Search

Abstract Systemic ALys amyloidosis is a debilitating protein misfolding disease that arises from the formation of amyloid fibrils from C-type lysozyme. We here present a 2.8 Å cryo-electron microscopy structure of an amyloid fibril, which was isolated from the abdominal fat tissue of a patient who expressed the D87G variant of human lysozyme. We find that the fibril possesses a stable core that is formed by all 130 residues of the fibril precursor protein. There are four disulfide bonds in each fibril protein that connect the same residues as in the globularly folded protein. As the conformation of lysozyme in the fibril is otherwise fundamentally different from native lysozyme, our data provide a structural rationale for the need of protein unfolding in the development of systemic ALys amyloidosis.

This mixed-methods study investigates the adoption of generative AI among orthopaedic surgeons, employing a Unified Theory of Acceptance and Use of Technology (UTAUT) based survey (n = 177) and follow-up interviews (n = 7). The research reveals varying levels of AI familiarity and usage patterns, with higher adoption in research and professional development compared to direct patient care. A significant generational divide in perceived ease of use highlights the need for tailored training approaches. Qualitative insights uncover barriers to adoption, including the need for more evidence-based support, as well as concerns about maintaining critical thinking skills. The study exposes a complex interplay of individual, technological, and organisational factors influencing AI adoption in orthopaedic surgery. The findings underscore the need for a nuanced approach to AI integration that considers the unique aspects of orthopaedic surgery and the diverse perspectives of surgeons at different career stages. This provides valuable insights for educational institutions and healthcare organisations in navigating the challenges and opportunities of AI adoption in specialised medical fields.

Abstract Systemic AL amyloidosis is one of the most frequently diagnosed forms of systemic amyloidosis. It arises from mutational changes in immunoglobulin light chains. To explore whether these mutations may affect the structure of the formed fibrils, we determine and compare the fibril structures from several patients with cardiac AL amyloidosis. All patients are affected by light chains that contain an IGLV3-19 gene segment, and the deposited fibrils differ by the mutations within this common germ line background. Using cryo-electron microscopy, we here find different fibril structures in each patient. These data establish that the mutations of amyloidogenic light chains contribute to defining the fibril architecture and hence the structure of the pathogenic agent.

Streptococcus (S.) equi subspecies (subsp.) zooepidemicus is an important facultative pathogen in horses and can cause severe infections in other species including humans. Facing the post-antibiotic era, novel antimicrobials are needed for fighting bacterial infections. Bacteriophages (phages) are the natural predators of bacteria and discussed as a promising antimicrobial treatment option. The objective of this study was to isolate and characterize S. equi subsp. zooepidemicus-specific phages for the first time and to evaluate their efficacy in vitro and ex vivo. In total, 13 phages with lytic activity were isolated and host ranges were determined. Two phages with broad host ranges and high efficiency of plating (vB_SeqZP_LmqsRe26-2 (lytic activity: 30/37 bacterial isolates) and vB_SeqZP_LmqsRe26-3 (lytic activity: 29/37 bacterial isolates)) and one phage with relatively low efficiency of plating (vB_SeqZP_LmqsRe26-1) were selected for further characterization, including electron microscopy and whole genome sequencing. In in vitro planktonic killing assays at two tested multiplicities of infection (MOI 1 and MOI 10), significant bacterial growth reduction was observed when the phages vB_SeqZP_LmqsRe26-2 and vB_SeqZP_LmqsRe26-3 were added. These phages were subsequently co-incubated with clinical S. equi subsp. zooepidemicus isolates in an equine endometrial explant model but did not achieve bacterial growth reduction at MOI 1 and MOI 10. However, helium ion microscopy revealed presence of particles adherent to the bacteria on the explant after incubation (25 h), suggesting possible phage-bacteria interactions. In conclusion, phages against S. equi subsp. zooepidemicus were successfully isolated and characterized. Promising results were observed in in vitro but no significant reduction was detected in ex vivo experiments, requiring additional investigations. However, after further adaptations (e.g., optimization of MOIs and phage administration or use of phage-antibiotic combination), phages could be a potential antimicrobial tool for future therapeutic use in S. equi subsp. zooepidemicus infections, although the available results do not currently support the therapeutic usage.

Abstract Noether invariance in statistical mechanics provides fundamental connections between the symmetries of a physical system and its conservation laws and sum rules. The latter are exact identities that involve statistically averaged forces and force correlations and they are derived from statistical mechanical functionals. However, the implications for more general observables and order parameters are unclear. Here, we demonstrate that thermally averaged classical phase space functions are associated with exact hyperforce sum rules that follow from translational Noether invariance. Both global and locally resolved identities hold and they relate the mean gradient of a phase-space function to its negative mean product with the total force. Similar to Hirschfelder’s hypervirial theorem, the hyperforce sum rules apply to arbitrary observables in equilibrium. Exact hierarchies of higher-order sum rules follow iteratively. As applications we investigate via computer simulations the emerging one-body force fluctuation profiles in confined liquids. These local correlators quantify spatially inhomogeneous self-organization and their measurement allows for the development of stringent convergence tests and enhanced sampling schemes in complex systems.

