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Chemical mutagenesis-driven forward genetic screens are pivotal in unveiling gene functions, yet identifying causal mutations behind phenotypes remains laborious, hindering their high-throughput application. Here, we reveal a non-uniform mutation rate caused by Ethyl Methane Sulfonate (EMS) mutagenesis in the C. elegans genome, indicating that mutation frequency is influenced by proximate sequence context and chromatin status. Leveraging these factors, we developed a machine learning enhanced pipeline to create a comprehensive EMS mutagenesis probability map for the C. elegans genome. This map operates on the principle that causative mutations are enriched in genetic screens targeting specific phenotypes among random mutations. Applying this map to Whole Genome Sequencing (WGS) data of genetic suppressors that rescue a C. elegans ciliary kinesin mutant, we successfully pinpointed causal mutations without generating recombinant inbred lines. This method can be adapted in other species, offering a scalable approach for identifying causal genes and revitalizing the effectiveness of forward genetic screens. Author summary: Exploring gene functions through chemical mutagenesis-driven genetic screens is pivotal, yet the cumbersome task of identifying causative mutations remains a bottleneck, limiting their high-throughput potential. In this investigation, we uncovered a non-uniform mutation pattern induced by Ethyl Methane Sulfonate (EMS) mutagenesis in the C. elegans genome, highlighting the influence of proximate sequence context and chromatin status on mutation frequency. Leveraging these insights, we engineered a machine learning enhanced pipeline to construct a comprehensive EMS mutagenesis probability map for the C. elegans genome. This map operates on the principle that causative mutations are selectively enriched in genetic screens targeting specific phenotypes amid the backdrop of random mutations. Applying this mapping tool to Whole Genome Sequencing (WGS) data derived from genetic suppressors rescuing a C. elegans ciliary kinesin mutant, we achieved precise identification of causal mutations without resorting to the conventional generation of recombinant inbred lines. Our work not only advances understanding of mutation dynamics but also revitalizes the efficacy of forward genetic screens, contributing to the refinement of genetic exploration methodologies with implications for various organisms. [ABSTRACT FROM AUTHOR]

Dynamic properties are essential for microtubule (MT) physiology. Current techniques for in vivo imaging of MTs present intrinsic limitations in elucidating the isotype-specific nuances of tubulins, which contribute to their versatile functions. Harnessing the power of the AlphaFold2 pipeline, we engineered a strategy for the minimally invasive fluorescence labeling of endogenous tubulin isotypes or those harboring missense mutations. We demonstrated that a specifically designed 16-amino acid linker, coupled with sfGFP11 from the split-sfGFP system and integration into the H1-S2 loop of tubulin, facilitated tubulin labeling without compromising MT dynamics, embryonic development, or ciliogenesis in Caenorhabditis elegans. Extending this technique to human cells and murine oocytes, we visualized MTs with the minimal background fluorescence and a pathogenic tubulin isoform with fidelity. The utility of our approach across biological contexts and species set an additional paradigm for studying tubulin dynamics and functional specificity, with implications for understanding tubulin-related diseases known as tubulinopathies. GFP-tagged tubulins have been widely used to monitor and visualize tubulin isotypes, but this is known to interfere with their cellular functions. This study describes a functional labeling strategy, guided by AlphaFold2 structural predictions, to label endogenous tubulins across diverse species while preserving functionality. [ABSTRACT FROM AUTHOR]

The regulation of PD‐L1 is the key question, which largely determines the outcome of the immune checkpoint inhibitors (ICIs) based therapy. However, besides the transcription level, the protein stability of PD‐L1 is closely correlated with its function and has drawn increasing attention. In this study, EZH2 inhibition enhances PD‐L1 expression and protein stability, and the deubiquitinase ubiquitin‐specific peptidase 22 (USP22) is identified as a key mediator in this process. EZH2 inhibition transcriptionally upregulates USP22 expression, and upregulated USP22 further stabilizes PD‐L1. Importantly, a combination of EZH2 inhibitors with anti‐PD‐1 immune checkpoint blockade therapy improves the tumor microenvironment, enhances sensitivity to immunotherapy, and exerts synergistic anticancer effects. In addition, knocking down USP22 can potentially enhance the therapeutic efficacy of EZH2 inhibitors on colon cancer. These findings unveil the novel role of EZH2 inhibitors in tumor immune evasion by upregulating PD‐L1, and this drawback can be compensated by combining ICI immunotherapy. Therefore, these findings provide valuable insights into the EZH2‐USP22‐PD‐L1 regulatory axis, shedding light on the optimization of combining both immune checkpoint blockade and EZH2 inhibitor‐based epigenetic therapies to achieve more efficacies and accuracy in cancer treatment. [ABSTRACT FROM AUTHOR]

