| 1. |
- 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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| 2. |
- Berg, Gunnar, et al.
(författare)
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Skolans kommunalisering och de professionellas frirum : Rapport utarbetad på uppdrag av utredningen om skolans kommunalisering
- 2014
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Rapport (övrigt vetenskapligt/konstnärligt)abstract
- FörordAllt fler är överens om att lärarna är skolans viktigaste resurs. Diskussionen om skolan handlar därför i hög grad om hur vi ska ge bästa tänkbara förutsättningar för lärarna att bedriva en bra undervisning. Man brukar säga att lärare och rektorer är exempel på professioner eller, lite beroende på definitionen, semiprofessioner. För alla grupper av professionella är det av yttersta vikt med autonomi i yrket –så även för lärare och rektorer. När effekterna av skolans kommunalisering ska bedömas är det därför relevant att analysera på vilket sätt lärarnas och rektorernas handlingsutrymme för självständiga professionella handlingar i den svenska skolan förändrats från tiden närmast före skolans kommunalisering och framåt. I denna rapport visas bland annat att staten vid olika tidsperioder tillerkänt skolans professionella varierande grad av handlingsutrymme för professionella handlingar i skolans vardagsarbete. I rapporten talas om att skolan före kommunaliseringen kännetecknades av central regelstyrning, som i samband med kommunaliseringen ersattes av en decentraliserad målstyrning. Numera utmärksstyrningen av centraliserad resultatstyrning. Vidare framhålls i rapporten att lärarna och rektorerna av olika skäl inte alltid använt det frirum för professionella handlingar som staten medgett.Undersökningen har genomförts efter ett uppdrag från denna utredning till professor Gunnar Berg, som utarbetat rapporten i samverkan med en grupp forskare bestående av Fia Andersson, Göran Bostedt, Judit Novak, Jan Perselli, Frank Sundh och Christer Wede. Forskarna ansvarar själva för innehållet i rapporten.Utredningen om skolans kommunalisering (U 2012:09)Leif Lewin Särskild utredare
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| 3. |
- Mischak, Harald, et al.
(författare)
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Comprehensive human urine standards for comparability and standardization in clinical proteome analysis
- 2010
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Ingår i: Proteomics - Clinical Applications. - : Wiley. - 1862-8346 .- 1862-8354. ; 4:4, s. 464-478
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Tidskriftsartikel (refereegranskat)abstract
- Purpose: Urine proteomics is emerging as a powerful tool for biomarker discovery. The purpose of this study is the development of a well-characterized "real life" sample that can be used as reference standard in urine clinical proteomics studies. Experimental design: We report on the generation of male and female urine samples that are extensively characterized by different platforms and methods (CE-MS, LC-MS, LC-MS/MS, 1-D gel analysis in combination with nano-LC MS/MS (using LTQ-FT ultra), and 2-DE-MS) for their proteome and peptidome. In several cases analysis involved a definition of the actual biochemical entities, i.e. proteins/peptides associated with molecular mass and detected PTMs and the relative abundance of these compounds. Results: The combination of different technologies allowed coverage of a wide mass range revealing the advantages and complementarities of the different technologies. Application of these samples in "inter-laboratory" and "inter-platform" data comparison is also demonstrated. Conclusions and clinical relevance: These well-characterized urine samples are freely available upon request to enable data comparison especially in the context of biomarker discovery and validation studies. It is also expected that they will provide the basis for the comprehensive characterization of the urinary proteome.
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| 4. |
- Mischak, Harald, et al.
(författare)
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Implementation of proteomic biomarkers : making it work
- 2012
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Ingår i: European Journal of Clinical Investigation. - : Wiley. - 0014-2972 .- 1365-2362. ; 42:9, s. 1027-1036
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Tidskriftsartikel (refereegranskat)abstract
- While large numbers of proteomic biomarkers have been described, they are generally not implemented in medical practice. We have investigated the reasons for this shortcoming, focusing on hurdles downstream of biomarker verification, and describe major obstacles and possible solutions to ease valid biomarker implementation. Some of the problems lie in suboptimal biomarker discovery and validation, especially lack of validated platforms with well-described performance characteristics to support biomarker qualification. These issues have been acknowledged and are being addressed, raising the hope that valid biomarkers may start accumulating in the foreseeable future. However, successful biomarker discovery and qualification alone does not suffice for successful implementation. Additional challenges include, among others, limited access to appropriate specimens and insufficient funding, the need to validate new biomarker utility in interventional trials, and large communication gaps between the parties involved in implementation. To address this problem, we propose an implementation roadmap. The implementation effort needs to involve a wide variety of stakeholders (clinicians, statisticians, health economists, and representatives of patient groups, health insurance, pharmaceutical companies, biobanks, and regulatory agencies). Knowledgeable panels with adequate representation of all these stakeholders may facilitate biomarker evaluation and guide implementation for the specific context of use. This approach may avoid unwarranted delays or failure to implement potentially useful biomarkers, and may expedite meaningful contributions of the biomarker community to healthcare.
