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Abstract Over the past five decades, a large number of wild animals have been individually identified by various observation systems and/or temporary tracking methods, providing unparalleled insights into their lives over both time and space. However, so far there is no comprehensive record of uniquely individually identified animals nor where their data and metadata are stored, for example photos, physiological and genetic samples, disease screens, information on social relationships. Databases currently do not offer unique identifiers for living, individual wild animals, similar to the permanent ID labelling for deceased museum specimens. To address this problem, we introduce two new concepts: (1) a globally unique animal ID (UAID) available to define uniquely and individually identified animals archived in any database, including metadata archived at the time of publication; and (2) the digital ‘home’ for UAIDs, the Movebank Life History Museum (MoMu), storing and linking metadata, media, communications and other files associated with animals individually identified in the wild. MoMu will ensure that metadata are available for future generations, allowing permanent linkages to information in other databases. MoMu allows researchers to collect and store photos, behavioural records, genome data and/or resightings of UAIDed animals, encompassing information not easily included in structured datasets supported by existing databases. Metadata is uploaded through the Animal Tracker app, the MoMu website, by email from registered users or through an Application Programming Interface (API) from any database. Initially, records can be stored in a temporary folder similar to a field drawer, as naturalists routinely do. Later, researchers and specialists can curate these materials for individual animals, manage the secure sharing of sensitive information and, where appropriate, publish individual life histories with DOIs. The storage of such synthesized lifetime stories of wild animals under a UAID (unique identifier or ‘animal passport’) will support basic science, conservation efforts and public participation.

Abstract Background Network theory is largely applied in real-world systems to assess landscape connectivity using empirical or theoretical networks. Empirical networks are usually built from discontinuous individual movement trajectories without knowing the effect of relocation frequency on the assessment of landscape connectivity while theoretical networks generally rely on simple movement rules. We investigated the combined effects of relocation sampling frequency and landscape fragmentation on the assessment of landscape connectivity using simulated trajectories and empirical high-resolution (1 Hz) trajectories of Alpine ibex (Capra ibex). We also quantified the capacity of commonly used theoretical networks to accurately predict landscape connectivity from multiple movement processes. Methods We simulated forager trajectories from continuous correlated biased random walks in simulated landscapes with three levels of landscape fragmentation. High-resolution ibex trajectories were reconstructed using GPS-enabled multi-sensor biologging data and the dead-reckoning technique. For both simulated and empirical trajectories, we generated spatial networks from regularly resampled trajectories and assessed changes in their topology and information loss depending on the resampling frequency and landscape fragmentation. We finally built commonly used theoretical networks in the same landscapes and compared their predictions to actual connectivity. Results We demonstrated that an accurate assessment of landscape connectivity can be severely hampered (e.g., up to 66% of undetected visited patches and 29% of spurious links) when the relocation frequency is too coarse compared to the temporal dynamics of animal movement. However, the level of landscape fragmentation and underlying movement processes can both mitigate the effect of relocation sampling frequency. We also showed that network topologies emerging from different movement behaviours and a wide range of landscape fragmentation were complex, and that commonly used theoretical networks accurately predicted only 30–50% of landscape connectivity in such environments. Conclusions Very high-resolution trajectories were generally necessary to accurately identify complex network topologies and avoid the generation of spurious information on landscape connectivity. New technologies providing such high-resolution datasets over long periods should thus grow in the movement ecology sphere. In addition, commonly used theoretical models should be applied with caution to the study of landscape connectivity in real-world systems as they did not perform well as predictive tools.

