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Abstract In this community effort, we compare measurements between 34 laboratories from 19 countries, utilizing mixtures of labelled authentic synthetic standards, to quantify by mass spectrometry four clinically used ceramide species in the NIST (National Institute of Standards and Technology) human blood plasma Standard Reference Material (SRM) 1950, as well as a set of candidate plasma reference materials (RM 8231). Participants either utilized a provided validated method and/or their method of choice. Mean concentration values, and intra- and inter-laboratory coefficients of variation (CV) were calculated using single-point and multi-point calibrations, respectively. These results are the most precise (intra-laboratory CVs ≤ 4.2%) and concordant (inter-laboratory CVs < 14%) community-derived absolute concentration values reported to date for four clinically used ceramides in the commonly analyzed SRM 1950. We demonstrate that calibration using authentic labelled standards dramatically reduces data variability. Furthermore, we show how the use of shared RM can correct systematic quantitative biases and help in harmonizing lipidomics. Collectively, the results from the present study provide a significant knowledge base for translation of lipidomic technologies to future clinical applications that might require the determination of reference intervals (RIs) in various human populations or might need to estimate reference change values (RCV), when analytical variability is a key factor for recall during multiple testing of individuals.

Abstract Mitochondrial oxidative phosphorylation (OXPHOS) fuels cellular ATP demands. OXPHOS defects lead to severe human disorders with unexplained tissue specific pathologies. Mitochondrial gene expression is essential for OXPHOS biogenesis since core subunits of the complexes are mitochondrial-encoded. COX14 is required for translation of COX1, the central mitochondrial-encoded subunit of complex IV. Here we describe a COX14 mutant mouse corresponding to a patient with complex IV deficiency. COX14M19I mice display broad tissue-specific pathologies. A hallmark phenotype is severe liver inflammation linked to release of mitochondrial RNA into the cytosol sensed by RIG-1 pathway. We find that mitochondrial RNA release is triggered by increased reactive oxygen species production in the deficiency of complex IV. Additionally, we describe a COA3Y72C mouse, affected in an assembly factor that cooperates with COX14 in early COX1 biogenesis, which displays a similar yet milder inflammatory phenotype. Our study provides insight into a link between defective mitochondrial gene expression and tissue-specific inflammation.

The rapid increase in lipidomic studies has led to a collaborative effort within the community to establish standards and criteria for producing, documenting, and disseminating data. Creating a dynamic easy-to-use checklist that condenses key information about lipidomic experiments into common terminology will enhance the field's consistency, comparability, and repeatability. Here, we describe the structure and rationale of the established Lipidomics Minimal Reporting Checklist to increase transparency in lipidomics research.

Synchronization is widespread in animals, and studies have often emphasized how this seemingly complex phenomenon can emerge from very simple rules. However, the amount of flexibility and control that animals might have over synchronization properties, such as the strength of coupling, remains underexplored. Here, we studied how pairs of marmoset monkeys coordinated vigilance while feeding. By modeling them as coupled oscillators, we noted that (1) individual marmosets do not show perfect periodicity in vigilance behaviors, (2) nevertheless, marmoset pairs started to take turns being vigilant over time, a case of anti-phase synchrony, (3) marmosets could couple flexibly; the coupling strength varied with every new joint feeding bout, and (4) marmosets could control the coupling strength; dyads showed increased coupling if they began in a more desynchronized state. Such flexibility and control over synchronization require more than simple interaction rules. Minimally, animals must estimate the current degree of asynchrony and adjust their behavior accordingly. Moreover, the fact that each marmoset is inherently non-periodic adds to the cognitive demand. Overall, our study provides a mathematical framework to investigate the cognitive demands involved in coordinating behaviors in animals, regardless of whether individual behaviors are rhythmic or not.

