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Halting the loss of grassland biodiversity and restoring degraded ecosystems are high priority tasks in the EU Biodiversity Strategy. Sowing low-diversity seed mixtures is widely used in grassland restoration because of its high predictability and fast, promising results. Generally, the sown perennial grasses establish within a few years and form a dense sward, which effectively suppresses weeds. Unfortunately, these grasslands are often species-poor because the sown grasses hamper the colonisation of target grassland forbs. Our aim was to test a novel approach to increase the diversity of species-poor grasslands. We selected eight 8-year-old grasslands restored by low-diversity seed sowing where we created 32 establishment gaps by breaking up the grass sward and sowing a high-diversity seed mixture (35 native species). Altogether, we established three grazed gaps (1m × 1m, 2m × 2m and 4m × 4m) and one fenced gap (4m × 4m) per site and monitored the presence and abundance of sown and non-sown species within a time frame of two years. We asked the following questions: (1) Which target species establish most successfully? (2) What is the effect of establishment gap size on the establishment success of target species and weeds? (3) What is the effect of management (grazed versus not managed) on the species composition of the establishment gaps? Our results showed that by creating establishment gaps and sowing diverse seed mixtures, we were able to overcome microsite and propagule limitation, successfully introducing target species into the species-poor grasslands. We found that all sown species established in the gaps, and the majority of the species maintained or even increased their first-year cover in the second year. Smaller gaps were characterised by lower cover of sown species and a quite stochastic development compared to the larger ones. Weed cover was moderate in the first year and decreased significantly in the second year, regardless of gap size. Therefore, in restoration practice, the use of larger establishment gaps is recommended. We found that the cover of sown species and weeds were similar in the grazed and unmanaged gaps during our study. However, management by extensive grazing might be crucial in the long-term because livestock can disperse target species propagules and create microsites. Our study shows that establishment gaps can serve as biodiversity hotspots. Further studies need to clarify to what extent they can improve the restoration success across the entire grassland.
EDTA is commonly used as an efficient chelator of metal ion enzyme cofactors. It is highly soluble, optically inactive and does not interfere with most chemicals used in standard buffers making EDTA a common choice to generate metal-free conditions for biochemical and biophysical investigations. However, the controversy in the literature on metal-free enzyme activities achieved using EDTA or by other means called our attention to a putative effect of EDTA beyond chelation. Here, we show that EDTA competes for the nucleotide binding site of the nucleotide hydrolase dUTPase by developing an interaction network within the active site similar to that of the substrate. To achieve these findings, we applied kinetics and molecular docking techniques using two different dUTPases. Furthermore, we directly measured the binding of EDTA to dUTPases and to two other dNTPases, the Taq polymerase and MutT using isothermal titration calorimetry. EDTA binding proved to be exothermic and mainly enthalpy driven with a submicromolar dissociation constant considerably lower than that of the enzyme:substrate or the Mg:EDTA complexes. Control proteins, including an ATPase, did not interact with EDTA. Our findings indicate that EDTA may act as a selective inhibitor against dNTP hydrolyzing enzymes and urge the rethinking of the utilization of EDTA in enzymatic experiments.
Background & Aims: Spontaneous portosystemic shunts (SPSS) frequently develop in liver cirrhosis. Recent data suggested that the presence of a single large SPSS is associated with complications, especially overt hepatic encephalopathy (oHE). However, the presence of >1 SPSS is common. This study evaluates the impact of total cross-sectional SPSS area (TSA) on outcomes in patients with liver cirrhosis.
Methods: In this retrospective international multicentric study, CT scans of 908 cirrhotic patients with SPSS were evaluated for TSA. Clinical and laboratory data were recorded. Each detected SPSS radius was measured and TSA calculated. One-year survival was the primary endpoint and acute decompensation (oHE, variceal bleeding, ascites) was the secondary endpoint.
Results: A total of 301 patients (169 male) were included in the training cohort. Thirty percent of all patients presented with >1 SPSS. A TSA cut-off of 83 mm2 was used to classify patients with small or large TSA (S-/L-TSA). Patients with L-TSA presented with higher model for end-stage liver disease score (11 vs. 14) and more commonly had a history of oHE (12% vs. 21%, p <0.05). During follow-up, patients with L-TSA experienced more oHE episodes (33% vs. 47%, p <0.05) and had lower 1-year survival than those with S-TSA (84% vs. 69%, p <0.001). Multivariate analysis identified L-TSA (hazard ratio 1.66; 95% CI 1.02–2.70, p <0.05) as an independent predictor of mortality. An independent multicentric validation cohort of 607 patients confirmed that patients with L-TSA had lower 1-year survival (77% vs. 64%, p <0.001) and more oHE development (35% vs. 49%, p <0.001) than those with S-TSA.
Conclusion: This study suggests that TSA >83 mm2 increases the risk for oHE and mortality in patients with cirrhosis. Our results support the clinical use of TSA/SPSS for risk stratification and decision-making in the management of patients with cirrhosis.
Lay summary: The prevalence of spontaneous portosystemic shunts (SPSS) is higher in patients with more advanced chronic liver disease. The presence of more than 1 SPSS is common in advanced chronic liver disease and is associated with the development of hepatic encephalopathy. This study shows that total cross-sectional SPSS area (rather than diameter of the single largest SPSS) predicts survival in patients with advanced chronic liver disease. Our results support the clinical use of total cross-sectional SPSS area for risk stratification and decision-making in the management of SPSS.