Adeno-associated viruses (AAV) are widely used as delivery vectors in clinical trials for in vivo gene therapy due to their unique features. Göttingen minipigs are a well-established animal model for several diseases and can be used for the efficacy and safety testing of AAV-based gene therapy. Pre-existing antibodies against AAV may influence the results of testing and, therefore, the animals should be tested for the presence of antibodies against relevant AAV serotypes. The detection of AAVs in pigs may be also important for the virus safety of xenotransplantation. In this study, we screened Göttingen minipigs from Ellegaard Göttingen Minipigs A/S, Denmark, and Marshall BioResources, USA, for antibodies against AAV1, AAV2, AAV6, AAV9 serotypes. Of the 20 animals tested, 18 had no neutralizing antibodies for all AAVs tested, none had antibodies against AAV9, only one had antibodies against AAV6, and the titers of antibodies against AAV1 and AAV2 were less than 1:100, with two exceptions. For total binding IgG, more individuals showed positivity for all the tested serotypes but, in general, the levels were low or zero. Three animals had no antibodies at all against the AAVs tested. Therefore, Göttingen minipigs could be considered an attractive animal model for gene therapy studies. Since some animals were negative for all AAVs tested, these may be selected and used as donor animals for xenotransplantation.

A new Cl–Cl···−O–N+ halogen-bonded paradigm has been demonstrated, using dichlorine as a halogen bond (XB) donor and N-oxide as an XB acceptor. Their crystalline complexes were formed during the warm-up process from −196 °C to −80 °C for X-ray diffraction analysis. They exhibit high instability in the crystalline state, even at these low temperatures, leading to rapid decomposition and the formation of Cl⋯H–O–N hydrogen-bonded complexes. The normalized XB interaction ratio (RXB) of Cl⋯O interactions in the solid-state demonstrates affinity comparable to traditional I⋯O interactions observed in I–I···−O–N+ halogen-bonded systems. The Cl–Cl⋯O XB angles vary from 172° to 177°, manifesting the structure-guiding influence of the electronegative chlorine atom's σ-hole on these XB interactions.

Sulfur-containing polymers, such as thioesters and thiocarbonates, offer sustainability advantages, including enhanced degradability and chemical recyclability. However, their synthesis remains underdeveloped compared to that of their oxygen-containing counterparts. Although catalytic ring-opening copolymerization (ROCOP) can provide access to sulfur-containing polymers, these materials often exhibit uncontrolled microstructures and unpredictable properties. A comprehensive model that elucidates the factors determining selectivity in these catalytic reactions is still lacking, despite its central importance for advancing these polymerizations into widely applicable methodologies. In this study, we investigate the factors that lead to selectivity in sulfurated ROCOP across various monomer combinations, including thioanhydrides or carbon disulfide with epoxides, thiiranes, and oxetanes. We find that unwanted by-products primarily arise from backbiting reactions of catalyst-bound alkoxide chain ends, which can be mitigated by (i) selecting monomers that form primary alkoxide of thiolate chain ends, (ii) maximizing ring strain in the backbiting step, and (iii) timely termination of the polymerization. By applying these strategies, the selectivity of the catalytic ROCOP can be controlled and we successfully synthesized perfectly alternating poly(esters- alt -thioesters) from various oxetanes and the highly industrially relevant ethylene oxide. Our study thereby shifts the focus for achieving selectivity from catalyst to monomer choice providing valuable mechanistic insights for the development of future selective polymerizations, paving the way for sulfurated polymers as potential alternatives to current commodity materials.