Mycobacterium leprae, the pathogen that causes human leprosy, has a unique affinity for infecting and persisting inside Schwann cells, the principal glia of the peripheral nervous system. Several studies have focused on this intricate host-pathogen interaction as an attempt to advance the current knowledge of the mechanisms governing nerve destruction and disease progression. However, during the chronic course of leprosy neuropathy, Schwann cells can respond to and internalize both live and dead M. leprae and bacilli-derived antigens, and this may result in divergent cellular pathobiological responses. This may also distinctly contribute to tissue degeneration, failure to repair, inflammatory reactions, and nerve fibrosis, hallmarks of the disease. Therefore, the present study systematically searched for published studies on M. leprae-Schwann cell interaction in vitro to summarize the findings and provide a focused discussion of Schwann cell dynamics following challenge with leprosy bacilli.

Abstract For successful infection of host cells and virion production, enveloped viruses, including Zika virus (ZIKV), extensively rely on cellular lipids. However, how virus protein–lipid interactions contribute to the viral life cycle remains unclear. Here, we employ a chemo-proteomics approach with a bifunctional cholesterol probe and show that cholesterol is closely associated with the ZIKV structural protein prM. Bioinformatic analyses, reverse genetics alongside with photoaffinity labeling assays, and atomistic molecular dynamics simulations identified two functional cholesterol binding motifs within the prM transmembrane domain. Loss of prM–cholesterol association has a bipartite effect reducing ZIKV entry and leading to assembly defects. We propose a model in which membrane-resident M facilitates cholesterol-supported lipid exchange during endosomal entry and, together with cholesterol, creates a platform promoting virion assembly. In summary, we identify a bifunctional role of prM in the ZIKV life cycle by mediating viral entry and virus assembly in a cholesterol-dependent manner.

Dolichyl phosphates (DolP) are ubiquitous lipids that are present in almost all eukaryotic membranes. They play a key role in several protein glycosylation pathways and the formation of glycosylphosphatidylinositol anchors. These lipids constitute only ~0.1% of total phospholipids, and their analysis by reverse phase (RP) liquid chromatography–high-resolution mass spectrometry (LC–HRMS) is challenging due to their high lipophilicity (log P > 20), poor ionization efficiency, and relatively low abundance. To overcome these challenges, we have introduced a new approach for DolP analysis by combining trimethylsilyldiazomethane (TMSD)-based phosphate methylation and HRMS analysis. The analytical method was validated for its reproducibility, sensitivity, and accuracy. The established workflow was successfully applied for the simultaneous characterization and quantification of DolP species with different isoprene units in lipid extracts of HeLa and Saccharomyces cerevisiae cells.

iCLiPSpy is a simple assay to identify residues important for substrate binding and activity of intramembrane-cleaving proteases (I-CLiPs) in vivo and can be adapted to a screening assay for I-CLiP inhibitor or activator discovery.

Abstract Mitochondria are central for cellular metabolism and energy supply. Barth syndrome (BTHS) is a severe disorder, due to dysfunction of the mitochondrial cardiolipin acyl transferase tafazzin. Altered cardiolipin remodeling affects mitochondrial inner membrane organization and function of membrane proteins such as transporters and the oxidative phosphorylation (OXPHOS) system. Here, we describe a mouse model that carries a G197V exchange in tafazzin, corresponding to BTHS patients. TAZG197V mice recapitulate disease‐specific pathology including cardiac dysfunction and reduced oxidative phosphorylation. We show that mutant mitochondria display defective fatty acid‐driven oxidative phosphorylation due to reduced levels of carnitine palmitoyl transferases. A metabolic switch in ATP production from OXPHOS to glycolysis is apparent in mouse heart and patient iPSC cell‐derived cardiomyocytes. An increase in glycolytic ATP production inactivates AMPK causing altered metabolic signaling in TAZG197V. Treatment of mutant cells with AMPK activator reestablishes fatty acid‐driven OXPHOS and protects mice against cardiac dysfunction.