We use supervised machine learning together with the concepts of classical density functional theory to investigate the effects of interparticle attraction on the pair structure, thermodynamics, bulk liquid-gas coexistence, and associated interfacial phenomena in many-body systems. Local learning of the one-body direct correlation functional is based on Monte Carlo simulations of inhomogeneous systems with randomized thermodynamic conditions, randomized planar shapes of the external potential, and randomized box sizes. Focusing on the prototypical Lennard-Jones system, we test predictions of the resulting neural attractive density functional across a broad spectrum of physical behavior associated with liquid-gas phase coexistence in bulk and at interfaces. We analyze the bulk radial distribution function g(r) obtained from automatic differentiation and the Ornstein-Zernike route and determine (i) the Fisher-Widom line, i.e., the crossover of the asymptotic (large distance) decay of g(r) from monotonic to oscillatory, (ii) the (Widom) line of maximal correlation length, (iii) the line of maximal isothermal compressibility, and (iv) the spinodal by calculating the poles of the structure factor in the complex plane. The bulk binodal and the density profile of the free liquid-gas interface are obtained from density functional minimization and the corresponding surface tension from functional line integration. We also show that the neural functional describes accurately the phenomena of drying at a hard wall and of capillary evaporation for a liquid confined in a slit pore. Our neural framework yields results that improve significantly upon standard mean-field treatments of interparticle attraction. Comparison with independent simulation results demonstrates a consistent picture of phase separation even when restricting the training to supercritical states only. We argue that phase coexistence and its associated signatures can be discovered as emerging phenomena via functional mappings and educated extrapolation.

The reversible reaction Ca(OH)2 + 104.4 kJ/mol ⇌ CaO + H2O offers several advantages as energy storage system. For example, it possesses a higher energy density than lead-acid or nickel–cadmium batteries. In addition, it has proven cyclability, low cost and worldwide availability. For this reason, it is suitable for seasonal heat storage applications. However, the raw powder material displays properties that represent a major challenge for the design of reactors that decouple power from capacity e.g. low thermal conductivity, cohesivity and tendency to form agglomerates. In order to overcome these drawbacks, different approaches to stabilize the Ca(OH)2/CaO particles have been investigated e.g. shaping, micro encapsulation, macroencapsulation, etc. The assessment of the stabilized products, however, has limitations in terms of amount of mass and reaction conditions as it is carried out in TA (thermal analyzer). Furthermore, it does not allow to analyze the structural decay and agglomeration of stabilized particles in-situ. For this reason, a more comprehensive assessment of a bulk of material under reactor conditions is necessary. In this work, a reaction chamber is developed to enable the observation of a bulk of storage material (0.1 L) during thermal cycling. Thus, two samples were subject of 20 cycles of dehydration and rehydration. The experiments were carried out at a temperature range of 350 °C – 500 °C and water vapor pressure of 0 – 1 bar. The analysis of images and post experiment tests (e.g. thermogravimetric analysis (TGA), dynamometry, X-ray diffraction (XRD) analysis) prove that the mechanical strength and structural integrity resulted enhanced for both samples. In addition, it allows to further understand the bulk behavior of Ca(OH)2 granules and thus important implications for the design of technical scale reactors can be derived.

Abstract The transactive response DNA-binding protein-43 (TDP-43) is a multi-facet protein involved in phase separation, RNA-binding, and alternative splicing. In the context of neurodegenerative diseases, abnormal aggregation of TDP-43 has been linked to amyotrophic lateral sclerosis and frontotemporal lobar degeneration through the aggregation of its C-terminal domain. Here, we report a cryo-electron microscopy (cryo-EM)-based structural characterization of TDP-43 fibrils obtained from the full-length protein. We find that the fibrils are polymorphic and contain three different amyloid structures. The structures differ in the number and relative orientation of the protofilaments, although they share a similar fold containing an amyloid key motif. The observed fibril structures differ from previously described conformations of TDP-43 fibrils and help to better understand the structural landscape of the amyloid fibril structures derived from this protein.

Abstract Systemic ATTR amyloidosis is an increasingly important protein misfolding disease that is provoked by the formation of amyloid fibrils from transthyretin protein. The pathological and clinical disease manifestations and the number of pathogenic mutational changes in transthyretin are highly diverse, raising the question whether the different mutations may lead to different fibril morphologies. Using cryo-electron microscopy, however, we show here that the fibril structure is remarkably similar in patients that are affected by different mutations. Our data suggest that the circumstances under which these fibrils are formed and deposited inside the body - and not only the fibril morphology - are crucial for defining the phenotypic variability in many patients.

Long-term storage of seasonally available solar energy and its provision to balance heating energy demand can contribute significantly to the sustainable use of energy resources. Thermochemical energy storage is a suitable process for this purpose, offering the possibility of loss-free long-term energy storing and heat supply. In order to develop suitable technical solutions for the use of this technology, novel reactor concepts and scientific questions regarding material and technology development are being investigated. In this publication, the energy storage process of a long-term energy storage system based on a ploughshare reactor is experimentally investigated under various technically relevant operating conditions. One specific aspect of this technology is related to the release of water vapour during the charging process. Therefore, this work focusses, in particular, on the possibility of technically utilizing the latent heat of the released water vapour in the range of 45 °C to 80 °C, which covers the operating requirements of common heating systems in households. The experiments have shown that the dehydration process enables the separation of two heat fluxes: the chemically bound energy for long-term storage and the physically (sensible and latent) stored energy for short-term applications. However, the limitation of gas transport was also identified as the most important influencing parameter for optimising the performance of the process.

