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Advancements in synthetic biology (SynBio) and other emerging and converging technologies, such as artificial intelligence (AI) additive manufacturing (3D printing), and nanotechnology are driving progress at an unprecedented pace. However, these promising and groundbreaking advances could also lead to novel biological risks, including the potential development of SynBio-enabled bioweapons (BW). Conducting a Delphi process, we consulted 13 experts from diverse relevant sectors. The multi-stage process included insights from literature reviews, expert interviews, two rounds of expert surveys, and two workshops. We identified consistent biological threat prioritizations and established consensus-driven policy recommendations. Based on this, we developed a novel hybrid governance framework. Our key proposal includes a multifaceted and integrative approach involving four sequential, iterative components: raising awareness; establishing robust training and monitoring systems to improve biosecurity measures; developing and implementing agile governance frameworks; and strengthening international treaties, such as the Biological Weapons Convention (BWC). We consider these integral, interconnected components to be interdependent and equally important. In an era of SynBio, AI-driven bioengineering, and democratization of biotechnology, implementing these recommendations will better safeguard against the potential misuse of these advancements in the context of the development and proliferation of BW.

The bodies of animals host millions of microbial communities collectively known as the microbiome. The microbiome plays a crucial role in various processes related to the host's health and well-being. Although our understanding of the microbiome's importance to host functioning is growing rapidly, many aspects remain poorly understood. One such aspect is the role of the microbiome in chemical communication. To address this question, we used the sand lizard (Lacerta agilis), a reptile with well-developed chemosensory abilities and commonly distributed in Central Europe. Our first goal was to characterize the bacterial microbiome associated with different body parts potentially involved in chemical signalling (e.g., femoral glands, cloaca, and skin). Additionally, we examined sex-related differences in the microbiome that could be connected to intraspecific communication. Over two years, a total of 274 samples were collected. Amplicon sequencing of the 16S rRNA V3-V4 region revealed significant variation in microbial diversity across body parts, with the skin hosting the most diverse and balanced communities. In contrast, the cloaca and femoral glands contained less diverse but more specialised assemblages. No differences in microbial diversity between sexes were observed, but the year of sampling was an important factor, suggesting a highly dynamic microbiome in sand lizards. There was minimal overlap in the number of unique operational taxonomic units (OTUs) between body parts, indicating a small core microbiome (∼1% of shared taxa). Sex differences in tissue-specific bacteria were more pronounced in the cloaca, supporting the idea that the cloacal microbiome is highly specialised. Our findings suggest that microbial communities vary significantly among body parts, with strong tissue specificity, indicating that each region provides a distinct ecological niche. This study offers promising directions for future research into how host-associated microbiomes could influence chemical communication in vertebrates.

Growing evidence suggests that plant proteomes contain numerous proteins that specifically bind abscisic acid (ABA). Many of them are complex multidomain proteins where specific ABA-binding can cause biochemical and physiological changes. Here we show that the Arabidopsis thaliana K+ transporter AtKUP5 contains both a functional cytoplasmic N-terminal adenylate cyclase (AC) enabling the synthesis of 3',5'-cAMP from ATP and a C-terminal phosphodiesterase (PDE) that hydrolyses 3',5'-cAMP to 5'-AMP. We found that ABA binds in a ligand-specific manner to the catalytic center of the PDE thereby causing a reduction of 3',5'-cAMP hydrolysis in vitro. The hydrolytic activity of the PDE is ABA concentration-dependent, biphasic and requires the presence of an intact ABA-binding site similar to the one in the canonical Pyrabactin resistance 1/PYR-like/Abscisic acid receptors, with Vmax of 1.19 pmole min-1 μg-1 in the absence of ABA, increasing to 1.58 pmole min-1 μg-1 at 2 nM ABA, and decreasing to 0.75 pmole min-1 μg-1 at 50 nM ABA. These findings are therefore consistent with a direct role of ABA in PDE activity modulations and form a functional link between 3',5'-cAMP signaling and K+ flux. Furthermore, we predict that a growing number of such receptor-like proteins that specifically and directly interact with ABA will be discovered thereby uncovering complex and ancient layers of signaling and metabolic regulation.

