Maintenance Of Stability Research Articles

Global coupling of R-loopdynamics with RNA polymerase II modulates gene expression and early development of Drosophila.

R-loops are involved in many biological processes in cells, yet the regulatory principles for R-loops in vivo and their impact on development remain to be explored. Here, we modified the CUT&Tag strategy to profile R-loops in Drosophila at multiple developmental stages. While high GC content promotes R-loop formation in mammalian cells, it is not required in Drosophila. In contrast, RNAPII abundance appears to be a universal inducing factor for R-loop formation, including active promoters and enhancers, and H3K27me3 decorated repressive regions and intergenic repeat sequences. Importantly, such a regulatory relationship is dynamically maintained throughout development, and development-related transcription factors may regulate RNAPII activation and R-loop dynamics. By ablating Spt6, we further showed the global R-loop induction coupled with RNAPII pausing. Importantly, depending on the gene length, genes underwent up- or down-regulation, both of which were largely reversed by rnh1 overexpression, suggesting that R-loops play a significant role in the divergent regulation of transcription by Spt6 ablation. DNA damage, defects in survival, and cuticle development were similarly alleviated by rnh1 overexpression. Altogether, our findings indicate that dynamic R-loop regulation is dictated by RNAPII pausing and transcription activity, and plays a feedback role in gene regulation, genome stability maintenance, and Drosophila development.

Unraveling the Molecular Mechanisms of SIRT7 in Angiogenesis: Insights from Substrate Clues

Angiogenesis, a vital physiological or pathological process regulated by complex molecular networks, is widely implicated in organismal development and the pathogenesis of various diseases. SIRT7, a member of the Sirtuin family of nicotinamide adenine dinucleotide + (NAD+) dependent deacetylases, plays crucial roles in cellular processes such as transcriptional regulation, cell metabolism, cell proliferation, and genome stability maintenance. Characterized by its enzymatic activities, SIRT7 targets an array of substrates, several of which exert regulatory effects on angiogenesis. Experimental evidence from in vitro and in vivo studies consistently demonstrates the effects of SIRT7 in modulating angiogenesis, mediated through various molecular mechanisms. Consequently, understanding the regulatory role of SIRT7 in angiogenesis holds significant promise, offering novel avenues for therapeutic interventions targeting either SIRT7 or angiogenesis. This review delineates the putative molecular mechanisms by which SIRT7 regulates angiogenesis, taking its substrates as a clue, endeavoring to elucidate experimental observations by integrating knowledge of SIRT7 substrates and established angiogenenic mechanisms.

Chronic interferon-stimulated gene transcription promotes oncogene-induced breast cancer.

The MRE11 complex (comprising MRE11, RAD50, and NBS1) is integral to the maintenance of genome stability. We previously showed that a hypomorphic Mre11 mutant mouse strain (Mre11 ATLD1/ATLD1 ) was highly susceptible to oncogene-induced breast cancer. Here we used a mammary organoid system to examine which MRE11-dependent responses are tumor-suppressive. We found that Mre11 ATLD1/ATLD1 organoids exhibited an elevated interferon-stimulated gene (ISG) signature and sustained changes in chromatin accessibility. This Mre11 ATLD1/ATLD1 phenotype depended on DNA binding of a nuclear innate immune sensor, IFI205. Ablation of Ifi205 in Mre11 ATLD1/ATLD1 organoids restored baseline and oncogene-induced chromatin accessibility patterns to those observed in WT. Implantation of Mre11 ATLD1/ATLD1 organoids and activation of the oncogene led to aggressive metastatic breast cancer. This outcome was reversed in implanted Ifi205 -/- Mre11 ATLD1/ATLD1 organoids. These data reveal a connection between innate immune signaling and tumor development in the mammary epithelium. Given the abundance of aberrant DNA structures that arise in the context of genome instability syndromes, the data further suggest that cancer predisposition in those contexts may be partially attributable to chronic innate immune transcriptional programs.