In recent years, there has been rapid advancement in new energy technologies aimed at mitigating greenhouse gas emissions stemming from fossil fuels. Nonetheless, uncertainties persist in both the power output of new energy sources and load. To effectively harness the economic and operational potential of an Integrated Energy System (IES), this paper introduces an enhanced uncertainty set. This set incorporates N-1 contingency considerations and the nuances of source–load distribution. This framework is applied to a robust optimization model for an Electric Vehicle Integrated Energy System (EV-IES), which includes Electric Vehicle Battery Swapping Station (EVBSS). Firstly, this paper establishes an IES model of the EVBSS, and then proceeds to classifies and schedules the large-scale battery groups within these stations. Secondly, this paper proposes an enhanced uncertainty set to account for the operational status of multiple units in the system. It also considers the output characteristics of both new energy sources and loads. Additionally, it takes into consideration the N-1 contingency state and multi-interval distribution characteristics. Subsequently, a multi-time-scale optimal scheduling model is established with the objective of minimizing the total cost of the IES. The day-ahead robust optimization fully considers the multivariate uncertainty of the IES. The solution employs the Nested Column and Constraint Generation (C&CG) algorithm, based on the distribution characteristics of multiple discrete variables in the model. The intraday optimal scheduling reallocates the power of each unit based on the robust optimization results from the day-ahead scheduling. Finally, the simulation results demonstrate that the proposed method effectively reduces the conservatism of the uncertainty set, ensuring economic and stable operation of the EV-IES while meeting the demands of electric vehicle users. [ABSTRACT FROM AUTHOR]

Field tests of full-scale screw micropiles with a diameter varying from 76 to 114 mm and a length varying from 1.6 to 3 m were undertaken to investigate the axial pile capacities, load-transfer mechanism, and end installation torques of the piles in cohesive soils. Forty tests were performed on piles subjected to axial compressive and tensile loads. Six tests were instrumented with strain gauges on the pile shaft. Results showed the piles reached the limit state before the displacement exceeded 10% of the shaft diameter. The majority of axial load was transferred to the threaded segment. The adhesion coefficient of the top smooth shaft at the limit state was less than 0.1 The failure mode along the cylindrical threaded shaft was cylindrical shearing along the edge of the threads; the threads increased the axial capacities of the segment. Axial capacities of the threaded tapered segment were 43% on average greater than that of a cylindrical segment with the equivalent volume. Compressive capacities of all test piles were estimated and the results agreed reasonably well with the measured capacities. A theoretical torque model was proposed to estimate the end installation torques based on the cone penetration test results; the theoretical results matched the measured end torques very well. Key words: field test, screw micropile, tapered, axial behaviour, installation torque. Des essais in situ de micropieux a vis a pleine echelle d'un diametre variant de 76 a 114 mm et une longueur variant de 1, 6 a 3 m ont ete entrepris pour etudier les capacites axiales d'un pieu, le mecanisme de transfert de charge, et les couples d'extremites d'installation des pieux dans les sols coherents. Quarante essais ont ete realises sur des pieux soumis a des charges en traction et en compression axiale. Six essais ont ete instrumentes avec des extensometres sur l'arbre de pieu. Les resultats ont montre que les pieux ont atteint l'etat limite avant que le deplacement ait depasse 10 % du diametre de l'arbre. La majorite de la charge axiale a ete transferee au segment filete. Le coefficient d'adherence de la partie superieure de l'arbre lisse a l'etat limite etait inferieur a 0, 1. Le mode de rupture le long de l'arbre filete cylindrique etait le cisaillement cylindrique le long du bord du filete; les filets ont augmente la capacite axiale du segment. Les capacites axiales du segment conique filete etaient 43 % en moyenne plus elevee que celle d'un segment cylindrique avec un volume equivalent. Les capacites de compression de tous les pieux d'essai ont ete estimees et les resultats concordent assez bien avec les mesures de capacite. Un modele de couple theorique a ete propose pour estimer les couples d'extremites d'installation basee sur les resultats d'essai de penetration du cone; les resultats theoriques correspondent tres bien aux couples mesures. [Traduit par la Redaction] Mots-cles: essai sur le terrain, micropieu a vis, conique, comportement axial, couple d'installation., Introduction Micropiles are piles with small diameters, normally ranging between 100 and 300 mm. Micropiles were introduced to underpin historic buildings damaged during World War II in Italy (Bruce 1988). [...]