Search Results (307)

Recently, there has been great interest in AI-based approaches for de novo design of novel drug candidates. However, the generation of useful lead drug candidate compounds requires more than predicting engagement with the desired protein target. Candidate molecules must also be anchored in the real world of medicinal chemistry for their synthesis and modification as well as satisfying multiple drug development-related criteria. Here, we present Nevermore, an AI target-conditioned, database-grounded workflow for prioritizing candidate ligands from large compound libraries. Nevermore uses a geometry-aware protein–ligand affinity oracle to score target-specific binding and perform sparse integer edits in count-based Morgan fingerprint space. Nevermore then retrieves the most structurally similar molecules from public chemical databases. This design enables multi-objective search over predicted affinity and absorption, distribution, metabolism, excretion, and toxicity (ADMET) proxies while keeping all candidates anchored to valid database compounds. We evaluated Nevermore’s performance across three biologically distinct targets: Menin, a protein-interaction target relevant to leukemia; SARS-CoV-2 M^pro, a viral cysteine protease relevant to antiviral discovery; and epidermal growth factor receptor (EGFR), a kinase-superfamily oncology target with extensive experimentally tested compounds. Nevermore retrieved candidate sets with favorable predicted affinity–property trade-offs. These results support database-grounded fingerprint steering as a practical computational strategy for lead prioritization and for generating testable molecular hypotheses, although the prioritized candidates remain predictions, requiring follow-up experimental validation. Full article

Cyanobacteria are increasingly recognised as photosynthetic chassis for sustainable metabolic engineering because oxygenic photosynthesis generates ATP and NADPH via the photosynthetic electron transport chain, which drive CO₂ fixation through the Calvin–Benson–Bassham cycle into carbon intermediates that can be redirected toward engineered heterologous pathways. Their genetic tractability, CO₂-fixing capacity, ecological adaptability, and relatively simple cellular organisation make them attractive platforms for developing low-carbon biotechnological processes. This review explores recent progress in engineering cyanobacteria for heterologous pathway construction, critically evaluating genetic tools including transformation methods, genome integration strategies, promoter systems, and CRISPR-based editing, with specific emphasis on challenges of direct relevance to phototrophic chassis: host–pathway metabolic compatibility, precursor supply, cofactor balancing between photosynthetic output and heterologous pathway demand, and achieving genetic stability in polyploid cyanobacterial genomes. The review also addresses key limitations with mechanistic context: metabolic burden from multi-gene pathway expression reduces growth rate and selects against producing cells; polyploidy delays complete chromosomal segregation of engineered constructs; slow photoautotrophic growth constrains volumetric productivity; native regulatory networks resist carbon flux redirection; and cultivation constraints—including light attenuation in dense cultures and mismatches between photosynthetic ATP/NADPH supply and heterologous pathway demand—further limit achievable yields. Full article

Generative large language models (LLMs) are increasingly used to support writing feedback. However, the pedagogical safety and usefulness of LLM feedback for primary students remains under-evaluated. This study reports an educator-centered evaluation of GPT-4 Turbo for Year 5 narrative and persuasive writing in the context of an established online tutoring program. Using authentic students’ drafts paired with tutor feedback, we generated parallel LLM feedback via rubric-aligned prompting and compared the two feedback sources in a blinded, within-script design. Four experienced English specialists co-designed a six-dimensional rubric (clarity, specificity, helpfulness, feasibility, relevance, and overall effectiveness) and rated tutor versus LLM feedback for each script; their written reflections were analyzed thematically to surface boundary conditions and risk perceptions. Across dimensions, tutor feedback received slightly higher mean ratings, with the clearest descriptive advantage in perceived helpfulness; however, none of the differences remained statistically significant after Holm-Bonferroni correction. LLM feedback was often rated similarly for clarity and feasibility but was frequently characterized as generic, surface-focused, and occasionally misaligned with the student draft, which increased verification effort and posed a risk of misleading learners if used without mediation. Synthesizing ratings and educator reflections, we identify conditions under which LLM feedback is most appropriate as rapid first-pass support for routine structure and surface revision, and least appropriate for developmental judgment and context-sensitive guidance. We translate these findings into design requirements for teacher-in-the-loop primary writing feedback systems, including alignment to explicit pedagogical constructs, editable workflows, and safeguards that reduce unsupported feedback before release to students. Full article

