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Reconstituting transcription–translation-coupled DNA replication within complex in vitro biological systems

Correction: Informal care in different European care systems: Effects of caregiving on mental health over time

Divergent synthesis of Haplomitrium diterpenoids via a late-stage biomimetic skeletal reorganization approach

Development of a competition assay to assess the in vitro fitness of dengue virus serotypes using an optimized serotype-specific qRT-PCR

Background Comparing the in vitro fitness of dengue virus (DENV) isolates is a pivotal approach to assess the contribution of DENV strains’ replicative fitness to epidemiological contexts, including serotype replacements. Competition assays are the gold standard to compare the in vitro replicative fitness of viral strains. Implementing competition assays between DENV serotypes requires an experimental setup and an appropriate read-out to quantify the viral progeny of strains belonging to different serotypes. Methods In the current study, we optimized an existing serotyping qRT-PCR by adapting primer/probe design and multiplexing the serotype-specific qRT-PCR reactions, allowing to accurately detect and quantify all four DENV serotypes. We next developed an in vitro competition assay to compare the replicative fitness of two DENV serotypes in the human hepatic cell line HuH7. Findings The qRT-PCR was specific, and had a limit of detection below 7.52, 1.19, 3.48 and 1.36 genome copies/µL, an efficiency of 1.993, 1.975, 1.902, 1.898 and a linearity (R²) of 0.99975, 0.99975, 0.99850, 0.99965 for DENV-1, −2, −3 and −4, respectively. Challenge of this multiplex serotype-specific qRT-PCR on mixes of viral supernatants containing known concentrations of strains from two serotypes evidenced an accurate quantification of the amount of genome copies of each serotype. Quantification of the viral progeny of each serotype in the inoculum and the supernatant of competition assays using the serotype-specific multiplex qRT-PCR unveiled an enrichment of the supernatant in DENV-1 genome copies, uncovering the enhanced replicative fitness of this DENV-1 isolate. Conclusions This optimized qRT-PCR combined with a relevant cellular model allowed to accurately quantify the viral progeny of two DENV strains belonging to two different serotypes in a competition assay, allowing to determine which strain had a replicative advantage. This reliable experimental setup is adaptable to the comparative study of the replicative fitness of any DENV serotypes.

Author Correction: Strong crystalline thermal insulation induced by extended antibonding states

Development of a floxed Gabbr2 gene allows for widespread conditional disruption of GABBR2 and recapitulates the phenotype of germline Gabbr2 knockout mice

GABBR1 and GABBR2 are widely expressed in the brain and genetic inhibition of their function leads to neurologic dysfunction and premature death in mice. Given that GABBR1 and GABBR2 heterodimerize to form a functional receptor, global knockout of GABBR1 or GABBR2 results in a similar phenotype, characterized by spontaneous epileptiform activity, hyperlocomotor activity, hyperalgesia, impaired memory and premature death. Both GABBR1 and GABBR2 are expressed in a variety of tissues outside the nervous system and GABA-B receptors have been shown to heterodimerize with other class C GPCRs. However, the neurologic consequences of global GABBR1 or GABBR2 knockout mice have made it difficult to study the effects of loss of GABBR function in other organs. While a conditional knockout for GABBR1 is available, the Gabbr2 gene had not been “floxed”. Therefore, we used CRISPR to insert loxP sites into the Gabbr2 locus in mice. These mice are normal at baseline but when bred with mice expressing Cre-recombinase under the control of the ubiquitously expressed Actin gene promoter, Gabbr2 lox/lox mice recapitulate the phenotype of global GABBR2 knockout mice demonstrating alterations throughout the brain, including the cortex, hippocampus and cerebellum. We document abnormal neurological function, increased neuronal cell death, changes in neuronal architecture, and premature death. These mice should be useful tools to study cell type-specific loss of GABBR2 function in the brain and other organs.

