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Fibre-reinforced epoxy composites are well established in regard to load-bearing applications in the aerospace, automotive and wind power industries, owing to their light weight and high durability. These composites are based on thermoset resins embedding glass or carbon fibres1. In lieu of viable recycling strategies, end-of-use composite-based structures such as wind turbine blades are commonly landfilled1–4. Because of the negative environmental impact of plastic waste5,6, the need for circular economies of plastics has become more pressing7,8. However, recycling thermoset plastics is no trivial matter1–4. Here we report a transition-metal-catalysed protocol for recovery of the polymer building block bisphenol A and intact fibres from epoxy composites. A Ru-catalysed, dehydrogenation/bond, cleavage/reduction cascade disconnects the C(alkyl)–O bonds of the most common linkages of the polymer. We showcase the application of this methodology to relevant unmodified amine-cured epoxy resins as well as commercial composites, including the shell of a wind turbine blade. Our results demonstrate that chemical recycling approaches for thermoset epoxy resins and composites are achievable.

Zusammenfassung Einleitung Das Erscheinungsbild des Bauchnabels nach DIEP-Lappenplastik oder Abdominoplastik spielt eine wesentliche Rolle bei der ästhetischen Wahrnehmung (1). Obgleich der Nabel keine Funktion hat, besteht kein Zweifel an der Bedeutung, die die Nabelform für das Selbstwertgefühl der Patientinnen gerade nach einer Brustkrebserkrankung hat (2). In der vorliegenden Studie haben wir zwei der bevorzugten in der Literatur beschriebenen Techniken an 72 Patientinnen hinsichtlich des ästhetischen Ergebnisses, der Komplikationen und der Sensibilität verglichen: das kaudale Läppchen (kuppelförmige Form) und die querovale Form des Nabels. Patienten und Methode Zweiundsiebzig Patientinnen, die zwischen Januar 2016 und Juli 2018 eine DIEP-Lappenplastik zur Brustrekonstruktion erhielten, wurden in dieser Studie retrospektiv erfasst. Es wurden zwei Techniken zur Nabelrekonstruktion verglichen: die querovale Nabelform und die Umbilikoplastik unter Verwendung eines kaudalen Läppchens, welche in einer kuppelförmigen Form des Bauchnabels resultiert. Um die ästhetischen Ergebnisse zu vergleichen, wurde eine Evaluation durch die Patientinnen und eine Bewertung durch drei unabhängige Plastische Chirurg*innen mindestens 6 Monate postoperativ durchgeführt. Patientinnen und Chirurg*innen wurden gebeten, das allgemeine Erscheinungsbild des Nabels, einschließlich der Narbenbildung und der Form, mit einer Punktzahl zwischen 1 und 6 zu bewerten (1=Sehr gut, 2=gut, 3=befriedigend, 4=ausreichend, 5=mangelhaft, 6=ungenügend). Des Weiteren wurde die Sensibilität des Nabels durch Befragung der Patientinnen und das Auftreten von Wundheilungsstörungen untersucht. Ergebnisse Beide Techniken zeigten ähnliche Werte der ästhetischen Zufriedenheit (p=0,49) im Rahmen der Selbsteinschätzung durch die Patientinnen. Die Plastischen Chirurg*innen bewerteten die Technik mit kaudalem Läppchen als signifikant besser gegenüber der Nabelplastik mit querovaler Form (p=0,042). Beim kaudalen Läppchen traten mehr Wundheilungsstörungen auf (11,1%) im Vergleich zum querovalen Nabel. Dies war jedoch nicht signifikant (p=0,16). Eine operative Revision war nicht erforderlich. Beim Nabel mit kaudalem Läppchen zeigte sich eine Tendenz zur Verbesserung der Sensibilität (60% gegenüber 45%), die jedoch nicht signifikant war (p=0,19). Schlussfolgerung Die Patientenzufriedenheit zeigte ähnliche Ergebnisse für die beiden Methoden der Umbilikoplastik. Beide Techniken wurden im Durchschnitt mit einem guten Ergebnis bewertet. Andererseits bewerteten die Chirurg*innen die Umbilikoplastik mit kaudalem Läppchen als ästhetisch ansprechender.

In the era of blood group genomics, reference collections of complete and fully resolved blood group gene alleles have gained high importance. For most blood groups, however, such collections are currently lacking, as resolving full-length gene sequences as haplotypes (ie, separated maternal/paternal origin) remains exceedingly difficult with both Sanger and short-read next-generation sequencing. Using the latest third-generation long-read sequencing, we generated a collection of fully resolved sequences for all 6 main ABO allele groups: ABO∗A1/A2/B/O.01.01/O.01.02/O.02. We selected 77 samples from an ABO genotype data set (n = 25 200) of serologically typed Swiss blood donors. The entire ABO gene was amplified in 2 overlapping long-range polymerase chain reactions (covering ∼23.6 kb) and sequenced by long-read Oxford Nanopore sequencing. For quality validation, 2 samples per ABO group were resequenced using Illumina and Pacific Biosciences technology. All 154 full-length ABO sequences were resolved as haplotypes. We observed novel, distinct sequence patterns for each ABO group. Most genetic diversity was found between, not within, ABO groups. Phylogenetic tree and haplotype network analyses highlighted distinct clades of each ABO group. Strikingly, our data uncovered 4 genetic variants putatively specific for ABO∗A1, for which direct diagnostic targets are currently lacking. We validated A1-diagnostic potential using whole-genome data (n = 4872) of a multiethnic cohort. Overall, our sequencing strategy proved powerful for producing high-quality ABO haplotypes and holds promise for generating similar collections for other blood groups. The publicly available collection of 154 haplotypes will serve as a valuable resource for molecular analyses of ABO, as well as studies about the function and evolutionary history of ABO.

