poor. Third, the validity of the logs/diaries was more favourable compared to the validity of questionnaires, with little improvement in validity of questionnaires when including multiple questions. Fourth, a moderate-to-good reliability was found for questionnaires and logs/diaries, with the quality of these studies being largely fair-to-good. Taken together, logs and diaries are recommended to validly and reliably assess SB when only self-report measures are available. However, considering limitations pertaining to logs and diaries , one may prefer using questionnaires in larger scaled observations.Time spent in SB has markedly increased over the last few decades and is expected to continue to increase even further . Since SThis meta-analysis showed that the overall validity for instruments to assess SB characteristics was moderate to low. These observations raise the question whether these results relate to the poor validity of methods to assess SB per se or the poor quality of the studies that were included. Excluding studies with lower quality from our meta-analyses reinforced the poor-to-moderate validity of the various methods, suggesting measures of SB possess poor validity. It is important to indicate that questionnaires examining physical activity show similarly poor level of validity . This hiFactors explaining the poorer variation in validity of the questionnaires versus diaries/logs may relate to differences in qualitative attributes . For example, diaries/logs typically adopt a short recall period (e.g. every 15–30 min), whilst questionnaires are often filled in covering a longer recall period . Consequently, diaries and logs are less reliant on long-term recall and can more accurately capture sporadic and intermittent behaviours. This fits with the higher validity of diaries/logs versus questionnaires. Unfortunately, this approach of using diaries/logs comes with the cost of high participant burden (in time), which subsequently may limit the response and compliance rate and introduce reporting bias. Another potential limitation of logs/diaries is that repeatedly filling in SBs may influence participants’ behaviour and cause (unwanted) adjustment of SB. These factors should be considered when deciding on the preferred way to assess SB in a future study.Previous work-related poor validity of questionnaires to systematic and random error, specifically reporting and recall bias which may lead to a low agreement with over- and underestimation Table . For exaDespite the significant heterogeneity in validity of the various measures to assess SB, the reliability of the questionnaires and diaries or logs were moderate-to-good. Importantly, these conclusions are based on studies with a fair-to-good quality. A central question pertaining to the reliability of questionnaires is whether differences are present in reliability for weekdays versus weekend days or for workdays versus non-workdays, especially given the marked differences in (sedentary) behaviour that exist between these days . Indeed,Logs and diaries have a higher validity compared to the questionnaires, are less reliant on long-term recall and can more accurately capture sporadic and intermittent behaviours. Therefore, we recommend logs and dairies as self-reported measurement tools. However, important limitations such as time constrains, lack of resources and the potential to influence participants’ behaviour, make them less useful for large-scale observational studies and/or intervention studies. Within the spectrum of questionnaires, there is no obvious preference for a single questionnaire. In fact, the most appropriate tool seems to depend on the nature of the study, especially since this review showed large variety in both validity and questionnaire characteristics (i.e. type and context of SBs). Therefore, some studies will benefit from questionnaires focusing on specific domains of SB, whilst others will benefit from a reliable estimate of total sedentary time or distribution of SB. Furthermore, when performing an intervention study, measures will benefit from the ability to measure changes across time. Since this ability was not examined within this review, we cannot make specific recommendations related to this type of studies. Nonetheless, these characteristics should be taken into account when planning such studies. Ultimately, and when feasible, a combination of objective and subjective assessments is preferred to provide valid and reliable insight into SB.This review identified the widespread (and rapidly growing) use of a large range of self-reported measures of SB, which significantly differ in type, extensiveness, complexity and duration. Our results indicated that the criterion validity of subjective measures ranged between poor and excellent, whereas the quality of most studies was poor. The validity of the logs/diaries was significantly higher compared to the questionnaires, with little improvement in criterion validity of questionnaires when increasing items to assess SB. Therefore, when only self-report measures are feasible, logs and diaries are recommended to validly and reliably assess SB, but due to time constraints and resources related to logs and diaries, 1-item questionnaires may be preferred in large-scale studies when showing similar validity and reliability compared to longer questionnaires. Whenever feasible, the combination of objective and subjective assessments will provide the most valid and reliable method to assess SB.Additional file :1 Table S1. Search strategy. Table S2. Assessing the quality of studies examining the criterion validity. Table S3. Assessing the quality of studies examining the reliability.