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Abstract Impairment of protein translation can cause stalling and collision of ribosomes and is a signal for the activation of ribosomal surveillance and rescue pathways. Despite clear evidence that ribosome collision occurs stochastically at a cellular and organismal level, physiologically relevant sources of such aberrations are poorly understood. Here we show that a burst of the cellular signaling molecule nitric oxide (NO) reduces translational activity and causes ribosome collision in human cell lines. This is accompanied by activation of the ribotoxic stress response, resulting in ZAKα-mediated activation of p38 and JNK kinases. In addition, NO production is associated with ZNF598-mediated ubiquitination of the ribosomal protein RPS10 and GCN2-mediated activation of the integrated stress response, which are well-described responses to the collision of ribosomes. In sum, our work implicates a novel role of NO as an inducer of ribosome collision and activation of ribosomal surveillance mechanisms in human cells.

Abstract Background Sporadic late onset nemaline myopathy is a rare, progressive muscle disease, presenting in adulthood, mainly affecting proximal limb and bulbar muscles. Muscle biopsies show characteristic nemaline rods. The putative mechanism is considered immune-related. Other manifestations aside from neuromuscular symptoms have not been described previously. Case presentation We present a case with atypical sporadic late onset nemaline myopathy (SLONM) of a non-HIV, non-MGUS subtype, where skin manifestations preceded neuromuscular symptoms, and a residual thymus with the histology of thymic follicular hyperplasia was detected during the diagnostic workup. Thorough dermatological investigations could not explain the skin presentations. Muscle biopsy revealed variation in fiber diameter, ragged-red and COX-negative fibers associated with discrete fibrosis. Electron microscopy detected atrophic muscle fibres with disorganization of the myofibrils, nemaline rods and abnormal mitochondria. Single-fiber EMG suggested signs of a neuromuscular transmission defect, EMG showed signs of myopathy. Analyses of antibodies associated with myasthenia gravis were negative. The patient showed improvement after intravenous immunoglobulin treatment regarding both the skin and the muscle symptoms. Conclusions Our case highlights the heterogeneity of SLONM with its varied spectrum of presentation. A unique combination of dermatological symptoms and SLONM could be seen with skin lesions as primary presenting symptoms. An association can be considered between the different manifestations, presumably based on immune etiology, where immunosuppressive therapy has been beneficial.

Repeated blood sampling is required in certain clinical and research settings, which is currently performed by drawing blood from venous catheters requiring manual handling of each sample at the time of collection. A novel body-worn device for repeated serial samples, Fluispotter®, with automated extraction, collection, and storage of up to 20 venous dried blood spot samples over the course of 20 h may overcome problems with current methods for serial sampling. The purpose of this study was to assess the performance and safety of Fluispotter for the first time in healthy subjects. Fluispotter consists of a cartridge with tubing, a reservoir for flushing solution, pumps and filter paper, and a multi-lumen catheter placed in the brachial vein. We recruited healthy subjects for testing in an in-hospital setting. Fluispotter was attached by an anesthesiologist to 22 healthy subjects of which 9/22 (40.9%) participants had all 20 samples taken, which was lower than the goal of complete sampling in 80% of the subjects (P = 0.02). The main reason for sample failure was clogging of blood flow which was observed in 11/22 (50%) of the participants. No serious adverse events occurred, and the participants rated the pain from the insertion and the removal of catheter as very low. A cortisol profile showed nadir values at midnight and highest values at 05:00 h. Although full sampling was not successful in all participants, the Fluispotter technology proved safe and highly acceptable to the participants producing the expected cortisol profile without the requirement of staff during sample collection.

During the CO2 stunning of pigs, a variation in their reaction to the gas and the duration of the induction period is observed. The stunning process can be affected by several conditions, such as stressful events and previous experiences, but the stocking density in the gondola may also have an impact. The objective was to investigate the effect of stocking density on the stunning process under commercial conditions. To quantify the pigs’ reactions under industrial settings with a stocking density of up to eight pigs pr. Gondola (3.91 m2), the activity level was measured using an AI solution. Compared with a simulation of the expected induction period, a significantly longer induction period was found in gondolas containing seven and eight pigs (p < 0.001) but not when the gondolas contained three or four pigs. Both high and mean activity levels were significantly higher when stocking density was increased from three or four pigs to seven or eight pigs. The stunning process was thus negatively affected when increasing the stocking density. More knowledge is needed to explain this effect and to make statements on optimal stocking density. The measured activity levels may be a useful tool for obtaining information under commercial conditions and for documenting animal welfare.