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Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system. It is characterized by inflammation, areas of demyelination and axonal loss called plaques, recruitment of lymphocytes and monocytes, and bursts of focal blood-brain barrier leakage. Treatment strategies for MS focus on delaying disease progression and increasing patients' quality of life. However, most therapies have inconsistent efficacies and are associated with various side effects. Recently, long non-coding RNAs have been found to play a major role in the pathogenesis and development of several diseases. Several long non-coding RNAs have been correlated with MS. We focus on the role of AFAP1-AS1 in regulating the function of M2 macrophages, one of the immune cells believed to attenuate MS. Assessing this long non-coding RNA will improve our understanding of the molecular mechanics of immune cells in MS. We observe the impact of AFAP1-AS1 silencing in M2 macrophages on essential effector and regulatory proteins like MMP9, CCL5 and CXCL10 in MS patients receiving different treatments (Fingolimod, Interferon beta-1a, Interferon beta-1b, Teriflunomide or Dimethyl fumarate). Our results reported an upstream regulatory effect of AFAP1-AS1 on MMP9, CCL5, and CXCL10 in differently treated patients. By measuring the levels of proteins upon silencing of AFAP1-AS1, it was confirmed that this lncRNA has varying effects on the expression of these proteins depending on the treatment the patient is undergoing. These data shed light on the potential role of manipulating the anti-inflammatory activity of M2 cells making it a possible therapeutic target for certain MS patients.

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Allergic rhinitis displays a relevant impact on quality of life. Medications used in the treatment of rhinitis have been assessed on their impact on rhinoconjunctivitis-related quality of life, but not on generic health-related quality of life metrics, such as utilities or EQ-5D visual analogue scale (VAS) levels. This study aimed to compare different medication classes and individual medications on utilities and EQ-5D VAS levels using data from a mobile app. We conducted an observational study using direct patient data from the MASK-air mobile application, collected between May 2015 and December 2024. We compared rhinitis medication classes and individual medications on health utilities (computed from the EQ-5D-5L questionnaire) and the EQ-5D VAS. To account for confounding, we employed inverse probability treatment weighting based on propensity scores, adjusting for demographics, baseline symptom control, and asthma status. The study analysed 69,973 observations with EQ-5D VAS data and 842 observations with utility data. At the medication class level, fixed combinations of intranasal antihistamines and corticosteroids were associated with improvements in EQ-5D VAS (mean difference = 1.900; 95% CI = 1.316-2.484) and utilities (mean difference = 0.022; 95% CI = -0.015 to 0.059) compared with oral antihistamines (OAH). Intranasal antihistamines were associated with lower EQ-5D VAS and utility scores than other intranasal treatments. For individual medications, mometasone was associated with a lower EQ-5D VAS than budesonide and fluticasone furoate, while fexofenadine and levocetirizine tended to be associated with lower VAS values than other OAH. Fixed combinations of intranasal antihistamines and corticosteroids were associated with better quality-of-life than oral antihistamines and intranasal antihistamines. These findings could support future cost-effectiveness analyses.

Plant Cullin RING Ubiquitin E3 ligases (CRLs) play a critical role in targeted protein degradation, essential for physiological development and stress adaptation. The deneddylase activity of the COP9 signalosome (CSN) tightly regulates the cellular balance of neddylated cullins, which is crucial for maintaining the full spectrum of CRL functions. Although selective inositol polyphosphates (InsPs) act as cofactors in plant responses that involve ubiquitylation of negative regulators, their connection to CSN-CRL activities has remained unclear. In this study, we reveal that the two Arabidopsis thaliana InsP-kinases, IPK1 and ITPK1, physically interact and orchestrate the metabolic regulation of the CSN holo-complex activity. Notably, ITPK1 deficiency lowers Nedd8 processing rates, elevates the cellular ratios of neddylated cullins, and disturbs the dissociation equilibrium of CSN5 and CUL1 from the holo-complex. These findings uncover a novel autoregulatory switch in CSN functions, governed by deneddylation activity. Furthermore, we demonstrate that the phosphate starvation response (PSR), induced in phosphate-limited wild-type plants and constitutively active in the InsP-kinase mutants, is partly regulated by reduced deneddylation rates, which affect the stability of SPX4, a key negative regulator of PSR. Pharmacological inhibition of cullin neddylation stabilizes SPX4 and impairs PSR, thereby linking CSN-CRL dynamics to phosphate sensing. Conversely, pharmacologically inhibiting CSN5 deneddylase activity causes wild-type plants to exhibit PSR phenotypes similar to those of the InsP-kinase mutants. Collectively, these results reveal that specific InsP-kinases are partly involved in modulating plant PSR by fine-tuning the coordination between CRL and CSN activities.