Mesocrystal growth through oriented sliding and attachment of nanoplates

Abstract Oriented attachment is a critical, yet poorly understood, crystal growth pathway based on the self-assembly of nanocrystals. During oriented attachment, solvent-separated particles align and coalesce through forces that enable precise rotation and translation. While prior studies emphasized intragap forces driving crystallographic alignment, the forces enabling uniform stacking and superlattice formation remain unclear. Here, we demonstrate how macroscopic gibbsite mesocrystals emerge from nanoplates guided into staggered positions by directional sliding. Electron microscopy and X-ray scattering reveal the monoclinic superlattice structure, based on nanoplate stacking with a uniform ≈50° stagger along the gibbsite [010] direction. In situ liquid-cell TEM captures preferential sliding along the gibbsite [010] direction, decelerating with increasing particle overlap. Molecular dynamics simulations reveal that this staggered arrangement corresponds to a global free-energy minimum, rather than full alignment. The simulations also confirm that sliding along the [010] direction is energetically favored and provide insight into the role of interfacial water in achieving long-range ordered assemblies. These insights highlight the energy landscape’s role in oriented attachment, with implications for material synthesis and hierarchical structures in nature.

Lifetime physical intimate partner violence (pIPV) among Mozambican women: Individual and contextual level factors driving its prevalence

Background Intimate partner violence (IPV) remains a significant public health issue in Mozambique. This study uses data from the 2022–2023 Mozambique Demographic and Health Survey (DHS) to examine the prevalence and sociodemographic determinants of Lifetime IPV among women. Specifically, this study focuses on physical IPV (pIPV), as measured in the Mozambique Demographic and Health Survey (DHS), which does not capture sexual or emotional violence. Methods A nationally representative sample of 4,813 women aged 15–49 was analyzed to assess the prevalence of Lifetime pIPV. Lifetime pIPV was defined as physical violence perpetrated by a current or former partner, by DHS definitions. Logistic regression models were used to identify individual- and contextual-level factors associated with Lifetime IPV. Results Nearly 1 in 4 women (21.92%) reported experiencing physical abuse from a current or former partner in their lifetime. Marital status emerged as a key individual-level determinant, with cohabitating and separated women being at significantly higher odds of experiencing pIPV compared to women who had never been in a union. Educational attainment and current employment were also associated with increased odds of pIPV. Similarly, women who justified physical abuse had higher odds of experiencing pIPV. Additionally, husbands/partners’ alcohol consumption was one of the strongest predictors, nearly tripling the odds of Lifetime pIPV. Finally, the effect modification between marital status and education showed that the effect of marital status on pIPV was further modified by women educational attainment. At the contextual level, provincial disparities were observed, with Cabo Delgado and Manica showing the highest pIPV prevalence, while Inhambane and Gaza had the lowest. Conclusion This study provides updated data on the prevalence of Lifetime pIPV in Mozambique and highlights key individual and contextual factors contributing to pIPV. The findings underscore the need for targeted interventions addressing socio-cultural norms, improving educational opportunities, mitigating male partner’s alcohol consumption, and implementing province-specific strategies to reduce pIPV and enhance women’s safety across Mozambique.

Complete structures of the YenTc holotoxin prepore and pore reveal the evolutionary basis for chitinase incorporation into ABC toxins

A multi-factor dynamic time series measure for stock correlation analysis

Existing similarity measures in stock correlation analysis often overlook the multidimensional nature of stock data and the dynamics of the time-lag effect (TLE) in phase differences. To address these limitations, this paper proposes a novel method, called Multi-Factor Dynamic Temporal Similarity Measure (MFDTSM). The method introduces an enhanced eXtreme Gradient Boosting (XGBoost) model based on Shapley Additive exPlanations (SHAP) to comprehensively evaluate the influence of stock factors. The proposed method effectively categorizes stocks and reveals the heterogeneity of factor influence by clustering the SHAP values of stock factors. Furthermore, MFDTSM is able to successfully quantify the dynamic rate of phase differences in TLE by constructing the cumulative distance matrix and analyzing the optimal alignment paths of time series data, thereby significantly improving the accuracy of the similarity measure. Empirical analysis is performed using 102 stocks from the communication and financial industries, including 12 key stock factors. The experimental results demonstrate that MFDTSM improves the accuracy of the analysis of industry correlation, linear correlation, and stock correlation pricing by 10%, 16%, and 5%, respectively, over existing methods, which highlight the efficiency and stability of MFDTSM in analyzing complex stock market dynamics.

