Synthesis, screening, and sequencing of cysteine-rich one-bead one-compound peptide libraries.

Cysteine-rich peptides are valued as tags for biarsenical fluorophores and as environmentally important reagents for binding toxic heavy metals. Due to the inherent difficulties created by cysteine, the power of one-bead one-compound (OBOC) libraries has never been applied to the discovery of short cysteine-rich peptides. We have developed the first method for the synthesis, screening, and sequencing of cysteine-rich OBOC peptide libraries. First, we synthesized a heavily biased cysteine-rich OBOC library, incorporating 50% cysteine at each position (Ac-X8-KM-TentaGel). Then, we developed conditions for cysteine alkylation, cyanogen bromide cleavage, and direct MS/MS sequencing of that library at the single bead level. The sequencing efficiency of this library was comparable to a traditional cysteine-free library. To validate screening of cysteine-rich OBOC libraries, we reacted a library with the biarsenical FlAsH and identified beads bearing the known biarsenical-binding motif (CCXXCC). These results enable OBOC libraries to be used in high-throughput discovery of cysteine-rich peptides for protein tagging, environmental remediation of metal contaminants, or cysteine-rich pharmaceuticals

Structure calculation, refinement and validation using CcpNmr Analysis

CcpNmr Analysis provides a streamlined pipeline for both NMR chemical shift assignment and structure determination of biological macromolecules. In addition, it encompasses tools to analyse the many additional experiments that make NMR such a pivotal technique for research into complex biological questions. This report describes how CcpNmr Analysis can seamlessly link together all of the tasks in the NMR structure-determination process. It details each of the stages from generating NMR restraints [distance, dihedral,hydrogen bonds and residual dipolar couplings (RDCs)],exporting these to and subsequently re-importing them from structure-calculation software (such as the programs CYANA or ARIA) and analysing and validating the results obtained from the structure calculation to, ultimately, the streamlined deposition of the completed assignments and the refined ensemble of structures into the PDBe repository. Until recently, such solution-structure determination by NMR has been quite a laborious task, requiring multiple stages and programs. However, with the new enhancements to CcpNmr Analysis described here, this process is now much more intuitive and efficient and less error-prone

Blockchain Technology in Business and Information Systems Research

Peer reviewe

Identification of challenges to the availability and accessibility of opioids in twelve European countries:conclusions from two ATOME six-country workshops

Background: Access to many controlled medicines is inadequate in a number of European countries. This leads to deficits in the treatment of moderate to severe pain as well as in opioid agonist therapy. Objective: The study objective was to elaborate the reasons for this inadequacy. The work plan of the Access to Opioid Medication in Europe (ATOME) project included two six-country workshops. These workshops comprised a national situational analysis, drafting tailor-made recommendations for improvement and developing action plans for their implementation. Methods: In total, 84 representatives of the national Ministries of Health, national controlled substances authorities, experts representing regulatory and law enforcement authorities, leading health care professionals, and patient representatives from 13 European countries participated in either one of the workshops. The delegates used breakout sessions to identify key common challenges. Content analysis was used for the evaluation of protocols and field notes. Results: A number of challenges to opioid accessibility in the countries was identified in the domains of knowledge and educational, regulatory, legislative, as well as public awareness and training barriers that limit opioid prescription. In addition, short validity of prescriptions and bureaucratic practices resulting in overregulation impeded availablity of some essential medicines. Stigmatization and criminalisation of people who use drugs remained the major impediment to increasing opioid agonist program coverage. Conclusions: The challenges identified during outcomes of the workshops were used as the basis for subsequent dissemination and implementation activities in the ATOME project, and in some countries the workshop proceedings already served as a stepping-stone for the first changes in regulations and legislation

Probing interactions in mesoscopic gold wires

We have measured in gold wires the energy exchange rate between quasiparticles, the phase coherence time of quasiparticles and the resistance vs. temperature, in order to probe the interaction processes which are relevant at low temperatures. We find that the energy exchange rate is higher than expected from the theory of electron-electron interactions, and that it has a different energy dependence. The dephasing time is constant at temperatures between 8 K and 0.5 K, and it increases below 0.5 K. The magnetoresistance is negative at large field scales, and the resistance decreases logarithmically with increasing temperatures, indicating the presence of magnetic impurities, probably Fe. Whereas resistivity and phase coherence measurements can be attributed to magnetic impurities, the question is raised whether these magnetic impurities could also mediate energy exchanges between quasiparticles.Comment: latex pothier.tex, 12 files, 15 pages in: Proceedings of the NATO Advanced Research Workshop on Size Dependent Magnetic Scattering, Pesc, Hungary, May 28 - June 1st, 2000 Chandrasekhar V., Van Haesendonck C. eds (Kluwer, 2001) [SPEC-S00/083

European farmers' experiences with precision livestock farming systems

In the public debate about modern animal production methods, the voice of the farmer is rarely heard. Little is known about the daily work and economic pressure the single farmer is exposed to and what he thinks and feels about the increasingly complex production systems with demanding new control and monitoring technologies such as PLF. In a limited survey based on 21 farm visits (nine pig, five broiler, and seven dairy farms) in 10 EU countries, the knowledge of farmers on options and opportunities of precision livestock farming (PLF) technologies in modern animal production systems was investigated. The farmers were asked by personal free-format interviews face to face on their farms. Most pig and poultry farms were visited in 2014, just after the installation of the PLF technology in the farms and 2016 again after 2 yr of experience. The dairy farms could be visited only once in 2016. All farmers who get sufficient support from the providers developed a positive to very positive attitude to the real-time monitoring PLF systems. This applies for pig, broiler, and cow farms. Broiler farmers were more open to PLF than pig farmers. All farmers emphasized with few exceptions that the personal contact to the animals cannot be replaced by video cameras, but the PLF systems can be of great help in daily life. They enable the farmer to recognize problems significantly earlier than with conventional methods. These techniques are not only helpful and animal friendly, they may also assist to bridge the presently existing gap between producers and consumers by transparency of production. Drawbacks are the relative high prices for PLF equipment, sometimes poor maintenance service by the delivering companies, and the lack of broader experience with the systems in practice. Although one farmer responded after 2 yr of experience with his PLF system that he would not miss it anymore and that he understands his animals much better since he uses PLF monitoring, there is an urgent need for more and wider experiences. It is recommended to further test and develop PLF technologies in demonstration farms under practical conditions. It seems that PLF technologies can play an important role in the development of a future-oriented, sustainable, animal-friendly, and efficient livestock production with healthy animals