La crisi familiare e le giurisdizioni religiose negli Stati Uniti: tensioni tra separatismo e pluralismo nel sistema delle ADR

Familiar crisis and religious jurisdictions the United States: tensions between separatism and pluralism in the ADR system ABSTRACT: In the United States, religious denominations play a central role in the out-of-court resolution of disputes. In fact, religious groups provide negotiation facilitation, mediation and arbitration services, according to models of increasing procedural complexity and binding nature. The contribution will illustrate case law guidelines about law enforcement of Jewish and Islamic judgements in familiar matters. I will focus on attempts of secular courts to mediate between opposing demands: the security of legal statuses, the maintenance of social stability and public order, the protection of rights of the individual, the deflation of civil litigation, the greater confidence of litigants towards the minister of religion acting as judge. In the conclusion, I will evaluate the impact of these trends on the intercultural paradigm that characterizes the American pluralist legal system. SOMMARIO: 1. Le ADR religiose negli Stati Uniti tra luci e ombre - 2. Pactum suprema lex esto? Il riconoscimento dei lodi arbitrali emessi dai tribunal rabbinici in sede di divorzio - 3. Il diritto familiare musulmano tra ostracismo e neutralità - 4. E pluribus unum? Il molteplice e il singolo di fronte allegiurisdizioni religiose nella società americana

Flexural Behavior of 3D-Printed Carbon Fiber-Reinforced Nylon Lattice Beams

This study investigates the flexural behavior of 3D-printed multi-topology lattice beams, with a specific emphasis on octet and cube lattice geometries created through fused deposition modeling (FDM). The mechanical properties of these beams were evaluated through quasi-static three-point bending tests. A comparative analysis of load-carrying capacity, energy absorption, and specific energy absorption (SEA) indicates that octet lattice beams exhibit superior performance to cube lattice beams. The octet lattice beam in the triple-layer double-column (TL-DC) arrangement absorbed 14.99 J of energy, representing a 38% increase compared to the 10.86 J absorbed by the cube lattice beam in the same design. The specific energy absorption (SEA) of the octet beam was measured at 0.39 J/g, which exceeds the 0.29 J/g recorded for the cube beam. Two distinct types of deformations were identified for the struts and the beam layers. Octet struts exhibit enhanced performance in stretch-dominated zones, whereas the cube system demonstrates superior efficacy in compressive-dominated regions. The results highlight the enhanced efficacy of octet lattice structures in energy absorption and mechanical stability maintenance. The investigation of sandwich lattice topologies integrating octet and cube structures indicates that while hybrid designs may exhibit efficiency, uniform octet structures yield superior performance. This study provides valuable insights into the structural design and optimization of lattice systems for applications requiring high-energy absorption and mechanical robustness.

Vine Tea Extract Enhanced the Fermentation of Skimmed Milk by Lacticaseibacillus casei

ABSTRACTVine tea extract (VTE), from the traditional Chinese herbal tea, was added to reconstituted skimmed milk; the mixture was fermented with Lacticaseibacillus casei, and fermentation characteristics, flavonoid content, antioxidant capacity (AOC), and viability of L. casei were measured. 2 mg/mL VTE promoted L. casei growth and 8 mg/mL VTE inhibited growth, an effect consistent with observed pH changes. Total flavonoid content and AOC increased with increasing VTE dosage. Dihydromyricetin was partially metabolized during fermentation and accounted for most of the antioxidant function of VTE. 2 mg/mL VTE was optimal for maintenance of probiotic culture and pH stability during cold storage and improved AOC during product shelf life. VTE has the potential to increase the health benefits of probiotic dairy products, and the resulting mixture may be suitable to use as a daily milk‐based health drink.