The stomatopod is a fascinating animal that uses its weaponized appendage dactyl clubs for breaking mollusc shells. Dactyl clubs are a well studied example of biomineralized hierarchical structures. Most research has focused on the regions close to the action, namely the impact region and surface composed of chitin and apatite crystallites. Further away from the site of impact, the club has lower mineralization and more amorphous phases; these areas have not been as actively studied as their highly mineralized counterparts. This work focuses on the side of the club, in what is known as the periodic and striated regions. A combination of laboratory micro-computed tomography, synchrotron X-ray diffraction mapping and synchrotron X-ray fluorescence mapping has shown that the mineral in this region undergoes the transition from an amorphous to a crystalline phase in some, but not all, clubs. This means that this side region can be mineralized by either an amorphous phase, calcite crystallites or a mixture of both. It was found that when larger calcite crystallites form, they are organized (textured) with respect to the chitin present in this biocomposite. This suggests that chitin may serve as a template for crystallization when the side of the club is fully mineralized. Further, calcite crystallites were found to form as early as 1 week after moulting of the club. This suggests that the side of the club is designed with a significant safety margin that allows for a variety of phases, i.e. the club can function independently of whether the side region has a crystalline or amorphous mineral phase. Published version We thank the Danish Agency for Science, Technology and Innovation for funding the instrument center DanScatt. We acknowledge DESY (Hamburg, Germany), a member of the Helmholtz Association HGF, for the provision of experimental facilities. Beam time was allocated for proposal I20180923EC. The research leading to this result has been supported by the project CALIPSOplus from the EU Framework Programme for Research and Innovation HORIZON 2020 (grant no. 730872). We thank Alexander Bernthz Jensen, Kristine Hjerrild Neldeborg, Jonas Palle and Maja Østergaard for help with measuring data on P06 at Petra III. We acknowledge the MAX IV Laboratory for time on Beamline DanMAX (proposal no. 20210507). Research conducted at MAX IV, a Swedish national user facility, is supported by the Swedish Research council (contract no. 2018-07152), the Swedish Governmental Agency for Innovation Systems (contract no. 2018-04969) and Formas (contract no. 2019-02496). DanMAX is funded by NUFI (grant no. 4059- 00009B). We thank Peter Alling Strange Vibe for assistance with measuring data at DanMAX. Use of the Novo Nordisk Foundation research infrastructure AXIA (grant no. NNF19OC0055801) is gratefully acknowledged. We acknowledge support from the ESS lighthouse on hard materials in 3D, SOLID, funded by the Danish Agency for Science and Higher Education (grant no. 8144-00002B).

Purpose The new Medical Licensing Regulations 2025 (Ärztliche Approbationsordnung, ÄApprO) require the development of competence-oriented teaching formats. In addition, there is a great need for high-quality teaching in the field of radiation oncology, which manifests itself already during medical school. For this reason, we developed a simulation-based, hands-on medical education format to teach competency in performing accelerated partial breast irradiation (APBI) with interstitial multicatheter brachytherapy for early breast cancer. In addition, we designed realistic breast models suitable for teaching both palpation of the female breast and implantation of brachytherapy catheters. Methods From June 2021 to July 2022, 70 medical students took part in the hands-on brachytherapy workshop. After a propaedeutic introduction, the participants simulated the implantation of single-lead catheters under supervision using the silicone-based breast models. Correct catheter placement was subsequently assessed by CT scans. Participants rated their skills before and after the workshop on a six-point Likert scale in a standardized questionnaire. Results Participants significantly improved their knowledge-based and practical skills on APBI in all items as assessed by a standardized questionnaire (mean sum score 42.4 before and 16.0 after the course, p Conclusion The simulation-based medical education course for multicatheter brachytherapy can improve self-assessed technical competence. Residency programs should provide resources for this essential component of radiation oncology. This course is exemplary for the development of innovative practical and competence-based teaching formats to meet the current reforms in medical education.

Zusammenfassung Abstrakt Einleitung Tranexamsäure (TXA) ist ein antifibrinolytisches Medikament, das zur Therapie von Blutungen als auch zur Blutungsprophylaxe verwendet werden kann. Die Anwendung in der plastischen und rekonstruktiven Chirurgie ist bisher äußerst begrenzt. Unseres Wissens fehlen bisher veröffentlichte Daten zur topischen Anwendung von TXA bei subkutaner Mastektomie transidenter Patienten. Methoden Es erfolgte eine retrospektive Datenanalyse von Patienten, die eine subkutanen Mastektomie bei Frau-zu-Mann Transidentität erhielten. Insgesamt wurden 22 Patienten mit topisch applizierter TXA behandelt, während 29 Patienten als Vergleichsgruppe dienten. Primärer Endpunkt war die Menge der Wundsekretion in den ersten 48 h nach der Operation. Sekundäre Endpunkte waren die Dauer der Drainage und das Auftreten von Nachblutungen. Ergebnisse Innerhalb der ersten 48 h zeigte sich in der Gruppe der TXA Patienten eine durchschnittliche Menge von 97±50 ml im Vergleich zu 180±111 ml in der Kontrollgruppe (p Diskussion Es zeigte sich eine signifikante Reduktion der Fördermenge über die Drainage innerhalb der ersten 48 h sowie über den Gesamtzeitraum nach topischer Applikation von TXA. Des Weiteren kam es zu einer Verkürzung des Zeitraums mit einliegender Drainage bei den mit TXA behandelten Patienten. Die vorliegende Studie konnte einen signifikanten Vorteil für die topische Anwendung von TXA bei subkutaner Mastektomie bei Frau-zu-Mann transidenten Patienten zeigen.