Adoptive T cell therapy holds great promise for the treatment of solid tumors but remains constrained by tumor heterogeneity, inefficient neoantigen targeting, and the complexity of T cell manufacturing. Here, we present a patient-specific, broadly applicable platform using physically inactivated tumor organoids (PIOs) to generate tumor-specific cytotoxic T cells ex vivo. Derived from droplet-engineered tumor organoids (DEOs), PIOs preserve the full antigenic repertoire of the patient's tumor without requiring synthetic peptides, antigen-presenting cells, or neoantigen prediction. Using matched tumor tissue and PBMCs from colorectal and liver cancer patients, we show that PIOs activate and expand tumor-specific T cells with enhanced infiltration, selective cytotoxicity, and robust secretion of IFN-γ and IL-2. Multi-round PIO stimulation achieves 80-400-fold expansion of CD8+CD137+ T cells within two weeks. Transcriptomic and epigenetic profiling suggest that PIOs modulate T cell programs linked to migration and persistence. This work redefines tumor organoids as immunotherapeutic materials and establishes a rapid, cost-effective platform for personalized T cell manufacturing. Our findings provide a new translational route for adoptive cell therapy in solid tumors using patient-derived materials.

Pulmonary fibrosis (PF) is an irreversible chronic lung disease in which dysregulation of tissue repair leads to excessive deposition of extracellular matrix (ECM). Rosa roxburghii Tratt (RRT), which has anti-inflammatory and antioxidant properties, exhibits the potential to attenuate organ fibrosis. In this study, we evaluated the bioactive content and antioxidant capacity of different polar extracts of RRT (RRTEs), and explored the multi-target mechanism of the optimal extract in alleviating PF. The ethyl acetate extract of RRT (EAE) exhibited the highest bioactive content and the strongest antioxidant capacity. EAE intervention reshaped the gut microbiota composition in PF mice by enriching beneficial bacteria and reducing pathogenic taxa. Metabolomic analysis identified 11 potential serum biomarkers associated with PF, which were involved in 7 metabolic pathways. Notably, EAE attenuated the disruption of L-tryptophan metabolism, primarily through modulation of serotonin. Moreover, EAE was found to alleviate epithelial-mesenchymal transition (EMT) and inhibit inflammatory cytokine via the JAK2/STAT3 pathway. In conclusion, EAE may exert anti-PF effects associated with the structure of the gut microbiota, correction of amino acid metabolic disorders (notably tryptophan metabolism) and modulating JAK2/STAT3 signaling pathway in the lung. These findings provide new insights into the therapeutic potential of EAE against PF.

