Combinatorial biosynthesis in yeast leads to over 200 diterpenoids.

Diterpenoids form a diverse group of natural products, many of which are or could become pharmaceuticals or industrial chemicals. The modular character of diterpene biosynthesis and the promiscuity of the enzymes involved make combinatorial biosynthesis a promising approach to generate libraries of diverse diterpenoids. Here, we report on the combinatorial assembly in yeast of ten diterpene synthases producing (+)-copalyl diphosphate-derived backbones and four cytochrome P450 oxygenases (CYPs) in diverse combinations. This resulted in the production of over 200 diterpenoids. Based on literature and chemical database searches, 162 of these compounds can be considered new-to-Nature. The CYPs accepted most substrates they were given but remained regioselective with few exceptions. Our results provide the basis for the systematic exploration of the diterpenoid chemical space in yeast using sequence databases.

Selected ice nanoparticle accelerator hypervelocity impact mass spectrometer (SELINA-HIMS): features and impacts of charged particles.

The selected ice nanoparticle accelerator, SELINA, was used to prepare beams of single ice particles with positive or negative charge. Positively charged particles were prepared from deionized water and 0.05-0.2 molar solutions of sodium chloride in water, and negatively charged ice particles were generated from water without salt. Depending on the electrospray source configuration, the measured particles vary from 50 to 1000 nm in diameter. The kinetic energy per charge for all particles was set to 200 eV by the collisional equilibration in quadrupoles, which resulted in primary velocities up to 600 m/s for the lowest m/z particles. The electrospray ionization and thus particle formation from SELINA become less efficient with increasing salt concentration, resulting in a lower detected particle frequency and size. Good instrument operation is achievable for concentrations below 0.2 M. After we have verified and characterized positively and negatively charged ice particles, we have combined SELINA with a target and time-of-flight spectrometer for a 'proof-of-principle' post acceleration of 120 nm particles towards hypervelocity (v ~ 3000 m/s) and detection of fragments from the particle impact (SELINA-HIMS). General conditions are discussed for the acceleration of particles between 50 and 1000 nm to velocities well above 3000 m/s with SELINA-HIMS instrument. This article is part of the theme issue 'Dust in the Solar System and beyond'.

The DESTINY+ Dust Analyser - a dust telescope for analysing cosmic dust dynamics and composition.

The DESTINY+(Demonstration and Experiment of Space Technology for INterplanetary voYage with Phaethon fLyby and dUst Science) Dust Analyser (DDA) is a state-of-the-art dust telescope for the in situ analysis of cosmic dust particles. As the primary scientific payload of the DESTINY+ mission, it serves the purpose of characterizing the dust environment within the Earth-Moon system, investigating interplanetary and interstellar dust populations at 1 AU from the Sun and studying the dust cloud enveloping the asteroid (3200) Phaethon. DDA features a two-axis pointing platform for increasing the accessible fraction of the sky. The instrument combines a trajectory sensor with an impact ionization time-of-flight mass spectrometer, enabling the correlation of dynamical, physical and compositional properties for individual dust grains. For each dust measurement, a set of nine signals provides the surface charge, particle size, velocity vector, as well as the atomic, molecular and isotopic composition of the dust grain. With its capabilities, DDA is a key asset in advancing our understanding of the cosmic dust populations present along the orbit of DESTINY+. In addition to providing the scientific context, we are presenting an overview of the instrument's design and functionality, showing first laboratory measurements and giving insights into the observation planning. This article is part of a theme issue 'Dust in the Solar System and beyond'.

Macromolecular organic compounds from the depths of Enceladus.

Saturn's moon Enceladus harbours a global water ocean 1 , which lies under an ice crust and above a rocky core 2 . Through warm cracks in the crust 3 a cryo-volcanic plume ejects ice grains and vapour into space4-7 that contain materials originating from the ocean8,9. Hydrothermal activity is suspected to occur deep inside the porous core10-12, powered by tidal dissipation 13 . So far, only simple organic compounds with molecular masses mostly below 50 atomic mass units have been observed in plume material6,14,15. Here we report observations of emitted ice grains containing concentrated and complex macromolecular organic material with molecular masses above 200 atomic mass units. The data constrain the macromolecular structure of organics detected in the ice grains and suggest the presence of a thin organic-rich film on top of the oceanic water table, where organic nucleation cores generated by the bursting of bubbles allow the probing of Enceladus' organic inventory in enhanced concentrations.