Abstract The application of supervised machine learning methods to identify behavioural modes from inertial measurements of bio-loggers has become a standard tool in behavioural ecology. Several design choices can affect the accuracy of identifying the behavioural modes. One such choice is the inclusion or exclusion of segments consisting of more than a single behaviour (mixed segments) in the machine learning model training data. Currently, the common practice is to ignore such segments during model training. In this paper we tested the hypothesis that including mixed segments in model training will improve accuracy, as the model would perform better in identifying them in the test data. We test this hypothesis using a series of data simulations on four datasets of accelerometer data coupled with behaviour observations, obtained from four study species (Damaraland mole-rats, meerkats, olive baboons, polar bears). Results show that when a substantial proportion of the test data are mixed behaviour segments (above ~ 10%), including mixed segments in machine learning model training improves the accuracy of classification. These results were consistent across the four study species, and robust to changes in segment length, sample size, and degree of mixture within the mixed segments. However, we also find that in some cases (particularly in baboons) models trained with mixed segments show reduced accuracy in classifying test data containing only single behaviour (pure) segments, compared to models trained without mixed segments. Based on these results, we recommend that when the classification model is expected to deal with a substantial proportion of mixed behaviour segments (> 10%), it is beneficial to include them in model training, otherwise, it is unnecessary but also not harmful. The exception is when there is a basis to assume that the training data contains a higher rate of mixed segments than the actual (unobserved) data to be classified—such a situation may occur particularly when training data are collected in captivity and used to classify data from the wild. In this case, excess inclusion of mixed segments in training data should probably be avoided.

Global hydrological models enhance our understanding of the Earth system and support the sustainable management of water, food and energy in a globalized world. They integrate process knowledge with a multitude of model input data (e.g., precipitation, soil properties, and the location and extent of surface waterbodies) to describe the state of the Earth. However, they do not fully utilize observations of model output variables (e.g., streamflow and water storage) to reduce and quantify model output uncertainty through processes like parameter estimation. For a pilot region, the Mississippi River basin, we assessed the suitability of three ensemble-based multi-variable approaches to amend this: Pareto-optimal calibration (POC); the generalized likelihood uncertainty estimation (GLUE); and the ensemble Kalman filter, here modified for joint calibration and data assimilation (EnCDA). The paper shows how observations of streamflow (Q) and terrestrial water storage anomaly (TWSA) can be utilized to reduce and quantify the uncertainty of model output by identifying optimal and behavioral parameter sets for individual drainage basins. The common first steps in all approaches are (1) the definition of drainage basins for which calibration parameters are uniformly adjusted (CDA units), combined with the selection of observational data; (2) the identification of potential calibration parameters and their a priori probability distributions; and (3) sensitivity analyses to select the most influential model parameters per CDA unit that will be adjusted by calibration. Data assimilation with the ensemble Kalman filter was modified, to our knowledge, for the first time for a global hydrological model to assimilate both TWSA and Q with simultaneous parameter adjustment. In the estimation of model output uncertainty, we considered the uncertainties of the Q and TWSA observations. Applying the global hydrological model WaterGAP, we found that the POC approach is best suited for identifying a single “optimal” parameter set for each CDA unit. This parameter set leads to an improved fit to the monthly time series of both Q and TWSA as compared to the standard WaterGAP variant, which is only calibrated against mean annual Q, and can be used to compute the best estimate of WaterGAP output. The GLUE approach is almost as successful as POC in increasing WaterGAP performance and also allows, with a comparable computational effort, the estimation of model output uncertainties that are due to the equifinality of parameter sets given the observation uncertainties. Our experiment reveals that the EnCDA approach performs similarly to POC and GLUE in most CDA units during the assimilation phase but is not yet competitive for calibrating global hydrological models; its potential advantages remain unrealized, likely due to its high computational burden, which severely limits the ensemble size, and the intrinsic nonlinearity in simulating Q. Partitioning the whole Mississippi River basin into five CDA units (sub-basins) instead of only one improved model performance in terms of the Nash–Sutcliffe efficiency during the calibration and validation periods. Diverse parameter sets achieved comparable fits to observations, narrowing the range for at least three parameters. Low coverage of observation uncertainty bands by GLUE-derived model output bands is attributed to model structure uncertainties, especially regarding artificial reservoir operations, the location and extent of small wetlands, and the lack of representation of rivers that may lose water to the subsurface. These uncertainties are also likely to be responsible for significant trade-offs between optimal fits to Q and TWSA. Calibration performed exclusively against TWSA in regions without Q observations may worsen the Q simulation as compared to the uncalibrated model variant. We recommend that modelers improve the realism of the output of global hydrological models by calibrating them against observations of multiple output variables, including at least Q and TWSA. Further work on improving the numerical efficiency of the EnCDA approach is necessary.