Background: Therapeutic climbing (TC) has been increasingly implemented in the context of mental health care, with promising effects. While most evaluation studies focus on symptom reduction, the specific elements and mechanisms that make climbing effective compared to other therapeutic sports remain unclear. This study explores the subjective experiences of participants of a climbing therapy program in order to explore the underlying processes and the subjectively perceived effects. Method: The study took place at an inpatient rehabilitation facility in southern Germany, where a weekly group TC program was part of the treatment for individuals with mental disorders. Data from 265 participants with ICD-10 mental disorders collected between 2010 and 2022 was analyzed via questionnaires quantitatively assessing physical, psychological, and social improvements with rating items. Additionally open answers on subjectively relevant psychological experiences were analyzed using a qualitative content analysis. Results: Climbing therapy was generally perceived as relevant to their rehabilitation and rated as influential in terms of improving physical and psychological condition as well as sociability. The qualitative analysis identified five main categories being “Successful coping and positive mood”, “Challenge and coping strategies”, “Self-reflection and self-perception”, “Social experiences and belaying” and “Letting go of worries and being in the moment”. Conclusion: The results support previous findings on the benefits of TC and highlight its relevance as a therapeutic tool. Participants with mental disorders reported various positive experiences, emphasizing the unique demands, motivational aspects and social context of climbing. Further high-quality research is needed to compare climbing’s potential with other therapeutic sports across different target groups.

Background

Until now, scholarship on innovation processes in healthcare systems lack an in-depth appreciation of tensions. Tensions often revolve around barriers and result from individual assessments and prioritizations that guide actions to eventually overcome these barriers. In order to develop a more differentiated understanding of tensions’ role in healthcare innovation processes, this paper aims to shed light on the multifaceted ways in which tensions emerge, are being dealt with, and how they hinder or, at times, facilitate innovation processes.

Methods

A systematic review of published and grey literature was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guideline. The review involved searching three databases for original research articles and manually searching citations. Twenty-nine original full texts were identified, evaluated, and coded. These include papers on innovation in healthcare systems that investigated innovation-related organizational tensions. The findings were synthesized into different types of tensions in healthcare system innovation and the descriptions of the conflicting elements. We also analyzed the investigated innovations by type, process stages, and across different countries and healthcare systems.

Results

A total of forty-two tensions were identified and grouped into nine categories. Organizing tensions were predominant, followed by learning/belonging, performing, and performing/organizing tensions. Tensions most frequently occurred in the implementation phase and in the form of a dilemma. Included studies were conducted mainly in government-funded healthcare systems.

Conclusion

Our data suggest that innovation processes in healthcare systems are impaired by conflicts between contradictory elements, working cultures, and convictions and the organizational and regulatory context. Since the majority of the tensions we collected in our study can be addressed, future policy-making and research should take advantage of this fact and develop strategies that significantly influence the successful management of tensions and thus improve the implementation of innovations.

Collaborative online learning became a necessity for universities during the COVID-19 pandemic. Even though it is known from research that online collaboration is an effective way of learning, digital interaction can be challenging for learners. Group members have to create a high-quality interaction to ensure the success of the collaborative learning process. Based on a theoretical model of collaborative learning, high-quality interaction can be determined with regard to cognitive group activities (prior knowledge activation, transactivity), meta-cognitive group activities (organization of the work process), and relational group activities (group climate, participation and task-related communication). Our study aims to examine how students manage a self-directed collaborative learning setting, how they perceive the process quality of digital interaction and how the interaction quality is related to self-reported outcomes (learning gain and satisfaction). We use a newly developed questionnaire to assess the quality of digital interaction in terms of the aforementioned dimensions. Furthermore, we focus on associations with the beliefs about web-based learning and the ability of perspective-taking at the individual level as well as the sense of community at the group level. We conducted a quantitative study within online university courses that were implemented asynchronously due to the COVID-19 pandemic. N = 298 undergraduate students in teacher education rated the quality of a digital collaborative learning settings (response rate of 72%). The students worked on collaborative tasks autonomously without any guidance from the teacher. We find differences between (meta-)cognitive and relational factors of interaction quality, and differences in the strength of the associations with outcomes and individual and group-related factors. Our study provides insights into students´ collaborative online learning and examines the relationships between different dimensions of group interaction quality and the input and outcome variables. Limitations and areas for further research are discussed.