Effects of Balance-Based Recreational Therapy on Postural Stability in Those Diagnosed with Parkinson’s Disease

Individuals diagnosed with Parkinson’s disease (PD) have reported increased incidence rates of falling in compari-son to undiagnosed peers. Falls in this population have been found to be related to increasing symptom intensity as well as decreases in overall quality of life. The follow-ing examines the effectiveness of balance-based Recre-ational Therapy within the community setting as part of a multidiscipline treatment team known as the Stillwater Communication, Arts, Nutrition, & Exercise program. A combination of previous data analysis from this longitudinal study have found that participants improved or main-tained their physical functioning and reported less occurrences of falling. Treatment protocol focused on strength, balance, and gait-utilizing modalities such as exercise, obstacle courses, boxing training, and tai chi. Data collection of postural stability, or static balance, occurred throughout the 8-month treatment period on a monthly ba-sis and the month 1 and month 8 mean differences were analyzed for statistical and clinical significance. Assessments revealed the successful increase and/or maintenance of overall postural stability through the 8-month treatment period. Results were not statistically significant, yet provide clinical evidence to the effectiveness of RT com-munity-based balance training for individuals diagnosed with PD. The following study builds upon previous evidence that recreational therapy can be an effective treatment option for those who have been diagnosed with PD and potentially others who are ex-periencing issues with their balance. Implications for future studies and current prac-tice are provided.

Enhanced composite controller for PV/PMSG/PEMFC and BESS‐based DC microgrids voltage regulation: Integrating integral terminal sliding mode controller and recursive backstepping controller

AbstractThe variability of renewable energy sources due to weather patterns often leads to a mismatch between power generation and consumption within microgrids (MGs). This challenge is exacerbated when integrating bio‐renewable units, complicating stability maintenance in MGs. This research work introduces a novel solution to address this issue: a composite controller merging an integral terminal sliding mode controller with a recursive backstepping controller for direct current MGs (DCMGs). The proposed DCMG incorporates solar photovoltaic units, wind farms based on permanent magnet synchronous generators, proton exchange membrane fuel cells fuelled by hydrogen gas, an electrolyser, battery energy storage systems, and DC loads. First, a comprehensive mathematical model for the components within the DCMG is developed to design the proposed composite controller. This controller not only overcomes the inherent limitations and convergence issues of conventional SMCs but also ensures stable DC‐bus voltage and maintains power balance across various operational conditions. Moreover, a fuzzy logic‐based energy management system is introduced to regulate power flow, considering factors like battery state of charge and renewable energy sources' total power output. The control Lyapunov function confirms the DCMG system's asymptotic stability. Finally, the proposed controller's effectiveness is validated through simulations on both MATLAB/Simulink and Arduino Mega 2650 processor‐in‐the‐loop platforms under various operational conditions. In both platforms, the proposed controller surpasses an existing controller in terms of settling time, overshoot, and tracking error of the DC‐bus voltage.

Depth-dependent effects of forest diversification on soil functionality and microbial community characteristics in subtropical forests

Soil microbes are critical to the maintenance of forest ecosystem function and stability. Forest diversification, such as monocultures versus mixed forests stands, can strongly influence microbial community patterns and processes, as well as their role in soil ecosystem multifunctionality, such as in subtropical forest ecosystems. However, less is known about these patterns and processes vary with soil depth. Here, we investigated the results of an eight-year forest diversification field experiment comparing the soil ecosystem multifunctionality, bacterial and fungal community assembly, and network patterns in mixed versus monoculture plantations along vertical profiles (0–80 cm depth) in a subtropical region. We found that the introduction of broadleaf trees in coniferous monocultures led to enhanced synergies between multiple functions, thus improving soil multifunctionality. The effects of mixed plantations on the functional potential in top soils were greater than in deep soils, especially for carbon degradation genes (apu, xylA, cex, and glx). Microbial community assembly in the top layer, particularly in mixed plantations, was dominated by stochastic processes, whereas deterministic were more important in the deep layer. Soil microbial network complexity and stability were higher in the top layer of mixed plantations, but in the deep layer was monoculture. Interestingly, the changes in microbial communities and multifunctionality in the top layer were mainly related to variation in nutrients, whereas those in the deep were more influenced by soil moisture. Overall, we reveal positive effects of mixed forest stands on soil microbial characteristics and functionality compared to that of monocultures. Our findings highlighted the importance of enhancing functional diversity through the promotion of tree species diversity, and managers can better develop forest management strategies to promote soil health under global change scenarios.