Abstract Biologging has proven to be a powerful approach to investigate diverse questions related to movement ecology across a range of spatiotemporal scales and increasingly relies on multidisciplinary expertise. However, the variety of animal‐borne equipment, coupled with little consensus regarding analytical approaches to interpret large, complex data sets presents challenges and makes comparison between studies and study species difficult. Here, we present a combined hardware and analytical approach for standardizing the collection, analysis, and interpretation of multisensor biologging data. Here, we present (i) a custom‐designed integrated multisensor collar (IMSC), which was field tested on 71 free‐ranging wild boar (Sus scrofa) over 2 years; (ii) a machine learning behavioral classifier capable of identifying six behaviors in free‐roaming boar, validated across individuals equipped with differing collar designs; and (iii) laboratory and field‐based calibration and accuracy assessments of animal magnetic heading measurements derived from raw magnetometer data. The IMSC capacity and durability exceeded expectations, with a 94% collar recovery rate and a 75% cumulative data recording success rate, with a maximum logging duration of 421 days. The behavioral classifier had an overall accuracy of 85% in identifying the six behavioral classes when tested on multiple collar designs and improved to 90% when tested on data exclusively from the IMSC. Both laboratory and field tests of magnetic compass headings were in precise agreement with expectations, with overall median magnetic headings deviating from ground truth observations by 1.7° and 0°, respectively. Although multisensor equipment and sophisticated analyses are now commonplace in biologging studies, the IMSC hardware and analytical framework presented here provide a valuable tool for biologging researchers and will facilitate standardization of biologging data across studies. In addition, we highlight the potential of additional analyses available using this framework that can be adapted for use in future studies on terrestrial mammals.

Abstract Many animals return to their home areas (i.e., ‘homing’) after translocation to sites further away. Such translocations have traditionally been used in behavioral ecology to understand the orientation and migration behavior of animals. The movement itself can then be followed by marking and recapturing animals or by tracking, for example, using GPS systems. Most detailed studies investigating this behavior have been conducted in smaller vertebrates (e.g., birds, amphibians, and mice), whereas information on larger mammals, such as red deer, is sparse. We conducted GPS-assisted translocation experiments with red deer at two sites in the Czech Republic. Individuals were translocated over a distance of approximately 11 km and their home journey was tracked. Circular statistics were used to test for significant homeward orientation at distances of 100, 500, 1000, and 5000 m from the release site. In addition, we applied Lavielle trajectory segmentation to identify the different phases of homing behavior. Thirty-one out of 35 translocations resulted in successful homing, with a median time of 4.75 days (range 1.23–100 days). Animals were significantly oriented towards home immediately after release and again when they came closer to home; however, they did not show a significant orientation at the distances in between. We were able to identify three homing phases, an initial ‘exploratory phase’, followed by a ‘homing phase’ which sometimes was again followed by an ‘arrival phase’. The ‘homing phase’ was characterized by the straightest paths and fastest movements. However, the variation between translocation events was considerable. We showed good homing abilities of red deer after translocation. Our results demonstrate the feasibility of conducting experiments with environmental manipulations (e.g., to impede the use of sensory cues) close to the release site. The homing behavior of red deer is comparable to that of other species, and might represent general homing behavior patterns in animals. Follow-up studies should further dissect and investigate the drivers of the individual variations observed and try to identify the sensory cues used during homing.

Abstract Foraging is a key driver of animal movement patterns, with specific challenges for predators which must search for mobile prey. These patterns are increasingly impacted by global changes, principally in land use and climate. Understanding the degree of flexibility in predator foraging and social strategies is pertinent to wildlife conservation under global change, including potential top-down effects on wider ecosystems. Here we propose key future research directions to better understand foraging strategies and social flexibility in predators. In particular, rapid continued advances in biologging technology are helping to record and understand dynamic behavioural and movement responses of animals to environmental changes, and their energetic consequences. Data collection can be optimised by calibrating behavioural interpretation methods in captive settings and strategic tagging decisions within and between social groups. Importantly, many species’ social systems are increasingly being found to be more flexible than originally described in the literature, which may be more readily detectable through biologging approaches than behavioural observation. Integrating the effects of the physical landscape and biotic interactions will be key to explaining and predicting animal movements and energetic balance in a changing world.