Inflammatory activation is a major cause to nasal diseases, such as chronic rhinosinusitis and allergic rhinitis. However, in vitro research model to mimic the process of olfactory inflammation and to screen new therapeutic target is still lacking. We established three inflammatory models based on olfactory epithelium (OE) organoids, using lipopolysaccharide (LPS), TNFα treatment and doxycycline induction. The efficacy of these models was evaluated by immunostaining, RNA sequencing, qPCR, and functional assays. These inflammatory organoid models mimicked impairment in cell proliferation and neuronal genesis, and showed upregulation of inflammation-related signaling pathway and downregulation of cell cycle-related pathway. We identified that DNA damage inducible transcript 3 (Ddit3) was upregulated in all inflammatory organoid models. Ddit3 downregulation counteracted apoptosis, alleviated cell proliferation and neuronal differentiation, and recovered the functional response to odor stimulation in all three inflammatory organoid models. Ddit3 deficiency counteracted effect of LPS instillation by promoting cell proliferation, recovering neurogenesis, attenuating inflammation, and improving electrophysiological response to odor mixes in the OE. Single-cell RNA sequencing analysis showed that Ddit3 upregulation in mature olfactory sensory neurons of inducible inflammation model and patients with aging-related olfactory dysfunction correlated with endoplasmic reticulum stress and neuron apoptotic process. We established olfactory inflammation organoid models, and made use of these models to identify Ddit3 as a potential therapeutic target against inflammation-related olfactory neuronal loss and functional deficit.

Fc-containing GLP-1 therapeutics exhibit complex post-translational modification (PTM) heterogeneity, necessitating advanced analytical methods for quality control (QC) and process analytical technology (PAT). We developed a reverse-phase liquid chromatography (RP-LC) method for the PTM-specific profiling of these biologics. Using dulaglutide (IgG4-Fc) as a model, critical parameters─including mobile-phase additives, acid concentration, and shallow gradients─were optimized to resolve PTM variants (e.g., hydroxylation, N-terminal truncation, disulfide reduction, glycosylation) within 40 min. Mass spectrometry (MS) compatibility was enabled by adopting difluoroacetic acid (DFA) as an alternative ion-pairing reagent to support intact-mass characterization of variants. This enabled the identification of additional PTMs not readily resolved under the initial RP-LC conditions, including site-specific HyK-Gal-Glc O-glycosylation and process-dependent truncations. The method also allowed for the direct quantification of critical impurities and the detection of process-induced variants across biosimilar clones. The method was further demonstrated on the IgG2-subtype GLP-1-Fc-fusion (supaglutide), showing applicability across the two Fc subtypes examined without additional optimization. This robust, MS-compatible RP-LC platform provides a rapid, accurate, and comprehensive (RAC) means of conducting PTM-specific QC for Fc-GLP-1 therapeutics, supporting PAT implementation and accelerating biosimilar and next-generation drug development.

ARL15, coding for a small GTPase, was identified as a non-HLA susceptibility gene in rheumatoid arthritis (RA) through a GWAS in a North Indian cohort, with serum adiponectin and ARL15 levels higher in RA patients with the associated genotype. This study aimed to delineate the functional role of ARL15 in RA pathobiology. Differential transcriptomics in both ex vivo RA synovial fibroblasts and in vitro MH7A cells using a gene knockdown (KD) approach and standard analyses pipeline were performed to obtain insights into ARL15's role. In RASF, ARL15 KD led to downregulation of COMP-an extracellular matrix stabilizer linked to severe RA-alongside upregulation of adiponectin and IFN response genes like IFI6 and USP18. Furthermore, upregulation of NPTX1 and MX1, previously associated with disease modulation and treatment response, was observed. Downregulation of CTGF, CD248, and PTX3 suggested involvement of ARL15 in inflammation and RA-associated cardiovascular risk. Conversely, ARL15 KD in MH7A cells displayed distinct signatures with upregulated cytokines (IL1A, IL8, CXCLs) and downregulated inflammatory regulators (DOCK2, TLR4, TGFB2), reflecting an inflammatory bias distinct from patient-derived RASF. The dual-system approach, despite its divergent differential expression, underscores the multifaceted role of ARL15 in regulating connective tissue architecture, inflammation, and immune response. Limitations of immortalized cell models in capturing patient heterogeneity and disease complexity are apparent, but the key findings position ARL15 as a promising therapeutic target, warranting further investigation in RA animal models and genomic medicine. Taken together, this work provides a compelling rationale to pursue ARL15 targeted interventions in RA management.