Inhaltsanalysen über die Darstellung von Konflikten in den Medien werden häufig durchgeführt, um den Vorwurf der Einseitigkeit empirisch zu überprüfen. Dies wirft die normative Frage auf, wie die Forderung nach einer ausgewogenen Bewertung, d. h. einer Gleichverteilung positiver und negativer Bewertungen zwischen Kontrahenten, als Qualitätsmaßstab rechtfertigt werden kann und wie dies fallbezogen, d. h. kontextabhängig geschehen muss. Diese Frage wird am Beispiel von Inhaltsanalysen zum russischen Angriffskrieg gegen die Ukraine diskutiert. In einer qualitativen Metastudie von 22 Inhaltsanalysen wurden zunächst induktiv acht Annahmen zur Rechtfertigung der Anwendung des Maßstabs für diesen Fall ermittelt. Vier davon beziehen sich auf das Verhältnis der Medien zu ihrer Umwelt: zur journalistischen Profession, der Medien untereinander sowie zum Publikum und zur Politik. Vier weitere Begründungen sind vom Verhältnis der Medien zum Krieg als Thema abgeleitet: zum normativ „richtigen“ und „falschen“ Handeln der Kontrahenten, zur Antizipation negativer Folgen, zum Framing des Kriegs als Machtkonflikt (statt als Konflikt um Werte) und aus der Position des Werterelativismus. Die Forderung nach einer ausgewogenen Bewertung war in den Studien häufig mit der Kritik an einer angeblich zu negativen Sicht der russischen Seite verknüpft. Die Diskussion der Fundstellen zeigt, dass die Argumente zur Rechtfertigung einer ausgewogenen Bewertung für den vorliegenden Fall nur begrenzt oder gar nicht tauglich sind. Damit kann die Studie zeigen, dass das Problem der „False Balance“ nicht nur im Bereich des Wissens, sondern auch des Wertens zu finden ist. Außerdem ist „False Balance“ damit nicht nur ein Phänomen im Journalismus, sondern auch in der Wissenschaft.

Mountains harbour one third of the world’s biodiversity and much of it is under increasing anthropogenic pressure. Yet, global assessments of the occurrence, and threat status of most mountain taxa, especially elusive ones are lacking, thereby hindering conservation and research prioritisation. In this study, we synthesise the distribution and conservation status of bats, a species rich taxon on mountains. By using data on geographical and elevational distribution ranges from the International Union for Conservation of Nature (IUCN), we examined bat species richness on mountains, species that predominantly occur in mountains (‘mountain dwelling species’), and those restricted to upper montane and alpine regions within mountains (‘highland dwelling species’). We also used published trait datasets to investigate the traits that are associated with mountain dwelling in bats. Globally, we identified 148 mountain dwelling and 46 highland dwelling bat species. Bat diversity is highest in the Northern Andes and Guiana Highlands. The mountain dwelling nature of bat species was found to be significantly associated with biogeographic realm. Importantly, our results show that mountain dwelling species are proportionately more data deficient than species that predominantly occur in lowlands. Additionally, highland dwelling species are proportionately more threatened than lowland species. Our results highlight a significant dearth of knowledge on mountain dwelling bat species. We conclude that more research is needed for bats specialised on mountain ecosystems. Our results draw attention towards improving the knowledge and protection of bat species that occur predominantly at high elevations across the world.

Background and Objective

Voriconazole (VRC), a broad-spectrum antifungal drug, exhibits nonlinear pharmacokinetics (PK) due to saturable metabolic processes, autoinhibition and metabolite-mediated inhibition on their own formation. VRC PK is also characterised by high inter- and intraindividual variability, primarily associated with cytochrome P450 (CYP) 2C19 genetic polymorphism. Additionally, recent in vitro findings indicate that VRC main metabolites, voriconazole N-oxide (NO) and hydroxyvoriconazole (OHVRC), inhibit CYP enzymes responsible for VRC metabolism, adding to its PK variability. This variability poses a significant risk of therapeutic failure or adverse events, which are major challenges in VRC therapy. Understanding the underlying processes and sources of these variabilities is essential for safe and effective therapy. This work aimed to develop a whole-body physiologically-based pharmacokinetic (PBPK) modelling framework that elucidates the complex metabolism of VRC and the impact of its metabolites, NO and OHVRC, on the PK of the parent, leveraging both in vitro and in vivo clinical data in a middle-out approach.