Unravelling variations: an examination of entry point selection in proximal femoral cephalomedullary nailing.

BACKGROUND: The exact positioning of the cephalomedullary (CM) nail entry point for managing femoral fractures remains debatable, with significant implications for fracture reduction and postoperative complications. This study aimed to explore the variability in the selection of the entry point among trauma surgeons, hypothesizing potential differences and their association with surgeon experience. METHODS: In this prospective multicenter study, 16 participants, ranging from residents to senior specialists, partook in a simulation wherein they determined the optimal entry point for the implantation of a proximal femoral nail antirotation (PFN-A; DePuy Synthes) in various femora. The inter- and intra-observer variability was calculated, along with comprehensive descriptive statistical analysis, to assess the variability in entry point selection and the impact of surgeon experience. RESULTS: In this study, the mean distance from the selected entry points to the calculated mean entry point was 3.98 mm, with a smaller distance observed among surgeons with more than 500 implantations (ANOVA, p = 0.050). Intra-surgeon variability for identical femora averaged at 5.14 mm, showing no significant differences across various levels of surgical experience or training. Notably, 13.6% of selected entry points would not allow a proper intramedullary positioning of the implant, thereby rendering anatomical repositioning unfeasible. Among these impossible entry points, a significant skew towards anterior placement was observed (70.6% of the impossible entry points), with a smaller fraction being overly lateral (27.5%) or medial (13.7%). On a patient level, the impossibility rate varied widely from 0 to 35% among the different femora examined, with a significantly higher rate seen in younger patients (mean age 55.02 versus 60.32; t-test for independent samples, p = 0.04). CONCLUSIONS: Significant variations exist in surgeons' selection of entry points for proximal femoral nailing, underscoring the task's complexity. Experience does not prevent the choice of unfeasible entry points, emphasizing the inadequacy of a universal approach and pointing towards the necessity for a patient-specific strategy for improved outcomes. TRIAL REGISTRATION NUMBER: DRKS00032465.

Characterization of patterns and variability in the dynamics of outdoor aquatic mesocosms: exploring the capabilities and challenges in data supporting aquatic system models.

Aquatic mesocosms are complex test systems used within regulatory risk assessment of plant protection products. These model ecosystems allow researchers to capture interactions of multiple species under realistic environmental conditions. They enable assessment of direct and indirect effects of stressors at all trophic levels (i.e., from primary producers to secondary consumers) and impacts on ecosystem functions. Due to the limited ability to test the multitude of potential exposure scenarios, cross-linking aquatic mesocosm studies with virtual mesocosms, i.e., aquatic system models (ASMs), can serve to meet the demand for more environmental realism and ecological relevance in risk assessment. In this study, full control data sets from seven aquatic mesocosm studies conducted at a single test facility under GLP were analysed graphically and using descriptive statistics. Thereby, not only a comprehensive data base but also an insight into the species present, their dynamics over time, and variability in unchallenged mesocosms was observed. While consistency in dynamics could be discerned for physical and chemical parameters, variability was evident for several biological endpoints. This variability points to amplification of small differences over time as well as to stochastic processes. The outline of existing gaps and uncertainties in data leads to the estimation of what can be expected to be captured and predicted by ASMs.

High-Throughput Virtual Screening and Validation of a SARS-CoV-2 Main Protease Noncovalent Inhibitor.