INTRODUCTION: Differentiated thyroid cancer (DTC) in adolescents rare but with a favorable outcome, despite higher rates of cervical lymph node and pulmonary metastasis compared to adults. The aim of this study was to critically evaluate treatment of adolescents with DTC at a single center. METHODS: Patients receiving postoperative radioiodine treatment (RAIT) for DTC between 2005 and 2020 at our institution were screened to identify adolescents according to the World Health Organization definition (10-19 years of age). Demographics, clinico-pathological characteristics, treatment and outcome were analyzed. RESULTS: Among 1,897 DTC patients, 23 (1.3%) were adolescents with a median (range) age of 16 (10-18) years. The female to male ratio was 3.6: 1. Sixty percent had classic papillary thyroid cancer, with follicular variant in 40%, which was higher than previously reported (15-25%) for this age group. pT-status was pT1 in 9 (39.2%), pT2 in 8 (34.8%), pT3 in 3 (13%) and pT4 in 3 (13%) patients. In 19 (82.6%) patients, central lymphadenectomy was performed and metastasis was seen in 57%. All patients received RAIT with initial activities of 1.2 (n=1, 4.3%), 2 (n=12, 52.2%) or 3.7 GBq (n=10, 43.5%). Eighteen (78.2%) patients were free of biochemical and radiologic disease at a median follow-up of 60.7 months. Second-line surgery for lymph node relapse was necessary in 3 (13%) cases. There was one disease-associated death. DISCUSSION AND CONCLUSION: Despite high rates of metastasis, most patients were cured, and second-line surgery was rarely required. Further prospective studies are needed to determine whether less aggressive surgical management or omitting adjuvant RAIT are feasible in patients with limited stages at diagnosis.

Abstract Type I modular polyketide synthases (PKSs) are multi-domain enzymes functioning like assembly lines. Many engineering attempts have been made for the last three decades to replace, delete and insert new functional domains into PKSs to produce novel molecules. However, inserting heterologous domains often destabilize PKSs, causing loss of activity and protein misfolding. To address this challenge, here we develop a fluorescence-based solubility biosensor that can quickly identify engineered PKSs variants with minimal structural disruptions. Using this biosensor, we screen a library of acyltransferase (AT)-exchanged PKS hybrids with randomly assigned domain boundaries, and we identify variants that maintain wild type production levels. We then probe each position in the AT linker region to determine how domain boundaries influence structural integrity and identify a set of optimized domain boundaries. Overall, we have successfully developed an experimentally validated, high-throughput method for making hybrid PKSs that produce novel molecules.

Introduction Epilepsy is a common neurological disorder characterised by recurrent seizures. Almost half of patients who have an unprovoked first seizure (UFS) have additional seizures and develop epilepsy. No current predictive models exist to determine who has a higher risk of recurrence to guide treatment. Emerging evidence suggests alterations in cognition, mood and brain connectivity exist in the population with UFS. Baseline evaluations of these factors following a UFS will enable the development of the first multimodal biomarker-based predictive model of seizure recurrence in adults with UFS.Methods and analysis 200 patients and 75 matched healthy controls (aged 18–65) from the Kingston and Halifax First Seizure Clinics will undergo neuropsychological assessments, structural and functional MRI, and electroencephalography. Seizure recurrence will be assessed prospectively. Regular follow-ups will occur at 3, 6, 9 and 12 months to monitor recurrence. Comparisons will be made between patients with UFS and healthy control groups, as well as between patients with and without seizure recurrence at follow-up. A multimodal machine-learning model will be trained to predict seizure recurrence at 12 months.Ethics and dissemination This study was approved by the Health Sciences and Affiliated Teaching Hospitals Research Ethics Board at Queen’s University (DMED-2681-22) and the Nova Scotia Research Ethics Board (1028519). It is supported by the Canadian Institutes of Health Research (PJT-183906). Findings will be presented at national and international conferences, published in peer-reviewed journals and presented to the public via patient support organisation newsletters and talks.Trial registration number NCT05724719.

Abstract Background Investigations into the growth and self-organization of plant roots is subject to fundamental and applied research in various areas such as botany, agriculture, and soil science. The growth activity of the plant tissue can be investigated by isotope labeling experiments with heavy water and subsequent detection of the deuterium in non-exchangeable positions incorporated into the plant biomass. Commonly used analytical methods to detect deuterium in plants are based on mass-spectrometry or neutron-scattering and they either suffer from elaborated sample preparation, destruction of the sample during analysis, or low spatial resolution. Confocal Raman micro-spectroscopy (CRM) can be considered a promising method to overcome the aforementioned challenges. The substitution of hydrogen with deuterium results in the measurable shift of the CH-related Raman bands. By employing correlative approaches with a high-resolution technique, such as helium ion microscopy (HIM), additional structural information can be added to CRM isotope maps and spatial resolution can be further increased. For that, it is necessary to develop a comprehensive workflow from sample preparation to data processing. Results A workflow to prepare and analyze roots of hydroponically grown and deuterium labeled Zea mays by correlative HIM-CRM micro-analysis was developed. The accuracy and linearity of deuterium detection by CRM were tested and confirmed with samples of deuterated glucose. A set of root samples taken from deuterated Zea mays in a time-series experiment was used to test the entire workflow. The deuterium content in the roots measured by CRM was close to the values obtained by isotope-ratio mass spectrometry. As expected, root tips being the most actively growing root zone had incorporated the highest amount of deuterium which increased with increasing time of labeling. Furthermore, correlative HIM-CRM analysis allowed for obtaining the spatial distribution pattern of deuterium and lignin in root cross-sections. Here, more active root zones with higher deuterium incorporation showed less lignification. Conclusions We demonstrated that CRM in combination with deuterium labeling can be an alternative and reliable tool for the analysis of plant growth. This approach together with the developed workflow has the potential to be extended to complex systems such as plant roots grown in soil.