Methods

A coupled parent-metabolite PBPK model for VRC, NO and OHVRC was developed in a stepwise manner using PK-Sim® and MoBi®. Based on available in vitro data, NO formation was assumed to be mediated by CYP2C19, CYP3A4, and CYP2C9, while OHVRC formation was attributed solely to CYP3A4. Both metabolites were assumed to be excreted via renal clearance, with hepatic elimination also considered for NO. Inhibition functions were implemented to describe the complex interaction network of VRC autoinhibition and metabolite-mediated inhibition on each CYP enzyme.

Results

Using a combined bottom-up and middle-out approach, incorporating data from multiple clinical studies and existing literature, the model accurately predicted plasma concentration-time profiles across various intravenous dosing regimens in healthy adults, of different CYP2C19 genotype-predicted phenotypes. All (100%) of the predicted area under the concentration–time curve (AUC) and 94% of maximum concentration (Cmax) values of VRC met the 1.25-fold acceptance criterion, with overall absolute average fold errors of 1.12 and 1.14, respectively. Furthermore, all predicted AUC and Cmax values of NO and OHVRC met the twofold acceptance criterion.

Conclusion

This comprehensive parent-metabolite PBPK model of VRC quantitatively elucidated the complex metabolism of the drug and emphasised the substantial impact of the primary metabolites on VRC PK. The comprehensive approach combining bottom-up and middle-out modelling, thereby accounting for VRC autoinhibition, metabolite-mediated inhibition, and the impact of CYP2C19 genetic polymorphisms, enhances our understanding of VRC PK. Moreover, the model can be pivotal in designing further in vitro experiments, ultimately allowing for extrapolation to paediatric populations, enhance treatment individualisation and improve clinical outcomes.

In April 2022, Ukrainian President Volodymyr Zelenskyy argued that post-war Ukraine could resemble a “big Israel” in Europe regarding its security policy. But what implications would this have on the European security order? By comparing Kyiv’s and Jerusalem’s security circumstances, this paper shows that Ukraine cannot view itself as a “European Israel”, although some parallels exist between both states. However, Israel’s foreign policy could serve as a template for Kyiv.

This study presents a new approach for predicting forest aboveground biomass (AGB) from airborne laser scanning (ALS) data: AGB is predicted from sequences of images depicting vertical cross-sections through the ALS point clouds. A 3D version of the VGG16 convolutional neural network (CNN) with initial weights transferred from pre-training on the ImageNet dataset was used. The approach was tested on datasets from Canada, Poland, and the Czech Republic. To analyse the effect of training sample size on model performance, different-sized samples ranging from 10 to 375 ground plots were used. The CNNs were compared with random forest models (RFs) trained on point cloud metrics. At the maximum number of training samples, the difference in RMSE between observed and predicted AGB of CNNs and RFs ranged from −2 t/ha to 5 t/ha, and the difference in squared Pearson correlation coefficient ranged from −0.05 to 0.06. Additional pre-training on synthetic data derived from virtual laser scanning of simulated forest stands could only improve the prediction performance of the CNNs when only a few real training samples (10–40) were available. While 3D CNNs trained on cross-section images derived from real data showed promising results, RFs remain a competitive alternative.

The fabrication of stable perovskite nanofilm patterns is important for the development of functional optical devices. However, current production approaches are limited by the requirement for strict inert gas protection and long processing times. Here, a confined flash printing synthesis method is presented to generate perovskite nanofilms under ambient conditions, combining precursor transfer, perovskite synthesis, crystallization, and polymer protection in a single step within milliseconds. A laser simultaneously prints and induces the flash synthesis, confined in a polymer nanofilm, under normal ambient conditions. Due to its simplicity and flexibility, the method enables the combination and screening of many different perovskite precursor materials on various substrates. Besides for the development of novel perovskite materials and devices, the nanofilms can be applied for biodetection. The unique H2O2-responsive property of the ultrathin perovskite quantum dot film is applied for biomolecule detection based on oxidase-catalyzed enzymatic reactions.