Despite the recent availability of vaccines against the acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the search for inhibitory therapeutic agents has assumed importance especially in the context of emerging new viral variants. In this paper, we describe the discovery of a novel noncovalent small-molecule inhibitor, MCULE-5948770040, that binds to and inhibits the SARS-Cov-2 main protease (Mpro) by employing a scalable high-throughput virtual screening (HTVS) framework and a targeted compound library of over 6.5 million molecules that could be readily ordered and purchased. Our HTVS framework leverages the U.S. supercomputing infrastructure achieving nearly 91% resource utilization and nearly 126 million docking calculations per hour. Downstream biochemical assays validate this Mpro inhibitor with an inhibition constant (Ki) of 2.9 µM (95% CI 2.2, 4.0). Furthermore, using room-temperature X-ray crystallography, we show that MCULE-5948770040 binds to a cleft in the primary binding site of Mpro forming stable hydrogen bond and hydrophobic interactions. We then used multiple µs-time scale molecular dynamics (MD) simulations and machine learning (ML) techniques to elucidate how the bound ligand alters the conformational states accessed by Mpro, involving motions both proximal and distal to the binding site. Together, our results demonstrate how MCULE-5948770040 inhibits Mpro and offers a springboard for further therapeutic design.

[Outcome Quality of Medical Rehab in the Mirror of Singular and Multiple Outcome Criteria - Where is Psychosomatic Rehab when Comparing Indications?] / Ergebnisqualität medizinischer Rehabilitation im Spiegel singulärer und multipler Ergebniskriterien ­ Wo steht die Psychosomatische Rehabilitation im Indikationenvergleich?

BACKGROUND: A series of studies proves a good outcome quality of psychosomatic rehabilitation. However, outcome-related comparisons with other indications are hardly available. METHODS: As part of a multicenter study, n=6608 rehabilitants from the indications psychosomatics, cardiology, neurology, oncology and orthopedics were checked regarding starting features and longer-term outcome quality (one-point survey 1 year after the end of the rehab). With a generic measurement and evaluation approach, direct and quasi-indirect change measurements and status measurements were made. In addition to comparing singular and multiple outcome criteria ("Patient Reported Outcomes", PRO), outcome criteria from the rehab statistics database (RSD) have also been checked. RESULTS: The 5 indication groups differ in both starting and process characteristics as well as in the short and longer-term outcome criteria. However, the effect sizes of the associations are mostly low. In all indications, there are positive changes in the field of health-related characteristics. The highest pre-post effect sizes are mostly found in psychosomatics, the least in neurology. In all indications, social security contributions in the first year after rehab are a bit declining - least in oncology, most clearly in neurology. Despite the biggest pre-post effects sizes in the health-related features, the rehabilitants of psychosomatics are less satisfied with the rehab and evaluate the benefits of rehab less positive. At the level of multiple outcome criteria, the indications - except neurology - are relatively little different. The multiple outcome criterion can be predicted to 28% from starting and process characteristics. Best predictor is the user sided rating regarding the job-related orientation of the rehab. CONCLUSION: The study once again proves a good longer-term outcome quality of psychosomatic rehab. However, it also shows that the longer-term outcome quality of all major indications measured by means of multiple outcome criteria is at a similar level (except neurology).Possible limitations of the study result from the one-point measurement and the resulting mode of change measurement.

[Rehabilitation Outcome and Therapeutic Alliance of Inpatients of Psychosomatic Rehabilitation with Pension Request]. / Psychosomatische Rehabilitation bei Patientinnen und Patienten mit Rentenüberlegungen: Reha-Erfolg und therapeutische Beziehung.

OBJECTIVE: Inpatients of psychosomatic rehabilitation with a latent or manifest desire for a pension are a risk group with regard to successful participation in rehabilitation and return to work. Reaching this group of patients, is both a legal requirement (rehabilitation before retirement), and important from an economic and health perspective. This study examines patients who are considering applying for a reduced earning capacity. The questions are whether these patients differ in socio-demographic variables and the success of rehabilitation from patients without pension considerations, and which patients benefitted from their rehabilitation despite pension considerations. Particular attention is paid to the therapeutic relationship. METHODS: Data are routinely collected from patients of 2 rehabilitation clinics who were undergoing inpatient psychosomatic treatment between October 2012 and February 2017 (n=8503). RESULTS: Inpatients with pension considerations were slightly older, more often incapacitated for work and more often receiving unemployment benefits. Both groups reported a significant reduction in symptoms over the course of the rehabilitation. Within the group considering retirement, patients who reported an improvement in symptoms reported a more positive relationship satisfaction. DISCUSSION/CONCLUSION: Patients who are thinking about retirement often have difficult socio-demographic and health-related conditions for successful rehabilitation. Despite these conditions, many report an improvement in psychosomatic symptoms and a positive therapeutic relationship.