Purpose The aim of this study was to analyze the heart dose for left-sided breast cancer that can be achieved during daily practice in patients treated with multicatheter brachytherapy (MCBT) accelerated partial-breast irradiation (APBI) and deep-inspiration breath-hold (DIBH) whole-breast irradiation (WBI) using a simultaneous integrated tumor bed boost (SIB)—two different concepts which nonetheless share some patient overlap. Materials and methods We analyzed the nominal average dose (Dmean) to the heart as well as the biologically effective dose (BED) and the equivalent dose in 2‑Gy fractions (EQD2) for an α/β of 3 in 30 MCBT-APBI patients and 22 patients treated with DIBH plus SIB. For further dosimetric comparison, we contoured the breast planning target volume (PTV) in each of the brachytherapy planning CTs according to the ESTRO guidelines and computed tangential field plans. Mean dose (Dmean), EQD2 Dmean, and BED Dmean for three dosing schemes were calculated: 50 Gy/25 fractions and two hypofractionated regimens, i.e., 40.05 Gy/15 fractions and 26 Gy/5 fractions. Furthermore, we calculated tangential field plans without a boost for the 22 cases treated with SIB with the standard dosing scheme of 40.05 Gy/15 fractions. Results MCBT and DIBH radiation therapy both show low-dose exposure of the heart. As expected, hypofractionation leads to sparing of the heart dose. Although MCBT plans were not optimized regarding dose to the heart, Dmean differed significantly between MCBT and DIBH (1.28 Gy vs. 1.91 Gy, p p Conclusion In daily practice, both DIBH radiation therapy as well as MCBT show a very low heart exposure and may thus reduce long term cardiac morbidity as compared to currently available long-term clinical data of patients treated with conventional tangential field plans in free breathing. Our analysis confirms particularly good cardiac sparing with MCBT-APBI, so that this technique should be offered to patients with left-sided breast cancer if the tumor-associated eligibility criteria are fulfilled.

International audience; Bone is a complex, biological tissue made up primarily of collagen fibrils and biomineral nanoparticles. The importance of hierarchical organization in bone was realized early on, but the actual interplay between structural features and the properties on the nanostructural and crystallographic level is still a matter of intense discussion. Bone is the only mineralized tissue that can be remodeled and, at the start of the formation of new bone during this process, a structure called a cement line is formed on which regular bone grows. Here, the orientational relationship of nanostructural and crystallographic constituents as well as the structural properties of both nanostructural and crystallographic constituents around cement lines and the Haversian system in human lamellar bone are investigated. A combination of small- and wide-angle X-ray scattering tensor tomography is employed together with diffraction tomography and synchrotron computed tomography to generate a multi-modal image of the sample. This work shows that the mineral properties vary as a function of the distance to the Haversian canal and, importantly, shows that the cement line has differing mineral properties from the surrounding lamellar bone, in particular with respect to crystallite size and degree of orientation. Cement lines make up a significant portion of the bone matrix despite their small size, hence the reported findings on an altered mineral structure, together with the spatial modulation around the Haversian canal, have implications for the formation and mechanics of bone.

Abstract An injured skin is rapidly restored in a manner of wound healing. We have previously shown that intact insulin signaling and glucose uptake are fundamental to proper wound closure. Consequently, under exacerbated inflammation, compromised insulin action and glucose uptake lead to impaired healing. However, in spite of the increased attention to cell metabolism during tissue regeneration, metabolic mediators that govern cellular and physiological processes throughout skin repair remained largely elusive. Through assessment of mRNA using real-time PCR and protein blot analysis, we report that healing of cutaneous wounds comprise a boosted expression of genes involved in glycolysis, oxidative phosphorylation, pentose phosphate shunt, and glutamine anaplerosis. We further focused on the functional role of pyruvate kinase M (PKM) isoenzymes that catalyze the final and rate-limiting step of glycolysis. Whereas the expression of the metabolic constitutively active Pkm1 isozyme remained almost unchanged, Pkm2 is augmented during the inflammatory phase of healing. The immunohistochemistry and RNA in situ hybridization analysis showed a confined Pkm2 expression to keratinocytes of the hyperproliferative epithelium and, to a lesser extent, infiltrating neutrophils and monocytes as well as later on in macrophages. Notably, the expression of Pkm2 in keratinocytes facing the wound bed side colocalized with VEGF expression. The in vitro knockdown of PKM2 in HaCaT keratinocytes using small interfering (si) RNA confirmed an acute role for PKM2 in facilitating the complete induction of VEGF mRNA and protein expression in keratinocytes; this function is mainly HIF-1α independent. Key messages • Wound healing involves activation of glycolysis, oxidative phosphorylation, pentos-phosphate shunt, and replenishment of tri-carboxylic acid (TCA) cycle through glutamine anaplerosis. • The pyruvate kinase M2 (PKM2) isoform is upregulated during the inflammatory phase of cutaneous healing, mainly in keratinocytes of hyperproliferative epithelia. • In vivo, the expression of VEGF in wound keratinocytes is colocalized with PKM2. • PKM2 is required for full induction of VEGF in HaCaT keratinocytes in vitro.

