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I pædagogisk praksis må vi forholde os til den tid og det samfund vi lever i, og i den antropocæne tidsalder betyder det, at klima- og biodiversitetskrise nu og i fremtiden kommer til at præge vores tilværelse. For at afsøge muligheder for at udvikle fremtidens bæredygtighedspædagogik har vi anvendt participatoriske tilgange til at udvikle et fremtidsværkstedsformat der tager udgangspunkt i deltagernes perspektiver og ønsker i projektet ’Børnepanelet’. Perspektiver på gode børne- og ungeliv nu og i fremtiden, og hvilken rolle bæredygtighed spiller i disse forestillinger, er her skabt af en gruppe 14-15- årige. Vores teoretiske udgangspunkt er posthumanistiske perspektiver, der muliggør intra-aktive tilblivelser af forholdemåder. Vi viser, hvordan et pædagogisk greb, fremtidsværkstedsformat med brug af kreative metoder, skaber rum for fælles udforskning af mulige, bedre fremtider. Abstract The Children’s panel – a participatory workshop approach to sustainable futures We are living in the Anthropocene, an era where climate crisis and biodiversity crisis are deeply entangled with the human way of living. In pedagogy we need to find new ways to integrate perspectives on the Anthropocene in our everyday practice. In the project the Children’s Panel we work with a group of 14–15-year-old children to investigate how Participatory Design-inspired methods can be employed to study children’s perceptions of sustainability and their wishes for learning about sustainability. With a theoretical onset in posthumanist perspectives we develop a situated approach to becoming together for sustainability-related intra-actions and connections. The methods enable a common articulation of possible futures, exploring possibilities for living better lives in times of climate crisis.

This protocol paper outlines an innovative multimodal and multilevel approach to studying the emergence and evolution of how children build social bonds with their peers, and its potential application to improving social artificial intelligence (AI). We detail a unique hyperscanning experimental framework utilizing functional near-infrared spectroscopy (fNIRS) to observe inter-brain synchrony in child dyads during collaborative tasks and social interactions. Our proposed longitudinal study spans middle childhood, aiming to capture the dynamic development of social connections and cognitive engagement in naturalistic settings. To do so we bring together four kinds of data: the multimodal conversational behaviors that dyads of children engage in, evidence of their state of interpersonal rapport, collaborative performance on educational tasks, and inter-brain synchrony. Preliminary pilot data provide foundational support for our approach, indicating promising directions for identifying neural patterns associated with productive social interactions. The planned research will explore the neural correlates of social bond formation, informing the creation of a virtual peer learning partner in the field of Social Neuroergonomics. This protocol promises significant contributions to understanding the neural basis of social connectivity in children, while also offering a blueprint for designing empathetic and effective social AI tools, particularly for educational contexts.

The demand for alternative sources of food proteins is increasing due to the limitations and challenges associated with conventional food production. Advances in biotechnology have enabled the production of proteins using microorganisms, thus prompting the exploration of attractive microbial hosts capable of producing functional proteins in high titers. Corynebacterium glutamicum is widely used in industry for the production of amino acids and has many advantages as a host organism for recombinant protein production. However, its performance in this area is limited by low yields of target proteins and high levels of native protein secretion. Despite representing a challenge for heterologous protein production, the C. glutamicum secretome has not been fully characterized. In this study, state-of-the-art mass spectrometry-based proteomics was used to identify and analyze the proteins secreted by C. glutamicum. Both the wild-type strain and a strain that produced and secreted a recombinant β-lactoglobulin protein were analyzed. A total of 427 proteins were identified in the culture supernatants, with 148 predicted to possess a secretion signal peptide. MS-based proteomics on the secretome enabled a comprehensive characterization and quantification (based on abundance) of the secreted proteins through label-free quantification (LFQ). The top 12 most abundant proteins accounted for almost 80% of the secretome. These are uncharacterized proteins of unknown function, resuscitation promoting factors, protein PS1, Porin B, ABC-type transporter protein and hypothetical membrane protein. The data can be leveraged for protein production by, e.g., utilizing the signal peptides of the most abundant proteins to improve secretion of heterologous proteins. In addition, secretory stress can potentially be alleviated by inactivating non-essential secreted proteins. Here we provide targets by identifying the most abundant, secreted proteins of which majority are of unknown function. The data from this study can thus provide valuable insight for researchers looking to improve protein secretion and optimize C. glutamicum as a host for secretory protein production.