Ontology-based computer aid for the automation of HAZOP studies.

Hazard and Operability (HAZOP) studies are conducted to identify and assess potential hazards which originate from processes, equipment, and process plants. These studies are human-centered processes that are time and labor-intensive. Also, extensive expertise and experience in the field of process safety engineering are required. There have been several attempts by different research groups to (semi-)automate HAZOP studies in the past. Within this research, a knowledge-based framework for the automatic generation of HAZOP worksheets was developed. Compared to other approaches, the focus is on representing semantic relationships between HAZOP relevant concepts under consideration of the degree of abstraction. In the course of this, expert knowledge from the process and plant safety (PPS) domain is embedded within the ontological model. Based on that, a reasoning algorithm based on semantic reasoners is developed to identify hazards and operability issues in a HAZOP similar manner. An advantage of the proposed method is that by modeling causal relationships between HAZOP concepts, automatically generated but meaningless scenarios can be avoided. The results of the enhanced causation model are high quality extended HAZOP worksheets. The developed methodology is applied within a case study that involves a hexane storage tank. The quality and quantity of the automatically generated results agree with the original worksheets. Thus the ontology-based reasoning algorithm is well-suited to identify hazardous scenarios and operability issues. Node-based analyses involving multiple process units can also be carried out by a slight adjustment of the method. The presented method can help to support HAZOP study participants and non-experts in conducting HAZOP studies.

Changes in health in the countries of the UK and 150 English Local Authority areas 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016.

BACKGROUND: Previous studies have reported national and regional Global Burden of Disease (GBD) estimates for the UK. Because of substantial variation in health within the UK, action to improve it requires comparable estimates of disease burden and risks at country and local levels. The slowdown in the rate of improvement in life expectancy requires further investigation. We use GBD 2016 data on mortality, causes of death, and disability to analyse the burden of disease in the countries of the UK and within local authorities in England by deprivation quintile. METHODS: We extracted data from the GBD 2016 to estimate years of life lost (YLLs), years lived with disability (YLDs), disability-adjusted life-years (DALYs), and attributable risks from 1990 to 2016 for England, Scotland, Wales, Northern Ireland, the UK, and 150 English Upper-Tier Local Authorities. We estimated the burden of disease by cause of death, condition, year, and sex. We analysed the association between burden of disease and socioeconomic deprivation using the Index of Multiple Deprivation. We present results for all 264 GBD causes of death combined and the leading 20 specific causes, and all 84 GBD risks or risk clusters combined and 17 specific risks or risk clusters. FINDINGS: The leading causes of age-adjusted YLLs in all UK countries in 2016 were ischaemic heart disease, lung cancers, cerebrovascular disease, and chronic obstructive pulmonary disease. Age-standardised rates of YLLs for all causes varied by two times between local areas in England according to levels of socioeconomic deprivation (from 14 274 per 100 000 population [95% uncertainty interval 12 791-15 875] in Blackpool to 6888 [6145-7739] in Wokingham). Some Upper-Tier Local Authorities, particularly those in London, did better than expected for their level of deprivation. Allowing for differences in age structure, more deprived Upper-Tier Local Authorities had higher attributable YLLs for most major risk factors in the GBD. The population attributable fractions for all-cause YLLs for individual major risk factors varied across Upper-Tier Local Authorities. Life expectancy and YLLs have improved more slowly since 2010 in all UK countries compared with 1990-2010. In nine of 150 Upper-Tier Local Authorities, YLLs increased after 2010. For attributable YLLs, the rate of improvement slowed most substantially for cardiovascular disease and breast, colorectal, and lung cancers, and showed little change for Alzheimer's disease and other dementias. Morbidity makes an increasing contribution to overall burden in the UK compared with mortality. The age-standardised UK DALY rate for low back and neck pain (1795 [1258-2356]) was higher than for ischaemic heart disease (1200 [1155-1246]) or lung cancer (660 [642-679]). The leading causes of ill health (measured through YLDs) in the UK in 2016 were low back and neck pain, skin and subcutaneous diseases, migraine, depressive disorders, and sense organ disease. Age-standardised YLD rates varied much less than equivalent YLL rates across the UK, which reflects the relative scarcity of local data on causes of ill health. INTERPRETATION: These estimates at local, regional, and national level will allow policy makers to match resources and priorities to levels of burden and risk factors. Improvement in YLLs and life expectancy slowed notably after 2010, particularly in cardiovascular disease and cancer, and targeted actions are needed if the rate of improvement is to recover. A targeted policy response is also required to address the increasing proportion of burden due to morbidity, such as musculoskeletal problems and depression. Improving the quality and completeness of available data on these causes is an essential component of this response. FUNDING: Bill & Melinda Gates Foundation and Public Health England.