Hypoxia induces massive changes in alternative splicing (AS) to adapt cells to the lack of oxygen. Here, we identify the splicing factor SRSF6 as a key factor in the AS response to hypoxia. The SRSF6 level is strongly reduced in acute hypoxia, which serves a dual purpose: it allows for exon skipping and triggers the dispersal of nuclear speckles. Our data suggest that cells use dispersal of nuclear speckles to reprogram their gene expression during hypoxic adaptation and that SRSF6 plays an important role in cohesion of nuclear speckles. Down-regulation of SRSF6 is achieved through inclusion of a poison cassette exon (PCE) promoted by SRSF4. Removing the PCE 3′ splice site using CRISPR/Cas9 abolishes SRSF6 reduction in hypoxia. Aberrantly high SRSF6 levels in hypoxia attenuate hypoxia-mediated AS and impair dispersal of nuclear speckles. As a consequence, proliferation and genomic instability are increased, while the stress response is suppressed. The SRSF4–PCE–SRSF6 hypoxia axis is active in different cancer types, and high SRSF6 expression in hypoxic tumors correlates with a poor prognosis. We propose that the ultra-conserved PCE of SRSF6 acts as a tumor suppressor and that its inclusion in hypoxia is crucial to reduce SRSF6 levels. This may prevent tumor cells from entering the metastatic route of hypoxia adaptation.

Background: Under certain clinical and experimental conditions hematopoiesis occurs in other site than bone marrow (BM), such as the liver. Here, we develop a 3D organoid that mimics several components of the hematopoietic niche present during liver extramedullary hematopoiesis. Aim: To evaluate the capacity of a 3D hematopoietic organoid (3D-HO) to function as a hematopoietic like-niche allowing for blood cell production outside of the BM Methods: The 3D-HO is constituted by liver sinusoidal endothelial cells (LSEC) as the stromal component, BM isolated from 5-FU treated mice (FU-BMCs), collagen microspheres and plasma clot as scaffolds. The ability of the 3D-HO to support the survival and functionality of FU-BMCs was investigated by using confocal microscopy, histology analysis, flow cytometry, and clonogenic assays. Results: After 15 and 30 days, post-ectopic implantation, histological studies of the 3D-HO showed the presence of cells with myeloid and lymphoid lineage morphology. Flow cytometry analysis of these cells showed the presence of cells expressing hematopoietic stem progenitor cells (HSPC) (Sca-1+/c-Kit+), myeloid (Gr-1+) and lymphoid (B220+ and CD19+) markers. Clonogenic assays showed that cells from the 3D-HO formed hematopoietic colonies. Expression of the Sry gene by cells from the 3D-HO, implanted for 30 days in female mice, indicated that male donor cells persist in this model of extramedullary hematopoiesis. Conclusions: The 3D-HO constitutes an extramedullary hematopoietic-like niche which supports the survival and functionality of FU-BMCs. It may constitute an efficient model for investigating, in vitro and in vivo, those factors that control hematopoiesis outside BM.

One of the many ecosystem services provided by dung beetles is that of secondary seed dispersal. This paper experimentally evaluates the effectiveness of this service using the chimpanzee–dung beetle seed dispersal system in Taï National Park, Côte d’Ivoire. The study focussed on the germination rate and success of four species of seeds contained in the faeces ofPan troglodytes verus: Dacryodes klaineana(Pierre) H.J. Lam,Diospyros manniiHiern,Pycnanthus angolensis(Welw.) Warb., andUapaca guineensisMuell. Arg. For each species, 600 seeds, half from chimpanzee faeces and half from mother trees, were sown in nurseries at depths of 0, 2, 5, 10, and 15 cm (i.e. 120 seeds per depth; 60 from chimp and 60 from mother trees). After germination, only the seeds ofUapaca guineensisandDiospyros manniisown at 2 and 5 cm had a >14% rate (between 14.2 and 30.8%) of germination and seedling establishment, regardless of seed origin. An increase in the depth of seed burial appears to negatively affect the probability of seedling emergence and establishment. This study shows that dung beetles have positive impacts on seed fate. However, for these plant species, the chimpanzee’s role is limited to that of primary seed dispersal.

Introduction To help combat the SARS-CoV-2 (COVID-19) pandemic, elective inpatient procedures have been reduced. The authors hypothesized that a nationwide lockdown would negatively affect the postoperative outcome after total knee arthroplasty (TKA) due to reduced physiotherapy as well as restrictions in external facilities of physiotherapy and rehabilitation. Materials and methods We conducted a retrospective, comparative study including 41 patients who had undergone primary TKA during the first lockdown of the COVID-19 pandemic from March 2020 to April 2020 and a comparable control group consisting of 47 patients with a minimum follow-up of 6 months before the COVID-19 pandemic from 2019. Relevant end points were the visual analogue scale (VAS) for pain, Knee Society Function Score (KSS), Oxford Knee Score (OKS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and range of motion (ROM). Results The lockdown group had a significantly worse outcome compared to the control group 6 months after TKA regarding WOMAC (p = 0.001), KSS (p p p p = 0.132), KSFS (p = 0.933), VAS at rest (p = 0.9.22), and exercise (p = 0.304). Conclusion The COVID-19 pandemic negatively affected early clinical outcome parameters of elective primary TKA at 6 months of follow-up due to restrictions in postoperative care. We believe that standardized protocols for physiotherapy will improve clinical outcomes for TKA in the event of future lockdowns and underline the importance of appropriate postoperative care during this pandemic.