ABSTRACT: Somatic cell count (SCC) in milk is an essential indicator for defining and managing udder health. However, analyzing differential SCC (dSCC) can be helpful in determining the type or evolution stage of mastitis. A high abundance of polymorphonuclear cells (PMN) is associated with acute mastitis; however, the status of a chronic disease is less well characterized. A method capable of analyzing SCC and dSCC can prove to be a helpful tool for monitoring the status of evolution of mastitis disease in a better way. Therefore, a new direct-flow cytometry method was developed to count and differentiate somatic cells in milk without the steps of centrifugation or washing, avoiding variabilities that occur due to enrichment or loss of specific cell types. In this new method, SCC is analyzed using the method of DNA staining with Hoechst stain, whereas dSCC are analyzed using specific antibodies targeting 2 main cell types associated with mastitis: PMN cells and antigen-presenting cells, which are associated with innate and adaptive immunity. Equivalent SCC values were obtained between the new method and the routine ISO 13366-2 method in a comparison of 240 raw milk samples. Furthermore, dSCC results were confirmed by microscopy after May-Gründwald-Giemsa staining in 165 quarter milk samples from healthy and diseased cows. The method was verified with fluorescence microscopy on the 2 targeted cell types and in raw milk samples. The newly developed method is independent of any instrument and can be further designed to differentiate other cell types and animal species by selecting appropriate antibodies.

A comprehensive characterisation of Omicron-BA.1 and VOC Delta in numerous in vitro and in vivo models uncovers the factors that led to its global dominance.

Minimally invasive methods in pilonidal sinus disease (PSD) surgery are becoming standard. Although long-term results are available for some techniques, long-term outcome data of patients after pit picking is lacking. We aimed at investigating perioperative and long-term outcomes of patients undergoing pit picking, Limberg flap or primary open surgery to treat PSD.

When Daniel Dayan and Elihu Katz published their seminal book Media Events: The Live Broadcasting of History in 1992, television occupied centre stage, whereas computer networks were only beginning to be used. Since the late 1990s, television and digital media have co-existed and co-evolved in still more entangled ways. In this article, I ask how the supplementing of television by a new media form, the Web, has affected the ways media events as understood by Dayan and Katz can unfold and be conceptualised. Based on a medium theory perspective where focus is more on “media” and less on “event”, I introduce the article by tracing how Dayan and Katz understand television as a medium. Then follows a brief account of the vast literature about Media Events, with a particular focus on how digital media are conceptualised. With these two sections as a stepping stone, the Web's digital features are outlined, followed by a historical analysis of the interplay of the development of the Web and a concrete media event: the Olympics from 1996 to 2016. Finally, this web historical outline is used to re-evaluate Dayan and Katz's conceptualisation of media events. The analysis is guided by three themes – liveness, control, and participation – pivotal for Dayan and Katz's understanding of media events as well as the history of the Web.

Summary: Replenishing nicotinamide adenine dinucleotide (NAD) via supplementation of nicotinamide riboside (NR) has been shown to confer neuroprotective effects in models of aging and neurodegenerative diseases, including Parkinson’s disease (PD). Although generally considered safe, concerns have been raised that NR supplementation could impact methylation dependent reactions, including DNA methylation, because of increased production and methylation dependent breakdown of nicotinamide (NAM). We investigated the effect of NR supplementation on DNA methylation in a double blinded, placebo-controlled trial of 29 human subjects with PD, in blood cells and muscle tissue. Our results show that NR had no impact on DNA methylation homeostasis, including individuals with common pathogenic mutations in the MTHFR gene known to affect one-carbon metabolism. Pathway and methylation variance analyses indicate that there might be minor regulatory responses to NR. We conclude that short-term therapy with high-dose NR for up to 30 days has no deleterious impact on methylation homeostasis.

