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Incorporation of noncanonical amino acids (ncAAs) with bioorthogonal reactive groups by amber suppression allows the generation of synthetic proteins with desired novel properties. Such modified molecules are in high demand for basic research and therapeutic applications such as cancer treatment and in vivo imaging. The positioning of the ncAA-responsive codon within the protein's coding sequence is critical in order to maintain protein function, achieve high yields of ncAA-containing protein, and allow effective conjugation. Cell-free ncAA incorporation is of particular interest due to the open nature of cell-free systems and their concurrent ease of manipulation. In this study, we report a straightforward workflow to inquire ncAA positions in regard to incorporation efficiency and protein functionality in a Chinese hamster ovary (CHO) cell-free system. As a model, the well-established orthogonal translation components Escherichia coli tyrosyl-tRNA synthetase (TyrRS) and tRNATyr(CUA) were used to site-specifically incorporate the ncAA p-azido-l-phenylalanine (AzF) in response to UAG codons. A total of seven ncAA sites within an anti-epidermal growth factor receptor (EGFR) single-chain variable fragment (scFv) N-terminally fused to the red fluorescent protein mRFP1 and C-terminally fused to the green fluorescent protein sfGFP were investigated for ncAA incorporation efficiency and impact on antigen binding. The characterized cell-free dual fluorescence reporter system allows screening for ncAA incorporation sites with high incorporation efficiency that maintain protein activity. It is parallelizable, scalable, and easy to operate. We propose that the established CHO-based cell-free dual fluorescence reporter system can be of particular interest for the development of antibody-drug conjugates (ADCs).
Law smells
(2022)
Building on the computer science concept of code smells, we initiate the study of law smells, i.e., patterns in legal texts that pose threats to the comprehensibility and maintainability of the law. With five intuitive law smells as running examples-namely, duplicated phrase, long element, large reference tree, ambiguous syntax, and natural language obsession-, we develop a comprehensive law smell taxonomy. This taxonomy classifies law smells by when they can be detected, which aspects of law they relate to, and how they can be discovered. We introduce text-based and graph-based methods to identify instances of law smells, confirming their utility in practice using the United States Code as a test case. Our work demonstrates how ideas from software engineering can be leveraged to assess and improve the quality of legal code, thus drawing attention to an understudied area in the intersection of law and computer science and highlighting the potential of computational legal drafting.
Deep lipidomics in human plasma: cardiometabolic disease risk and effect of dietary fat modulation
(2022)
Background: In blood and tissues, dietary and endogenously generated fatty acids (FAs) occur in free form or as part of complex lipid molecules that collectively represent the lipidome of the respective tissue. We assessed associations of plasma lipids derived from high-resolution lipidomics with incident cardiometabolic diseases and subsequently tested if the identified risk-associated lipids were sensitive to dietary fat modification. Methods: The EPIC Potsdam cohort study (European Prospective Investigation into Cancer and Nutrition) comprises 27 548 participants recruited within an age range of 35 to 65 years from the general population around Potsdam, Germany. We generated 2 disease-specific case cohorts on the basis of a fixed random subsample (n=1262) and all respective cohort-wide identified incident primary cardiovascular disease (composite of fatal and nonfatal myocardial infarction and stroke; n=551) and type 2 diabetes (n=775) cases. We estimated the associations of baseline plasma concentrations of 282 class-specific FA abundances (calculated from 940 distinct molecular species across 15 lipid classes) with the outcomes in multivariable-adjusted Cox models. We tested the effect of an isoenergetic dietary fat modification on risk-associated lipids in the DIVAS randomized controlled trial (Dietary Intervention and Vascular Function; n=113). Participants consumed either a diet rich in saturated FAs (control), monounsaturated FAs, or a mixture of monounsaturated and n-6 polyunsaturated FAs for 16 weeks. Results: Sixty-nine lipids associated (false discovery rate<0.05) with at least 1 outcome (both, 8; only cardiovascular disease, 49; only type 2 diabetes, 12). In brief, several monoacylglycerols and FA16:0 and FA18:0 in diacylglycerols were associated with both outcomes; cholesteryl esters, free fatty acids, and sphingolipids were largely cardiovascular disease specific; and several (glycero)phospholipids were type 2 diabetes specific. In addition, 19 risk-associated lipids were affected (false discovery rate<0.05) by the diets rich in unsaturated dietary FAs compared with the saturated fat diet (17 in a direction consistent with a potential beneficial effect on long-term cardiometabolic risk). For example, the monounsaturated FA-rich diet decreased diacylglycerol(FA16:0) by 0.4 (95% CI, 0.5-0.3) SD units and increased triacylglycerol(FA22:1) by 0.5 (95% CI, 0.4-0.7) SD units. Conclusions: We identified several lipids associated with cardiometabolic disease risk. A subset was beneficially altered by a dietary fat intervention that supports the substitution of dietary saturated FAs with unsaturated FAs as a potential tool for primary disease prevention.