Abstract Amyloid fibrils have emerged as innovative tools to enhance the transduction efficiency of retroviral vectors in gene therapy strategies. In this study, we used cryo-electron microscopy to analyze the structure of a biotechnologically engineered peptide fibril that enhances retroviral infectivity. Our findings show that the peptide undergoes a time-dependent morphological maturation into polymorphic amyloid fibril structures. The fibrils consist of mated cross-β sheets that interact by the hydrophobic residues of the amphipathic fibril-forming peptide. The now available structural data help to explain the mechanism of retroviral infectivity enhancement, provide insights into the molecular plasticity of amyloid structures and illuminate the thermodynamic basis of their morphological maturation.

The status of the Steppe Eagle (Aquila nipalensis) was unclear in Turkey, which forms the south-western border of the species’ breeding distribution. The status of the species in Turkey has never been clear, as historically only two nesting sites were found and documented in the wider area of Tuz Gölü, which were proved to be active only in some years between 1969 and 1980. One nest site in Tuz Gölü was found in some years between 1969 and 1975, while another nest was found in Bolluk Gölü in 1975, and was active also in 1980. There were no published breeding records since 1980 until 2003, when a Steppe Eagle brood was found accidentally in central Anatolia by Hungarian birdwatchers. There were no other published breeding records of Steppe Eagles in Turkey in the twentieth century, although adult birds have been observed in a few occasions in Central and Eastern Anatolia during the breeding season between 1998 and 2015.

The Central European Population of the Eastern Imperial Eagle (Aquila heliaca) is located on its most western global distribution edge. After a strong decline in the last two centuries the population recovered in the past decades not least due to extensive protection measurements. Conservation and scientific programs on international and national level were implemented in several countries especially in Hungary, Slovakia, Austria, Czech Republic and Serbia. In frame of these activities imperial eagle were tagged by satellite transmitter since 2003 in several projects by different organisations of the region. In the current presentation we show preliminary results of an analysis of the natal dispersal movements of the immature birds of this residential population of Eastern Imperial Eagles.

The Imperial Eagle (Aquila heliaca, IE) reaches the western limit of its global breeding in range in Austria. As a result of targeted conservations measures in the Pannonian region, namely Hungary and Slovakia, the species repopulated Austria in 1999, nearly 190 years after its extirpation. Since then, a strong population increase took place and was monitored closely through different species conservation programmes of BirdLife Austria. Apart from the population monitoring, the programmes comprised measures to prevent illegal persecution and the disturbance of breeding sites, the satellite tagging of young IEs and the collection of all mortality data available.

Abstract Catalytic amyloid fibrils are novel types of bioinspired, functional materials that combine the chemical and mechanical robustness of amyloids with the ability to catalyze a certain chemical reaction. In this study we used cryo-electron microcopy to analyze the amyloid fibril structure and the catalytic center of amyloid fibrils that hydrolyze ester bonds. Our findings show that catalytic amyloid fibrils are polymorphic and consist of similarly structured, zipper-like building blocks that consist of mated cross-β sheets. These building blocks define the fibril core, which is decorated by a peripheral leaflet of peptide molecules. The observed structural arrangement differs from previously described catalytic amyloid fibrils and yielded a new model of the catalytic center.

Abstract Fast assessment of the composition of amyloid fibril samples from cryo-EM data poses a serious challenge to existing image analysis tools. We develop a method for automated segmentation of single fibrils requiring only little user input during the training process. This is achieved by combining a binary segmentation based on a convolutional neural network with preprocessing steps to allow for easy manual generation of training data. Subsequent skeletonization turns the binary segmentation into a single-object segmentation. Then, we compute properties of shape and texture of each segmented fibril, including an estimation of the fibril width. We discuss the composition of the sample based on the distributions of these computed properties and outline how a classification of fibril morphologies might be performed using these properties.

Pseudomonas putida has been engineered to produce medium chain length oleochemicals, including medium chain length free fatty acids and their ester forms, compounds which may be useful as biodiesel blending agents.

Abstract Background Human pluripotent stem cell‐derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. Methods Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three‐dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient‐specific induced pluripotent stem cell line was compared to a CRISPR/Cas9‐edited isogenic control line. Results The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two‐dimensional and three‐dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration‐competent satellite‐like cell pool. Tissue‐engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms (P

This study reports the cryo-EM structures of AA amyloid fibrils from two patients with vascular AA amyloidosis. The findings imply that different disease variants in systemic amyloidosis are associated with different fibril structures.

Noether’s theorem allows to find conservation laws corresponding to the symmetries of the system. The authors theoretically derive an exact relation of the variance of external force in statistical mechanics, using Noether invariance, providing a powerful way of looking at fluctuations.

Tajikistan's agricultural sector, primarily dominated by cotton cultivation, has experienced significant changes since the Soviet era. Although farmers introduced food crops into agricultural production to ensure food security after the collapse of the Soviet Union in 1991, the government still mandates that cotton cultivation occupy a considerable portion of Tajikistan's limited irrigated land. However, following the recent Covid-19 pandemic crisis, farmers have encountered the need to reassess their agricultural practices, given constrained governmental assistance and a range of socioeconomic considerations. This research adopts resilience as a conceptual framework to examine the coping strategies of smallholder farmers in the Khatlon region of Tajikistan, with a specific emphasis on the period spanning from 2019 to 2022. At the outset, our research employed the snowballing technique to increase respondent participation, later transitioning to a representative sample size that facilitated the collection of qualitative data from around 100 semi-structured interviews, 10 focus groups, and personal visits to agricultural fields over the course of four years. The paper not only demonstrates the overall creativity of Tajikistani farmers in growing and selecting non-cotton crops in this most recent period of crisis, but also points to wealthier farmers’ generally greater willingness to adopt technological innovations and gain new knowledge to apply to them. Although these strategies have been taken in order to address farmers’ immediate needs in troubled times against the backdrop of the existing state cotton policy, their sustainability remains uncertain. While the qualitative focus of this study bears certain inherent limitations, the data collected nevertheless show that this period of crisis has been generative for many farmers as they seek out new methods of subsistence.