Synthesis of stable isotope-labeled chloroquine and amodiaquine and their metabolites.

Anti-malaria drugs chloroquine and amodiaquine and their metabolites were synthesized to incorporate 13 C and 15 N starting from U-13 C-labeled benzene to give M + 7 isotopomers. Chloroquine and its metabolites were prepared from 7-chloro-1,2,3,4-tetrahydroquinolin-4-one through an aryl substitution with the corresponding amines; and the amodiaquine and its metabolites were prepared from 4,7-dichloroquinoline in a similar fashion.

Size distribution of particles in Saturn's rings from aggregation and fragmentation.

Saturn's rings consist of a huge number of water ice particles, with a tiny addition of rocky material. They form a flat disk, as the result of an interplay of angular momentum conservation and the steady loss of energy in dissipative interparticle collisions. For particles in the size range from a few centimeters to a few meters, a power-law distribution of radii, ~r(-q) with q ≈ 3, has been inferred; for larger sizes, the distribution has a steep cutoff. It has been suggested that this size distribution may arise from a balance between aggregation and fragmentation of ring particles, yet neither the power-law dependence nor the upper size cutoff have been established on theoretical grounds. Here we propose a model for the particle size distribution that quantitatively explains the observations. In accordance with data, our model predicts the exponent q to be constrained to the interval 2.75 ≤ q ≤ 3.5. Also an exponential cutoff for larger particle sizes establishes naturally with the cutoff radius being set by the relative frequency of aggregating and disruptive collisions. This cutoff is much smaller than the typical scale of microstructures seen in Saturn's rings.

Eye blink detection for different driver states in conditionally automated driving and manual driving using EOG and a driver camera.

In this article, we examine the performance of different eye blink detection algorithms under various constraints. The goal of the present study was to evaluate the performance of an electrooculogram- and camera-based blink detection process in both manually and conditionally automated driving phases. A further comparison between alert and drowsy drivers was performed in order to evaluate the impact of drowsiness on the performance of blink detection algorithms in both driving modes. Data snippets from 14 monotonous manually driven sessions (mean 2 h 46 min) and 16 monotonous conditionally automated driven sessions (mean 2 h 45 min) were used. In addition to comparing two data-sampling frequencies for the electrooculogram measures (50 vs. 25 Hz) and four different signal-processing algorithms for the camera videos, we compared the blink detection performance of 24 reference groups. The analysis of the videos was based on very detailed definitions of eyelid closure events. The correct detection rates for the alert and manual driving phases (maximum 94%) decreased significantly in the drowsy (minus 2% or more) and conditionally automated (minus 9% or more) phases. Blinking behavior is therefore significantly impacted by drowsiness as well as by automated driving, resulting in less accurate blink detection.

[Testing the Psychometric Properties of the Helping Alliance Questionnaire (HAQ) in Different Health Care Settings]. / Psychometrische Überprüfung des Helping Alliance Questionnaire (HAQ) in unterschiedlichen Versorgungssettings.

Introduction The Helping Alliance Questionnaire developed by Luborsky was psychometricly examined for the first time and translated into German by Bassler et al. in the mid-1990s. It consists of 11 Items, which are summarized to the scales "relation to the therapist" and "satisfaction with therapeutic outcome". HAQ is now one of the most used instruments to measure therapeutic alliance. The goal of this study was to test the psychometric properties based on three large patient samples in different treatment settings. Material and methods Analyses were conducted based on 2 samples of patients in inpatient psychosomatic/psychotherapeutic rehabilitation (n=655, n=2494) and one sample in outpatient psychotherapy (n=1477). Exploratory factor analyses and for verification confirmatory factor analyses were applied. Furthermore reliability and validity analysis were conducted. Results The 2-factorial structure found in literature was replicated with an increasing stability at the end of treatment. Item 2 and 3 had inconsistent factor loadings at different points of measurement and study. Reliability and validity indices were satisfying to good. The fit of the model, on the other hand, was less satisfying and suggests a solution without Item 2 and 3. For reasons of content as well as for reasons of dissemination, it is pleaded for the maintenance of the existing item assignments in the HAQ for the present.