The IUCN now recognizes the savannah Loxodonta africana and forest Loxodonta cyclotis elephants to be separate species. Despite ecological, behavioural and morphological differences, and different habitat ranges, genetic studies confirm that the two species and hybrids coexist in forest–savannah ecotones. However, the hybrid phenotypes have not yet been described. In this survey we examined whether the phenotypes of the two species and of hybrids can be distinguished. In the first step, we used a machine learning algorithm (K-nearest neighbours) to compare 296 reference images of African elephants from five forest areas and six savannah areas where hybrids have not been recorded, confirming that six morphological criteria can be used to distinguish the species with more than 90% confidence. In the second step, we analysed 1,408 videos of elephants from 14 camera traps in Sebitoli, in Kibale National Park, Uganda, part of the main hybridization area. We used a multiple correspondence analysis and a species assignment key, highlighting the presence of three categories of phenotypes. Compared to the savannah and forest phenotypes (36.8 and 12.1%, respectively), the intermediate phenotypes, which could include hybrids, were more frequent (51.1%). Further studies combining morphology and genetics of the same individuals will be necessary to refine this species assignment key to characterize phenotypes confidently. This non-invasive, fast and inexpensive phenotypical-based method could be a valuable tool for conservation programmes.

Abstract Background Physical activity (PA) at moderate intensity is recommended for healthy pregnant women. The three-arm FitMum randomised controlled trial showed that it was possible to increase PA level during pregnancy with structured supervised exercise training (EXE) compared to standard care. Motivational counselling on PA (MOT) did not increase PA. This process evaluation aims to understand the implementation and mechanisms of impact of EXE and MOT. Methods A mixed methods process evaluation was conducted using the UK Medical Research Council’s process evaluation framework by assessing implementation (reach, fidelity, and dose) and mechanisms of impact of the two interventions provided to pregnant women in FitMum. Data was collected both quantitatively (n = 220) and qualitatively (n = 20). Results The FitMum trial reached educated pregnant women (80% having an educational level ≥ bachelor’s degree) with high autonomy of everyday life. Most participants (58%) were recruited at their first-trimester ultrasonic scan. Reasons to participate were personal (91%) and altruistic (56%). The intervention dose was delivered as intended with high fidelity in the original physical intervention setup and in the altered online setup during the COVID-19 restrictions. A low dose received in EXE (1.3 [95% CI, 1.1; 1.5] sessions/week) was partly explained by the pre-scheduled EXE sessions favouring participants with a flexible everyday life and a supportive social network. Dose received in EXE increased during online intervention delivery. Participants in MOT received 5.2 [4.7; 5.7] of 7 sessions. Mechanisms of impact comprised a perception of intervention commitment among participants in EXE due to the scheduled EXE sessions, whereas participants in MOT considered themselves as PA self-determined. PA was considered as constrained activities in EXE and included in daily activities in MOT. Conclusion The FitMum interventions was delivered with high fidelity. During COVID-19, the dose received in EXE increased compared to the previous physical setup. Mechanisms of impact as commitment, perception of empowerment and perception of PA as well as the paradox between prioritising PA and family and the need of a flexible everyday life need to be considered when offering pregnant women PA interventions. Future interventions should consider a combination of physical and online exercise training for pregnant women.

Lympho-epithelial interactions between intestinal T resident memory cells (Trm) and the epithelium have been associated with inflammatory bowel disease (IBD) activity. We developed ex vivo autologous organoid-mucosal T cell cocultures to functionally assess lymphoepithelial interactions in Crohn’s Disease (CD) patients compared to controls. We demonstrate the direct epithelial cell death induced by autologous mucosal T cells in CD patients but not in controls. These findings were positively correlated with T cell infiltration of the organoids. This potential was inhibited by limiting lympho-epithelial interactions through CD103 and NKG2D blocking antibodies. These data directly demonstrate for the first time the direct deleterious effect of mucosal T cells on the epithelium of CD patients. Such ex-vivo models are promising techniques to unravel the pathophysiology of these diseases and the potential mode of action of current and future therapies.