Introduction Muscular strength can be viewed as a product of force and velocity. Increases in both maximal force and velocity will ultimately result in an increase in power (Suchomel et al., 2016), thus contributing to performance in tasks such as athletic competition or police and military special operations. Testing isometric peak force is a reliable method to assess maximal strength (De Witt et al., 2018). Isometric mid-thigh pull tests and squat tests in various knee- and hip angles are well known. These tests, however, require force plates with computer and software, as well as a barbell that disallows displacement of the bar (usually a heavy equipment). Decentralized testing requires a reliable, lightweight and mobile measurement device comparable to standard isometric tests as described elsewhere (Maier et al., 2016). Therefore, a tension gauge (load cells) was attached to a board and a handle to perform isometric sumo stance deadlifts. The question is whether the mobile device produces comparable results to maximal isometric strength tests on force plates. Method Correlation analysis (Bayesian Pearson’s rho, Prior~Beta 1,1) and Equivalence Bayesian Paired Samples T-Tests (Lakens, 2017; Prior~Cauchy 0,0.707, a priori Limits of Agreement = +/-5%) of the measurements from the mobile test device and from force plates (CYCCESS) were performed using JASP Version 0.14.1. Healthy, trained subjects (n = 12) volunteered to perform maximal isometric deadlifts on each test device. Results were validated by additional 6 pairs of lifts on each device. Results Pearson’s rho was 0.969 (95% credible interval of the posterior distribution 0.824, 0.993). After adjusting the measures of the mobile device for the offset (mean of the differences of 416 N), Bayes factor favoring the hypothesis that the effect size lies inside the interval of +/-5% was 3.006. The additional measurements confirmed the substantial evidence for equivalence of the two measurements. Discussion Isometric tests are relatively simple, safe and time efficient to administer and possess high degrees of reliability (Grgic et al., 2022). The strong relationship between isometric peak force and deadlift 1RM (De Witt et al., 2018) makes them a valuable tool for exercise prescription. Our lightweight, mobile test device offers high precision (technical measurement error of the load cells = 0.5%) and safe testing procedure. Concurrent validity, as well as face validity, seems to be reasonably high: measurements with the mobile device do not differ from a isometric deadlift test on force plates, and lifting casualties and heavy loads could be well represented by force produced from the floor position (Bartolomei et al., 2022). References Bartolomei, S., Rovai, C., Lanzoni, I. M., & di Michele, R. (2022). Relationships between muscle architecture, deadlift performance, and maximal isometric force produced at the midthigh and midshin pull in resistance-trained individuals. Journal of Strength and Conditioniung Research, 36(2), 299-303. https://doi.org/10.1519/jsc.0000000000003455 De Witt, J. K., English, K. L., Crowell, J. B., Kalogera, K. L., Guilliams, M. E., Nieschwitz, B. E., Hanson, A. M., & Ploutz-Snyder, L. L. (2018). Isometric midthigh pull reliability and relationship to deadlift one repetition maximum. The Journal of Strength and Conditioning Research, 32(2), 528-533. https://doi.org/10.1519/jsc.0000000000001605 Grgic, J., Scapec, B., Mikulic, P., & Pedisic, Z. (2022). Test-retest reliability of isometric mid-thigh pull maximum strength assessment: A systematic review. Biology of Sport, 39(2), 407-414. https://doi.org/10.5114/biolsport.2022.106149 Lakens, D. (2017). Equivalence tests: A practical primer for t tests, correlations, and meta-analyses. Social Psychological and Personality Science, 8(4), 355-362. https://doi.org/10.1177/1948550617697177 Maier, T., Gross, M., Trösch, S., Steiner, T., Müller, B., Bourban, P., Schärer, C., Hübner, K., Wehrlin, J., Tschopp, M., Wilhelm, M., Clenin, G. E., Züst, P., & Seidel, R. (2016). Manual Leistungsdiagnostik [Performance diagnostics manual]. http://www.swissolympic.ch/Portaldata/41/Resources/03_sport/00_dokumente/Manual_Leistungsdiagnostik_(letzte_Aenderung_01.02.2016).pdf Suchomel, T. J., Nimphius, S., & Stone, M. H. (2016). The importance of muscular strength in athletic performance. Sports Medicine, 46(10), 1419-1449. https://doi.org/10.1007/s40279-016-0486-0