IntroductionWe developed a new approach for site index curve models that combines longitudinal height development patterns derived from state-space data with the broad environmental conditions covered by space-for-time data.MethodsFor this, we gathered dendrometry from both inventories and research plots. Concerning environmental variables, we included soil mapping data as well as atmospheric data, i.e., precipitation, temperature, and nitrogen deposition. The atmospheric data was included as a weighted mean over the stand life of the sums for the dynamically determined vegetation period or as yearly sums in the case of nitrogen deposition, respectively. As a weight, the values of a height increment function were used. Then, we derived the basic shape of a height development curve from research plot data and transferred said shape to a site index curve model.ResultsThe model represents a substantial advancement of a previous version and was fitted as a generalized additive model (GAM). All effects were of relevant size and showed biologically feasible patterns.DiscussionThough the model is biased for young ages, we could predict site index curves that, under constant environmental conditions, closely follow yield table curves and thus accurately depict stand height development. Moreover, the model does not require initial dendrometry, which broadens its applicability. Thus, the model represents a useful tool for forest management and planning under climate change.

The Natura 2000 network of protected areas is the backbone of species conservation in the European Union. We investigated whether Austrian-hatched white-tailed eagles (Haliaeetus albicilla) make particular use of this multinational network during their natal dispersal, and what habitats were of importance to the eagles. We analyzed the utilization distribution of 907,466 GPS locations from 38 dispersing white-tailed eagles using a dynamic Brownian Bridge Movement Model. Eagles ranged over a huge area of central-eastern Europe. Natura 2000 sites overlapped with 67% of the resulting 50% isopleth; i.e., a high probability of utilization of Natura 2000 areas by white-tailed eagles was found. White-tailed eagles used wetlands, waterbodies, and deciduous forests adjacent to wet habitats disproportionately often. Coniferous forests and settlements were avoided. Anthropogenically caused mortalities hardly occurred within Natura 2000 sites. Our study suggests that the Natura 2000 network is a crucial tool for conserving the white-tailed eagle. This top predator is an ambassador for the Natura 2000 idea during all life stages, and should continue to be a conservation priority of the network.

This manuscript reports the structure of a pathologically relevant wild type ATTR amyloid fibril from systemic non-hereditary transthyretin amyloidosis. The comparison of the wild type ATTR fibril with two previous published ex vivo V30M ATTR fibrils highlighted numerous similarities between these different reconstructions, pointing to a common underlying structure for ATTR fibrils despite coming from different mutants and patients.

Das Logion in Lk 11,34–36 bleibt im literarischen Zusammenhang des Evangeli-ums sperrig, weil der situative Kontext nicht tradiert wurde. Während Lk 11,34 den Einfluss der moralischen Disposition auf die anatomische Sehfähigkeit beschreibt, thematisiert Lk 11,35–36 die Kompensation der fehlenden Sehfähigkeit durch die richtige innere Haltung, zur Sprache gebracht als τὸ φῶς τὸ ἐν σοί. Unter Einbindung von Lk 11,21–22 und Lk 12,15 sowie Lk 11,29–30 lässt sich als möglicher situativer Kontext die Ansprache eines blinden Gegenübers annehmen, dessen vermeintliche Bedürftigkeit mit der Besitzstandswahrung ei-ner ökonomisch etablierten Gruppe kontrastiert wurde. Bedeutsam für die Rezeption könnte das Logion werden, wo es als Hinweis auf die Anerkenntnis einer körperlichen oder geistigen Andersartigkeit und die allen gemeinsame Verantwortung gewertet wird.

ABSTRACT Many disciplines have become increasingly interested in cyanobacteria, due to their ability to fix CO2 while using water and sunlight as electron and energy sources. Further, several species of cyanobacteria are also capable of fixing molecular nitrogen, making them independent of the addition of nitrate or ammonia. Thereby they hold huge potential as sustainable biocatalysts. Here, we look into a dual-species biofilm consisting of filamentous diazotrophic cyanobacteria Tolypothrix sp. PCC 7712 and heterotrophic bacteria Pseudomonas taiwanensis VLB 120 growing in a capillary biofilm reactor. Such systems have been reported to enable high cell densities continuous process operation. By combining confocal laser scanning and helium-ion microscopy with a proteomics approach, we examined these organisms’ interactions under two nitrogen-feeding strategies: N2-fixing and nitrate assimilation. Not only did Pseudomonas facilitate the biofilm formation by forming a carpet layer on the surface area but also did N2-fixing biofilms show greater attachment to the surface. Pseudomonas proteins related to surface and cell attachments were observed in N2-fixing biofilms in particular. Furthermore, co-localized biofilm cells displayed a resilient response to extra shear forces induced by segmented media/air flows. This study highlights the role of Pseudomonas in the initial attachment process, as well as the effects of different nitrogen-feeding strategies and operation regimes on biofilm composition and growth. IMPORTANCE Cyanobacteria are highly interesting microorganisms due to their ability to synthesize sugars from CO2 while using water and sunlight as electron and energy sources. Further, many species are also capable of utilizing molecular nitrogen, making them independent of artificial fertilizers. In this study, such organisms are cultivated in a technical system, which enables them to attach to the reactor surface, and form three-dimensional structures termed biofilms. Biofilms achieve extraordinarily high cell densities. Furthermore, this growth format allows for continuous processing, both being essential features in biotechnological process development. Understanding biofilm growth and the influence technical settings and media composition have on biofilm maturation and stability are crucial for reaction and reactor design. These findings will help to open up these fascinating organisms for applications as sustainable, resource-efficient industrial workhorses.