Background

Fibulin-2 (FBLN2) is a secreted extracellular matrix (ECM) glycoprotein and has been identified in the mouse mammary gland, in cap cells of terminal end buds (TEBs) during puberty, and around myoepithelial cells during early pregnancy. It is required for basement membrane (BM) integrity in mammary epithelium, and its loss has been associated with human breast cancer invasion. Herein, we attempted to confirm the relevance of FBLN2 to myoepithelial phenotype in mammary epithelium and to assess its expression in molecular subtypes of human breast cancer.

Methods

The relationship between FBLN2 expression and epithelial markers was investigated in pubertal mouse mammary glands and the EpH4 mouse mammary epithelial cell line using immunohistochemistry, immunocytochemistry, and immunoblotting. Human breast cancer mRNA data from the METABRIC and TCGA datasets from Bioportal were analyzed to assess the association of Fbln2 expression with epithelial markers, and with molecular subtypes. Survival curves were generated using data from the METABRIC dataset and the KM databases.

Results

FBLN2 knockdown in mouse mammary epithelial cells was associated with a reduction in KRT14 and an increase in KRT18. Further, TGFβ3 treatment resulted in the upregulation of FBLN2 in vitro. Meta-analyses of human breast cancer datasets from Bioportal showed a higher expression of Fbln2 mRNA in claudin-low, LumA, and normal-like breast cancers compared to LumB, Her2 +, and Basal-like subgroups. Fbln2 mRNA levels were positively associated with mesenchymal markers, myoepithelial markers, and markers of epithelial–mesenchymal transition. Higher expression of Fbln2 mRNA was associated with better prognosis in less advanced breast cancer and this pattern was reversed in more advanced lesions.

Conclusion

With further validation, these observations may offer a molecular prognostic tool for human breast cancer for more personalized therapeutic approaches.

The possibility of treating a molecular liquid in an open region at ab initio electronic resolution embedded in a classical reservoir of energy and particles, is investigated. Because of its challenging properties and its relevance in many field of current research, the system chosen as prototype of molecular liquid is water at room conditions. A numerical protocol based on the mathematical model of open particle system is applied and the results are compared with results of a full ab initio simulation of reference. The key conclusion is that one can claim the existence of a mandatory minimal size of the quantum region in which structural and electronic properties reproduce those of reference and, at the same time, the exchange of molecules with the environment takes place as expected. This work provides a proof of concept about the possibility to systematically define a physically well founded open quantum system embedded in a classical environment. In turn, the proof of concept is a key information for the design of numerically efficient algorithms for ab initio molecular dynamics simulations of open systems.

Cellular membrane homoeostasis is maintained via a tightly regulated membrane and cargo flow between organelles of the endocytic and secretory pathways. Adaptor protein complexes (APs), which are recruited to membranes by the small GTPase ARF1, facilitate cargo selection and incorporation into trafficking intermediates. According to the classical model, small vesicles would facilitate bi-directional long-range transport between the Golgi, endosomes and plasma membrane. Here we revisit the intracellular organization of the vesicular transport machinery using a combination of CRISPR-Cas9 gene editing, live-cell high temporal (fast confocal) or spatial (stimulated emission depletion) microscopy as well as correlative light and electron microscopy. We characterize tubulo-vesicular ARF1 compartments that harbour clathrin and different APs. Our findings reveal two functionally different classes of ARF1 compartments, each decorated by a different combination of APs. Perinuclear ARF1 compartments facilitate Golgi export of secretory cargo, while peripheral ARF1 compartments are involved in endocytic recycling downstream of early endosomes. Contrary to the classical model of long-range vesicle shuttling, we observe that ARF1 compartments shed ARF1 and mature into recycling endosomes. This maturation process is impaired in the absence of AP-1 and results in trafficking defects. Collectively, these data highlight a crucial role for ARF1 compartments in post-Golgi sorting.