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| 5. |
- Novak, Martin, et al.
(författare)
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Incentive Learning Underlying Cocaine-Seeking Requires mGluR5 Receptors Located on Dopamine D1 Receptor-Expressing Neurons
- 2010
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Ingår i: JOURNAL OF NEUROSCIENCE. - : Society for Neuroscience. - 0270-6474. ; 30:36, s. 11973-11982
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Tidskriftsartikel (refereegranskat)abstract
- Understanding the psychobiological basis of relapse remains a challenge in developing therapies for drug addiction. Relapse in cocaine addiction often occurs following exposure to environmental stimuli previously associated with drug taking. The metabotropic glutamate receptor, mGluR5, is potentially important in this respect; it plays a central role in several forms of striatal synaptic plasticity proposed to underpin associative learning and memory processes that enable drug-paired stimuli to acquire incentive motivational properties and trigger relapse. Using cell type-specific RNA interference, we have generated a novel mouse line with a selective knock-down of mGluR5 in dopamine D1 receptor-expressing neurons. Although mutant mice self-administer cocaine, we show that reinstatement of cocaine-seeking induced by a cocaine-paired stimulus is impaired. By examining different aspects of associative learning in the mutant mice, we identify deficits in specific incentive learning processes that enable a reward-paired stimulus to directly reinforce behavior and to become attractive, thus eliciting approach toward it. Our findings show that glutamate signaling through mGluR5 located on dopamine D1 receptor-expressing neurons is necessary for incentive learning processes that contribute to cue-induced reinstatement of cocaine-seeking and which may underpin relapse in drug addiction.
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| 6. |
- Regber, Susann, 1956-
(författare)
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Barriers and Facilitators of Health Promotion and Obesity Prevention in Early Childhood : A Focus on Parents, Results from the IDEFICS Study
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Childhood obesity has increased dramatically during the past thirty years. Parents are key persons in their children’s lives and their efforts to create healthy lifestyles are very important. However, social and economic determinants of health also affect parents’ opportunities to promote a healthy lifestyle.Aims: To explore barriers and facilitators in promoting healthy lifestyles and preventing childhood obesity, focusing on parental roles.Methods and main findings: Three studies originated from the Identification and Prevention of Dietary- and Lifestyle-induced health Effects in Children and InfantS (IDEFICS) study of determinants for two to nine-year-old children’s health in eight European countries. The fourth study was a qualitative interview study conducted in southwest Sweden.Paper I: In focus group discussions (20 focus groups with children and 36 with parents), parents described lack of time, financial constraints, availability and food marketing techniques as barriers for promoting healthy eating. School policies about food varied; only Sweden and Estonia provided free school lunches. Children described great variation in the availability of unhealthy foods and beverages in their homes.Paper II: Objectively measured Body Mass Index (BMI) of children (n=16 220) were compared to parents’ perception of and concern for their children’s health and weight status. In all weight categories and all countries, a substantial proportion of parents failed to accurately judge their child’s weight status. In general, parents considered their children to be healthy, irrespective of their weight status. Parents of children with overweight or obesity systematically underestimated their children’s weight status across eight European countries. Accurate parental weight perception in Europe differed according to geographic region.Paper III: Swedish IDEFICS participants (n=1825) were compared with an age- and sex-matched referent population (n=1825), using registers from Statistics Sweden and the Swedish Medical Birth Register. Longitudinal child growth data (n=3650) were collected from child health centers and school health services. Families with low income, less education, foreign background or single parenthood were underrepresented in the IDEFICS study. BMI at inclusion had no selection effect but, at eight years of age, the obesity prevalence was significantly greater among referents.Paper IV: A qualitative content analysis was used to interpret the findings from interviews with nurses (n=15) working at child health centers in the southwest of Sweden. The BMI Chart to identify overweight and obesity in children facilitated greater recognition but nurses used it inconsistently, a barrier to prevention. Other barriers were obesity considered a sensitive issue and that some parents wanted overweight children.Conclusion: Parents may not perceive their child’s growth trajectory from overweight to obesity, and the preschool years may pass without effort to change lifestyle. Therefore, objective measurement and information of children’s BMI weight status by healthcare professionals is of great importance. To reach all parents and avoid selection bias, health surveys or health promoting activities must be tailored. Health promoting activities at the family level as well as the societal level should start early in children’s lives to prevent childhood obesity.