Advances in research on the relationship between the LMNA gene and human diseases (Review).

The LMNA gene, which is responsible for encoding lamin A/C proteins, is recognized as a primary constituent of the nuclear lamina. This protein serves crucial roles in various cellular physiological activities, including the maintenance of cellular structural stability, regulation of gene expression, mechanosensing and cellular motility. A significant association has been established between the LMNA gene and several major human diseases. Mutations, dysregulated expression of the LMNA gene, and improper processing of its encoded protein can result in a spectrum of pathological conditions. These diseases, collectively termed laminopathies, are directly attributed to LMNA gene dysfunction. The present review examines the recent advancements in research concerning the LMNA gene and its association with human diseases, while exploring its pathological roles. Particular emphasis is placed on the current status of LMNA gene research in the context of tumors. This includes an analysis of the abundance of LMNA alterations in cancer and its interplay with various signaling pathways. The aim of the present review was to provide novel perspectives for studying the development of LMNA‑related diseases and additional theoretical insights for basic and clinical translational research in this field.

USP15 inhibits hypoxia-induced IL-6 signaling by deubiquitinating and stabilizing MeCP2.

Methyl-CpG binding protein 2 (MeCP2) is an important X-linked DNA methylation reader and a key heterochromatin organizer. The expression level of MeCP2 is crucial, as indicated by the observation that loss-of-function mutations of MECP2 cause Rett syndrome, whereas an extra copy spanning the MECP2 locus results in MECP2 duplication syndrome, both being progressive neurodevelopmental disorders. Our previous study demonstrated that MeCP2 protein expression is rapidly induced by renal ischemia-reperfusion injury (IRI) and protects the kidney from IRI through transcriptionally repressing the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 signaling pathway. However, the mechanisms underlying the upregulation of MeCP2 have remained elusive. Here, by using two hypoxia cell models, hypoxia and reoxygenation and cobalt chloride stimulation, we confirmed that the removal of lysine 48-linked ubiquitination from MeCP2 prevented its proteasome-dependent degradation under hypoxic conditions. Through unbiased screening based on a deubiquitinating enzymes library, we identified ubiquitin-specific protease 15 (USP15) as a stabilizer of MeCP2. Further studies revealed that USP15 could attenuate hypoxia-induced MeCP2 degradation by cleaving lysine 48-linked ubiquitin chains from MeCP2, primarily targeting its C-terminal domain. Consistently, USP15 inhibited hypoxia-induced signal transducer and activator of transcription 3 activation, resulting in reduced transcription of IL-6 downstream genes. In summary, our study reveals an important role for USP15 in the maintenance of MeCP2 stability and the regulation of IL-6 signaling.

Synthesis of dynamic modelling framework and optimal control strategy for virtually coupled train sets with guaranteed safety

Virtual coupling represents a novel railway operational mechanism, wherein several trains share state information and adjust their behaviours based on control objectives, which include safety (collision avoidance) assurance and stability (uniform velocity and constant spacing between trains) maintenance. However, the conflict between safety and stability requirements becomes more apparent, as the closer spacing between trains. Moreover, the hybrid nature of the train platoon operation, which involves both discrete events and continuous dynamics, also poses a significant barrier to the control strategy synthesis. This paper presents the Virtually Coupled Train Sets (VCTS) dynamic modelling framework and strategy synthesis approach to address these problems. A Hybrid Markov Decision Process (HMDP) is adopted for relative longitudinal dynamics modelling of trains, considering the influence of the uncertainty dynamic of trains ahead. A Stackelberg game-based strategy synthesis algorithm is applied for the safe guarantee by overcoming the uncertainty, and a Q-learning method is used for stability strategy selection from the safe strategy. The results showed a 17.95% expansion in the safe state space when compared to the general relative braking scheme (which assumes maximal acceleration during the delay horizon). In a four-train tracking operation scenario, compared with Q-learning without safe constraints, our approach gives similar stability performance; compared with Q-learning with general relative braking safe constraints, our approach not only assurance safety but has a better stability performance.

