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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.

StatementThe International Society for Cellular and Gene Therapies (ISCT) and the International Society for Extracellular Vesicles (ISEV) recognize the potential of extracellular vesicles (EVs, including exosomes) from mesenchymal stromal cells (MSCs) and possibly other cell sources as treatments for COVID-19. Research and trials in this area are encouraged. However, ISEV and ISCT do not currently endorse the use of EVs or exosomes for any purpose in COVID-19, including but not limited to reducing cytokine storm, exerting regenerative effects or delivering drugs, pending the generation of appropriate manufacturing and quality control provisions, pre-clinical safety and efficacy data, rational clinical trial design and proper regulatory oversight.

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]

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 FROM AUTHOR]

Abstract Oceanic circulation and mass‐field variability play important roles in exciting Earth's wobbles and length‐of‐day changes (ΔΛ), on time scales from days to several years. Modern descriptions of these effects employ oceanic angular momentum (OAM) series from numerical forward models or ocean state estimates, but nothing is known about how ocean reanalyses with sequential data assimilation (DA) would fare in that context. Here, we compute daily OAM series from three 1/4° global ocean reanalyses that are based on the same hydrodynamic core and input data (e.g., altimetry, Argo) but different DA schemes. Comparisons are carried out (a) among the reanalyses, (b) with an established ocean state estimate, and (c) with Earth rotation data, all focusing on the period 2006–2015. The reanalyses generally provide credible OAM estimates across a range of frequencies, although differences in amplitude spectra indicate a sensitivity to the adopted DA scheme. For periods less than 120 days, the reanalysis‐based OAM series explain ∼40%–50% and ∼30%–40% of the atmosphere‐corrected equatorial and axial geodetic excitation, similar to what is achieved with the state estimate. We find mixed performance of the reanalyses in seasonal excitation budgets, with some questionable mean ocean mass changes affecting the annual cycle in ΔΛ. Modeled excitations at interannual frequencies are more uncertain compared to OAM series from the state estimate and show hints of DA artifacts in one case. If users are to choose any of the tested reanalyses for rotation research, our study points to the Ocean Reanalysis System 5 as the most sensible choice.

Abstract Addressing ongoing biodiversity loss requires collaboration between conservation scientists and practitioners. However, such collaboration has proved challenging. Despite the potential importance of tracking animal movements for conservation, reviews of the tracking literature have identified a gap between the academic discipline of movement ecology and its application to biodiversity conservation. Through structured conversations with movement ecologists and conservation practitioners, we aimed to understand whether the identified gap is also perceived in practice, and if so, what factors hamper collaboration and how these factors can be remediated. We found that both groups are motivated and willing to collaborate. However, because their motivations differ, there is potential for misunderstandings and miscommunications. In addition, external factors such as funder requirements, academic metrics, and journal scopes may limit the applicability of scientific results in a conservation setting. Potential solutions we identified included improved communication and better presentation of results, acknowledging each other's motivations and desired outputs, and adjustment of funder priorities. Addressing gaps between science and implementation can enhance collaboration and support conservation action to address the global biodiversity crisis more effectively.

Abstract Few biodiversity indicators are available that reflect the state of broad-sense biodiversity—rather than of particular taxa—at fine spatial and temporal resolution. One such indicator, the Biodiversity Intactness Index (BII), estimates how the average abundance of the native terrestrial species in a region compares with their abundances in the absence of pronounced human impacts. We produced annual maps of modelled BII at 30-arc-second resolution (roughly 1 km at the equator) across tropical and subtropical forested biomes, by combining annual data on land use, human population density and road networks, and statistical models of how these variables affect overall abundance and compositional similarity of plants, fungi, invertebrates and vertebrates. Across tropical and subtropical biomes, BII fell by an average of 1.9 percentage points between 2001 and 2012, with 81 countries seeing an average reduction and 43 an average increase; the extent of primary forest fell by 3.9% over the same period. We did not find strong relationships between changes in BII and countries’ rates of economic growth over the same period; however, limitations in mapping BII in plantation forests may hinder our ability to identify these relationships. This is the first time temporal change in BII has been estimated across such a large region.

Abstract Background Understanding what animals do in time and space is important for a range of ecological questions, however accurate estimates of how animals use space is challenging. Within the use of animal-attached tags, radio telemetry (including the Global Positioning System, ‘GPS’) is typically used to verify an animal’s location periodically. Straight lines are typically drawn between these ‘Verified Positions’ (‘VPs’) so the interpolation of space-use is limited by the temporal and spatial resolution of the system’s measurement. As such, parameters such as route-taken and distance travelled can be poorly represented when using VP systems alone. Dead-reckoning has been suggested as a technique to improve the accuracy and resolution of reconstructed movement paths, whilst maximising battery life of VP systems. This typically involves deriving travel vectors from motion sensor systems and periodically correcting path dimensions for drift with simultaneously deployed VP systems. How often paths should be corrected for drift, however, has remained unclear. Methods and results Here, we review the utility of dead-reckoning across four contrasting model species using different forms of locomotion (the African lion Panthera leo, the red-tailed tropicbird Phaethon rubricauda, the Magellanic penguin Spheniscus magellanicus, and the imperial cormorant Leucocarbo atriceps). Simulations were performed to examine the extent of dead-reckoning error, relative to VPs, as a function of Verified Position correction (VP correction) rate and the effect of this on estimates of distance moved. Dead-reckoning error was greatest for animals travelling within air and water. We demonstrate how sources of measurement error can arise within VP-corrected dead-reckoned tracks and propose advancements to this procedure to maximise dead-reckoning accuracy. Conclusions We review the utility of VP-corrected dead-reckoning according to movement type and consider a range of ecological questions that would benefit from dead-reckoning, primarily concerning animal–barrier interactions and foraging strategies.