Affective Image Stylization (AIS) converts an emotional intent into executable artistic visual styles. Existing methods are often limited to discrete label settings and provide limited interpretability of how target emotions are realized. We propose LLaVA-Emo, an interpretable AIS framework built on multimodal Chain-of-Thought (CoT) reasoning. Our method decouples generation into two structured outputs: <reasoning> provides visual–affective causal explanations grounded in the input image evidence, and <style_prompt> expresses actionable, renderer-ready style instructions that directly condition a frozen diffusion renderer. We constructed a training set by screening ArtEmis’ sentiment interpretations and fine-tune LLaVA-1.5-7B with LoRA, where SFT mainly supervises the structured intermediate <reasoning> (and output format), while the true executability of <style_prompt> is enforced by our DPO stage via render-and-reward feedback. The rendering stage remains training-free, and we further apply DPO for preference optimization to align candidate outputs with both emotion fidelity and instruction executability. Experiments on the EmoEdit inference set demonstrate that LLaVA-Emo improves emotion alignment while providing stronger process interpretability. Full article

Digital transformation in education enables the integration of interactive tools that foster engagement, creativity, and sustainability awareness among young learners. GameOn! is a serious game using Minecraft Education Edition (MEE) to promote sustainability, inclusivity, and peace among primary school students aged 6–11. Grounded in Constructionism, Experiential Learning Theory, and Citizen Science (CS), it is designed to support connections between classroom experiences and real-world environmental actions. The project followed a co-design methodology involving international partners and educators to develop the GameOn! MEE world and a complementary teacher toolkit. The game was later tested in three Italian primary schools, involving 100 students through both guided and free play sessions. Findings show that 95% of students enjoyed the game, 89% learned new concepts, and teachers observed great focus and engagement during structured play. These results align with our observations: most children quickly engaged with the game, adapted to its mechanics, and demonstrated understanding of key tasks. Some usability challenges emerged, emphasizing the importance of facilitation. Overall, the findings suggest that GameOn!, consistent with the pedagogical potential of other serious games, could enhance sustainability literacy, active citizenship, and environmental awareness in early education. Future work will expand its implementation and further strengthen the integration of CS-based activities. Future work will expand implementation and strengthen the integration of CS-based activities. Full article

Mutations in gap junction protein β-2 (GJB2), encoding Connexin 26 (Cx26), are the most common genetic cause of hearing loss, responsible for up to 50% of inherited non-syndromic cases worldwide. This review covers Cx26 from three perspectives: protein structure, mutant disease mechanisms, and treatment approaches. Structurally, 12 Cx26 subunits assemble into a gap junction channel connecting neighboring cells, enabling exchange of ions and signaling molecules; activity is regulated by calcium, pH, and CO₂. In the cochlea, Cx26 channels are required for the development of sound-sensing hair cells, maintenance of the electrical gradient needed for hearing, and energy supply during sound processing. GJB2 mutations cause hearing loss through three mechanisms, complete loss of functional protein, failure of channel assembly or membrane delivery, and abnormal channel gating, that damage cochlear cells. Severity ranges from profound congenital deafness to gradual decline, depending on which mutations are inherited. Gene therapy, genome editing, and pharmacological approaches are under investigation; cochlear implantation remains the current standard of care. Full article