We combine power functional theory and machine learning to study non-equilibrium overdamped many-body systems of colloidal particles at the level of one-body fields. We first sample in steady state the one-body fields relevant for the dynamics from computer simulations of Brownian particles under the influence of randomly generated external fields. A neural network is then trained with this data to represent locally in space the formally exact functional mapping from the one-body density and velocity profiles to the one-body internal force field. The trained network is used to analyse the non-equilibrium superadiabatic force field and the transport coefficients such as shear and bulk viscosities. Due to the local learning approach, the network can be applied to systems much larger than the original simulation box in which the one-body fields are sampled. Complemented with the exact non-equilibrium one-body force balance equation and a continuity equation, the network yields viable predictions of the dynamics in time-dependent situations. Even though training is based on steady states only, the predicted dynamics is in good agreement with simulation results. A neural dynamical density functional theory can be straightforwardly implemented as a limiting case in which the internal force field is that of an equilibrium system. The framework is general and directly applicable to other many-body systems of interacting particles following Brownian dynamics.

Natural differences in the regional development of Russia are presented in many legislative acts dedicated to the Russian Far North. In contrast, the unique nature and complexity of mountainous and high-mountain territories are protected only by a few regional acts. The reason for this lies in the complexity and multicomponent criteria required for assigning these territories the status of protected areas and in the fact that their boundaries do not correspond with administrative boundaries. The main materials underlying the article are legal documents (regulations, laws, etc.) concerning the institutionalization of the northern and mountainous territories. A comparative analysis of regional policy in relation to mountainous and northern territories takes into account similar criteria, such as vegetation types and patterns, forest borders or crop frontiers. Almost two-thirds of the territory of Russia refers to the North and more than half of the territory is occupied by mountains. The first attempts to institutionalize the North were undertaken in the 1930s, while the programmes for the development of mountainous territories gained legal support only at the end of 20th century and only in some regions. The most important difference between the institutionalization of the North and the mountains is the fact that the state initiated the creation of special legal conditions for the North. In the case of the mountains, the initiator was the public, initially at the regional level. Currently, three constituent entities of the Russian Federation adopted laws on mountain areas, but so far there are no all-Russian laws. The main lobbyists are the North Caucasian regions, while the Siberian regions (with the exception of the Altai Republic) are rather passive in discussing mountain issues. The elaborated legislation for the North seems to be closely related to the potential and realised income from natural resource extraction. For this reason, corresponding legislation for the mountain regions is not expected particularly soon, due to the lack of legal resources. Efforts aimed to provide legal support for mechanisms that compensate the socio-economic discrepancies between mountainous areas and more developed “flat places” should take into account the experience of institutionalizing the Northern territories of the Russian Federation.

Here, the authors present the cryo-EM structure of in vitro amyloid fibrils from recombinant SAA1.1 protein that were formed by seeding with fibrils purified from systemic AA amyloidosis tissue. This in vitro fibril structure resembles the structure of the ex vivo fibrils but differs from unseeded in vitro fibrils. These findings show that fibril morphologies can be propagated in vitro by seeding.

Systemic AL amyloidosis is caused by misfolding of immunoglobulin light chains (LCs) but how post-translational modifications (PTMs) of LCs influence amyloid formation is not well understood. Here, the authors present the cryo-EM structure of an AL amyloid fibril derived from the heart tissue of a patient that is partially pyroglutamylated, N-glycosylated and contains an intramolecular disulfide bond. Based on their structure and biochemical experiments the authors conclude that the mutational changes, disulfide bond and glycosylation determine the fibril protein fold and that glycosylation protects the fibril core from proteolytic degradation.

[This corrects the article DOI: 10.1371/journal.pone.0242247.].

Structural and functional brain alterations are found in adults with depression. It is not known whether these changes are a result of illness or exist prior to disorder onset. Asymptomatic offspring of parents with depression offer a unique opportunity to research neural markers of familial risk to depression and clarify the temporal sequence between brain changes and disorder onset. We conducted a systematic review to investigate whether asymptomatic offspring at high familial risk have structural and functional brain changes like those reported in adults with depression. Our literature search resulted in 44 studies on 18,645 offspring ranging from 4 weeks to 25 years old. Reduced cortical thickness and white matter integrity, and altered striatal reward processing were the most consistent findings in high-risk offspring across ages. These alterations are also present in adults with depression, suggesting the existence of neural markers of familial risk for depression. Additional studies reproducing current results, streamlining fMRI data analyses, and investigating underexplored topics (i.e intracortical myelin, gyrification, subcortical shape) may be among the next steps required to improve our understanding of neural markers indexing the vulnerability to depression.