The identification of safe and easily-determined-inflight biomarkers to monitor the immune system of astronauts is mandatory to ensure their well-being and the success of the missions. In this report, we evaluated the relevance of two biomarkers whose determination could be easily implemented in a spacecraft in the near future by using bedridden volunteers as a ground-based model of the microgravity of spaceflight. Our data confirm the relevance of the neutrophil to lymphocyte ratio (NLR) and suggest platelet to lymphocyte ratio (PLR) monitoring to assess long-lasting immune diseases. We recommend coupling these ratios to other biomarkers, such as the quantification of cytokines and viral load measurements, to efficiently detect immune dysfunction, determine when countermeasures should be applied to promote immune recovery, prevent the development of disease, and track responses to treatment.

Abstract It is important to determine the sex of elephants from their samples—faeces from the field or seized ivory—for forensic reasons or to understand population demography and genetic structure. Molecular sexing methods developed in the last two decades have often shown limited efficiency, particularly in terms of sensitivity and specificity, due to the degradation of DNA in these samples. These limitations have also prevented their use with ancient DNA samples of elephants or mammoths. Here we propose a novel TaqMan-MGB qPCR assay to address these difficulties. We designed it specifically to allow the characterization of the genetic sex for highly degraded samples of all elephantine taxa (elephants and mammoths). In vitro experiments demonstrated a high level of sensitivity and low contamination risks. We applied this assay in two actual case studies where it consistently recovered the right genotype for specimens of known sex a priori. In the context of a modern conservation survey of African elephants, it allowed determining the sex for over 99% of fecal samples. In a paleogenetic analysis of woolly mammoths, it produced a robust hypothesis of the sex for over 65% of the specimens out of three PCR replicates. This simple, rapid, and cost-effective procedure makes it readily applicable to large sample sizes.

In Havelange (Belgium), two farms are experimenting an ecological transition. We aimed to evaluate the impact of their agricultural activities on insect pollinator communities. This article depicts the situation at the very early stage of the farm transition. This study supports the environmental benefits that the agricultural diversification with the maintenance of farm-level natural habitats can provide to the conservation of two important pollinator communities: wild bees and hoverflies.Over two years (2018-2019), by using nets and colored pan-traps, we collected 6301 of bee and hoverfly specimens among contrasted habitats within two farmsteads in ecological transition at Havelange (Belgium). In combination with the historical data of the region, we reported 101 bee species and morphospecies from 15 genera within 6 families and 31 hoverfly species and morphospecies from 18 genera. This list reinforces the national pollinator database including new distribution data in the region for extinction threatened species such as Andrena schencki Morawitz, 1866, Bombus campestris (Panzer, 1801), Eucera longicornis (L.), Halictus maculatus Smith, 1848 and Melitta tricincta Kirby, 1802 or for data deficiency species such as A. semilaevis Pérez, 1903, A. fulvata (Müller, 1766), A. trimmerana (Kirby, 1802) and Hylaeus brevicornis Nylander, 1852.

Alterations of the immune system could seriously impair the ability to combat infections during future long-duration space missions. However, little is known about the effects of spaceflight on the B-cell compartment. Given the limited access to astronaut samples, we addressed this question using blood samples collected from 20 healthy male volunteers subjected to long-duration bed rest, an Earth-based analog of spaceflight. Hematopoietic progenitors, white blood cells, total lymphocytes and B-cells, four B-cell subsets, immunoglobulin isotypes, six cytokines involved in inflammation, cortisone and cortisol were quantified at five time points. Tibia microarchitecture was also studied. Moreover, we investigated the efficiency of antioxidant supplementation with a cocktail including polyphenols, omega 3, vitamin E and selenium. Our results show that circulating hematopoietic progenitors, white blood cells, total lymphocytes and B-cells, and B-cell subsets were not affected by bed rest. Cytokine quantification suggested a lower systemic inflammatory status, supported by an increase in serum cortisone, during bed rest. These data confirm the in vivo hormonal dysregulation of immunity observed in astronauts and show that bed rest does not alter B-cell homeostasis. This lack of an impact of long-term bed rest on B-cell homeostasis can, at least partially, be explained by limited bone remodeling. None of the evaluated parameters were affected by the administration of the antioxidant supplement. The non-effectiveness of the supplement may be because the diet provided to the non-supplemented and supplemented volunteers already contained sufficient antioxidants. Given the limitations of this model, further studies will be required to determine whether B-cell homeostasis is affected, especially during future deep-space exploration missions that will be of unprecedented durations.

