High Stakes of the Small EU Partnership Instrument on the Eve of the 2017 Mid-Term Review

The European Union (EU)’s so-called Partnership Instrument (PI) captures the changing psychology of EU relations with rising powers, and has been designed to give more room for manoeuvre for EU external action. Two years of practice leading up to the 2017 Mid-Term Review have shown that the PI is instrument-driven rather than objective-driven, and that its strategic potential is undermined by lengthy committee-based procedures and sector-based programming. In an arena of vested interests, the PI’s significance is tied up with what EU stakeholders want to make of the European External Action Service (EEAS). National foreign ministries struggle to connect national objectives with EU interests, apart from the wider debate not addressed here as to where European interests come from when they are not based on national objectives. This article argues that national foreign ministries should consider the potentially positive effect on EU external action of this relatively small initiative, notably as the instrument to connect one European entity to other global powerhouses. EU Member States have so far been unable to link fully with the PI, and – like the EEAS and the European Parliament – they have too little clout to exert influence on the European Commission. The EU Global Strategy offers a window of opportunity for making the PI more political, expedient and flexible. FGGA – Publicaties zonder aanstelling Universiteit Leide

Natuurlijke immobilisatie van zware metalen in de Roeventerpeel

De waterbodem in Roeventerpeel is verontreinigd met cadmium en zink. Het is een voor de natuur potenteel belangrijk vengebied. Voor de ontwikkeling van dit gebied is verwijdering van de waterbodem van belang. De zware metalen in de waterbodem zijn via het grondwater aangevoerd en vastgelegd als sulfide. Naast de vastgelegde zware metalen is in de waterbodem een grote hoeveelheid ijzersulfide gevormd. Dit beperkt de mogelijkheden voor verwerking, omdat opsalg boven het grondwaterniveau zal leiden tot sterke verzuring Het principe van natuurlijke immobilisatie kan ook worden gebruikt voor de verwerking van de te verwijderen baggerspecie, zodat zowel de zware metalen als het ijzersulfide niet zullen zorgen voor schadelijke effecten. Het vengebied en de directe omgeving voldoen aan de te stellen randvoorwaarden

Definition of the σW regulon of Bacillus subtilis in the absence of stress

Bacteria employ extracytoplasmic function (ECF) sigma factors for their responses to environmental stresses. Despite intensive research, the molecular dissection of ECF sigma factor regulons has remained a major challenge due to overlaps in the ECF sigma factor-regulated genes and the stimuli that activate the different ECF sigma factors. Here we have employed tiling arrays to single out the ECF σW regulon of the Gram-positive bacterium Bacillus subtilis from the overlapping ECF σX, σY, and σM regulons. For this purpose, we profiled the transcriptome of a B. subtilis sigW mutant under non-stress conditions to select candidate genes that are strictly σW-regulated. Under these conditions, σW exhibits a basal level of activity. Subsequently, we verified the σW-dependency of candidate genes by comparing their transcript profiles to transcriptome data obtained with the parental B. subtilis strain 168 grown under 104 different conditions, including relevant stress conditions, such as salt shock. In addition, we investigated the transcriptomes of rasP or prsW mutant strains that lack the proteases involved in the degradation of the σW anti-sigma factor RsiW and subsequent activation of the σW-regulon. Taken together, our studies identify 89 genes as being strictly σW-regulated, including several genes for non-coding RNAs. The effects of rasP or prsW mutations on the expression of σW-dependent genes were relatively mild, which implies that σW-dependent transcription under non-stress conditions is not strictly related to RasP and PrsW. Lastly, we show that the pleiotropic phenotype of rasP mutant cells, which have defects in competence development, protein secretion and membrane protein production, is not mirrored in the transcript profile of these cells. This implies that RasP is not only important for transcriptional regulation via σW, but that this membrane protease also exerts other important post-transcriptional regulatory functions

Towards the development of Bacillus subtilis as a cell factory for membrane proteins and protein complexes

Background: The Gram-positive bacterium Bacillus subtilis is an important producer of high quality industrial enzymes and a few eukaryotic proteins. Most of these proteins are secreted into the growth medium, but successful examples of cytoplasmic protein production are also known. Therefore, one may anticipate that the high protein production potential of B. subtilis can be exploited for protein complexes and membrane proteins to facilitate their functional and structural analysis. The high quality of proteins produced with B. subtilis results from the action of cellular quality control systems that efficiently remove misfolded or incompletely synthesized proteins. Paradoxically, cellular quality control systems also represent bottlenecks for the production of various heterologous proteins at significant concentrations. Conclusion: While inactivation of quality control systems has the potential to improve protein production yields, this could be achieved at the expense of product quality. Mechanisms underlying degradation of secretory proteins are nowadays well understood and often controllable. It will therefore be a major challenge for future research to identify and modulate quality control systems of B. subtilis that limit the production of high quality protein complexes and membrane proteins, and to enhance those systems that facilitate assembly of these proteins.

