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Protein O-linked mannose (O-Man) glycosylation is an evolutionary conserved posttranslational modification that fulfills important biological roles during embryonic development. Three nonredundant enzyme families, POMT1/POMT2, TMTC1-4, and TMEM260, selectively coordinate the initiation of protein O-Man glycosylation on distinct classes of transmembrane proteins, including α-dystroglycan, cadherins, and plexin receptors. However, a systematic investigation of their substrate specificities is lacking, in part due to the ubiquitous expression of O-Man glycosyltransferases in cells, which precludes analysis of pathway-specific O-Man glycosylation on a proteome-wide scale. Here, we apply a targeted workflow for membrane glycoproteomics across five human cell lines to extensively map O-Man substrates and genetically deconstruct O-Man initiation by individual and combinatorial knockout of O-Man glycosyltransferase genes. We established a human cell library for the analysis of substrate specificities of individual O-Man initiation pathways by quantitative glycoproteomics. Our results identify 180 O-Man glycoproteins, demonstrate new protein targets for the POMT1/POMT2 pathway, and show that TMTC1-4 and TMEM260 pathways widely target distinct Ig-like protein domains of plasma membrane proteins involved in cell-cell and cell-extracellular matrix interactions. The identification of O-Man on Ig-like folds adds further knowledge on the emerging concept of domain-specific O-Man glycosylation which opens for functional studies of O-Man-glycosylated adhesion molecules and receptors., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: To improve breast radiographers' individual performance by using automated software to assess the correctness of breast positioning and compression in tomosynthesis screening., Materials and Methods: In this retrospective longitudinal analysis of prospective cohorts, six breast radiographers with varying experience in the field were asked to use automated software to improve their performance in breast compression and positioning. The software tool automatically analyzes craniocaudal (CC) and mediolateral oblique (MLO) views for their positioning quality by scoring them according to PGMI classifications (perfect, good, moderate, inadequate) and checking whether the compression pressure is within the target range. The positioning and compression data from the studies acquired before the start of the project were used as individual baselines, while the data obtained after the training were used to test whether conscious use of the software could help the radiographers improve their performance. The percentage of views rated perfect or good and the percentage of views in target compression were used as overall metrics to assess changes in performance., Results: Following the use of the software, all radiographers significantly increased the percentage of images rated as perfect or good in both CCs and MLOs. Individual improvements ranged from 7 to 14% for CC and 10 to 16% for MLO views. Moreover, most radiographers exhibited improved compression performance in CCs, with improvements up to 16%., Conclusion: Active use of a software tool to automatically assess the correctness of breast compression and positioning in breast cancer screening can improve the performance of radiographers., Clinical Relevance Statement: This study suggests that the use of a software tool for automatically evaluating correctness of breast compression and positioning in breast cancer screening can improve the performance of radiographers on these metrics, which may ultimately lead to improved screening outcomes., Key Points: • Proper breast positioning and compression are critical in breast cancer screening to ensure accurate diagnosis. • Active use of the software increased the quality of craniocaudal and mediolateral oblique views acquired by all radiographers. • Improved performance of radiographers is expected to improve screening outcomes., (© 2023. The Author(s).)

We report on laser cooling of a large fraction of positronium (Ps) in free flight by strongly saturating the 1^{3}S-2^{3}P transition with a broadband, long-pulsed 243 nm alexandrite laser. The ground state Ps cloud is produced in a magnetic and electric field-free environment. We observe two different laser-induced effects. The first effect is an increase in the number of atoms in the ground state after the time Ps has spent in the long-lived 2^{3}P states. The second effect is one-dimensional Doppler cooling of Ps, reducing the cloud's temperature from 380(20) to 170(20) K. We demonstrate a 58(9)% increase in the fraction of Ps atoms with v&#95;{1D}<3.7×10^{4}  ms^{-1}.