After a long-standing debate, African elephants are now considered by the IUCN as two distinct species: savannah elephants (Loxodonta africana), listed as endangered on the IUCN Red List of Threatened Species, and forest elephants (Loxodonta cyclotis), critically endangered. Both are severely threatened by forest loss, fragmentation and degradation due to agriculture expansion, as well as by illegal ivory trade. Although the two species have different habitat preferences, their range overlaps in some ecotones; despite an ancient separation between these two species, hybrids have been reported in five locations. The main hybrid hotspot is located on the Democratic Republic of Congo-Uganda border and still remains understudied. Using 15 microsatellites, we investigated this hybridization zone by determining the species and hybrid status of 177 fecal samples collected in the area of Sebitoli, at the extreme North of Kibale National Park. Surprisingly for a forest area, no pure forest elephants were detected. Out of the 91 individuals sampled, a very large proportion (81.3%) were hybrid individuals mainly from a second generation or more. Only 18.7% of pure savannah elephants were detected, all originating from the DRC-Uganda border. Further analyses are necessary to assess the age of this hybridization zone. Our results emphasize that hybrids and savannah elephants can successfully range in forested area. They also show that forest elephants are rare even in their native habitat. In the current context of high threat faced by African elephant species, it is crucial to strengthen conservation efforts for these species before it is too late.

Abstract Background Immunotherapies still fail to benefit colorectal cancer (CRC) patients. Relevant functional assays aimed at studying these failures and the efficacy of cancer immunotherapy in human are scarce. 3D tumor cultures, called tumor organoids or spheroids, represent interesting models to study cancer treatments and could help to challenge these issues. Methods We analyzed heterotypic cocultures of human colon tumor-derived spheroids with immune cells to assess the infiltration, activation and function of T and NK cells toward human colorectal tumors in vitro. Results We showed that allogeneic T and NK cells rapidly infiltrated cell line-derived spheroids, inducing immune-mediated tumor cell apoptosis and spheroid destruction. NKG2D, a key activator of cytotoxic responses, was engaged on infiltrating cells. We thus assessed the therapeutic potential of an antibody targeting the specific ligands of NKG2D, MICA and MICB, in this system. Anti-MICA/B enhanced immune-dependent destruction of tumor spheroid by driving an increased NK cells infiltration and activation. Interestingly, tumor cells reacted to immune infiltration by upregulating HLA-E, ligand of the inhibitory receptor NKG2A expressed by CD8 and NK cells. NKG2A was increased after anti-MICA/B treatment and, accordingly, combination of anti-MICA/B and anti-NKG2A was synergistic. These observations were ultimately confirmed in a clinical relevant model of coculture between CRC patients-derived spheroids and autologous tumor-infiltrating lymphocytes. Conclusions Altogether, we show that tumor spheroids represent a relevant tool to study tumor-lymphocyte interactions on human tissues and revealed the antitumor potential of immunomodulatory antibodies targeting MICA/B and NKG2A.

Understanding phosphorylation-mediated regulation of metabolic enzymes, pathways, and cell phenotypes under metabolic shifts represents a major challenge. The kinases associated with most phosphorylation sites and the link between phosphorylation and enzyme activity remain unknown. In this study, we performed stable isotope labeling by amino acids in cell culture (SILAC)-based proteome and phosphoproteome analysis of Escherichia coli ΔyeaG, a strain lacking a poorly characterized serine/threonine kinase YeaG, to decipher kinase-substrate interactions and the effects on metabolic phenotype during shifts from glucose to malate. The starting point of our analysis was the identification of physiological conditions under which ΔyeaG exhibits a clear phenotype. By metabolic profiling, we discovered that ΔyeaG strain has a significantly shorter lag phase than the wild type during metabolic shift from glucose to malate. Under those conditions, our SILAC analysis revealed several proteins that were differentially phosphorylated in the ΔyeaG strain. By focusing on metabolic enzymes potentially involved in central carbon metabolism, we narrowed down our search for putative YeaG substrates and identified isocitrate lyase AceA as the direct substrate of YeaG. YeaG was capable of phosphorylating AceA in vitro only in the presence of malate, suggesting that this phosphorylation event is indeed relevant for glucose to malate shift. There is currently not enough evidence to firmly establish the exact mechanism of this newly observed regulatory phenomenon. However, our study clearly exemplifies the usefulness of SILAC-based approaches in identifying proteins kinase substrates, when applied in physiological conditions relevant for the activity of the protein kinase in question.