As a high-value horticultural crop, melon cultivation requires substantial labor input for plant architecture management. Dwarf architecture is a desirable trait for melon breeding, as it simplifies plant management and enables higher planting density. In this study, we identified a spontaneous dwarf mutant, HMN-d, derived from a cantalupensis melon HMN. Compared to HMN, HMN-d exhibited a 70% reduction in plant height with unchanged node number. BSA-seq mapped the mutation to a single 1.86 Mb interval on chromosome 7 containing CmER, a known regulator of melon height. A novel loss-of-function allele, Cmer-2, introducing a premature stop codon, was identified in this region of HMN-d. Linkage analysis using 1455 F₂ individuals via KASP marker developed from the Cmer-2 variant revealed complete co-segregation with the dwarf phenotype. Allelism analysis further demonstrated that Cmer-2 is allelic to Cmer-1, a previously identified loss-of-function allele of CmER. CmER knockout lines generated by gene editing recapitulated the dwarf phenotype, directly confirming that loss of CmER function is sufficient to cause dwarfism. Collectively, these findings establish Cmer-2 as the causal variant underlying the dwarf phenotype and provide valuable genetic resources for melon plant architecture improvement and for dissecting the mechanisms of height regulation. Full article

Background: In the context of a multi-stakeholder program promoted by Regione Lombardia in collaboration with Fondazione The Bridge and the University of Pavia, the present study investigates the organization and availability of hospital-based vaccination services for high-risk patients. Framing hospitals as strategic hubs for vaccination delivery, the study aimed to map service availability, operational settings and dedicated pathways across the region. Methods: A structured questionnaire was administered in 2025 to 40 healthcare organizations, encompassing 114 hospital facilities, including Local Health and Social Care Authorities (ASSTs) and both public and private Scientific Institutes for Research, Hospitalization and Healthcare (IRCCSs). Descriptive and inferential statistical analyses were performed, and findings were compared with those from the 2023 and 2024 editions of the same survey, developed within the “Vaccination—an opportunity for high-risk patients” project, using Pearson’s chi-square test. Results: In 2025, 99 facilities (86.8% of respondents) reported providing vaccination services for at-risk individuals. Dedicated vaccination pathways were generally available in more than 50% of facilities for nearly two-thirds of the risk categories considered. Vaccination services for diabetic patients were available in 70.7% of facilities. Among healthcare workers, influenza (93%) and SARS-CoV-2 (89.5%) vaccines were the most frequently offered, with rates approximately ten percentage points higher than those of other vaccines. Conclusions: Overall, these findings indicate a regional model progressively consolidating hospital-based vaccination for high-risk groups, with a consistent upward trend in service availability from 2023 to 2025. Full article

This Editorial introduces the Special Issue “Spatial Planning and Land-Use Management: 2nd Edition” and discusses the eight articles published in it. Taken together, these contributions demonstrate that contemporary spatial planning and land-use management can no longer be understood as narrowly regulatory or sector-specific activities. Rather, they must be approached as integrative and adaptive practices capable of mediating between ecological integrity, territorial cohesion, infrastructure provision, social justice, public health, and participatory governance. The Special Issue brings together case studies from China, the United States, Australia, Iran, Portugal, Slovakia, and Belgium, as well as comparative evidence from peri-urban landscapes, and spans a wide range of spatial scales, from neighbourhoods and urban forests to metropolitan green belts, urban agglomerations, peri-urban territories, and ecoregions. Several major lines of inquiry emerge across the volume. First, the articles reaffirm the need for multiscale planning frameworks able to connect local action with regional and supra-regional structures. Second, they broaden the understanding of infrastructure by including not only transport and urban facilities, but also ecological, green, and even nocturnal infrastructures. Third, they show that many of today’s most difficult planning questions arise in spaces of transition and overlap, especially peri-urban areas, where conflicts among land uses, ecosystem services, development pressures, and governance arrangements become particularly acute across sectors and across spatial and temporal scales. Fourth, they underline that planning effectiveness increasingly depends on participation, co-design, and cooperation among diverse actors, including civic initiatives and local communities. Overall, the Special Issue highlights spatial planning as a strategic field of action through which societies can address land-use conflicts, reconcile environmental and social objectives, and design more sustainable, resilient, and liveable territories. Full article