Introduction: New EU regulations for increased sustainability and better utilization of side-streams are pushing the European mussel farming and processing industry to adapt to new regulations regarding their wastewater production, which leads to incentives for optimizing methods for better utilization of this side-stream resource. In this study, one such European company is faced with a choice of a filtration system out of three possible options, to invest in and achieve the option with the best balance between economic gain and environmentally sound production.Method: For this, a specialized Analytical Hieratical Processes (AHP) tool was utilized together with a supplementary Debias Your Decisions (DYD) tool to visualize the local best choice for the company.Results: Based on the generated user weights from the company it was apparent that the user weights were biased towards certain criteria in the AHP tool, which meant that half of the tool’s criteria had greatly diminished influence on the tool’s scores. This brought into question the objectivity of the Performance index (Pi) score results from the AHP tool when the user weights were applied, since these Pi scores deviated significantly from the expert Pi scores. The supplement tool DYD was found to be able to identify the dominant decision biases influencing the user weights in the current case, which enabled the implementation of relevant debiasing techniques to correct for the overweighted user weights from the company. With these corrections and the case company’s input data, the optimum alternative suggested for the case company is membrane filtration based on the final Pi scores for all weight sets.Discussion: This process and final debiased results further validated the AHP tools results as objectively grounded due to their convergence with the experts’ scoring results. This study also served as further validation for the new supplement DYD tool as the current sensitivity analysis showed the tool’s robustness to be high.

Objective: To describe 1) the proportion of patients with knee osteoarthritis (OA) undergoing guideline-adherent core treatments until six months after primary referral to an orthopaedic surgeon, 2) which specific treatment pathways these patients undertake and 3) the characteristics of patients choosing different treatment pathways. Design: This prospective cohort study consecutively invited patients referred to an orthopaedic surgeon due to knee OA at two Danish hospitals from October 2018 to December 2020. Before and six months after consulting the surgeon, patients answered a questionnaire reporting which treatments they had received for knee OA. The proportion receiving the combination of guideline-adherent treatments (i.e., exercise, education, and dietary weight management if needed) was determined. We evaluated the specific treatment usage before and until six months after the consultation and investigated characteristics of patients undertaking different pathways. Results: Out of 5251 eligible patients, 2574 (49%) had complete data and were included in analyses. 23% received guideline-adherent treatments, 10% had no treatment. Patients underwent 1143 unique treatment pathways, 62% including treatments not recommended/recommended against. Those who underwent guideline-adherent pathways had similar characteristics to those who did not but tended to be females, retired, had longer-lasting knee problems, have comorbidities, and higher education levels. Conclusions: Only one in four patients with knee OA received treatment adhering to clinical guidelines before and six months after consulting the surgeon. Patients used many different treatment pathways. There is a need for a structured effort to increase the use of guideline-adherent core treatments. Trial Identifiers: Registration: NCT03746184, Protocol: PMID: 34233992.

Traditional Chinese Medicine (TCM) is popular for the treatment of endometriosis, a complex gynecological disease that affects 10% of women globally. The growing market for TCMs has yielded a significant incentive for product adulteration, and although emerging technologies show promise to improve their quality control, many challenges remain. We tested the authenticity of two traditional Chinese herbal formulae used in women’s healthcare for the treatment of endometriosis, known as Gui Zhi Fu Ling Wan (FL) and Ge Xia Zhu Yu Tang (GX). Dual-locus DNA metabarcoding analysis coupled with high-performance thin-layer chromatography (HPTLC) were used to authenticate 19 FL and six GX commercial herbal products, as well as three ad hoc prepared artificial mixtures. HPTLC was able to detect most of the expected ingredients via comparative component analysis. DNA metabarcoding was able to detect an unexpected species diversity in the products, including 38 unexpected taxa. Chromatography has a resolution for all species indirectly through the identification of marker compounds for the different species ingredients. Metabarcoding on the other hand yields an overview of species diversity in each sample, but interpretation of the results can be challenging. Detected species might not be present in quantities that matter, and without validated quantification, some detected species can be hard to interpret. Comparative analysis of the two analytical approaches also reveals that DNA for species might be absent or too fragmented to amplify as the relevant chemical marker compounds can be detected but no amplicons are assigned to the same species. Our study emphasizes that integrating DNA metabarcoding with phytochemical analysis brings valuable data for the comprehensive authentication of Traditional Chinese Medicines ensuring their quality and safe use.

The aim was to conduct a meta-aggregation of qualitative studies on the interplay between playful learning, digital materials and physical activity in higher education. A literature search was performed across multiple databases and web pages up until May 2022. A critical appraisal following the JBI checklist for qualitative studies was conducted, and the GRADE-CERQual tool was used to evaluate confidence in the cumulative evidence. Three eligible studies were identified. We extracted 81 findings and 44 illustrations and synthesised them into six categories: (1) experience of playful approaches to learning; (2) interplay between play and learning; (3) experience with digital materials; (4) experience of collaboration; (5) experience with space significance; and (6) experience of getting a reward for participating in the activity. The synthesis showed that the benefits of interplay between playful learning, digital materials and physical activity in higher education were that students were motivated by gamified learning activities, including themes such as competitive spirits, receiving rewards, collaboration and creativity. Both students and educators experience that game-based learning strategies provide meaningful practice because they may facilitate the learning and retention of information by highlighting key information and breaking down information. The synthesis showed that the constraints of the interplay were time as a resource, frustration with using digital materials and that it challenges traditional learning strategies and learning spaces. Confidence in the evidence is low due to moderate concerns regarding methodological limitations and serious concerns regarding the adequacy of the data. Therefore, we highlight the need to expand the field both in practice and research.