Various studies have shown that video-based learning by explaining to a fictitious audience can be an effective learning strategy for promoting multiple knowledge domains such as memory, comprehension and knowledge transfer. However, field studies testing the effectiveness of this learning strategy in an applied setting are rare. The present study examines the effectiveness of the one-take video (OTV) approach on undergraduate students' learning performance. The OTV method involves users recording short oral presentations without any editing and with the support of handwritten visualisations (video-based learning by explaining). To test the learning outcomes, 218 undergraduate teaching students for special educational needs were randomly assigned to two test groups (OTV and explaining in writing). After that, they completed three study tasks throughout the semester, each followed by immediate and delayed knowledge tests. The results for the OTV group show that students achieved significantly better results in the immediate memory test the more handwritten visualisations they used, but not the more often they repeated the video recordings. Analyses of variance revealed that the OTV group outperformed the writing group in terms of memory performance in the immediate test but not in the delayed posttest. The OTV group also significantly outperformed the writing group in both posttests in the transfer domain. No significant differences were found in the comprehension test scores.

The spread of antimicrobial resistance (AMR) in animal husbandry is usually attributed to the use of antibiotics and poor hygiene and biosecurity. We therefore conducted experimental trials to improve hygiene management in weaned pig houses and assessed the impact on the spread. For each of the two groups examined, the experimental group (EG) and the control group (CG), three replicate batches of piglets from the same pig breeder, kept in pre-cleaned flat decks, were analyzed. In the flat decks of the experimental groups, the hygiene conditions (cleaning, disinfection, dust removal and fly control) were improved, while regular hygiene measures were carried out in the control groups. The occurrence and spread of AMR were determined in Escherichia coli (E. coli; resistance indicator) using cultivation-dependent (CFU) and -independent (qPCR) methods as well as whole genome sequencing of isolates in samples of various origins, including feces, flies, feed, dust and swabs. Surprisingly, there were no significant differences (p > 0.05) in the prevalence of resistant E. coli between the flat decks managed with conventional techniques and those managed with improved techniques. Selective cultivation delivered ampicillin- and sulfonamide-resistant E. coli proportions of up to 100% and 1.2%, respectively. While 0.5% E. coli resistant to cefotaxime and no ciprofloxacin resistance were detected. There was a significant difference (p

Abstract Cell culture‐conditioned medium (CCM) is a valuable source of extracellular vesicles (EVs) for basic scientific, therapeutic and diagnostic applications. Cell culturing parameters affect the biochemical composition, release and possibly the function of CCM‐derived EVs (CCM‐EV). The CCM‐EV task force of the Rigor and Standardization Subcommittee of the International Society for Extracellular Vesicles aims to identify relevant cell culturing parameters, describe their effects based on current knowledge, recommend reporting parameters and identify outstanding questions. While some recommendations are valid for all cell types, cell‐specific recommendations may need to be established for non‐mammalian sources, such as bacteria, yeast and plant cells. Current progress towards these goals is summarized in this perspective paper, along with a checklist to facilitate transparent reporting of cell culturing parameters to improve the reproducibility of CCM‐EV research.

Abstract Gallbladder cancer (GBC) is a rare, highly aggressive malignancy with a 5-year survival rate of 5–10% in advanced cases, highlighting the need for more effective therapies. The aim of this study was to identify potentially actionable therapeutic targets for GBC. Specimens and clinicopathological data of 642 GBC patients, diagnosed between 2000 and 2019 were collected using the Dutch Pathology Registry (PALGA) and the Netherlands Cancer Registry. All cases were histologically reviewed and a subset was subjected to a comprehensive next generation sequencing panel. We assessed mutations and gene amplifications in a panel of 54 actionable genes, tumor-mutational burden (TMB), and microsatellite instability (MSI). Additionally, the entire cohort was screened for HER2, PD-L1, pan-TRK, and p53 expression with immunohistochemistry. Histopathological subtypes comprised biliary-type adenocarcinoma (AC, 69.6%), intestinal-type AC (20.1%) and other subtypes (10.3%). The median total TMB was 5.5 mutations/Mb (range: 0–161.1) and 17.7% of evaluable cases had a TMB of >10 mutations/Mb. MSI was observed in two cases. Apart from mutations in TP53 (64%), tumors were molecularly highly heterogeneous. Half of the tumors (50%) carried at least one molecular alteration that is targetable in other tumor types, including alterations in CDKN2A (6.0% biallelically inactivated), ERBB2 (9.3%) and PIK3CA (10%). Immunohistochemistry results correlated well with NGS results for HER2 and p53: Pearson r = 0.82 and 0.83, respectively. As half of GBC patients carry at least one potentially actionable molecular alteration, molecular testing may open the way to explore targeted therapy options for GBC patients.