Abrupt maritime-energy disruption can generate system-wide production losses before firms and policymakers can adjust. Existing assessments usually emphasize direct exposure or long-run equilibrium responses, which makes them less suitable for short-run risk assessment in energy-dependent production systems. We develop a threshold-cascade framework that combines dual-track dependence topology, edge-level inventories, smooth operability bands, and a separate price-validation step to identify the blockade intensity at which a localized chokepoint shock becomes systemic production loss. The framework is evaluated against the March 2021 Suez blockage and the 2022 Russia–Ukraine producer-price episode, and then applied to a 2026 Strait of Hormuz stress scenario using the Organisation for Economic Co-operation and Development (OECD) Inter-Country Input-Output (ICIO) tables, 2025 edition, with the 2022 benchmark year. Under the baseline 150-day horizon, terminal loss first reaches 50% at about 32% blockade intensity, with a broader calibrated threshold band of 32–46%. Losses spread beyond the point of origin and become concentrated in East and Southeast Asian manufacturing supply chains and in downstream consumer markets after inventories at connected hubs are depleted. Policy experiments show that single-channel interventions shift the threshold only modestly, whereas an integrated package that relaxes logistics, inventories, and upstream scarcity moves the threshold to about 46% in this calibration. The analysis targets the weeks-to-months interval before substitution, contract renegotiation, and broader market adjustments dominate. Within that interval, the model identifies when buffers fail, how production losses spread, and which intervention packages delay systemic disruption. Full article

Lightweighting Building Information Modeling (BIM) models for digital-twin applications requires balancing aggressive geometric reduction with component-level engineering tolerances and mesh usability. Most geometric simplification pipelines apply uniform ratios or hand-tuned heuristics, which struggle to accommodate the strong heterogeneity of BIM components in functional role, geometric complexity, and detail distribution. End-to-end learning-based simplification can be adaptive, but it often entangles decision-making with geometric editing, making engineering constraints difficult to enforce and audit. We present Semantic-Geometric Co-driven Adaptive Budget Estimation and Reduction for BIM (SABER-BIM), which formulates lightweighting as a component-level face-budget allocation problem. Conditioned on Industry Foundation Classes (IFC) types and structure-sensitive geometric descriptors, SABER-BIM predicts target face counts for individual components and then meets a user-specified global budget through global scaling. The predicted budgets are executed by a robust geometric backend (e.g., quadric error metrics, QEM), yielding an auditable and easily deployable pipeline. To address the absence of direct supervision, we introduce an offline pseudo-ground-truth procedure that searches for the minimum feasible target face count for each component under semantic-aware tolerance and mesh-validity constraints. Experiments on the IFCNet dataset show that SABER-BIM allocates budgets more effectively under identical global constraints, improving stability in both geometric error control and engineering usability. Full article

Recent advances in gene therapy have highlighted the potential of CRISPR-based gene-editing systems combined with adeno-associated virus (AAV) vectors. However, the limited packaging capacity of AAV remains a significant challenge for the simultaneous expression of Cas effector proteins and guide RNAs within a single vector. To address this limitation, we developed a compact AAV vector that enables the co-expression of Acidaminococcus sp. Cas12a (AsCpf1) and CRISPR RNAs (crRNAs) using a single bidirectional promoter derived from the mouse H1 promoter. Our single bidirectional H1 promoter supported indel formation comparable to that achieved by dual-promoter systems and facilitated scalable genome editing with single-, dual-, and triple-target configurations. Genome editing was successfully accomplished both in vitro and in vivo following AAV delivery. This study shows that our engineered compact AAV vector platform is capable of simultaneously delivering AsCpf1 and multiplexed crRNAs. Full article