Chemotherapy-driven intestinal dysbiosis and indole-3-propionic acid rewire myelopoiesis to promote a metastasis-refractory state

Abstract The contribution of chemotherapy-induced tissue injury to individual susceptibility to metastasis remains largely unexplored. We report that chemotherapy indirectly prevents colorectal cancer (CRC) liver metastases by inducing a lasting systemic “chemomemory”. Chemotherapy-induced intestinal mucositis alters nutrient availability, promoting the expansion of tryptophan-metabolizing bacteria and production of the microbial metabolite indole-3-propionic acid (IPA). IPA reprograms bone marrow myelopoiesis by redirecting common myeloid progenitor fate toward the macrophage lineage, limiting generation of immunosuppressive Ly6C high CCR2 + monocytes. This shift enhances CD4 +  T cell antitumor function by promoting Th1 differentiation and spatially reorganizing CD8 + and CD4 +  T cell interactions within the metastatic microenvironment. In a subset of CRC patients, circulating IPA levels increase after chemotherapy and inversely correlate with monocyte abundance, while high monocyte levels were associated with reduced survival. Our findings reveal that chemotherapy-induced intestinal injury normalizes pathological myelopoiesis through a microbiota-derived metabolite and identify IPA as a potential adjuvant to counteract monocyte-driven immunosuppression and metastasis.

A novel algorithm for model uncertainty reduction in trapezoidal fuzzy fault tree risk assessment

Intelligent risk assessment in complex systems increasingly relies on methods like trapezoidal fuzzy fault trees. However, conventional techniques often struggle with accurately calculating top-event probabilities and handling model uncertainty, which undermines the reliability of risk evaluations. To address this, we propose a novel algorithm that effectively reduces model uncertainty in trapezoidal fuzzy fault tree analysis. By leveraging the cut-set theorem and operating directly on trapezoidal fuzzy numbers without defuzzification, our approach preserves the completeness of fuzzy information throughout the calculation. The algorithm accommodates three common fault tree logics: OR-only, AND-only, and mixed OR/AND gates. In a case study on liquid ammonia leakage, the method achieved a 45.25% reduction in model uncertainty—surpassing existing approaches, which reached only 36.65%. Results showed 99.40% consistency with benchmark literature, affirming the algorithm’s accuracy. Additional analysis using OREDA field data demonstrated the method’s stability across different system structures and risk levels. To further verify robustness under input uncertainty, we conducted 60 perturbation tests on representative basic events with high, medium, and low failure probabilities. The algorithm exhibited exceptional stability, with output variations remaining below ±1% even under ±15% input perturbations, confirming its suitability for real-world applications where data uncertainties are common. This work offers a reliable and scalable solution for risk assessment in high-stakes industries such as nuclear energy and chemical processing.

The potential of H5N1 viruses to adapt to bovine cells varies throughout evolution

Abstract Avian influenza H5N1 clade 2.3.4.4b viruses caused a global panzootic and, unexpectedly, widespread outbreaks in dairy cattle, therefore representing a pandemic threat. To inform control strategies, it is critical to determine whether the potential to adapt to bovine cells is a general feature of H5N1 viruses, is specific to viruses of clade 2.3.4.4b, or narrowly restricted to some genotypes within this clade. Using a large panel of recombinant viruses representing >60 years of H5N1 history and other IAVs for comparison, we demonstrate replicative fitness in bovine cells is: (i) highly variable across 2.3.4.4b genotypes, (ii) limited in viruses predating the global expansion of this clade, (iii) determined by the internal gene cassette, and (iv) not restricted to udder epithelial cells. Mutations in the PB2 polymerase subunit emerge as key determinants of adaptation, although their phenotypic effects are context dependent. Bovine B3.13 and some avian genotypes exhibit enhanced modulation of bovine interferon-induced antiviral responses, determined by at least PB2, nucleoprotein, and the non-structural protein NS1. Our results highlight the polygenic nature of IAV host range, and reveal that the replication fitness in bovine cells, and likely their potential to adapt to cattle, varies greatly during the evolutionary trajectory of H5N1 viruses.

Comparison of outcomes between minimally invasive percutaneous bunnell suture and traditional open modified Kessler technique for acute closed achilles tendon rupture: A single-center cohort study