The paper discusses the options for a conceptual framework to describe how the knowledge created and acquired in VET is changing due to technological innovation and the shift towards a knowledge economy. The discussion sets out from the question how vocational or professional knowledge may be distinguished from other forms of knowledge and what philosophical and epistemological traits may underpin this distinction. Building on an analysis of ‘intrinsic’ and ‘extrinsic’ conceptions of knowledge, the paper explores how appropriate descriptors for the classification of theoretical or propositional knowledge as well as practical knowledge may be identified. As a result, a proposal for structuring vocational knowledge is presented. The concluding part addresses the question how the proposed categories may serve to characterise changes in vocational knowledge and explores the requirements for applying the conceptual framework in empirical studies on the effects of technological innovation and societal developments.

Purpose The Working Group for Neuro-Oncology of the German Society for Radiation Oncology in cooperation with members of the Neuro-Oncology Working Group of the German Cancer Society aimed to define a practical guideline for the diagnosis and treatment of radiation-induced necrosis (RN) of the central nervous system (CNS). Methods Panel members of the DEGRO working group invited experts, participated in a series of conferences, supplemented their clinical experience, performed a literature review, and formulated recommendations for medical treatment of RN including bevacizumab in clinical routine. Conclusion Diagnosis and treatment of RN requires multidisciplinary structures of care and defined processes. Diagnosis has to be made on an interdisciplinary level with the joint knowledge of a neuroradiologist, radiation oncologist, neurosurgeon, neuropathologist, and neuro-oncologist. A multistep approach as an opportunity to review as many characteristics as possible to improve diagnostic confidence is recommended. Additional information about radiotherapy (RT) techniques is crucial for the diagnosis of RN. Misdiagnosis of untreated and progressive RN can lead to severe neurological deficits. In this practice guideline, we propose a detailed nomenclature of treatment-related changes and a multistep approach for their diagnosis.

ZusammenfassungTeil- oder Totalverluste der Nase führen zu ästhetischer Entstellung und zu hohem psychischen Leidensdruck sowie zu funktionellen Problemen im Bereich von Atmung und Geruchssinn. Aus rekonstruktiv-chirurgischer Sicht kann die Nase in drei Anteile geteilt werden: Innenauskleidung, Stützgerüst, Haut-Weichteil-Bedeckung. Im Rahmen der Rekonstruktion nasaler Defekte müssen alle drei Anteile bestmöglich wiederhergestellt werden. Je größer die Defekte sind, desto eher muss auf mikrovaskuläre Transplantate zurückgegriffen werden. Chondrokutane Transplantate von der Ohrhelix, gestielt an A. und V. temporalis superficialis, eignen sich sehr gut zur Rekonstruktion von Teildefekten der Nase. Osteoperiostale Transplantate vom medialen Femurkondylus, gestielt an A. descendens genus und Begleitvenen, eignen sich sehr gut zur Rekonstruktion des Nasenseptums oder des Nasenskeletts im Rahmen der Wiederherstellung großer Defekte. Der paramediane Stirnlappen stellt den Goldstandard zur Rekonstruktion großer Defekte der Haut-Weichteil-Bedeckung dar.

Driver drowsiness is one of the main factors leading to road fatalities and hazards in the transportation industry. Electroencephalography (EEG) has been considered as one of the best physiological signals to detect drivers' drowsy states, since it directly measures neurophysiological activities in the brain. However, designing a calibration-free system for driver drowsiness detection with EEG is still a challenging task, as EEG suffers from serious mental and physical drifts across different subjects. In this paper, we propose a compact and interpretable Convolutional Neural Network (CNN) to discover shared EEG features across different subjects for driver drowsiness detection. We incorporate the Global Average Pooling (GAP) layer in the model structure, allowing the Class Activation Map (CAM) method to be used for localizing regions of the input signal that contribute most for classification. Results show that the proposed model can achieve an average accuracy of 73.22% on 11 subjects for 2-class cross-subject EEG signal classification, which is higher than conventional machine learning methods and other state-of-art deep learning methods. It is revealed by the visualization technique that the model has learned biologically explainable features, e.g., Alpha spindles and Theta burst, as evidence for the drowsy state. It is also interesting to see that the model uses artifacts that usually dominate the wakeful EEG, e.g., muscle artifacts and sensor drifts, to recognize the alert state. The proposed model illustrates a potential direction to use CNN models as a powerful tool to discover shared features related to different mental states across different subjects from EEG signals. National Research Foundation (NRF) Submitted/Accepted version This research is supported by the National Research Foundation, Singapore under its International Research Centres in Singapore Funding Initiative.

Methane is a potent greenhouse gas that plays a significant role in the global climate system. High latitude methane is a particularly sensitive subject due to the uncertainty in future gas release due to multiple factors such as the thawing of permafrost and the evolution of wetland cover. Thus, these regions have the potential to significantly contribute to global warming. In this study, we present the results of the MAGIC2021 campaign, which was conducted in Lapland around Kiruna, Sweden in August 2021. The campaign included measurements with atmospheric air sampler AirCores on board weather balloons, three research aircraft equipped with in-situ sensors, and ground-based measurements of gas total columns using EM27/SUNs. We focus here on the combined measurements of 0-30 km profiles by AirCore and by ATR42 research aircraft to investigate sources of methane in the region. To this end, we employed back-trajectory Lagrangian models and conducted an in-depth comparison between model (ERA5, CAMS) and campaign data in a multi-species approach combining CH4, CO2 and CO. Our findings provide insight into the sources and transport of methane at high latitudes. They show that in order to properly study local sources of methane, it is mandatory to account for transported methane originating from regions as far as Northern Canada. Our work also highlights the importance of conducting in situ measurement campaigns like MAGIC2021, which provide valuable data for improving our understanding of atmospheric processes at high latitudes and informing the development of more accurate models and validate satellite retrievals. Plans for next campaigns will also be detailed.