Pseudomonas aeruginosa is the major cause of hospital-acquired infections and morbidity and mortality in individuals with burn wounds, due to the emergence of antibiotic resistance. As a result, some scientists are concentrating on research for alternative treatment, with phage therapy being one of the suggestions. However, a thorough description of the phages under consideration for use is necessary to optimize the treatment process. Thus, we show in this paper that the newly isolated phage vB_Pa_AN-12, member of the Pakpunavirus genus, is a perfect fit for phage therapy. It can infect several clinical strains of P. aeruginosa, including those resistant to multiple antibiotics. It is also able to decrease the viability of host cells strain by 5 logs in 1 h. Furthermore, it does not carry any harmful genes, and has efficient intracellular development with about 100 progeny virions per infected cell. Additionally, it did not affect the viability of cell lines that represented keratinocytes (HaCaT), fibroblasts (BJ), and monocytes (SC). These results suggest that usage of this phage, especially for skin infections, won't cause any side effects resulting from phage-human cell interactions. Nevertheless, given there is a possibility of phage resistance development, the action of isolated phage should be further investigated in combinations with other antimicrobials.

BIRC5 (survivin), an inhibitor of apoptosis protein, is overexpressed in most tumors and is associated with drug resistance, proliferation, and metastasis, while being largely undetectable in normal differentiated tissues. This unique expression pattern makes BIRC5 an exceptionally selective therapeutic target, offering the potential to maximize anticancer efficacy while minimizing systemic toxicity to healthy tissues. However, few BIRC5-targeted agents have advanced to late-stage clinical trials. We developed two nanodrug formulations using poly-L-lysine-modified NH2-Fe3O4 magnetite nanoparticles (PL-MNPs) for selective targeting of BIRC5-positive cancer cells. We further evaluated their anti-cancer efficacy in vitro and in vivo (zebrafish xenograft model), using cancer cell models that expressed BIRC5 and exhibited ABCB1-mediated drug resistance and IDO1-induced immune therapy insensitivity. The PL-MNPs delivered plasmids driven by the BIRC5 promoter (pBIRC5) encoding either antisense BIRC5 mRNA (As-BIRC5) or a dominant-negative BIRC5 protein (dN-BIRC5), for tumor-specific BIRC5 inhibition. These nanodrugs demonstrated robust in vitro and in vivo anti-cancer activity in multiple BIRC5-positive cell lines (MIA PaCa-2, NTUB1, NTU0.017, SK-OV-3, KB, and KB-TAX50). The activity was preserved across cancer types and independent of ABCB1-mediated drug resistance, while maintaining cancer cell specificity, and was not affected by IDO1 expression, a factor associated with poor responses to immune therapy. PL-MNP uptake was partially mediated by clathrin-dependent endocytosis, with acidic intracellular environments facilitating efficient plasmid release. Conjugation of nanoparticles with Herceptin® (trastuzumab) significantly increased cellular uptake and anticancer activity, especially in clathrin-deficient SK-BR-3 cells that overexpress ERBB2. These findings establish that the easily synthesized PL-MNP-pBIRC5/As-BIRC5 and PL-MNP-pBIRC5/dN-BIRC5 nanodrugs have strong potential to overcome BIRC5- and ABCB1-related drug resistance, representing a broadly applicable strategy against various malignancies. While the size of our nanodrug (~400 nm in hydrodynamic diameter) is compatible with reported effective nanoparticle sizes in some models, the extent to which the enhanced permeability and retention (EPR) effect contributes to tumor accumulation in human cancers remains uncertain and will require validation in more clinically relevant models and imaging modalities.

The distinctive aroma of fried flour-based products results from synergistic high-temperature reactions, primarily the Maillard reaction and lipid oxidation. However, these complex chemical pathways are prone to quality deterioration during processing and storage, including acrylamide formation, lipid rancidity, and flavor loss. This review systematically examines flavor formation mechanisms and establishes comprehensive control strategies across the entire industrial chain. It highlights modern flavor characterization techniques-including GC-MS, GC-O, GC-IMS, electronic nose, flavoromics, and artificial intelligence-based flavor analysis-that enable precise monitoring of volatile profiles and the degree of oxidation. The analysis covers the critical influence of ingredient ratios, oil selection, innovative pretreatments, novel frying methods, and post-frying processes on flavor precursors, reaction pathways, and product stability. Synergistic regulation via ingredient optimization and advanced technologies effectively enhances desirable aromas while suppressing harmful compounds and controlling oil absorption. This integrated approach provides a robust framework for advancing the sensory quality, food safety, and flavor stability of fried flour-based products in modern industrial manufacturing.