IntroductionOsteoarthritis (OA) affects a large percentage of the population worldwide. Current surgical and nonsurgical concepts for treating OA only result in symptom-modifying effects. However, there is no disease-modifying therapy available. Extracellular vesicles released by mesenchymal stem/stromal cells (MSC-EV) are promising agents to positively influence joint homeostasis in the osteoarthritic surroundings. This pilot study aimed to investigate the effect of characterized MSC-EVs on chondrogenesis in a 3D chondrocyte inflammation model with the pro-inflammatory cytokine TNFα.MethodsBovine articular chondrocytes were expanded and transferred into pellet culture at passage 3. TNFα, human MSC-EV preparations (MSC-EV batches 41.5-EVi1 and 84-EVi), EVs from human platelet lysate (hPL4-EV), or the combination of TNFα and EVs were supplemented. To assess the effect of MSC-EVs in the chondrocyte inflammation model after 14 days, DNA, glycosaminoglycan (GAG), total collagen, IL-6, and NO release were quantified, and gene expression of anabolic (COL-II, aggrecan, COMP, and PRG-4), catabolic (MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5), dedifferentiation (COL-I), hypertrophy (COL-X, VEGF), and inflammatory (IL-8) markers were analyzed; histological evaluation was performed using safranin O/Fast Green staining and immunohistochemistry of COL I and II. For statistical evaluation, nonparametric tests were chosen with a significance level of p < 0.05.ResultsTNFα supplementation resulted in catabolic stimulation with increased levels of NO and IL-6, upregulation of catabolic gene expression, and downregulation of anabolic markers. These findings were supported by a decrease in matrix differentiation (COL-II). Supplementation of EVs resulted in an upregulation of the chondrogenic marker PRG-4. All MSC-EV preparations significantly increased GAG retention per pellet. In contrast, catabolic markers and IL-8 expression were upregulated by 41.5-EVi1. Regarding protein levels, IL-6 and NO release were increased by 41.5-EVi1. Histologic and immunohistochemical evaluations indicated a higher differentiation potential of chondrocytes treated with 84-EVi.DiscussionMSC-EVs can positively influence chondrocyte matrix production in pro-inflammatory surroundings, but can also stimulate inflammation. In this study MSC-EV 41.5-EVi1 supplementation increased chondrocyte inflammation, whereas MSC-84-EVi supplementation resulted a higher chondrogenic potential of chondrocytes in 3D pellet culture. In summary, the selected MSC-EVs exhibited promising chondrogenic effects indicating their significant potential for the treatment of OA; however, the functional heterogeneity in MSC-EV preparations has to be solved.

Introduction Cardiovascular diseases are the most common cause of death in Germany and among the most frequent reasons for encounters in primary care. Most patients with cardiovascular risks (CVRs) have difficulties implementing health-promoting behavioural changes. In this study, a complex intervention containing evidence-based patient materials and structured follow-up consultations are intended to strengthen patients’ self-management to improve health behaviour.Methods and analysis In this cluster randomised controlled trial, we investigate the effects of the intervention Decision aid, action planning and follow-up support for patients to reduce the 10-year risk of cardiovascular diseases (DECADE) using a 2×2 design. All patients, including the control group (CG), receive a CVR calculation. Three intervention groups (IGs) receive one or both of two different components of the DECADE intervention: IG1 (patient materials), IG2 (follow-up consultations) and IG3 (patient materials and follow-up consultations). The study was planned to be conducted with 77 general practitioners in 3 German regions and a target sample size of 924 patients. The observation period for each patient amounts to 12 months with three patient surveys: baseline (t0), after 6 and 12 months (t1 and t2). The primary outcome is patient activation (Patient Activation Measure 13 (PAM13-D)) at t1. Secondary outcomes include PAM13-D at t2 and further patient-reported and clinical outcomes at t1 and t2. We will also analyse the cost-effectiveness of the intervention, the degree of usage and satisfaction with the intervention.Ethics and dissemination The study was first approved by the lead ethics committee of the University of Freiburg on 15 April 2021 (vote number: 21-1078) and subsequently by the other ethics committees in the study regions (Ethics committee of medical association Baden-Württemberg (B-F-2021-078), Ethics Committee of the Technische Universität Dresden, Dresden (BO-EK-251052021), Ethics Committee of the State Chamber of Physicians of Saxony (EK-BR-92/21-1), Ethics Committee of the Hamburg Medical Association (2021-200013-BO-bet)). Informed consent is required for patients to participate in the study. The results of this study will be published in peer-reviewed journals and presented at congresses by the DECADE team. The DECADE lead management will communicate the results to the funder of this study.Trial registration number German Clinical Trials Register, DRKS00025401 (registration date: 21 June 2021); International Clinical Trials Registry Platform, DRKS00025401.