The mpox virus (MPXV) is mutating at an exceptional rate for a DNA virus and its global spread is concerning, making genomic surveillance a necessity. With MpoxRadar, we provide an interactive dashboard to track virus variants on mutation level worldwide. MpoxRadar allows users to select among different genomes as reference for comparison. The occurrence of mutation profiles based on the selected reference is indicated on an interactive world map that shows the respective geographic sampling site in customizable time ranges to easily follow the frequency or trend of defined mutations. Furthermore, the user can filter for specific mutations, genes, countries, genome types, and sequencing protocols and download the filtered data directly from MpoxRadar. On the server, we automatically download all MPXV genomes and metadata from the National Center for Biotechnology Information (NCBI) on a daily basis, align them to the different reference genomes, generate mutation profiles, which are stored and linked to the available metainformation in a database. This makes MpoxRadar a practical tool for the genomic survaillance of MPXV, supporting users with limited computational resources. MpoxRadar is open-source and freely accessible at https://MpoxRadar.net.

BackgroundMany automatic approaches to brain tumor segmentation employ multiple magnetic resonance imaging (MRI) sequences. The goal of this project was to compare different combinations of input sequences to determine which MRI sequences are needed for effective automated brain metastasis (BM) segmentation.MethodsWe analyzed preoperative imaging (T1-weighted sequence ± contrast-enhancement (T1/T1-CE), T2-weighted sequence (T2), and T2 fluid-attenuated inversion recovery (T2-FLAIR) sequence) from 333 patients with BMs from six centers. A baseline 3D U-Net with all four sequences and six U-Nets with plausible sequence combinations (T1-CE, T1, T2-FLAIR, T1-CE+T2-FLAIR, T1-CE+T1+T2-FLAIR, T1-CE+T1) were trained on 239 patients from two centers and subsequently tested on an external cohort of 94 patients from four centers.ResultsThe model based on T1-CE alone achieved the best segmentation performance for BM segmentation with a median Dice similarity coefficient (DSC) of 0.96. Models trained without T1-CE performed worse (T1-only: DSC = 0.70 and T2-FLAIR-only: DSC = 0.72). For edema segmentation, models that included both T1-CE and T2-FLAIR performed best (DSC = 0.93), while the remaining four models without simultaneous inclusion of these both sequences reached a median DSC of 0.81-0.89.ConclusionsA T1-CE-only protocol suffices for the segmentation of BMs. The combination of T1-CE and T2-FLAIR is important for edema segmentation. Missing either T1-CE or T2-FLAIR decreases performance. These findings may improve imaging routines by omitting unnecessary sequences, thus allowing for faster procedures in daily clinical practice while enabling optimal neural network-based target definitions.

Cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl- channel and ABC transporter; its mutations cause the clinical picture of cystic fibrosis (CF). Of late, CFTR has emerged as an important regulator of platelet function, as CFTR dysfunction causes agonist-induced platelet hyperactivation. These findings are reminiscent of platelets from SARS-CoV-2 infected patients since thromboembolic complications represent hallmarks of severe COVID-19 that may critically contribute to morbidity and mortality. CFTR modulators have recently been introduced as a treatment for patients with various CFTR mutations, but have also been reported by us and others to enhance channel function of wild type CFTR. We therefore postulated that CFTR modulators may exert anti-coagulant effects on platelets of healthy donors (HD) and COVID-19 patients.We recruited 36 COVID-19 patients with moderate, and 34 COVID-19 patients with severe disease course (all w/o anti-platelet drugs), and 38 HDs. In line with our hypothesis, we observed significant reductions in platelet agonists adenosine diphosphate (ADP)- or thrombin receptor activating protein-6 (TRAP6)-induced CD62p/CD63 expression, Ca2+-mobilization, aggregation, and adhesion of platelets from HDs by pre-treatment with ivacaftor. In blood from COVID-19 patients, platelet activation correlates with disease severity, as demonstrated by a 5-fold and 8-fold increase in the proportion of CD62p+ platelets from patients with moderate and severe disease, respectively, relative to HDs. Similarly, the proportion of CD63+ platelets in patients with severe COVID-19 was 2-fold higher than in HDs. Retrospective analysis of clinical data from a total of 4,050 CF patients with COVID-19 receiving single or combination therapy of ivacaftor, lumacaftor, tezacaftor, or elexacaftor in comparison to an untreated cohort revealed that CF therapy reduced the relative risk to suffer thromboembolism-associated cardiovascular events such as heart attack or deep vein thrombosis by 50.0% or 61.1%, respectively, suggesting an anti-thrombotic effect of CFTR modulators in CF COVID-19 patients. In line with this observation, ex vivo pre-treatment of platelets from acute COVID-19 patients with ivacaftor reduced Ca2+ mobilization, adhesion, and aggregation of platelets .Our results demonstrate an anticoagulant effect of CFTR potentiators on platelets from HDs and severe COVID-19 patients and thus, suggest CFTR potentiators as a promising strategy to reduce the risk of thrombotic events in the clinical management of COVID-19 and similar pro-thrombotic disease states. F. Behrens received funding from the Berlin Institute of Health (BIH). L. Michalick reports grants from the BIH and the German Centre for Cardiovascular Research (DZHK). A. Haghikia is participant in the BIH-Charité Advanced Clinician Scientist Pilotprogram funded by the Charité - Universitätsmedizin Berlin and the BIH and reports a research grant within the BIH & MDC Focus Area Translational Vascular Biomedicine. R. Preissner reports partial funding of this work by the German Research Foundation (KFO339, TRR295). M. Witzenrath reports grants from the German Research Foundation (SFB-TR84 C06 and C09, SFB 1449 B02), and from the German Ministry of Education and Research (BMBF) in the framework of CAPSyS (01ZX1604B, 01ZX1304B), SYMPATH (01ZX1906A), PROVID (01KI20160A), Phage4Cure (16GW0141), MAPVAP (16GW0247) and NUM-NAPKON. W. M. Kuebler reports grants from the German Research Foundation (SFB-TR84 A2 and C9, SFB 1449 B1, SFB 1470 A4, KU1218/9-1, KU1218/11-1, and KU1218/12-1), the BMBF in the framework of SYMPATH (01ZX1906A) and PROVID (01KI20160A), and the DZHK. S. Simmons reports grants from the DZHK and the German Foundation for Heart Research (F-09-19). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The interaction of Eph receptor tyrosine kinases with their transmembrane ligands; the ephrins, is important for the regulation of cell-cell communication. Ephrin-Eph signaling is probably best known for the discrimination of arterial and venous territories by repulsion of venous endothelial cells away from those with an arterial fate. Ultimately, cell repulsion is mediated by initiating the collapse of the actin cytoskeleton in membrane protrusions. Here, we investigated the role of the Ena/VASP family of actin binding proteins in endothelial cell repulsion initiated by ephrin ligands. Human endothelial cells dynamically extended sheet-like lamellipodia over ephrin-B2 coated surfaces. While lamellipodia of control siRNA transfected cells rapidly collapsed, resulting in a pronounced cell repulsion from the ephrin-B2 surfaces, the knockdown of Ena/VASP proteins impaired the cytoskeletal collapse of membrane protrusions and the cells no longer avoided the repulsive surfaces. Mechanistically, ephrin-B2 stimulation elicited the EphB-mediated tyrosine phosphorylation of VASP, which abrogated its interaction with the focal adhesion protein Zyxin. Nck2 was identified as a novel VASP binding protein, which only interacted with the tyrosine phosphorylated VASP protein. Nck links Eph-receptors to the actin cytoskeleton. Therefore, we hypothesize that Nck-Ena/VASP complex formation is required for actin reorganization and/or Eph receptor internalization downstream of ephrin-Eph interaction in endothelial cells, with implications for endothelial navigation and pathfinding.