In seismic risk assessment, the sources of uncertainty associated with building exposure modelling have not received as much attention as other components related to hazard and vulnerability. Conventional practices such as assuming absolute portfolio compositions (i.e., proportions per building class) from expert-based assumptions over aggregated data crudely disregard the contribution of uncertainty of the exposure upon earthquake loss models. In this work, we introduce the concept that the degree of knowledge of a building stock can be described within a Bayesian probabilistic approach that integrates both expert-based prior distributions and data collection on individual buildings. We investigate the impact of the epistemic uncertainty in the portfolio composition on scenario-based earthquake loss models through an exposure-oriented logic tree arrangement based on synthetic building portfolios. For illustrative purposes, we consider the residential building stock of Valparaiso (Chile) subjected to seismic ground-shaking from one subduction earthquake. We have found that building class reconnaissance, either from prior assumptions by desktop studies with aggregated data (top-down approach), or from building-by-building data collection (bottom-up approach), plays a fundamental role in the statistical modelling of exposure. To model the vulnerability of such a heterogeneous building stock, we require that their associated set of structural fragility functions handle multiple spectral periods. Thereby, we also discuss the relevance and specific uncertainty upon generating either uncorrelated or spatially cross-correlated ground motion fields within this framework. We successively show how various epistemic uncertainties embedded within these probabilistic exposure models are differently propagated throughout the computed direct financial losses. This work calls for further efforts to redesign desktop exposure studies, while also highlighting the importance of exposure data collection with standardized and iterative approaches.
Hot, compact, hydrogen-deficient pre-white dwarfs (pre-WDs) with effective temperatures of Teff > 70 000 K and a surface gravity of 5.0 < logg < 7.0 are rather rare objects despite recent and ongoing surveys. It is believed that they are the outcome of either single star evolution (late helium-shell flash or late helium-core flash) or binary star evolution (double WD merger). Their study is interesting because the surface elemental abundances reflect the physics of thermonuclear flashes and merger events. Spectroscopically they are divided in three different classes, namely PG1159, O(He), or He-sdO. We present a spectroscopic analysis of five such stars that turned out to have atmospheric parameters in the range Teff = 70 000-80 000 K and logg = 5.2-6.3. The three investigated He-sdOs have a relatively high hydrogen mass fraction (10%) that is unexplained by both single (He core flash) and binary evolution (He-WD merger) scenarios. The O(He) star JL 9 is probably a binary helium-WD merger, but its hydrogen content (6%) is also at odds with merger models. We found that RL 104 is the 'coolest' (Teff = 80 000 K) member of the PG1159 class in a pre-WD stage. Its optical spectrum is remarkable because it exhibits C※ IV lines involving Rydberg states with principal quantum numbers up to n = 22. Its rather low mass (0.48-0.02+0.03 M·) is difficult to reconcile with the common evolutionary scenario for PG1159 stars due to it being the outcome of a (very) late He-shell flash. The same mass-problem faces a merger model of a close He-sdO plus CO WD binary that predicts PG1159-like abundances. Perhaps RL 104 originates from a very late He-shell flash in a CO/He WD formed by a merger of two low-mass He-WDs.
The benefits of counting butterflies: recommendations for a successful citizen science project
(2022)
Citizen science (CS) projects, being popular across many fields of science, have recently also become a popular tool to collect biodiversity data. Although the benefits of such projects for science and policy making are well understood, relatively little is known about the benefits participants get from these projects as well as their personal backgrounds and motivations. Furthermore, very little is known about their expectations. We here examine these aspects, with the citizen science project "German Butterfly Monitoring" as an example. A questionnaire was sent to all participants of the project and the responses to the questionnaire indicated the following: center dot Most transect walkers do not have a professional background in this field, though they do have a high educational level, and are close to retirement, with a high number of females; center dot An important motivation to join the project is to preserve the natural environment and to contribute to scientific knowledge; center dot Participants benefit by enhancing their knowledge about butterflies and especially their ability to identify different species (taxonomic knowledge); center dot Participants do not have specific expectations regarding the project beyond proper management and coordination, but have an intrinsic sense of working for a greater good. The willingness to join a project is higher if the project contributes to the solution of a problem discussed in the media (here, insect decline). Based on our findings from the analysis of the questionnaire we can derive a set of recommendations for establishing a successful CS project. These include the importance of good communication, e.g., by explaining what the (scientific) purpose of the project is and what problems are to be solved with the help of the data collected in the project. The motivation to join a CS project is mostly intrinsic and CS is a good tool to engage people during difficult times such as the COVID-19 pandemic, giving participants the feeling of doing something useful.