Aim Methods and Results Conclusion The term ‘intramucosal carcinoma’ in the colorectum is controversial when used as a synonym for high‐grade dysplasia. However, setting clear definitions for this diagnosis (i.e. unequivocal infiltrative growth in the lamina propria, which might be most easily recognized in cases with overt poor differentiation or formation of signet ring cells or tumour budding) allow us to investigate its relevance.We reviewed cases from our archive (1990–2024) and selected 14 true intramucosal carcinomas using our strict histological criteria, excluding high‐grade dysplasia and invasive carcinomas. These occur primarily in conventional adenomas and are frequently associated with microsatellite instability (50%). Our study shows a low estimated incidence of intramucosal carcinoma (0.01%) in population screening and highlights the low lymph node risk and the good outcome of patients with intramucosal carcinoma.The rare diagnosis of intramucosal colorectal carcinoma aids in identifying patients at increased colorectal cancer risk, notably those with hereditary syndromes. Standardizing this diagnosis is critical to prevent overdiagnosis and unnecessary treatment. [ABSTRACT FROM AUTHOR]

Aims: Traditionally, mismatch repair (MMR) status is determined by a panel of four antibodies (MLH1, PMS2, MSH2, MSH6). If all proteins are retained, cases are MMR proficient (pMMR), while loss of one or more proteins is indicative of MMR deficiency (dMMR). This approach has been challenged in favour of a two‐antibody approach, using PMS2 and MSH6 as a first screening. Their retainment is deemed sufficient to declare cases pMMR. In this study we aim to verify the validity of the two‐antibody approach. Methods and Results: We performed a nationwide study in colorectal cancer (CRC) and endometrial cancer (EC) diagnosed between 2016 and 2023, including 47,657 patients to evaluate the two‐antibody approach. In 0.17% and 0.4% of cases of CRC and EC, respectively, dMMR cases would be missed with the two‐antibody approach. Subgroup analyses pointed towards slightly increased miss rates in younger patients (under the age of 50 years) in both groups and identified special subtypes (signet ring cell carcinoma, medullary carcinoma, and mucinous carcinoma in CRC and clear cell carcinoma in EC) with increased miss rates. For these specific subgroups, a low threshold should be used for further testing. In case of ambiguous or heterogeneous staining patterns, four antibodies should be used. Conclusion: In general, the application of a two‐antibody MMR testing strategy does not lead to considerable failure of dMMR identification and saves costs. [ABSTRACT FROM AUTHOR]

This work is concerned with the modeling of a cold‐box sand (CBS), a composition of sand grains and a resin binder. To this end, experiments are performed which show the following characteristics: localization phenomena in form of a shear band, softening behavior in the force‐displacement curve, asymmetric behavior for compression and tension. These complex characteristics require a sophisticated material model. For this purpose a micropolar continuum, which is a special case of the micromorphic continuum, is used. In addition to the degrees of freedom of a classical continuum, the micropolar model has additional degrees of freedom in the form of micro‐rotations, which represent the grains of sand in our analyzed material. The macro‐part of this model represents the binder of the CBS. In order to make the micropolar model as flexible as possible, two flow functions are introduced; one for the macro‐part and one for the micro‐part. This approach is called double‐surface plasticity. The two flow functions can be implemented independently or coupled, depending on the application. For the coupled case, the radial‐return algorithm is no longer sufficient, so that the local iteration is calculated with the Fischer–Burmeister residual. Due to the lack of rotational symmetry in the experimental results, it is not possible to compute in two dimensions; instead, a 3D model is used for the simulation. [ABSTRACT FROM AUTHOR]