Summary: Objectives: The aim of this study was to quantify helmet use in skiing and snowboarding in Switzerland. Design: Regular cross-sectional surveys of skiers and snowboarders were conducted on Swiss slopes from the 2002/2003 to the 2021/2022 seasons. Method: Surveys included information on helmet use, sport, gender, age, skill level, and country of residence. Data from 15 single surveys were used to trace the evolution of helmet use in Switzerland. A logistic regression on data from the most recent survey was conducted to detect differences in helmet use among demographic groups. Results: In the 2002/2003 season, 14% of skiers (95% confidence interval [CI] ​= ​12%–18%) and 20% of snowboarders (CI 16%–26%) used a helmet. By the most recent season (2021/2022), wearing rates for both skiers and snowboarders had increased significantly (skiers: 95% [CI 93%–97%]; snowboarders: 92% [CI 89%–95%]). The regression results showed that skiers, females, younger people, and people in the German-speaking part of Switzerland were more likely to wear a helmet. Conclusions: The results show an impressive evolution of helmet use rates among skiers and snowboarders since the 2002/2003 season. Steep differences detected in earlier seasons have leveled out, and most people on the slopes today wear helmets. Nevertheless, the ongoing efforts for promoting helmet use should be maintained.

Double membrane vesicles (DMV) are used as replication organelles by several RNA viruses. Applying proteomics and lipidomics, Tabata and Prasad et al. find that two cellular acyltransferases (AGPAT1/2), responsible for synthesis of phosphatidic acid, play a role in the DMV-biogenesis of HCV and SARS-CoV-2, highlighting a common biogenesis mechanism for evolutionary distant positive-strand RNA viruses.

This article presents the project Language Traces in the City, developed at the Western Norway University of Applied Sciences (HVL) in 2021. The project aims to increase interest in and knowledge of language as intangible cultural heritage. To do this, students develop digital language routes in the cityscape. Central theoretical frameworks in this project are onomastics and sociolinguistics, and the posts disseminate knowledge on themes such as history of language, language variation, multilingualism, and language in relation to identity and power. We argue that inviting students to curate knowledge about language in the cityscape can offer creative and new perspectives on place names and the linguistic landscape.

Summary: Here, we present a protocol to analyze the T cell profiles of the neonatal ovine lung during respiratory syncytial virus (RSV) infection. The protocol delivers standardized multiparameter flow cytometry (FCM) analysis of CD4+, CD8+, regulatory, and γδ T cells isolated from lung, lymph nodes, and bronchoalveolar lavages (BALs). We detail the preparation of RSV and transtracheal inoculation of newborn lambs. We then describe tissue isolation and preparation of cell suspensions, followed by FCM acquisition to identify different T cell subsets.For complete details on the use and execution of this protocol, please refer to Démoulins et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Flinders Petrie’s first two seasons in Hawara, between 1888 and 1889, and their subsequent exhibitions in London, were arguably pivotal for the career of the British archaeologist. They also provide a wealth of documentation in his own hand to better 'understand' the man. But to better 'critique' him, this paper aims to reassess Petrie’s mindset, field work, and results in Hawara, first by the standards he began to craft for himself in the field, before briefly taking a modern perspective to complete the critical picture. To evaluate Petrie’s work by his own ambition, what could be more appropriate than to take him at his word? In the seminal 'Methods & Aims in Archaeology' of 1904, he would neatly set out his vision and the practicalities for the discipline. This assessment proffers to proceed along the original processual chapters of the book to examine how Petrie’s practices in Hawara in 1888–1889 already pioneer the theory he would consolidate 15 years later, while incorporating the latest research views. The main sources for this review are, by order of relevance, threefold: first, his original hand-written documentation from two so-called ‘Journals’ collected from letters, eight excavation ‘Notebooks’ and three ‘Day Diaries’; second, his publications for both seasons; and third, his autobiographical pieces. It appears that the ‘Father of Egyptian Archaeology’ did not entirely live up to his nascent ambition, leaving a contentious legacy to this day. The urge of the ‘salvage man’, trapped in contradictions, produced good results for the time but may also have led him astray in terms of aims and methods.