Design and stability performance optimization of a novel hybrid inspection robot walking device for smart grid applications

Inspecting in high-altitude windy environments requires maintaining a stable attitude to avoid significant oscillations, which is a crucial factor in advancing the use of hybrid power transmission line inspection robots (PTLIRs) in real-world scenarios for the electrical power and energy industry. We developed and optimized a novel walking device to improve the safety of hybrid PTLIRs operating in strong wind conditions. First, we investigate the operational conditions of transmission line inspection and design a walking device capable of transporting a flight device to be suspended under the line for inspection purposes. Second, a force equilibrium analysis is performed to explore the relationship between the key structural parameters of the walking device and the wind-induced deflection of the robot. A multiobjective optimization model is formulated to minimize the maximum deflection angle of the robot along three axes under wind loads. Finally, an optimization algorithm, using an improved non-dominated sorting particle swarm optimization (NSPSO) approach, is proposed to solve the model with good global search ability and fast convergence speed. A test platform is constructed to collect the wind deflection angle of a self-developed hybrid PTLIR under the transmission line. The test results demonstrate that the walking device can carry 30 kg to move along the transmission line. The average deflection angle of the robot around the Z axis is the highest under the influence of wind force level 7 (wind speed of 15.5 m/s) perpendicular to the windward surface of the robot. After optimization, the maximum deflection angle is limited to 10.38°, aligning within safety thresholds. The optimization algorithm effectively reduces the maximum deflection angles around the X, Z, and Y axes by 33.19 %, 39.21 %, and 13.23 %, with minimal average errors of 2.40 %, 3.87 %, and 2.13 %, respectively. The research in this paper can solve the bottleneck problem of robot practicality, which has important theoretical significance and practical application value for the safety, stability, and intelligent operation and maintenance management of power transmission lines.

A-209 Preanalytical Phase Errors Constitute the Vast Majority of Errors in Clinical Laboratory Testing

Abstract Background Laboratory errors can occur during the preanalytical, analytic, or post-analytic phase of testing and may pose a considerable threat to patient safety. Studies have estimated that pre-analytical error encapsulates ∼45-68% of laboratory errors and occurs in up to 1.52% of specimens. However, limitations of previously published studies include the use of manual error detection, reliance on manual transcription, small study sizes, and exclusion of common errors. Herein, we aim to identify the types and frequency of errors occurring in each phase of clinical laboratory testing using a large retrospective dataset across a broad range of errors detected in clinical laboratories. Methods This study was performed in the Barnes-Jewish Hospital core laboratory which provides testing for a 1,400-bed tertiary care hospital and included all tests performed for clinical chemistry, special chemistry, toxicology, automated hematology, blood gas testing, coagulation, non-infectious serology, infectious disease serology, urinalysis, and send out testing between January 2022 and May 2023. Errors were compiled from two separate sources. The first source of errors was captured from the laboratory information system (LIS, Cerner). These errors were flagged manually by laboratory staff in real time. The second source of error was a daily report manually curated by a trained medial laboratory scientist as part of routine, laboratory operating procedures and standard workflow. Results There were ∼40,000,000 individual results from ∼11,000,000 specimens during the study period and 87,317 errors observed (∼0.8% of specimens). Of these errors, specimens that were hemolyzed sufficiently to impact result but still reportable were the majority (41,047, 47.2%). When excluding minimal hemolysis, there were 46,270 errors documented. Among these, 44,847 (97.0%) were in the preanalytical phase, 451 (1.0%) were in the analytical phase, and 972 (2.0%) occurred in the postanalytical phase. Subcategorization of preanalytical errors revealed that most of the errors were related to hemolyzed samples requiring results to be suppressed (n=19,701), quantity not sufficient samples (n=8,068), clotted specimens (n=5,840), collection errors (e.g., wrong tube type, n=5,780), not on ice (n=1,369) and IV fluid contamination (n=1,122). In the analytical phase, the causes of error were interfering substances (n=279), QC out of range (n=65), instrument problems (n=58) and reagent issues (n=49). Among the postanalytical errors, 581 were comment errors, 216 were results entry errors, 131 were comments errors with potential impact on patient care, and 44 were dilution errors. Conclusions This study demonstrates that the vast majority of lab errors (∼97.0%) occurring during the preanalytical phase. The high incidence of preanalytical errors, especially errors related to sample integrity implies a need for enhanced tools and training to minimizing errors from specimen collections, maintenance of sample stability, transportation, and processing.