Percutaneous Bunnell repair and open modified Kessler repair remain debated options for acute Achilles tendon rupture (AATR). We retrospectively compared a minimally invasive percutaneous Bunnell technique (Group A) with an open modified Kessler repair (Group B) within a standardized early functional rehabilitation (EFR) protocol at a single center. Fifty-five adults with closed AATR treated between January 2021 and December 2022 were analyzed (Group A, n = 25; Group B, n = 30). Between-group comparisons used Welch t tests for continuous variables and χ² or Fisher exact tests for categorical variables; American Orthopaedic Foot & Ankle Society (AOFAS) and Achilles Tendon Total Rupture Score (ATRS) were assessed at 12 and 24 weeks, with Holm adjustment applied within each scale. Compared with Group B, Group A had shorter operative time (56.6 ± 15.1 vs 68.2 ± 23.2 minutes; mean difference −11.6; 95% CI −22.05 to −1.15; P = 0.030), less intraoperative blood loss (28.4 ± 8.4 vs 74.7 ± 19.4 mL; −46.3; 95% CI −54.22 to −38.38; P < 0.001), and a shorter length of stay (4.8 ± 2.0 vs 6.6 ± 2.9 days; −1.8; 95% CI −3.13 to −0.47; P = 0.009). At 12 weeks, AOFAS favored Group A (+1.36; 95% CI + 0.19 to +2.53; P = 0.047) and ATRS also favored Group A (+1.73; 95% CI + 0.48 to +2.98; P = 0.008). At 24 weeks, AOFAS was not different (+0.51; 95% CI −0.49 to +1.51; P = 0.312), whereas ATRS remained higher in Group A (+2.17; 95% CI + 0.88 to +3.46; P = 0.003). Overall complications were 12.0% vs 26.7% (risk difference −14.7%; 95% CI −34.10 to +7.20; P = 0.310); no sural nerve injuries occurred. Within an EFR framework, percutaneous Bunnell repair provided superior perioperative efficiency with comparable safety and modest, sub-MCID functional advantages, supporting confirmation in larger prospective studies.

Multi-target and ultra-high-speed optical wireless communication using a thin-film lithium niobate optical phased array

Abstract Optical wireless communication (OWC) effectively addresses challenges such as radio spectrum scarcity and signal attenuation by leveraging the properties of laser beams. A major advance in this field comes from the incorporation of optical phased arrays (OPAs), which enable inertial-free, high-speed beam steering with transformative potential. In this contribution, we propose and demonstrate a multi-target and ultra-high-speed OWC system based on a thin-film lithium niobate (TFLN) OPA. It enables real-time multi-target connection without mechanical components or lenses. This system can achieve OWC with a single-channel communication data rate of up to 320 Gbps in the modulation format of 16-Quadrature Amplitude Modulation (QAM), significantly exceeding the peak capabilities of 5 G and current 6 G proposals. System performance is further validated through the stable transmission of uncompressed high-definition video. This work establishes a new paradigm for fully solid-state, chip-scale OWC systems, combining unprecedented single-channel data throughput with dynamic multi-target support.

Correction: Targeted modulation of MMP9 and GRP78 via molecular interaction and in silico profiling of Curcuma caesia rhizome metabolites: A computational drug discovery approach for cancer therapy

Conformation-driven reversibility control in ring-opening metathesis polymerization of non-bicyclic cyclooctenes

A disproportionality analysis of FDA adverse event reporting system (FAERS) events for filgotinib

Background Although filgotinib, a selective Janus kinase 1 inhibitor, has been increasingly applied in the treatment of inflammatory diseases, its comprehensive safety profile remains insufficiently characterized. Using data from the FAERS database covering Q1 2014 to Q2 2024, this study attempts to analyze adverse event signals linked to filgotinib and provide guidance for the safe and sensible clinical usage of filgotinib. Methods From Q1 2014 to Q2 2024, information on adverse drug events (ADEs) associated with filgotinib was gathered. The reporting odds ratio (ROR), proportional reporting ratio (PRR), Bayesian confidence propagation neural network (BCPNN), and multi-item gamma Poisson shrinker (MGPS) were among the signal detection methods that were employed for analysis following data normalization. Results Filgotinib was shown to be the main suspected medication in ADE reports, exposing 103 preferred terms (PTs) in 17 system organ classes (SOCs). Infections, gastrointestinal disorders, and musculoskeletal and connective tissue disorders were the most commonly reported adverse effects. Additionally, atrial fibrillation, alopecia, elevated serum creatinine, blood creatinine increased, pulmonary embolism, epididymitis, respiratory failure, and osteopenia were identified as potential disproportionate reporting signals for filgotinib, although these were not listed in the official drug label. Notable significant signals included large intestine erosion (ROR 2186.05, 95%CI (ROR) : 1015.94–4703.86, PRR 2176.18, 95%CI (PRR) : 1014.64–4667.42), mesenteric arterial occlusion (ROR 1832.17, 95%CI (ROR) : 897.68–3739.48, PRR 1822.71, 95%CI (PRR) : 896.17–3707.20), repetitive strain injury (ROR 1149.27, 95%CI (ROR) : 363.16–3637.01, PRR 1147.05, 95%CI (PRR) : 363.24–3622.15), oligoarthritis (ROR 755.02, 95%CI (ROR) : 310.74–1834.54, PRR 752.59, 95%CI (PRR) : 310.60–1823.51), and periostitis (ROR 676.03, 95%CI (ROR) : 319.36–1431.06, PRR 672.98, 95%CI (PRR) : 318.97–1419.87). The subgroup analysis identified obvious sex and age-specific trends in filgotinib-related adverse reactions, emphasizing a higher risk of renal disorders in females, a preponderance of gastrointestinal events in males, and age-dependent trends involving mesenteric occlusion, increased serum creatinine, and immunoglobulin reduction. Conclusion While filgotinib demonstrates therapeutic efficacy, it is associated with a range of potential adverse events, underscoring the need for vigilant clinical monitoring. Particular attention should be given to gastrointestinal, cardiovascular, respiratory, and metabolic complications.