The design of proteolysis targeting chimeras (PROTACs) has become a promising technology for modifying a protein of interest (POI) through protein degradation. Herein, we describe the synthetic pathway to develop N4-(2-amino-4-fluorophenyl)-N1-(3-{2-[2-(3-{[2-(2,6-dioxo-3-piperidyl)-1,3-dioxoisoindolin-4-yl]amino}propoxy)ethoxy]ethoxy}propyl)terephthalamide, which was designed to work as a selective degrader of histone deacetylase-3 (HDAC3). The newly synthesized compounds were characterized by 1H-NMR, 13C-NMR, IR and HRMS. The title compound was tested in vitro against human class-I HDACs isoforms and showed IC50 = 3.4 µM against HDAC3; however, it did not show degradation for the targeted HDACs.

Sedimentation experiments have shown that gravity can deeply affect bulk phenomena and sedimentation of colloidal mixtures. Here, the authors present a theoretical approach based on sedimentation paths to predict the effect of gravity on the emergence of different sedimentation phases in mixtures of anisotropic colloids.

Noether’s Theorem relates symmetries to fundamental physical laws. Rather than applying the concept to an action integral in order to obtain conservation laws, here the authors consider Statistical Mechanical objects, such as the free energy and density and power functionals to derive exact force and torque sum rules.

Currently, contrast-enhanced MRI is the method of choice for treatment planning and follow-up in patients with meningioma. However, positron emission tomography (PET) imaging of somatostatin receptor subtype 2 (SSTR2) expression using 68Ga-DOTATATE may provide a higher sensitivity for meningioma detection, especially in cases with complex anatomy or in the recurrent setting. Here, we report on a patient with a multilocal recurrent atypical meningioma, in which 68Ga-DOTATATE PET was considerably helpful for treatment guidance and decision-making.

In about 20% of all cases, the fine needle aspiration cytology (FNAC) results are equivocal, delivering the two common categories of indeterminate cytology, Bethesda III and IV. The observed rates of malignancy vary widely among institutions, with the urgent need for a more precise risk stratification. 99mcTc methoxyisobutylisonitrile scintigraphy (MIBI) is less expensive than molecular testing and has been shown to have a high negative predictive value. For this reason, the results of MIBI scintigraphy in adult patients with indeterminate FNAC were collected, and correlated with the final pathology reports of surgical specimens. Patients receiving FNAC, MIBI scintigraphy and surgery for sonographic suspicious hypofunctional thyroid nodules between 2015 and 2019 at the University Hospital of Cologne, Germany were identified. Sensitivity, specificity, positive predictive and negative predictive value of MIBI scintigraphy were calculated. Seventy-eight patients with sonographic suspicious hypofunctional thyroid nodules underwent surgery after interdisciplinary case discussion of both FNAC and MIBI results. In 49 (62.5%) cases, FNAC consisted of Bethesda III and IV results. In 39 (79.6%) of these cases, MIBI scintigraphy resulted in mismatch and intermediary results, but in only 4 (10.2%) of these cases was a carcinoma diagnosed. The negative predictive value of MIBI scintigraphy was 90–100%, respectively. Relying on the negative predictive value of MIBI match results might have obviated the need for surgery in 20.4% cases; one papillary microcarcinoma, however, would have been missed. MIBI scintigraphy has an underused potential for improving the diagnostic precision of hypofunctional thyroid nodules.

Systemic AA amyloidosis is a protein misfolding disease caused by the formation of amyloid fibrils from serum amyloid A (SAA) protein. Here, the authors present the cryo-EM structures of AA amyloid fibrils isolated from mouse tissue and in vitro formed fibrils, which differ in their structures and they also show that the ex vivo fibrils are more resistant to proteolysis than the in vitro fibrils and propose that pathogenic amyloid fibrils might originate from proteolytic selection.

Systemic AL amyloidosis is a protein misfolding disease caused by the aggregation and fibrillation of immunoglobulin light chains (LCs). Here, the authors present the cryo-EM structures of λ3 LC-derived amyloid fibrils that were isolated from patient tissue and they observe structural breaks, where the two different fibril structures co-exist at different z-axial positions within the same fibril.

BackgroundHürthle cell carcinoma (HCC) of the thyroid is rare. There are contrasting data on its clinical behavior. The aim of this study was to describe clinic-pathological features and outcomes of HCC patients at our institution, in order to adapt our surgical management.MethodsWe retrospectively studied 51 cases of HCC treated at the interdisciplinary endocrine center of the University Hospital of Cologne, Germany between 2005 and 2020.ResultsPatients median age was 63 years (range 29-78) with 64.7% of cases being female. Primary treatment included surgery and postoperative radioiodine therapy with 3.7 GBq in all patients. Surgery consisted of total thyroidectomy in all cases and additional central lymphadenectomy in 90.2% of cases. The median number of harvested lymph nodes was 11 (range 2-31). Lymph node involvement was found in two (4.3%) pT4a tumors. In all other cases (95.7%), central lymphadenectomy was prophylactic and lymph nodes were free of metastasis in final histopathology. Twelve (23.5%) patients with incomplete biochemical response to primary treatment were diagnosed with structural relapse during the course of disease, for which seven (58.4%) underwent resection of isolated cervical metastasis. Histopathology revealed soft tissue implants in all cases and cervical surgery led to biochemical and radiologic cure in only two (28.5%) cases. Five (41.6%) patients developed metastatic disease, followed by systemic therapy in two patients. Vascular invasion of the primary tumor was significantly associated with relapse (p