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| 7. |
- Schmitt, Roland, et al.
(författare)
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The Combined Role of Galactose-Deficient IgA1 and Streptococcal IgA-Binding M Protein in Inducing IL-6 and C3 Secretion from Human Mesangial Cells: Implications for IgA Nephropathy.
- 2014
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Ingår i: Journal of Immunology. - : The American Association of Immunologists. - 1550-6606 .- 0022-1767. ; 193:1, s. 317-326
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Tidskriftsartikel (refereegranskat)abstract
- IgA nephropathy (IgAN) is characterized by mesangial cell proliferation and extracellular matrix expansion associated with immune deposits consisting of galactose-deficient polymeric IgA1 and C3. We have previously shown that IgA-binding regions of streptococcal M proteins colocalize with IgA in mesangial immune deposits in patients with IgAN. In the present study, the IgA-binding M4 protein from group A Streptococcus was found to bind to galactose-deficient polymeric IgA1 with higher affinity than to other forms of IgA1, as shown by surface plasmon resonance and solid-phase immunoassay. The M4 protein was demonstrated to bind to mesangial cells not via the IgA-binding region but rather via the C-terminal region, as demonstrated by flow cytometry. IgA1 enhanced binding of M4 to mesangial cells, but not vice versa. Costimulation of human mesangial cells with M4 and galactose-deficient polymeric IgA1 resulted in a significant increase in IL-6 secretion compared with each stimulant alone. Galactose-deficient polymeric IgA1 alone, but not M4, induced C3 secretion from the cells, and costimulation enhanced this effect. Additionally, costimulation enhanced mesangial cell proliferation compared with each stimulant alone. These results indicate that IgA-binding M4 protein binds preferentially to galactose-deficient polymeric IgA1 and that these proteins together induce excessive proinflammatory responses and proliferation of human mesangial cells. Thus, tissue deposition of streptococcal IgA-binding M proteins may contribute to the pathogenesis of IgAN.
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| 8. |
- Wada, Yoshinao, et al.
(författare)
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Comparison of methods for profiling O-glycosylation: Human Proteome Organisation Human Disease Glycomics/Proteome Initiative multi-institutional study of IgA1.
- 2010
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Ingår i: Molecular & cellular proteomics. - 1535-9484. ; 9:4, s. 719-727
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Tidskriftsartikel (refereegranskat)abstract
- The Human Proteome Organisation Human Disease Glycomics/Proteome Initiative recently coordinated a multi-institutional study that evaluated methodologies that are widely used for defining the N-glycan content in glycoproteins. The study convincingly endorsed mass spectrometry as the technique of choice for glycomic profiling in the discovery phase of diagnostic research. The present study reports the extension of the Human Disease Glycomics/Proteome Initiative's activities to an assessment of the methodologies currently used for O-glycan analysis. Three samples of IgA1 isolated from the serum of patients with multiple myeloma were distributed to 15 laboratories worldwide for O-glycomics analysis. A variety of mass spectrometric and chromatographic procedures representative of current methodologies were used. Similar to the previous N-glycan study, the results convincingly confirmed the pre-eminent performance of MS for O-glycan profiling. Two general strategies were found to give the most reliable data, namely direct MS analysis of mixtures of permethylated reduced glycans in the positive ion mode and analysis of native reduced glycans in the negative ion mode using LC-MS approaches. In addition, mass spectrometric methodologies to analyze O-glycopeptides were also successful.
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