Despite prolonged viral inhibition with combination antiretroviral therapy (ART), HIV-1 survives as genetically intact, replication-capable proviruses within durable CD4+ T-cell fractions, involving central memory, transitional memory, and stem cell-like memory populations, as well as within tissue-resident compartments including lymphoid follicles and gut-associated lymphoid tissue. Reservoir stability is preserved via clonal growth of infected cells and epigenetic processes that impose proviral transcriptional silencing. As a result, current therapeutic approaches seek to either directly alter proviral survival or to improve immune-driven elimination of infected cells. At the molecular level, investigational strategies such as CRISPR–Cas9 and CRISPR–Cas12 gene-editing systems are intended to remove or induce inactivating mutations inside embedded proviral DNA, as well as alter host entrance co-receptors such as CCR5 to provide cellular resistance to infection. In addition, pharmacologic latency regulation is being studied via histone deacetylase inhibitors, protein kinase C agonists, and bromodomain inhibitors to reverse latency, along with Tat inhibitors and other transcriptional repressors aimed to persistently silence proviral expression. Moreover, immunological techniques aim to counteract inefficient endogenous antiviral defenses. Broadly neutralizing antibodies with tailored Fc-driven effector functions are under examination for both neutralization and antibody-dependent cellular cytotoxicity. Therapeutic vaccine approaches seek to elevate polyfunctional HIV-specific CD8+ T-cell responses, while adoptive cellular approaches, involving CAR-T cells aiming HIV envelope epitopes, remain in early clinical research. Immune checkpoint blockade is also being investigated to reverse T-cell depletion inside reservoir-rich tissues. Nevertheless, the key obstacles continue to be the diverse reservoir composition, restricted tissue penetration, viral escape, and safety limitations. The molecular and translational obstacles that characterize attempts toward an HIV cure must be addressed through ongoing multidisciplinary research. Full article

Genome editing is widely used and conceptually simple, yet in practice, it is hindered by laborious workflows and high costs. These challenges stem from the difficulty of identifying and isolating cells that contain the desired user-defined modifications, a problem compounded by the wide variability in editing efficiencies across cell types. While homology-directed repair (HDR) provides a mechanism for precise genome modification following nuclease-induced double-strand breaks (DSBs), it is frequently outcompeted by the dominant mutagenic non-homologous end-joining (NHEJ) pathway in mammalian cells. Therefore, we developed a novel enrichment method, Essential HDRescue, to increase the frequency of HDR events at a target site by co-targeting an essential genomic locus. Using both intrinsic positive and negative selection at a common essential gene, we enabled enrichment of precise editing events at a second, unlinked target site. We demonstrated that co-targeting essential genes in cancer cell lines and iPSCs increased HDR rates without the need for an exogenous reporter or selective drug. Analysis of resulting clones revealed that Essential HDRescue produced up to a 6-fold increase in single-allele edits and an ~4-fold increase in homozygous edits relative to single-targeted controls. By harnessing the intrinsic cellular dependencies that arise from DSB repair at essential loci, Essential HDRescue offers a widely applicable method to improve precise genome editing outcomes in mammalian cells, leaving only a minimal, protein-silent scar at the essential gene. Full article

Global climate change is rapid and poses an alarming threat to agricultural production, significantly impacting economies. Modern agriculture has strongly emphasized improving nutrient availability in crops to address rising malnutrition and contribute to global food security. However, abiotic stresses, including warmer temperatures, drought, waterlogging stress, and elevated CO₂, have critical direct and indirect effects on nutrient availability in plants. This systematic review was conducted in accordance with the PRISMA guidelines. The literature survey followed a time period of 2–5 months, during which the conceptualization, analysis, writing, and editing of the article were conducted. In the present era, it is essential to adopt measures to improve the nutritional value [enhance Nutrient Use Efficiency (NUE)] and nutrient management of plant-based foods. Plant-associated microbiomes have co-evolved with their plant counterparts and perform essential functions in nutrient acquisition, including microbial sensing and cross-talk with the plant host, nutrient uptake and sharing, and signaling mechanisms. In natural and agricultural ecosystems, plant microbiomes offer major opportunities and can be harnessed to sustainably supply essential plant nutrients and improve NUE in crops of global importance. Crop-associated microbiomes can be precisely tailored to achieve targeted outcomes, enhancing nutrient acquisition and utilization via microbiome engineering. However, bridging knowledge gaps, understanding microbial colonization, plant–microbiome dynamics, and adopting precise editing approaches are crucial to boost sustainable outcomes and crop productivity. By elucidating plant microbiome crosstalk and microbe–microbe signaling, a better understanding of microbe-mediated nutrient acquisition in plants can be achieved, defining key implications in global food security. Full article