In this paper, we employ an experimental paradigm using insights from the psychology of reasoning to investigate the question whether certain modals generate and draw attention to alternatives. The article extends and builds on the methodology and findings of Mascarenhas & Picat (2019). Based on experimental results, they argue that the English epistemic modal might raises alternatives. We apply the same methodology to the English modal allowed to to test different hypotheses regarding the involvement of alternatives in deontic modality. We find commonalities and differences between the two modals we tested. We discuss theoretical consequences for existing semantic analyses of these modals, and argue that reasoning tasks can serve as a diagnostic tool to discover which natural language expressions involve alternatives.
So far, various studies have aimed at decomposing the integrated terrestrial water storage variations observed by satellite gravimetry (GRACE, GRACE-FO) with the help of large-scale hydrological models. While the results of the storage decomposition depend on model structure, little attention has been given to the impact of the way that vegetation is represented in these models. Although vegetation structure and activity represent the crucial link between water, carbon, and energy cycles, their representation in large-scale hydrological models remains a major source of uncertainty. At the same time, the increasing availability and quality of Earth-observation-based vegetation data provide valuable information with good prospects for improving model simulations and gaining better insights into the role of vegetation within the global water cycle. In this study, we use observation-based vegetation information such as vegetation indices and rooting depths for spatializing the parameters of a simple global hydrological model to define infiltration, root water uptake, and transpiration processes. The parameters are further constrained by considering observations of terrestrial water storage anomalies (TWS), soil moisture, evapotranspiration (ET) and gridded runoff ( Q) estimates in a multi-criteria calibration approach. We assess the implications of including varying vegetation characteristics on the simulation results, with a particular focus on the partitioning between water storage components. To isolate the effect of vegetation, we compare a model experiment in which vegetation parameters vary in space and time to a baseline experiment in which all parameters are calibrated as static, globally uniform values. Both experiments show good overall performance, but explicitly including varying vegetation data leads to even better performance and more physically plausible parameter values. The largest improvements regarding TWS and ET are seen in supply-limited (semi-arid) regions and in the tropics, whereas Q simulations improve mainly in northern latitudes. While the total fluxes and storages are similar, accounting for vegetation substantially changes the contributions of different soil water storage components to the TWS variations. This suggests an important role of the representation of vegetation in hydrological models for interpreting TWS variations. Our simulations further indicate a major effect of deeper moisture storages and groundwater-soil moisture-vegetation interactions as a key to understanding TWS variations. We highlight the need for further observations to identify the adequate model structure rather than only model parameters for a reasonable representation and interpretation of vegetation-water interactions.
Guidance of postinfarct myocardial remodeling processes by an epicardial patch system may alleviate the consequences of ischemic heart disease. As macrophages are highly relevant in balancing immune response and regenerative processes their suitable instruction would ensure therapeutic success. A polymeric mesh capable of attracting and instructing monocytes by purely physical cues and accelerating implant degradation at the cell/implant interface is designed. In a murine model for myocardial infarction the meshes are compared to those either coated with extracellular matrix or loaded with induced cardiomyocyte progenitor cells. All implants promote macrophage infiltration and polarization in the epicardium, which is verified by in vitro experiments. 6 weeks post-MI, especially the implantation of the mesh attenuates left ventricular adverse remodeling processes as shown by reduced infarct size (14.7% vs 28-32%) and increased wall thickness (854 mu m vs 400-600 mu m), enhanced angiogenesis/arteriogenesis (more than 50% increase compared to controls and other groups), and improved heart function (ejection fraction = 36.8% compared to 12.7-31.3%). Upscaling as well as process controls is comprehensively considered in the presented mesh fabrication scheme to warrant further progression from bench to bedside.