Changes in health in England, with analysis by English regions and areas of deprivation, 1990-2013: a systematic analysis for the Global Burden of Disease Study 2013.

BACKGROUND: In the Global Burden of Disease Study 2013 (GBD 2013), knowledge about health and its determinants has been integrated into a comparable framework to inform health policy. Outputs of this analysis are relevant to current policy questions in England and elsewhere, particularly on health inequalities. We use GBD 2013 data on mortality and causes of death, and disease and injury incidence and prevalence to analyse the burden of disease and injury in England as a whole, in English regions, and within each English region by deprivation quintile. We also assess disease and injury burden in England attributable to potentially preventable risk factors. England and the English regions are compared with the remaining constituent countries of the UK and with comparable countries in the European Union (EU) and beyond. METHODS: We extracted data from the GBD 2013 to compare mortality, causes of death, years of life lost (YLLs), years lived with a disability (YLDs), and disability-adjusted life-years (DALYs) in England, the UK, and 18 other countries (the first 15 EU members [apart from the UK] and Australia, Canada, Norway, and the USA [EU15+]). We extended elements of the analysis to English regions, and subregional areas defined by deprivation quintile (deprivation areas). We used data split by the nine English regions (corresponding to the European boundaries of the Nomenclature for Territorial Statistics level 1 [NUTS 1] regions), and by quintile groups within each English region according to deprivation, thereby making 45 regional deprivation areas. Deprivation quintiles were defined by area of residence ranked at national level by Index of Multiple Deprivation score, 2010. Burden due to various risk factors is described for England using new GBD methodology to estimate independent and overlapping attributable risk for five tiers of behavioural, metabolic, and environmental risk factors. We present results for 306 causes and 2337 sequelae, and 79 risks or risk clusters. FINDINGS: Between 1990 and 2013, life expectancy from birth in England increased by 5·4 years (95% uncertainty interval 5·0-5·8) from 75·9 years (75·9-76·0) to 81·3 years (80·9-81·7); gains were greater for men than for women. Rates of age-standardised YLLs reduced by 41·1% (38·3-43·6), whereas DALYs were reduced by 23·8% (20·9-27·1), and YLDs by 1·4% (0·1-2·8). For these measures, England ranked better than the UK and the EU15+ means. Between 1990 and 2013, the range in life expectancy among 45 regional deprivation areas remained 8·2 years for men and decreased from 7·2 years in 1990 to 6·9 years in 2013 for women. In 2013, the leading cause of YLLs was ischaemic heart disease, and the leading cause of DALYs was low back and neck pain. Known risk factors accounted for 39·6% (37·7-41·7) of DALYs; leading behavioural risk factors were suboptimal diet (10·8% [9·1-12·7]) and tobacco (10·7% [9·4-12·0]). INTERPRETATION: Health in England is improving although substantial opportunities exist for further reductions in the burden of preventable disease. The gap in mortality rates between men and women has reduced, but marked health inequalities between the least deprived and most deprived areas remain. Declines in mortality have not been matched by similar declines in morbidity, resulting in people living longer with diseases. Health policies must therefore address the causes of ill health as well as those of premature mortality. Systematic action locally and nationally is needed to reduce risk exposures, support healthy behaviours, alleviate the severity of chronic disabling disorders, and mitigate the effects of socioeconomic deprivation. FUNDING: Bill & Melinda Gates Foundation and Public Health England.

Low chromatic Fresnel lens for broadband attosecond XUV pulse applications.