Virulence traits of Acinetobacter baumannii isolates of the worldwide most prevalent international clonal lineage, IC2, remain largely unknown. In our study, multidrug-resistant IC2 clinical isolates differed substantially in their virulence potential despite their close genetic relatedness.

A targeted modulation of the endocannabinoid system is currently discussed as a promising strategy for cancer treatment. An important enzyme for the endocannabinoid metabolism is the monoacylglycerol lipase (MAGL), which catalyzes the degradation of 2-arachidonoylglycerol (2-AG) to glycerol and free fatty acids. In this study, we investigated the influence of MAGL inhibition on lung cancer cell invasion and metastasis. Using LC-MS, significantly increased 2-AG levels were detected in A549 cells treated with the MAGL inhibitor JZL184. In athymic nude mice, JZL184 suppressed metastasis of A549 cells in a dose-dependent manner, whereby the antimetastatic effect was cancelled by the CB1 receptor antagonist AM-251. In vitro, JZL184 induced a time- and concentration-dependent reduction of A549 cell invasion through Matrigel-coated membranes, which was likewise reversed by AM-251. An MAGL inhibition–associated reduction of free fatty acids as a cause of the anti-invasive effect could be excluded by add-back experiments with palmitic acid. Both JZL184 and the MAGL substrate 2-AG led to an increased formation of the tissue inhibitor of metalloproteinase-1 (TIMP-1), whereby a TIMP-1 knockdown using siRNA significantly attenuated the anti-invasive effects of both substances. Decreased invasion and TIMP-1 upregulation was also caused by the MAGL inhibitors JW651 and MJN110 or transfection with MAGL siRNA. A CB1- and TIMP-1–dependent anti-invasive effect was further confirmed for JZL184 in H358 lung cancer cells. In conclusion, MAGL inhibition led to a CB1-dependent decrease in human lung cancer cell invasion and metastasis via inhibition of 2-AG degradation, with TIMP-1 identified as a mediator of the anti-invasive effect.

Supplementary figures S1 - S6

SummaryCharacterizing trajectories of the composition and function of hominid gut microbiota across diverse environments and host species can help reveal specific properties of the human microbiota, with possible implications for host evolution and health. Using shotgun metagenomic sequencing, we investigated taxonomic and functional diversity in the gut microbiota of wild-living great apes, including two gorilla subspecies (Gorilla gorilla gorilla, Gorilla beringei beringei), three chimpanzee subspecies (Pan troglodytes verus, P.t. troglodytes, P.t. schweinfurthii), and bonobos (Pan paniscus), together with human samples from Africa and Europe. We identified microbial taxonomic and functional adaptations convergent with host phylogeny at both the community and microbial genomic levels. We could show that repeated horizontal gene transfer and gene loss are processes involved in these adaptations. We hypothesize, that these adaptation processes and changes in the microbiome predispose the host to chronic inflammatory disorders, such as type 2 diabetes via altered histidine metabolism and inflammatory bowel disease indicated by adaptation of microbes to aerobic conditions. Additionally, we find multiple lines of evidence suggesting a widespread loss of microbial diversity and evolutionary conserved clades in the human microbiota, especially in the European population. Lastly, we observed patterns consistent with codivergence of hosts and microbes, particularly for the bacterial familyDialisteraceae, though we find that overall, co-phylogeny patterns are frequently disrupted in humans.