In the picture-word interference paradigm, participants name pictures while ignoring a written or spoken distractor word. Naming times to the pictures are slowed down by the presence of the distractor word. The present study investigates in detail the impact of distractor and target word properties on picture naming times, building on the seminal study by Miozzo and Caramazza. We report the results of several Bayesian meta-analyses based on 26 datasets. These analyses provide estimates of effect sizes and their precision for several variables and their interactions. They show the reliability of the distractor frequency effect on picture naming latencies (latencies decrease as the frequency of the distractor increases) and demonstrate for the first time the impact of distractor length, with longer naming latencies for trials with longer distractors. Moreover, distractor frequency interacts with target word frequency to predict picture naming latencies. The methodological and theoretical implications of these findings are discussed.
The purpose of the present study was to investigate the moderating effect of technology use for friendship maintenance in the associations between self-isolation during the COVID-19 pandemic and friendship quality, measured 6 months later (Time 2). Participants were 1,567 seventh and eighth graders (51% female; 51% white; M-age = 13.47) from the United States. They completed questionnaires on friendship quality at Time 1, and self-isolation during COVID-19 and technology use for friendship maintenance and friendship quality at Time 2. The findings revealed that self-isolation during COVID-19 was related positively to technology use for friendship maintenance and negatively to Time 2 friendship quality. Higher technology use for friendship maintenance buffered against the negative impacts on friendship quality associated with self-isolation during COVID-19, while lower technology use had the opposite effects on Time 2 friendship quality.
Räuberischer Menschenraub
(2022)
Einen Straftatbestand mit dem Namen „Räuberischer Menschenraub“ gibt es im deutschen Strafrecht nicht. Was es aber gibt, sind wirkliche Straftaten, die wegen ihrer tatsächlichen Bestandteile und deren strafrechtlicher Bedeutung mit „Räuberischer Menschenraub“ zutreffend bezeichnet sind. Die Frage ist daher, welchem Straftatbestand des geltenden Strafrechts eine solche Tat zugeordnet werden kann. Eine neue Entscheidung des BGH gibt Antwort auf die Frage. Der zugrunde liegende Sachverhalt enthält so viele weitere interessante strafrechtliche Elemente, dass er fast unverändert als Strafrechtsaufgabe im Examen Verwendung finden könnte. Die Beschäftigung mit dem Fall ist daher zu empfehlen. Hier soll das Hauptaugenmerk auf die mit dem „Räuberischen Menschenraub“ zusammenhängenden Probleme gerichtet werden.
Vielleicht stellen sich auch andere Benutzer der Berliner S-Bahn hin und wieder die Frage, wer von den Mitfahrenden wohl ein gültiges Ticket dabei hat. Selbstverständlich soll hier niemand diskriminiert werden; aber bei manchem Mitreisenden, der um eine kleine Spende bittet, eine Obdachlosenzeitung offeriert oder musikalische Darbietungen gibt, habe ich manchmal Zweifel. Nicht ganz fernliegend ist dann wohl die Erwägung, dass es sich jedenfalls zum Teil um Mitbürger handelt, deren Einkommens- und Vermögenssituation schlecht ist. Das gibt Anlass zu der Frage, ob unter dieser Voraussetzung die strafrechtliche Beurteilung der – das sei hier des Themas wegen unterstellt – unbefugt erlangten unentgeltlichen Personenbeförderung zu einem anderen Ergebnis führt als bei einem „Schwarzfahrer“, der über genügend Finanzmittel verfügt, um die Fahrt zu bezahlen. Einige Gerichtsentscheidungen zu Fällen, in denen es zwischen einem Taxichauffeur und dem von ihm beförderten Fahrgast zu gewalttätigen Auseinandersetzungen über die – vom Fahrgast verweigerte − Fahrpreisentrichtung gekommen war, suggerieren eine Straflosigkeit des Täters, der keinerlei pfändbares Vermögen hat. Die Entscheidungen betrafen den Erpressungstatbestand (§§ 253, 255 StGB), sind aber vielleicht auch präjudiziell für § 263 StGB und für § 265 a StGB. Dem soll hier nachgegangen werden.
Objective
Problem-drinking among university students is common and poses serious health-related risks. Therefore, identifying and addressing associated factors is important.
Participants and methods
A large cross-sectional online-survey with 12,914 university students from Berlin was conducted from November 2016 to August 2017. Relative-risk- and correlation-analysis was used to identify factors associated with problem-drinking and regular heavy-drinking. Independent t-tests compared impulsivity and personality traits, chi-square-tests compared drinking motives between risk- and non-risk-drinkers.
Results
Male gender, tobacco-smoking, illegal substance use, impulsivity and various sociodemographic and psychosocial variables were significantly related to problem/heavy-drinking. Extraversion was a risk, conscientiousness and agreeableness were protective factors. Drinking-motives did not differ significantly between risk- and non-risk-drinkers. Generally, the main drinking-motives were to feel elated, relax and social purposes.