Two-way role of iron-carbon in biochemical reactions: microelectrolysis and enhanced activity of aerobic granular sludge for efficient refractory wastewater treatment

Industrial wastewater contained a large amount of refractory organics, and single treatment processes had limitations. This study investigated the mechanism of refractory organics removal using iron-carbon built-in coupled activated sludge (ICAS) and explored the role and function of iron-carbon (IC) within the ICAS system. The aerobic granular sludge (AGS) cultivated with IC exhibited a loose surface and a tight interior structure. Iron in the AGS concentrated near the outer layer to form a crust, which protected the inner microorganisms. IC promoted EPS secretion and regulated the abundance of positive and negative signaling molecules to maintain AGS stability. Experiments using quinoline as a model refractory organic showed that both physical adsorption by IC and biological adsorption by sludge rapidly fixed a large amount of pollutants, providing a buffer capacity for the system. The iron mineral crust on the AGS surface enhanced quinoline adsorption. Hydroxylation was the first step in quinoline degradation, with IC upregulating the genes iorA/B, qorB, and wrbA involved in this process, and the relative abundances of quinoline-degrading bacteria. Both pyridine ring opening and benzene ring cleavage occurred in the single IC system, and the microelectrolysis process produced •OH and [H], which made degradation pathway for quinoline through IC more complex than microbial degradation. Although the IC-mediated pathway accounted for only a small part of overall quinoline removal in the ICAS system, the ICAS system not only preserved the microelectrolysis process but also enhanced microbial metabolic activity. This work provided insights into the synergistic removal of pollutants and maintenance of AGS stability by the ICAS process, ensuring efficient treatment of refractory organic wastewater.

FANCD2 genome binding is nonrandom and is enriched at large transcriptionally active neural genes prone to copy number variation

Fanconi anemia (FA) is a rare genetic disease characterized by congenital abnormalities and increased risk for bone marrow failure and cancer. Central nervous system defects, including acute and irreversible loss of neurological function and white matter lesions with calcifications, have become increasingly recognized among FA patients, and are collectively referred to as Fanconi Anemia Neurological Syndrome or FANS. The molecular etiology of FANS is poorly understood. In this study, we have used a functional integrative genomics approach to further define the function of the FANCD2 protein and FA pathway. Combined analysis of new and existing FANCD2 ChIP-seq datasets demonstrates that FANCD2 binds nonrandomly throughout the genome with binding enriched at transcription start sites and in broad regions spanning protein-coding gene bodies. FANCD2 demonstrates a strong preference for large neural genes involved in neuronal differentiation, synapse function, and cell adhesion, with many of these genes implicated in neurodevelopmental and neuropsychiatric disorders. Furthermore, FANCD2 binds to regions of the genome that replicate late, undergo mitotic DNA synthesis (MiDAS) under conditions of replication stress, and are hotspots for copy number variation. Our analysis describes an important targeted role for FANCD2 and the FA pathway in the maintenance of large neural gene stability.

Application of ion modification for alteration of anode materials based on ZnO/CoZn nanostructures