Cholera has re-emerged as a major global public health threat. Orally administered attenuated or inactivated vaccines offer protection against enteric pathogens such as Vibrio cholerae, and several World Health Organization-prequalified oral cholera vaccines are used globally as part of international cholera prevention and control measures. However, vaccine effectiveness, particularly in young children in low- and middle-income countries, is often lower than in adults, leaving this vulnerable age group at greater risk of cholera. The heat-labile toxin (LT) of enterotoxigenic Escherichia coli (ETEC) has strong mucosal adjuvant properties, and in detoxified form (double-mutant LT), it has been shown to safely improve immune responses to protein and somatic lipopolysaccharide antigens in children in Bangladesh who received the oral inactivated whole-cell enterotoxigenic ETEC vaccine ETVAX. Subsequent field trials of ETVAX in children in Zambia and The Gambia have also further demonstrated safety and immunogenicity in this difficult-to-immunize age group. Current concerns about reduced effectiveness of oral cholera vaccines in children led us to re-examine data from an earlier unpublished study conducted to discern for the first time, the adjuvant effects of LT on an orally administered vaccine in humans. The study showed that native LT improved immune responses to a cholera vaccine in adult volunteers. More recent work with double-mutant LT administered with the ETEC whole-cell vaccine ETVAX supports the findings reported here with the oral cholera vaccine Dukoral. These data suggest that modern attenuated versions of native LT, such as double-mutant LT, should be evaluated for improving immune responses to cholera vaccines.

Pattern recognition receptor (PRR) ligands represent a promising class of immunostimulants. Here, we demonstrate that the covalent conjugation of PRR ligands enables coordinated receptor engagement and amplifies immune responses beyond those achievable with unlinked mixtures. We synthesized a focused panel of chimeric PRR ligands comprising defined pairwise agonist combinations targeting selected extracellular and intracellular PRRs. Using phenotypic screening in human peripheral blood mononuclear cells, we identified conjugates that induced distinct cytokine signatures and enhanced cytotoxic immune activity. Among these, chimeric TLR4/TLR7 and TLR7/RIG-I ligands elicited broad innate immune activation in vitro and enhanced antigen-specific immune responses in murine vaccination models. Notably, intratumoral administration of the conjugated TLR4/TLR7 ligand resulted in significant antitumor activity in a syngeneic B16F10 melanoma model. Collectively, these findings establish covalent PRR ligand conjugation as a powerful chemical strategy for modulating innate immune signaling and support the development of conjugated PRR ligands as next-generation vaccine adjuvants and immunotherapeutics.

The increasing demand for sustainable protein sources has intensified interest in edible insects as potential multifunctional food ingredients. In this study, we investigated whether solvent-dependent extraction influences the biological activities of edible insect larval extracts. Aqueous and ethanolic extracts prepared from the larvae of Protaetia brevitarsis seulensis (PBS), Tenebrio molitor (TM), and Allomyrina dichotoma (AD) were evaluated using multiple in vitro models. The aqueous extracts of AD and PBS exhibited strong antioxidant activity in HepG2 hepatocytes and human dermal fibroblast (HDF) cells. Most extracts suppressed nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages, indicating anti-inflammatory effects. In addition, all larval extracts inhibited β-hexosaminidase release in IgE/BSA-stimulated mast cells, suggesting anti-allergic potential. The protein-rich aqueous and ethanolic extracts showed relatively stronger angiotensin-converting enzyme (ACE) inhibitory activity, whereas polyphenol-rich ethanolic extracts more effectively reduced lipid accumulation in differentiated 3T3-L1 adipocytes. These findings highlight solvent-dependent functional activity profiles of edible insect larval extracts and support their potential as sustainable sources of bioactive ingredients.