Discovery of the Lanthipeptide Curvocidin†and Structural Insights into its Trifunctional Synthetase CuvL.

Lanthipeptides are ribosomally-synthesized natural products from bacteria featuring stable thioether-crosslinks and various bioactivities. Herein, we report on a new clade of tricyclic class-IV lanthipeptides with curvocidin from Thermomonospora curvata as its first representative. We obtained crystal structures of the corresponding lanthipeptide synthetase CuvL that showed a circular arrangement of its kinase, lyase and cyclase domains, forming a central reaction chamber for the iterative substrate processing involving nine catalytic steps. The combination of experimental data and artificial intelligence-based structural models identified the N-terminal subdomain of the kinase domain as the primary site of substrate recruitment. The ribosomal precursor peptide of curvocidin employs an amphipathic α-helix in its leader region as an anchor to CuvL, while its substrate core shuttles within the central reaction chamber. Our study thus reveals general principles of domain organization and substrate recruitment of class-IV and class-III lanthipeptide synthetases.

Effects of Four Different EEG-Neurofeedback Reinforcement Types in Healthy Individuals - A Pilot Study.

EEG neurofeedback (EEG-NFB) is a promising tool for the treatment of depressive disorders. However, many methods for the presentation of neurobiological reactions are available and it is widely unknown which of these feedback options are preferrable. Moreover, the influence of motivation on NFB training success is insufficiently studied. This study analyzed the efficacy of a novel EEG protocol (FC3/Pz) based on findings for NFB in depression. The role of four feedback options (Rumination, Anxiety, Meditation Master, Moving Art) from the NFB software "Brain Assistant" and motivation in EEG-based NFB performance was studied. Regarding "Anxiety" and "Rumination" visual feedback was used to evoke emotions; reinforcement (both negative and positive operant conditioning) was continuous. Regarding "Meditation Master" visual feedback was combined with continuous positive reinforcement. Regarding "Moving Art" 20-min calm nature films with neutral character were used; both visual and auditive feedback were applied. The reinforcement was positive and continuous. 13 healthy participants completed 15 EEG sessions over four months combining simultaneous frontal (aims: reduction of theta-, alpha- and high beta-activity, increase of low and mid beta-activity) and parietal training (aims: reduction of theta-, alpha 1-, mid and high beta-activity, increase of alpha 2- and low beta-activity). We observed significantly more pronounced percentage change in the expected direction for Anxiety than Moving Art (mean difference = 3.32; p = 0.003). The association between motivation and performance was non-significant. Based on these results we conclude that feedback with both negative and positive operant conditioning and emotion evoking effects should be preferred.

Numaswitch: an efficient high-titer expression platform to produce peptides and small proteins.

The production of peptides as active pharmaceutical ingredients (APIs) by recombinant technologies is of emerging interest. A reliable production platform, however, is still missing due the inherent characteristics of peptides such as proteolytic sensitivity, aggregation and cytotoxicity. We have developed a new technology named Numaswitch solving present limitations. Numaswitch was successfully employed for the production of diverse peptides and small proteins varying in length, physicochemical and functional characteristics, including Teriparatide, Linaclotide, human ß-amyloid and Serum amyloid A3. Additionally, the potential of Numaswitch for a cost-efficient commercial production is demonstrated yielding > 2 g Teriparatide per liter fermentation broth in a quality meeting API standard.

The Computer-based Health Evaluation Software (CHES): a software for electronic patient-reported outcome monitoring.

BACKGROUND: Patient-reported Outcomes (PROs) capturing e.g., quality of life, fatigue, depression, medication side-effects or disease symptoms, have become important outcome parameters in medical research and daily clinical practice. Electronic PRO data capture (ePRO) with software packages to administer questionnaires, storing data, and presenting results has facilitated PRO assessment in hospital settings. Compared to conventional paper-pencil versions of PRO instruments, ePRO is more economical with regard to staff resources and time, and allows immediate presentation of results to the medical staff.The objective of our project was to develop software (CHES - Computer-based Health Evaluation System) for ePRO in hospital settings and at home with a special focus on the presentation of individual patient's results. METHODS: Following the Extreme Programming development approach architecture was not fixed up-front, but was done in close, continuous collaboration with software end users (medical staff, researchers and patients) to meet their specific demands. Developed features include sophisticated, longitudinal charts linking patients' PRO data to clinical characteristics and to PRO scores from reference populations, a web-interface for questionnaire administration, and a tool for convenient creating and editing of questionnaires. RESULTS: By 2012 CHES has been implemented at various institutions in Austria, Germany, Switzerland, and the UK and about 5000 patients participated in ePRO (with around 15000 assessments in total). Data entry is done by the patients themselves via tablet PCs with a study nurse or an intern approaching patients and supervising questionnaire completion. DISCUSSION: During the last decade several software packages for ePRO have emerged for different purposes. Whereas commercial products are available primarily for ePRO in clinical trials, academic projects have focused on data collection and presentation in daily clinical practice and on extending cancer registries with PRO data. CHES includes several features facilitating the use of PRO data for individualized medical decision making. With its web-interface it allows ePRO also when patients are home. Thus, it provides complete monitoring of patients'physical and psychosocial symptom burden.