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- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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- Bossini-Castillo, Lara, et al.
(författare)
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A replication study confirms the association of TNFSF4 (OX40L) polymorphisms with systemic sclerosis in a large European cohort
- 2011
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Ingår i: Annals of the Rheumatic Diseases. - : BMJ. - 1468-2060 .- 0003-4967. ; 70:4, s. 638-641
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Tidskriftsartikel (refereegranskat)abstract
- Objectives The aim of this study was to confirm the influence of TNFSF4 polymorphisms on systemic sclerosis (SSc) susceptibility and phenotypic features. Methods A total of 8 European populations of Caucasian ancestry were included, comprising 3014 patients with SSc and 3125 healthy controls. Four genetic variants of TNFSF4 gene promoter (rs1234314, rs844644, rs844648 and rs12039904) were selected as genetic markers. Results A pooled analysis revealed the association of rs1234314 and rs12039904 polymorphisms with SSc (OR 1.15, 95% CI 1.02 to 1.31; OR 1.18, 95% CI 1.08 to 1.29, respectively). Significant association of the four tested variants with patients with limited cutaneous SSc (lcSSc) was revealed (rs1234314 OR 1.22, 95% CI 1.07 to 1.38; rs844644 OR 0.91, 95% CI 0.83 to 0.99; rs844648 OR 1.10, 95% CI 1.01 to 1.20 and rs12039904 OR 1.20, 95% CI 1.09 to 1.33). Association of rs1234314, rs844648 and rs12039904 minor alleles with patients positive for anti-centromere antibodies (ACA) remained significant (OR 1.23, 95% CI 1.10 to 1.37; OR 1.12, 95% CI 1.01 to 1.25; OR 1.22, 95% CI 1.07 to 1.38, respectively). Haplotype analysis confirmed a protective haplotype associated with SSc, lcSSc and ACA positive subgroups (OR 0.88, 95% CI 0.82 to 0.96; OR 0.88, 95% CI 0.80 to 0.96; OR 0.86, 95% CI 0.77 to 0.97, respectively) and revealed a new risk haplotype associated with the same groups of patients (OR 1.14, 95% CI 1.03 to 1.26; OR 1.20, 95% CI 1.08 to 1.35; OR 1.23, 95% CI 1.07 to 1.42, respectively). Conclusions The data confirm the influence of TNFSF4 polymorphisms in SSc genetic susceptibility, especially in subsets of patients positive for lcSSc and ACA.
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- De Vriendt, T., et al.
(författare)
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Reliability and validity of the Adolescent Stress Questionnaire in a sample of European adolescents--the HELENA study
- 2011
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Ingår i: BMC Public Health. - : Springer Science and Business Media LLC. - 1471-2458. ; 11, s. 717-
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Since stress is hypothesized to play a role in the etiology of obesity during adolescence, research on associations between adolescent stress and obesity-related parameters and behaviours is essential. Due to lack of a well-established recent stress checklist for use in European adolescents, the study investigated the reliability and validity of the Adolescent Stress Questionnaire (ASQ) for assessing perceived stress in European adolescents. METHODS: The ASQ was translated into the languages of the participating cities (Ghent, Stockholm, Vienna, Zaragoza, Pecs and Athens) and was implemented within the HELENA cross-sectional study. A total of 1140 European adolescents provided a valid ASQ, comprising 10 component scales, used for internal reliability (Cronbach alpha) and construct validity (confirmatory factor analysis or CFA). Contributions of socio-demographic (gender, age, pubertal stage, socio-economic status) characteristics to the ASQ score variances were investigated. Two-hundred adolescents also provided valid saliva samples for cortisol analysis to compare with the ASQ scores (criterion validity). Test-retest reliability was investigated using two ASQ assessments from 37 adolescents. RESULTS: Cronbach alpha-values of the ASQ scales (0.57 to 0.88) demonstrated a moderate internal reliability of the ASQ, and intraclass correlation coefficients (0.45 to 0.84) established an insufficient test-retest reliability of the ASQ. The adolescents' gender (girls had higher stress scores than boys) and pubertal stage (those in a post-pubertal development had higher stress scores than others) significantly contributed to the variance in ASQ scores, while their age and socio-economic status did not. CFA results showed that the original scale construct fitted moderately with the data in our European adolescent population. Only in boys, four out of 10 ASQ scale scores were a significant positive predictor for baseline wake-up salivary cortisol, suggesting a rather poor criterion validity of the ASQ, especially in girls. CONCLUSIONS: In our European adolescent sample, the ASQ had an acceptable internal reliability and construct validity and the adolescents' gender and pubertal stage systematically contributed to the ASQ variance, but its test-retest reliability and criterion validity were rather poor. Overall, the utility of the ASQ for assessing perceived stress in adolescents across Europe is uncertain and some aspects require further examination.
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