AimsAdvanced age is unequivocally linked to the development of cardiovascular disease, however, the mechanisms leading to loss of endothelial cell regenerative capacity during aging remain poorly understood. Here we aimed to investigate novel mechanisms involved in endothelial cell senescence, that impact on endothelial cell transcription and the vascular repair response upon injuryMethods and resultsRNA sequencing of a unique collection of native endothelial cells from young and aged individuals, showed that aging (20 vs. 80 years) is characterized by p53- mediated reprogramming to promote the expression of senescence-associate genes. Molecular analysis revelead that p53 accumulated and acetylated in the nucleus of aged human endothelial cells to suppress glycolysis. Metabolic flux analysis identified an associated reduction in glucose uptake and ATP availability that inhibited the assembly of the telomerase complex, which was essential for proliferation. Nuclear translocation of p53 in aged endothelial cells was attributed to the loss of the vasoprotective enzyme, cystathionine γ-lyase (CSE), which physically anchored p53 in the cytosol. In mice, loss of endothelial cell CSE activated p53 and arrested vascular repair upon injury, while the AAV9 mediated re-expression of an active CSE mutant retained p53 in the cytosol, maintained endothelial glucose metabolism and proliferation, and prevented endothelial cell senescence. Adenoviral overexpression of CSE in human native aged endothelial cells maintained low p53 activity and re-activated telomerase to revert endothelial cell senescence.ConclusionOur data identified the interaction between CSE and p53 as a promising target to preserve vascular regeneration during aging.Key QuestionTo identify the mechanisms that regulate endothelial cell senescence under native conditions and their impact on vascular repair in aging.Key FindingLack of a physical interaction between CSE and p53 metabolically reprogrammes endothelial cells to reduce telomerase activity and halt endothelial cell regeneration.Take home messageInterventions to increase CSE expression represent a novel therapy against p53-induced endothelial cell cycle arrest and senescenseTranslational perspectiveEndothelial rejuvenation strategies could serve as promising therapies against age-related cardiovascular diseases. By investigating human native endothelial cells from young and aged individuals, we identified that the age-related nuclear accumulation of p53 reprograms endothelial cell metabolism, regulates telomerase activity and inhibits endothelial cell regeneration. Nuclear localization of p53 resulted from a loss of its interaction with the cysteine catabolizing enzyme cystathionine γ-lyase in the cytoplasm. Enhancing the physical interaction of p53 with CSE by gene therapy could revert endothelial cell senescence and activate endothelial reparative responses.

Background This retrospective study investigated factors influencing time to treatment initiation (TTI) and the influence of TTI on overall survival (OS) of primary head and neck cancer (HNC) patients in cohorts from 2003, 2008 and 2013. Methods Two hundred and ninenty seven patients (78.8% men; median age: 62 years) were included. Kaplan–Meier analyses and multivariate Cox regression were performed to investigate OS. Results Mean times to treatment initiation (TTI) of 2003, 2008 and 2013 were 17.11 ± 18.00, 30.26 ± 30.08 and 17.30 ± 37.04 days, respectively. TTI for patients with T3/T4 tumors was higher than for T1/T2 (p = 0.010). In univariable analysis on OS, TTI > 5 days showed lower OS (p = 0.047). In multivariate analysis, longer TTI had no influence on lower OS [hazard ratio (HR) 1.236; 95% CI 0.852–1.791; p = 0.264], but male gender [HR 2.342; 95% CI 1.229–4.466; p = 0.010], increased age [HR 1.026; 95% CI 1.008–1.045; p = 0.005], M1 [HR 5.823; 95% CI 2.252–15.058; p = 0.003], hypopharynx tumor [HR 2.508; 95% CI 1.571–4.003; p 0.001] and oral cavity tumor [HR 1.712; CI 1.101–2.661; p = 0.017]. The year of treatment showed no significant effect on OS. Conclusion Median TTI seemed to be very short compared to other studies. There was no clear trend in the impact of TTI on OS from 2003 to 2013.

Bone is a complex hierarchical biomineralized material, which is special amongst biominerals because it is replete with cells, namely, osteocytes. While bone has been scrutinized for centuries, many questions remain open and new research hints that the ultrastructure of bone, encompassing both the bone matrix itself and the embedded cell network, is much more heterogeneous than hitherto realized. A number of these new findings have been made thanks to the enormous developments in X-ray imaging that have occurred in recent decades, and there is promise that they will also allow many of the remaining open questions to be addressed. X-ray absorption or phase imaging affords high three-dimensional (3D) resolution and allows traversing the length scales of bone all the way down to the fine details of the lacuno-canalicular network housing the osteocytes. Multimodal X-ray imaging provides combined information covering both the length scales defined by the size of the measured volume and tomographic resolution, as well as those probed by the signal that is measured. In X-ray diffraction computed tomography (XRD-CT), for example, diffraction signals can be reconstructed tomographically, which offers detailed information about the spatial variations in the crystallographic properties of the bone biomineral. Orientational information can be obtained by tensor tomography. The combination of both small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) tensor tomography gives information on the orientation of bone nanostructure and crystals, respectively. These new technical developments promise that great strides towards understanding bone structure can be expected in the near future. In this review, recent findings that have resulted from X-ray imaging are highlighted and speculation is given on what can be expected to follow.