During the conducted studies, it was established that the use of ion modification by irradiation with O+ and Ar+ ions makes it possible to elevate the degradation resistance of anode materials due to the effect of vacancy defect creation, the density of which varies with the irradiation fluence. At the same time, the analysis of changes in the band gap and the optical density value, expressing changes in structural distortions, revealed that ion irradiation leads to a rise in the stability of the preservation of electronic properties during long-term resource tests, which are inextricably linked with the degradation of ZnO/CoZn nanostructures due to oxidation processes as a result of lithiation. During assessment of changes in the parameters of the band gap and optical density of the samples after resource tests, it was found that the observed growth in these indicators is due to oxidation processes and partial amorphization due to the formation of oxide inclusions in the structure of nanowires, the presence of which is due to the interaction of nanostructures with the electrolyte over a long period of time during charging/discharging, which results in near-surface layer degradation due to the introduction of oxygen, and in the case of a long service life, to the formation of oxide inclusions that elevate the density of defects and vacancies in the structure. According to tests of synthesized ZnO/CoZn nanostructures as anode materials, it was found that the use of O+ and Ar+ ions not only leads to a growth in the degradation resistance of capacitive characteristics during long-term tests, but also to the stability maintenance of indicators with a reversible decrease in the charging rate at charging rate variation.

CDK-Dependent Phosphorylation Regulates PNKP Function in DNA Replication

Okazaki fragment maturation (OFM) stands as a pivotal DNA metabolic process, crucial for genome integrity and cell viability. Dysregulation of OFM leads to DNA single-strand breaks- accumulation, which is linked to various human diseases such as cancer and neurodegenerative disorders. Recent studies have implicated LIG3-XRCC1 acting in an alternative OFM pathway to the canonical FEN1-LIG1 pathway. Here, we reveal that polynucleotide kinase-phosphatase (PNKP) is another key participant in DNA replication, akin to LIG3-XRCC1. Through functional experiments, we demonstrate PNKP's enrichment at DNA replication forks and its association with PCNA, indicating its involvement in replication processes. Cellular depletion of PNKP mirrors defects observed in OFM-related proteins, highlighting its significance in replication fork dynamics. Additionally, we identify PNKP as a substrate for cyclin-dependent kinase 1/2 (CDK1/2), which phosphorylates PNKP at multiple residues. Mutation analysis of these phosphorylation sites underscores the importance of CDK2-mediated PNKP phosphorylation in DNA replication. Our findings collectively indicate a novel role for PNKP in facilitating Okazaki fragment joining, thus shedding light on its contribution to genome stability maintenance.

Development of an orally bioavailable CDK12/13 degrader and induction of synthetic lethality with AKT pathway inhibition

Cyclin-dependent kinases 12/13 play pivotal roles in orchestrating transcription elongation, DNA damage response, and maintenance of genomic stability. Biallelic CDK12 loss has been documented in various malignancies. Here, we develop a selective CDK12/13 PROTAC degrader, YJ9069, which effectively inhibits proliferation in subsets of prostate cancer cells preferentially over benign immortalized cells. CDK12/13 degradation rapidly triggers gene-length-dependent transcriptional elongation defects, leading to DNA damage and cell-cycle arrest. Invivo, YJ9069 significantly suppresses prostate tumor growth. Modifications of YJ9069 yielded an orally bioavailable CDK12/13 degrader, YJ1206, which exhibits comparable efficacy with significantly less toxicity. To identify pathways synthetically lethal upon CDK12/13 degradation, phosphorylation pathway arrays were performed using cell lines treated with YJ1206. Interestingly, degradation or genetic knockdown of CDK12/13 led to activation of the AKT pathway. Targeting CDK12/13 for degradation, in conjunction with inhibiting the AKT pathway, resulted in a synthetic lethal effect in preclinical prostate cancer models.

RNAi-Based Approaches to Control Mycotoxin Producers: Challenges and Perspectives.

Mycotoxin contamination of food and feed is a worldwide problem that needs to be addressed with highly efficient and biologically safe techniques. RNA interference (RNAi) is a natural mechanism playing an important role in different processes in eukaryotes, including the regulation of gene expression, maintenance of genome stability, protection against viruses and others. Recently, RNAi-based techniques have been widely applied for the purposes of food safety and management of plant diseases, including those caused by mycotoxin-producing fungi. In this review, we summarize the current state-of-the-art RNAi-based approaches for reducing the aggressiveness of key toxigenic fungal pathogens and mycotoxin contamination of grain and its products. The ways of improving RNAi efficiency for plant protection and future perspectives of this technique, including progress in methods of double-stranded RNA production and its delivery to the target cells, are also discussed.