Intestinal tissue-resident memory CD8 T cells (Trm) are non-recirculating effector cells ideally positioned to detect and react to microbial infections in the gut mucosa. There is an emerging understanding of Trm cell differentiation and functions, but their implication in inflammatory bowel diseases, such as Crohn's disease (CD), is still unknown. Here, we describe CD8 cells in the human intestine expressing KLRG1 or CD103, two receptors of E-cadherin. While CD103 CD8 T cells are present in high numbers in the mucosa of CD patients and controls, KLRG1 CD8 T cells are increased in inflammatory conditions. Mucosal CD103 CD8 T cells are more responsive to TCR restimulation, but KLRG1 CD8 T cells show increased cytotoxic and proliferative potential. CD103 CD8 T cells originate mostly from KLRG1 negative cells after TCR triggering and TGFβ stimulation. Interestingly, mucosal CD103 CD8 T cells from CD patients display major changes in their transcriptomic landscape compared to controls. They express Th17 related genes including CCL20, IL22, and IL26, which could contribute to the pathogenesis of CD. Overall, these findings suggest that CD103 CD8 T cells in CD induce a tissue-wide alert increasing innate immune responses and recruitment of effector cells such as KLRG1 CD8 T cells.

A great deal of research exists in the use of multimedia communications in online classrooms as a means of furthering student engagement. However, little research exists that examines the perceptions of students when such technologies are used. Additionally, it is unclear that students are likely to engage in the use of such technologies when available. This research explores the perceptions of 69 students taking both online and hybrid undergraduate project management courses. Specifically, the study seeks to explore how students experienced the use of multimedia by their instructor and classmates in both online announcements and discussions, as well as whether these same students used or would be likely to use multimedia for similar communications. Finally, student perceptions of social presence, the degree to which one is perceived as a real person in computer-mediated communication (Gunawardena, 1995), are examined. The results of the study indicate that while students overwhelming enjoy the instructor’s use of multimedia communication, they are unlikely to engage in using these technologies themselves. A discussion of these results and recommendations for further research complete this paper.

In eukaryotes many players in the DNA-damage response (DDR) catalyze protein sumoylation or ubiquitylation. Emphasis has been placed on how these modifications orchestrate the sequential recruitment of repair factors to sites of DNA damage or stalled replication forks. Here, we shed light on a pathway in which sumoylated factors are eliminated through the coupled action of Sumo-targeted ubiquitin ligases (STUbLs) and the ubiquitin-fusion degradation protein 1 (Ufd1). Ufd1 is a subunit of the Cdc48-Ufd1-Npl4 complex implicated in the sorting of ubiquitylated substrates for degradation by the proteasome. We find that in fission yeast, Ufd1 interacts physically and functionally with the Sumo-targeted ubiquitin ligase (STUbL) Rfp1, homologous to human RNF4, and with the Sumo E3 ligase Pli1, homologous to human PIAS1. Deleting a C-terminal domain of Ufd1 that mediates the interaction of Ufd1 with Rfp1, Pli1, and Sumo (ufd1ΔCt(213-342) ) lead to an accumulation of high-molecular-weight Sumo conjugates and caused severe genomic instabilities. The spectrum of sensitivity of ufd1ΔCt(213-342) cells to genotoxins, the epistatic relationships of ufd1ΔCt(213-342) with mutations in DNA repair factors, and the localization of the repair factor Rad22 in ufd1ΔCt(213-342) cells point to ufd1ΔCt(213-342) cells accumulating aberrant structures during replication that require homologous recombination (HR) for their repair. We present evidence that HR is however often not successful in ufd1ΔCt(213-342) cells and we identify Rad22 as one of the high-molecular-weight conjugates accumulating in the ufd1ΔCt(213-342) mutant consistent with Rad22 being a STUbL/Ufd1 substrate. Suggesting a direct role of Ufd1 in the processing of Sumo-conjugates, Ufd1 formed nuclear foci colocalizing with Sumo during the DDR, and Sumo-conjugates accumulated in foci in the ufd1ΔCt(213-342) mutant. Broader functional relationships between Ufd1 and STUbLs conceivably affect numerous cellular processes beyond the DDR.