Verspreiden van bagger op het land in klei- en veengebieden

De meeste baggerspecie, inclusief licht verontreinigde baggerspecie die vrijkomt bij onderhoud van watergangen, wordt verspreid op landbouwpercelen. Het in dit rapport beschreven onderzoek is opgezet om op praktijkschaal na te gaan of op dit moment de praktijk van het verspreiden van licht verontreinigde bagger meetbare effecten heeft veroorzaakt en aan te geven wat er in de toekomst kan worden verwacht bij voortzetting van de huidige praktijk. De belangrijkste conclusie van het onderzoek is dat in de onderzochte klei- en veengebieden de praktijk van verspreiden van baggerspecie niet heeft geleid tot een sterke achteruitgang van de bodem. Achteruitgang is echter wel mogelijk als de praktijk van verspreiden meer opschuift in de richting van de kwaliteit die maximaal is toegestaan. Op basis van het onderzoek worden diverse aanbevelingen voor het beleid gemaakt

Pneumococcal Gene Complex Involved in Resistance to Extracellular Oxidative Stress

Streptococcus pneumoniae is a Gram-positive bacterium which is a member of the normal human nasopharyngeal flora but can also cause serious disease such as pneumonia, bacteremia, and meningitis. Throughout its life cycle, S. pneumoniae is exposed to significant oxidative stress derived from endogenously produced hydrogen peroxide (H2O2) and from the host through the oxidative burst. How S. pneumoniae, an aerotolerant anaerobic bacterium that lacks catalase, protects itself against hydrogen peroxide stress is still unclear. Bioinformatic analysis of its genome identified a hypothetical open reading frame belonging to the thiol-specific antioxidant (TlpA/TSA) family, located in an operon consisting of three open reading frames. For all four strains tested, deletion of the gene resulted in an approximately 10-fold reduction in survival when strains were exposed to external peroxide stress. However, no role for this gene in survival of internal superoxide stress was observed. Mutagenesis and complementation analysis demonstrated that all three genes are necessary and sufficient for protection against oxidative stress. Interestingly, in a competitive index mouse pneumonia model, deletion of the operon had no impact shortly after infection but was detrimental during the later stages of disease. Thus, we have identified a gene complex involved in the protection of S. pneumoniae against external oxidative stress, which plays an important role during invasive disease.

Oral ribose supplementation in dystroglycanopathy:A single case study

Three forms of muscular dystrophy-dystroglycanopathies are linked to the ribitol pathway. These include mutations in the isoprenoid synthase domain-containing protein (ISPD), fukutin-related protein (FKRP), and fukutin (FKTN) genes. The aforementioned enzymes are required for generation of the ribitol phosphate linkage in the O-glycan of alpha-dystroglycan. Mild cases of dystroglycanopathy present with slowly progressive muscle weakness, while in severe cases the eyes and brain are also involved. Previous research showed that ribose increased the intracellular concentrations of cytidine diphosphate-ribitol (CDP-ribitol) and had a therapeutic effect. Here, we report the safety and effects of oral ribose supplementation during 6 months in a patient with limb girdle muscular dystrophy type 2I (LGMD2I) due to a homozygous FKRP mutation. Ribose was well tolerated in doses of 9 g or 18 g/day. Supplementation with 18 g of ribose resulted in a decrease of creatine kinase levels of 70%. Moreover, metabolomics showed a significant increase in CDP-ribitol levels with 18 g of ribose supplementation (p &lt; 0.001). Although objective improvement in clinical and patient-reported outcome measures was not observed, the patient reported subjective improvement of muscle strength, fatigue, and pain. This case study indicates that ribose supplementation in patients with dystroglycanopathy is safe and highlights the importance for future studies regarding its potential effects.</p

Oral ribose supplementation in dystroglycanopathy:A single case study

Three forms of muscular dystrophy-dystroglycanopathies are linked to the ribitol pathway. These include mutations in the isoprenoid synthase domain-containing protein (ISPD), fukutin-related protein (FKRP), and fukutin (FKTN) genes. The aforementioned enzymes are required for generation of the ribitol phosphate linkage in the O-glycan of alpha-dystroglycan. Mild cases of dystroglycanopathy present with slowly progressive muscle weakness, while in severe cases the eyes and brain are also involved. Previous research showed that ribose increased the intracellular concentrations of cytidine diphosphate-ribitol (CDP-ribitol) and had a therapeutic effect. Here, we report the safety and effects of oral ribose supplementation during 6 months in a patient with limb girdle muscular dystrophy type 2I (LGMD2I) due to a homozygous FKRP mutation. Ribose was well tolerated in doses of 9 g or 18 g/day. Supplementation with 18 g of ribose resulted in a decrease of creatine kinase levels of 70%. Moreover, metabolomics showed a significant increase in CDP-ribitol levels with 18 g of ribose supplementation (p &lt; 0.001). Although objective improvement in clinical and patient-reported outcome measures was not observed, the patient reported subjective improvement of muscle strength, fatigue, and pain. This case study indicates that ribose supplementation in patients with dystroglycanopathy is safe and highlights the importance for future studies regarding its potential effects.</p