The ionic currents passing through nanopores can be used to sequence DNA and identify molecules at the single-molecule level. Recently, researchers have started using nanopores for the detection and analysis of proteins, providing a new platform for single-molecule enzymology studies and more efficient biomolecular sensing applications. For this approach, the homo-oligomeric Cytolysin A (ClyA) nanopore has been demonstrated as a powerful tool. Here, we describe a simple protocol allowing the production of ClyA nanopores. Monomers of ClyA are expressed in Escherichia coli and oligomerized in the presence of detergent. Subsequently, different oligomer variants are electrophoretically resolved and stored in a gel matrix for long-term use.Biological nanopores are an emerging class of biosensors with high-end precision owing to their reproducible fabrication at the nanometer scale. Most notably, nanopore-based DNA sequencing applications are currently being commercialized, while nanopore-based proteomics may become a reality in the near future.Although membrane proteins often prove to be difficult to purify, we describe a straightforward protocol for the preparation of Fragaceatoxin C (FraC) nanopores, which may have applications for DNA analysis and nanopore-based proteomics. Recombinantly expressed FraC nanopores are purified via two rounds of Ni-NTA affinity chromatography before and after oligomerization on sphingomyelin-containing liposomes. Starting from a plasmid vector containing the FraC gene, our method allows the production of purified nanopores within a week. Afterward, the FraC nanopores can be stored at +4 °C for several months, or frozen.Plexiform neurofibroma is mainly associated with neurofibromatosis type 1 and is seldom observed in the liver. Its occurrence in the liver without neurofibromatosis type 1 is even rarer. We report an extremely rare case of plexiform neurofibroma of the liver diagnosed by laparoscopic biopsy in a patient without neurofibromatosis type 1. The patient was a 35-year-old man who had neither clinical signs nor any family history of neurofibromatosis type 1. Abdominal ultrasonography, as part of a health screening, had detected a hepatic tumor. Subsequent contrast ultrasonography, computed tomography, and magnetic resonance imaging showed the tumor extending from the retroperitoneal space around the aorta to the hepatic hilum and distal portal branches in the right hepatic lobe, gallbladder, and left hepatic lobe. 18F-fluorodeoxyglucose positron emission tomography showed no abnormal accumulation. Histopathological examination of the tumor obtained laparoscopically led to a diagnosis of plexiform neurofibroma. Because the patient was asymptomatic with no features of malignancy, he was only monitored and managed. At follow-up 10 years later, computed tomography showed a decrease in tumor size. It is important to recognize that, while rare, plexiform neurofibroma can occur without neurofibromatosis type 1. We recommend follow-up instead of unreasonable surgery in such cases.
  Kidney stones are a common condition in the general population, however, high-quality evidence for its management is scarce. We propose the creation of an international network with the aim of sharing practice patterns and patient data towards an improvement of our knowledge of the disease.
 
  Cross-sectional survey.
 
  An online survey was circulated through several scientific societies. Items were grouped into six domains. Each center's overall score (OS) was also calculated.
 
  Chi square and Mann-Whitney tests were performed for differences across centers.
 
  The countries that contributed most were Italy (8.6%), Turkey (6.6%), France and Spain (6.1%). Some type of nutritional work-up was implemented in 62% of centers. A DEXA scan was performed by 46% of centers, whereas some kind of acidification test was performed by 25% of centers. Most centers (80%) implemented blood investigations at least at baseline. With regard to 24-h urine exams, 7 out of 16 were performed by at least 50% of centers. Information on stone composition was collected by 58% of centers. The OS was significantly higher among higher-volume centers compared with lower-volume centers (p = 0.002). Significant differences between EU and non-EU centers were found.
 
  Cross-sectional design; no validation on information.
 
  Our survey highlights the potential for the creation of a network of centers that could share information in a common database for observational research and for enrollment of patients in interventional trials.
 Our survey highlights the potential for the creation of a network of centers that could share information in a common database for observational research and for enrollment of patients in interventional trials.Nordic water bodies face multiple stressors due to human activities, generating diffuse loading and climate change. The 'green shift' towards a bio-based economy poses new demands and increased pressure on the environment. Bioeconomy-related pressures consist primarily of more intensive land management to maximise production of biomass. These activities can add considerable nutrient and sediment loads to receiving waters, posing a threat to ecosystem services and good ecological status of surface waters. The potential threats of climate change and the 'green shift' highlight the need for improved understanding of catchment-scale water and element fluxes. Here, we assess possible bioeconomy-induced pressures on Nordic catchments and associated impacts on water quality. We suggest measures to protect water quality under the 'green shift' and propose 'road maps' towards sustainable catchment management. We also identify knowledge gaps and highlight the importance of long-term monitoring data and good models to evaluate changes in water quality, improve understanding of bioeconomy-related impacts, support mitigation measures and maintain ecosystem services.Constructed wetlands (CWs) are one of the main countermeasures to reduce diffuse phosphorus (P) losses, but there is still a lack of systematic guidance accounting for spatially variable effects of hydraulic and P load on P retention. We present a three-step modelling approach for determining suitable placement of CWs in four different size groups (0.1-1.0 ha), based on incoming hydraulic and P load. The modelled hypothetical CW area was only 17% of that previously estimated and area of efficient CWs is even lower. The mean area-specific P retention increased with CW size.  https://www.selleckchem.com/products/bexotegrast.html  However, the spatial variation in retention was large for all size groups and largest (6-155 kg ha-1 year-1) for the smallest CWs due to highly variable incoming P loads, showing the possible benefits of targeted placement of CWs. The presented modelling approach has also flexibility to include and account for possible future changes in land cover and management.