Despite recent improvements, complete motor recovery occurs in

Tractography enables identifying and evaluating the healthy and diseased brain's white matter pathways from diffusion-weighted magnetic resonance imaging data. As previous evaluation studies have reported significant false-positive estimation biases, recent microstructure-informed tractography algorithms have been introduced to improve the trade-off between specificity and sensitivity. However, a major limitation for characterizing the performance of these techniques is the lack of ground truth brain data. In this study, we compared the performance of two relevant microstructure-informed tractography methods, SIFT2 and COMMIT, by assessing the subject specificity and reproducibility of their derived white matter pathways. Specifically, twenty healthy young subjects were scanned at eight different time points at two different sites. Subject specificity and reproducibility were evaluated using the whole-brain connectomes and a subset of 29 white matter bundles. Our results indicate that although the raw tractograms are more vulnerable to the presence of false-positive connections, they are highly reproducible, suggesting that the estimation bias is subject-specific. This high reproducibility was preserved when microstructure-informed tractography algorithms were used to filter the raw tractograms. Moreover, the resulting track-density images depicted a more uniform coverage of streamlines throughout the white matter, suggesting that these techniques could increase the biological meaning of the estimated fascicles. Notably, we observed an increased subject specificity by employing connectivity pre-processing techniques to reduce the underlaying noise and the data dimensionality (using principal component analysis), highlighting the importance of these tools for future studies. Finally, no strong bias from the scanner site or time between measurements was found. The largest intraindividual variance originated from the sole repetition of data measurements (inter-run).

S-acylation is the post-translational covalent attachment of fatty acids (FA) onto cysteines. Nuskova et al. find that exposure of cells to different FAs affects which FA is used to S-acylate GNAI proteins, thereby altering GNAI function and EGFR pathway activation, linking metabolism to signaling.

The interplay between non-vesicular lipid transport, calcium signaling, and membrane dynamics is unclear. Here, the authors report a function for inositol triphosphate hydrolysis by the inositol 5-phosphatase INPP5A in controlling lipid exchange at interorganelle membrane contact sites between the endoplasmic reticulum and Golgi.

Quantitative magnetic resonance imaging (qMRI) can increase the specificity and sensitivity of conventional weighted MRI to underlying pathology by comparing meaningful physical or chemical parameters, measured in physical units, with normative values acquired in a healthy population. This study focuses on multi-echo T2 relaxometry, a qMRI technique that probes the complex tissue microstructure by differentiating compartment-specific T2 relaxation times. However, estimation methods are still limited by their sensitivity to the underlying noise. Moreover, estimating the model's parameters is challenging because the resulting inverse problem is ill-posed, requiring advanced numerical regularization techniques. As a result, the estimates from distinct regularization strategies are different. In this work, we aimed to investigate the variability and reproducibility of different techniques for estimating the transverse relaxation time of the intra- and extra-cellular space (T2IE) in gray (GM) and white matter (WM) tissue in a clinical setting, using a multi-site, multi-session, and multi-run T2 relaxometry dataset. To this end, we evaluated three different techniques for estimating the T2 spectra (two regularized non-negative least squares methods and a machine learning approach). Two independent analyses were performed to study the effect of using raw and denoised data. For both the GM and WM regions, and the raw and denoised data, our results suggest that the principal source of variance is the inter-subject variability, showing a higher coefficient of variation (CoV) than those estimated for the inter-site, inter-session, and inter-run, respectively. For all reconstruction methods studied, the CoV ranged between 0.32 and 1.64%. Interestingly, the inter-session variability was close to the inter-scanner variability with no statistical differences, suggesting that T2IE is a robust parameter that could be employed in multi-site neuroimaging studies. Furthermore, the three tested methods showed consistent results and similar intra-class correlation (ICC), with values superior to 0.7 for most regions. Results from raw data were slightly more reproducible than those from denoised data. The regularized non-negative least squares method based on the L-curve technique produced the best results, with ICC values ranging from 0.72 to 0.92.

Summary: We present a protocol to generate an advanced ex vivo model of human placenta. We use a vibrating tissue slicer to obtain precision-cut slices representative of the entire thickness of human placenta. This approach delivers standardized cultures with a preserved microstructure and cellular composition comparable to the native tissue. We applied this system to study SARS-CoV-2 infection at the maternal-fetal interface. Moreover, this system can be used to investigate the basic functions of the human placenta in health and disease.For complete details on the use and execution of this protocol, please refer to Fahmi et al. (2021).