Pinus sylvestris trees are known to efficiently defend themselves against eggs of the herbivorous sawfly Diprion pini. Their direct defense against eggs is primable by prior exposure to the sex pheromones of this species and their indirect defense involves attraction of egg parasitoids by egg-induced pine needle odor. But it is unknown whether exposure of pine to D. pini sex pheromones also affects pine indirect defense against sawfly eggs. In this study, we investigated the influence of exposure of P. sylvestris trees to the sex pheromones of D. pini on indirect defense mediated by egg parasitoids. Behavioral assays with Closterocerus ruforum, a key parasitoid of sawfly eggs, revealed no significant attraction to odor from egg-free pines pre-exposed to pheromones. Chemical analyses of odor from egg-free pines showed no pheromone-induced change in the emission rates of the known key terpenoids promoting parasitoid attraction. Further comparative analyses of odor from egg-laden pines pre-exposed to the sex pheromones and of odor from egg-laden pines unexposed to pheromones neither revealed significant differences in the emission rates of terpenoids relevant for parasitoid attraction. The results suggest that a pheromone-induced or pheromone-primed, egg-induced pine indirect defense seems to be redundant in addition to the known pheromone-primable pine direct defense against the eggs and the known egg-inducible indirect defense.

The growing utilization of remote sensing data in lake studies provides crucial spatial insights into biogeochemistry and biology. However, clarity regarding the development and intended use of remote sensing products is often lacking. This letter aims to elucidate the tradeoffs for the utilization of remote sensing data in limnological studies with an example of based on the estimation of chlorophyll a due to its importance as a water quality indicator. The analysis initiates with a meticulous product selection, requiring an evaluation of its capacity to address the optical complexity of freshwater systems. Assessing atmospheric correction and product limitations ensures alignment with the study's objectives. Subsequently, rigorous validation of remote sensing products is essential, accompanied by a cautious interpretation of the data. This letter advocates for the use of remote sensing data, offering key strategies for their optimal utilization in lake studies.

Precipitation nowcasting refers to the prediction of precipitation intensity in a local region and in a short timeframe up to 6 hours. The evaluation of spatial and temporal information still challenges state-of-the-art numerical weather prediction models. The increasing possibilities to store and evaluate data combined with the advancements in the developments of artificial intelligence algorithms make it natural to use these methods to improve precipitation nowcasting. In this work, a Trajectory Gated Recurrent Unit (TrajGRU) is applied to radar data of the German Weather Service. The impact of finetuning a network pretrained at a different location and for several precipitation intensity thresholds with respect to the training time is evaluated. In cases with little availability of training data at the target location, for example when heavy rainfall is rare, the finetuned model can benefit from the original model performance at the pretraining location. Furthermore, the skill scores for the different thresholds are shown for a prediction time up to 100 minutes. The results highlight promising regional extrapolation capabilities for such neural networks for precipitation nowcasting.

Structural diversity of diterpenes is mediated by the enigmatic family of diterpene synthases. The overall enzymatic contribution hereby lies in a carefully concerted chemistry of highly reactive carbocation intermediates mainly guided by aromatic and polar amino acid side chains and the pyrophosphate cofactor. To date several studies aimed to shed light on the mechanism underlining terpene synthases chemistry. Specifically, the diterpene synthase CotB2 serves as model enzyme for detailed mutagenesis studies. Here we investigate the catalytic mechanism of CotB2 variant V80L in a holistic, biochemical, structural, and computational biology approach. We were able to identify an altered product profile compared to CotB2WT for the substrates geranylgeranyl diphosphate and farnesyl diphosphate. Moreover, we solved the crystal structure, and shed further light on terpene synthase chemistry by modelling of the substrate and intermediate binding.