Conclusion
The identified markers and related problem behaviors may serve as a tool to enhance the identification of student subgroups at risk for problem/heavy-drinking, and hence improve targeted health-intervention-programs.
Objectives To compare image quality of deep learning reconstruction (AiCE) for radiomics feature extraction with filtered back projection (FBP), hybrid iterative reconstruction (AIDR 3D), and model-based iterative reconstruction (FIRST). Methods Effects of image reconstruction on radiomics features were investigated using a phantom that realistically mimicked a 65-year-old patient's abdomen with hepatic metastases. The phantom was scanned at 18 doses from 0.2 to 4 mGy, with 20 repeated scans per dose. Images were reconstructed with FBP, AIDR 3D, FIRST, and AiCE. Ninety-three radiomics features were extracted from 24 regions of interest, which were evenly distributed across three tissue classes: normal liver, metastatic core, and metastatic rim. Features were analyzed in terms of their consistent characterization of tissues within the same image (intraclass correlation coefficient >= 0.75), discriminative power (Kruskal-Wallis test p value < 0.05), and repeatability (overall concordance correlation coefficient >= 0.75). Results The median fraction of consistent features across all doses was 6%, 8%, 6%, and 22% with FBP, AIDR 3D, FIRST, and AiCE, respectively. Adequate discriminative power was achieved by 48%, 82%, 84%, and 92% of features, and 52%, 20%, 17%, and 39% of features were repeatable, respectively. Only 5% of features combined consistency, discriminative power, and repeatability with FBP, AIDR 3D, and FIRST versus 13% with AiCE at doses above 1 mGy and 17% at doses >= 3 mGy. AiCE was the only reconstruction technique that enabled extraction of higher-order features. Conclusions AiCE more than doubled the yield of radiomics features at doses typically used clinically. Inconsistent tissue characterization within CT images contributes significantly to the poor stability of radiomics features.
Applications of advanced cathode materials with well-designed chemical components and/or optimized nanostructures promoting the sulfur redox kinetics and suppressing the shuttle effect of polysulfides are highly valued. However, in the case of actual lithium-sulfur (Li-S) batteries under practical working conditions, one long-term obstacle still exists, which is mainly due to the difficulties in massive synthesis of such nanomaterials with low cost and ease of control on the nanostructure. Herein, we develop a facile synthesis of carbon coated Ti4O7 hollow nanoparticles (Ti4O7) using spherical polymer electrolyte brush as soft template, which is scalable via utilizing a minipilot reactor. The C Ti4O7 hollow nanoparticles provide strong chemical adsorption to polysulfides through the large polar surface and additional physical confinement by rich micro- & mesopores and have successfully been employed as an efficient sulfur host for multilayer pouch cells. Besides, the sluggish kinetics of the sulfur and lithium sulfide redox mechanism can be improved by the highly conductive Ti4O7 via catalyzation of the conversion of polysulfides. Consequently, the C-Ti4O7 based pouch cell endows a high discharge capacity of 1003 mAhg(-1) at 0.05 C, a high-capacity retention of 83.7% after 100 cycles at 0.1 C, and a high Coulombic efficiency of 97.5% at the 100th cycle. This work proposes an effective approach to transfer the synthesis of hollow Ti4O7 nanoparticles from lab- to large-scale production, paving the way to explore a wide range of advanced nanomaterials for multilayer Li-S pouch cells.
Immense uncertainty and the need for drastic interventions cause politicians to rely heavily on scientific advice for underpinning or legitimating their COVID-19 decision-making. This paper explores the role of scientific advice in this policy field in Germany, Italy, the Netherlands and the UK. It shows that scientific advice is based on the disciplinary, mainly medical, backgrounds of advisors but is also influenced by social and economic values, which are core to what politicians find important. During the pandemic a growing gap between scientific advice and political decisions is observed.
We report generation of ultra-broadband longitudinal acoustic coherent phonon wavepackets in SrTiO3 (STO) with frequency components extending throughout the first Brillouin zone. The wavepackets are efficiently generated in STO using femtosecond infrared laser excitation of an atomically flat 1.6 nm-thick epitaxial SrRuO3 film. We use femtosecond x-ray diffraction at the European X-Ray Free Electron Laser Facility to study the dispersion and damping of phonon wavepackets. The experimentally determined damping constants for multi-THz frequency phonons compare favorably to the extrapolation of a simple ultrasound damping model over several orders of magnitude.