A large-scale trial in Chennai, India shows effective reduction of Anopheles stephensi populations by the biolarvicide Natular

Biolarvicides present an environmentally safer alternative to chemical larvicides, demonstrating selectivity against target species. However, the available options for biolarvicide use in mosquito control remain limited. This study aimed to assess the effectiveness of three formulations of the biolarvicide Natular, namely the 20.6% Emulsifiable Concentrate (EC), 2.5% G30 granular, and 7.48% tablet (DT) for direct application, against Anopheles stephensi in large-scale field trials conducted in Chennai, India. When the 7.48% DT formulation was applied at a rate of 1 tablet per 200 L in overhead tanks with An. stephensi breeding sites, on an average, led to a > 90–100% reduction in An. stephensi larval and pupal density for a duration of 8–9 weeks in rainy, winter and summer seasons. Similarly, the 2.5% G30 formulation, at a concentration of 0.074 mg (AI)/L in overhead tanks, impressively suppressed An. stephensi larval and pupal density by 91–100% for 4–5 weeks in three seasons. The application of the 20.6% EC formulation at 0.073 mg (AI)/L in overhead tanks achieved a remarkable 100% reduction in larval and pupal density for 2 weeks in all seasons. The Natular 7.48% DT formulation is well-suited for An. stephensi larval control at 1 tablet per 200 L with an 8-week application interval. The 2.5% G30 formulation is recommended at 0.074 mg (AI)/L at 4-week intervals. Meanwhile, the 20.6% EC formulation can effectively control An. stephensi at 0.073 mg (AI)/L with fortnightly application intervals. These Natular formulations serve as potent biolarvicides, offering a valuable tool for An. stephensi control within the Integrated Vector Management (IVM) program.

Gender and mental health of adolescents: A conceptual framework developed in a Delphi study

Objective Adolescence is one of the most neglected developmental phases, yet there are few phases in the life course when gender socialisation is as intense. So far, no framework or theory exists that theoretically conceptualises gender norms, gender attitudes and mental health outcomes. To address this gap, we aim to develop a conceptual framework that illustrates the interplay of key constructs in the gendered pathways of health, using the Delphi technique. Methods We carried out a Delphi study enrolling an international panel of 21 experts from different disciplines, with 43% of the respondents completing all three survey rounds. We asked the experts to identify core constructs (first round), suggest operationalisations of the constructs (second round), and to hypothesise about their relationships (third round). Items were included, excluded or adapted according to experts’ feedback. A 70% threshold was used for consensus. Results The panelists consented on the following core constructs: gender norms (social environment), sex assigned at birth, gender attitudes, gender roles, competencies, and gender identity to the mental health of adolescents. They further consented on gender approaches, namely the intersectional, multidimensional and multilevel approach as well as the power relations lens. The operationalisation of the constructs in round two led to the inclusion of variables forming an intersectional lens and four social environment levels: the household, community, political and digital level. In round three, the experts formed 14 hypotheses about the relationships between the core constructs. Conclusion The derived conceptual framework links the interplay of six key constructs, four gender approaches and four social environment levels to adolescent mental health. Future research should validate and apply the hypothesised relationships between the constructs to disentangle the gendered pathways to (mental) health in adolescence.