Fresnel zone plates show a great potential in achieving high spatial resolution imaging or focusing for XUV and soft/hard X-ray radiation, however they are usually strictly monochromatic due to strong chromatic dispersion and thus do not support broad radiation spectra, preventing their application to attosecond XUV pulses. Here we report on the design and theoretical simulations based on the design of an achromatic hybrid optics combining both, a refractive and diffractive lens in one optical element. We are able to show by calculation that the chromatic dispersion along the optical axis can be greatly reduced compared to a standard Fresnel zone plate while preserving the temporal structure of the attosecond XUV pulses at focus.

Structure and Absolute Configuration of Pseudohygrophorones A(12) and B(12), Alkyl Cyclohexenone Derivatives from Hygrophorus abieticola (Basidiomycetes).

Pseudohygrophorones A(12) (1) and B(12) (2), the first naturally occurring alkyl cyclohexenones from a fungal source, and the recently reported hygrophorone B(12) (3) have been isolated from fruiting bodies of the basidiomycete Hygrophorus abieticola Krieglst. ex Gröger & Bresinsky. Their structures were assigned on the basis of extensive one- and two-dimensional NMR spectroscopic analysis as well as ESI-HRMS measurements. The absolute configuration of the three stereogenic centers in the diastereomeric compounds 1 and 2 was established with the aid of (3)JH,H and (4)JH,H coupling constants, NOE interactions, and conformational analysis in conjunction with quantum chemical CD calculations. It was concluded that pseudohygrophorone A(12) (1) is 4S,5S,6S configured, while pseudohygrophorone B(12) (2) was identified as the C-6 epimer of 1, corresponding to the absolute configuration 4S,5S,6R. In addition, the mass spectrometric fragmentation behavior of 1-3 obtained by the higher energy collisional dissociation method allows a clear distinction between the pseudohygrophorones (1 and 2) and hygrophorone B(12) (3). The isolated compounds 1-3 exhibited pronounced activity against phytopathogenic organisms.

Chilenopeptins A and B, Peptaibols from the Chilean Sepedonium aff. chalcipori KSH 883.

The Chilean Sepedonium aff. chalcipori strain KSH 883, isolated from the endemic Boletus loyo Philippi, was studied in a polythetic approach based on chemical, molecular, and biological data. A taxonomic study of the strain using molecular data of the ITS, EF1-α, and RPB2 barcoding genes confirmed the position of the isolated strain within the S. chalcipori clade, but also suggested the separation of this clade into three different species. Two new linear 15-residue peptaibols, named chilenopeptins A (1) and B (2), together with the known peptaibols tylopeptins A (3) and B (4) were isolated from the semisolid culture of strain KSH 883. The structures of 1 and 2 were elucidated on the basis of HRESIMS(n) experiments in conjunction with comprehensive 1D and 2D NMR analysis. Thus, the sequence of chilenopeptin A (1) was identified as Ac-Aib(1)-Ser(2)-Trp(3)-Aib(4)-Pro(5)-Leu(6)-Aib(7)-Aib(8)-Gln(9)-Aib(10)-Aib(11)-Gln(12)-Aib(13)-Leu(14)-Pheol(15), while chilenopeptin B (2) differs from 1 by the replacement of Trp(3) by Phe(3). Additionally, the total synthesis of 1 and 2 was accomplished by a solid-phase approach, confirming the absolute configuration of all chiral amino acids as l. Both the chilenopeptins (1 and 2) and tylopeptins (3 and 4) were evaluated for their potential to inhibit the growth of phytopathogenic organisms.

Generation of circularly polarized high harmonic radiation using a transmission multilayer quarter waveplate.

High harmonic radiation is meanwhile nearly extensively used for the spectroscopic investigation of electron dynamics with ultimate time resolution. The majority of high harmonic beamlines provide linearly polarized radiation created in a gas target. However, circular polarization greatly extends the spectroscopic possibilities for high harmonics, especially in the analysis of samples with chirality or prominent spin polarization. We produced a free-standing multilayer foil as a transmission EUV quarter waveplate and applied it for the first time to high harmonic radiation. We measured a broadband (4.6 eV FWHM) ellipticity of 75% at 66 eV photon energy with a transmission efficiency of 5%. The helicity is switchable and the ellipticity can be adjusted to lower values by angle tuning. As a single element it can be easily integrated in any existing harmonic beamline without major changes.