Scanning helium-ion microscopy (HIM) is an imaging technique with sub-nanometre resolution and is a powerful tool to resolve some of the tiniest structures in biology. In many aspects, the HIM resembles a field-emission scanning electron microscope (FE-SEM), but the use of helium ions rather than electrons provides several advantages, including higher surface sensitivity, larger depth of field, and a straightforward charge-compensating electron flood gun, which enables imaging of non-conductive samples, rendering HIM a promising high-resolution imaging technique for biological samples. Starting with studies focused on medical research, the last decade has seen some particularly spectacular high-resolution images in studies focused on plants, microbiology, virology, and geomicrobiology. However, HIM is not just an imaging technique. The ability to use the instrument for milling biological objects as small as viruses offers unique opportunities which are not possible with more conventional focused ion beams, such as gallium. Several pioneering technical developments, such as methods to couple secondary ion mass spectrometry (SIMS) or ionoluminescence with the HIM, also offer the possibility for new and exciting research on biological materials. In this review, we present a comprehensive overview of almost all currently published literature which has demonstrated the application of HIM for imaging of biological specimens. We also discuss some technical features of this unique type of instrument and highlight some of the new advances which will likely become more widely used in the years to come.

PurposeIt is not mandatory to report lymphatic vessel invasion in pathology reports of papillary thyroid cancer (PTC) according to the current Union for International Cancer Control (UICC) TNM (tumor, nodes, and metastases) classification. However, there is some evidence for its correlation with lymph node metastasis (LNM) and prognosis. The aim of this study was to explore the clinical implication of lymphatic vessel invasion documentation of PTC because pathology reports play a pivotal role in postsurgical clinical decision-making in endocrine tumor boards.MethodsPatients undergoing postoperative radioiodine treatment for PTC at the University Hospital of Cologne, Germany between December 2015 and March 2020 were identified. Pathology reports were screened for documentation of lymphatic vessel invasion. Demographics and clinicopathologic data of patients documented, including lymphatic vessel invasion and lymph nodal involvement were analyzed.ResultsA total of 578 patients were identified and included. Lymphatic vessel invasion was reported in pathology reports of 366 (63.3%) and omitted in 112 (36.7%) patients. Positive lymphatic vessel invasion (L1) was diagnosed in 67 (18.3%) of 366 patients and was documented as absent (L0) in 299 (81.7%) patients. Lymph nodal (N) status was positive (N+) in 126 (45.6%) and negative (N0) in 150 (54.3%) of these patients. In 54 (80.6%) L1 cases N+ status and in 137 (65.6%) L0 cases N0 status was diagnosed. In 13 (19.4%) cases with L1 status, there were no LNMs (L1 N0). In total, 72 (34.4%) patients had LNM despite L0 status (L0 N+). The sensitivity and specificity of LVI reporting for LNM were 0.42 and 0.91, respectively.ConclusionIn routine pathology reports of PTC used for indication to postoperative radioiodine treatment by a German endocrine tumor board, lymphatic vessel invasion was found to be reported inconsistently and mostly as L0. L1 diagnoses, however, reliably correlated with reported LNM and might, thus, be relevant for clinical decision-making. For this reason, we advocate for standardized pathologic reassessment of lymphatic vessel invasion, in particular for cases where lymph nodes are not included in the pathologic specimen and if L0 is documented.

AA amyloidosis is one of the most prevalent forms of systemic amyloidosis and affects both humans and other vertebrates. In this study, we compare MAS solid-state NMR data with a recent cryo-EM study of fibrils involving full-length murine SAA1.1. We address the question whether the specific requirements for the reconstitution of an amyloid fibril structure by cryo-EM can potentially yield a bias towards a particular fibril polymorph. We employ fibril seeds extracted from in to vivo material to imprint the fibril structure onto the biochemically produced protein. Sequential assignments yield the secondary structure elements in the fibril state. Long-range DARR and PAR experiments confirm largely the topology observed in the ex-vivo cryo-EM study. We find that the β-sheets identified in the NMR experiments are similar to the β-sheets found in the cryo-EM study, with the exception of amino acids 33–42. These residues cannot be assigned by solid-state NMR, while they adopt a stable β-sheet in the cryo-EM structure. We suggest that the differences between MAS solid-state NMR and cryo-EM data are a consequence of a second conformer involving residues 33–42. Moreover, we were able to characterize the dynamic C-terminal tail of SAA in the fibril state. The C-terminus is flexible, remains detached from the fibrils, and does not affect the SAA fibril structure as confirmed further by molecular dynamics simulations. As the C-terminus can potentially interact with other cellular components, binding to cellular targets can affect its accessibility for protease digestion.

In order to partially mimic the efficient lignocellulose pretreatment process performed naturally in the gut system of Pachnoda marginata larvae, two wheat straw pretreatments were evaluated: a mechanical pretreatment via cutting the straw into two different sizes and an alkaline pretreatment with calcium hydroxide. After pretreatment, gut enrichment cultures on wheat straw at alkaline pH were inoculated and kept at mesophilic conditions over 45 days. The methanogenic community was composed mainly of the Methanomicrobiaceae and Methanosarcinaceae families. The combined pretreatment, size reduction and alkaline pretreatment, was the best condition for methane production. The positive effect of the straw pretreatment was higher in the midgut cultures, increasing the methane production by 192%, while for hindgut cultures the methane production increased only by 149% when compared to non-pretreated straw. Scanning electron microscopy (SEM) showed that the alkaline pretreatment modified the surface of the wheat straw fibers, which promoted biofilm formation and microbial growth. The enrichment cultures derived from larva gut microbiome were able to degrade larger 1 mm alkaline treated and smaller 250 µm but non-pretreated straw at the same efficiency. The combination of mechanical and alkaline pretreatments resulted in increased, yet not superimposed, methane yield.