Bone fracture healing is a multifaceted process that involves different stages and intercellular interactions. In this study, we aimed to investigate the effect of Taohong Siwu decoction (TSD) on bone fracture healing and the underlying mechanisms.

The addition of CDK4/6 inhibitors to endocrine therapy has significantly improved outcomes in HR+/HER2- breast cancer (BC). However, variable patient responses and acquired resistance remain a clinical challenge. We therefore defined the comprehensive molecular response to palbociclib, the most clinically used CDK4/6 inhibitor. Global analyses of gene expression, protein abundance, splicing, and chromatin accessibility revealed broad patterns and specific changes that result from CDK4/6-inhibition in BC cells. We uncovered unexpected feedback between CDK4/6 and estrogen-response signaling, which has clear clinical implications. We also revealed a widespread alternative splicing program that partially overlapped with genes whose expression is regulated, and which is expected to impact protein function. These molecular changes nominated combination therapies that interfere with the activation of CDKs or ERα. Accordingly, co-targeting CDK7, which regulates CDK2, CDK4/6, and ERα, additively impacted cell fitness. Collectively, these data reveal a complex, multitiered response to CDK4/6 inhibition, with implications for therapeutic efficacy.

Inhibiting telomerase could lead to telomere shortening, chromosomal instability, and eventually cell death in cancer cells. The use of steroids in cancer treatment is significant; besides, drug repurposing can be a quicker and less expensive method of drug discovery than de novo drug development. Accordingly, diverse 280 steroidal drugs-following a pharmacophore-based rationale-were docked against telomerase receptor and compared to the co-crystallized inhibitor (BIBR1532) as a reference standard. Additionally, a molecular dynamics simulation for 200 ns was performed to confirm the docking results. Moreover, the six steroidal drugs (Betamethasone disproportionate, Beclomethasone disproportionate, Clobetasone butyrate, Desonide, diflucortolone valerate, and Hydrocortisone butyrate) were selected for further in vitro investigation. The antitumor activities of the six steroidal drugs against H1299, HuH7, HCT116, A549, MDA-MB-231, MCF7, PC3, MG63, and A375 cancer cell lines, besides OEC and HSF normal cell lines, were evaluated and compared to doxorubicin (Dox) as a reference positive standard. Especially, Clobetasone butyrate represented the better cytotoxicity against seven cancer cell lines (H1299, HuH7, HCT116, MDA-MB-231, PC3, MG63, and A375) with IC50 values of 12.56, 13.95, 12.00, 17.51, 11.69, 20.64, and 20.21 µg/mL, respectively. The outstanding antitumor six steroidal drugs were further investigated for their telomerase downregulatory potentials. All analogs recorded outstanding downregulatory results against telomerase, especially Clobetasone butyrate, Desonide, Diflucortolone valerate, and Hydrocortisone butyrate, where the protein expression for telomerase was downregulated up to 0.66-, 0.76-, 0.56-, and 0.73-fold change for clobetasone butyrate, desonide, diflucortolone valerate, and hydrocortisone butyrate, respectively, compared to the control.

Despite mounting evidence that commensal microbes enhance host defenses, whether and how they directly suppress pathogen virulence remains elusive. Here, we investigate metabolites from the gut microbiota of infection‑resistant Tibetan chickens for their ability to reduce Salmonella virulence gene expression and elucidate the molecular mechanism by which these compounds inhibit the LuxS/AI‑2 quorum‑sensing system.

Sedum plumbizincicola is an efficient cadmium (Cd) hyperaccumulator with strong potential for phytoremediation. Although SpCTP3 has been identified as a key Cd-responsive gene, its functional contribution to Cd tolerance remains partially characterized. Here, SpCTP3-overexpressing (OE) plants displayed increased Cd accumulation, improved growth, and enhanced photochemical efficiency under Cd exposure, providing phenotypic evidence supporting a positive role of SpCTP3 in Cd response. Transcriptomic profiling further indicated that SpCTP3 overexpression modulates pathways related to redox homeostasis and stress signaling. Moreover, we identified a Cd-inducible transcription factor, SpMYB108, that binds to and activates the SpCTP3 promoter. Yeast one-hybrid and dual-luciferase assays confirmed this direct regulatory interaction. Collectively, these results provide integrated phenotypic, transcriptional, and regulatory insights into the role of SpCTP3 in Cd hyperaccumulation.

Transforming growth factor (TGF)-β1 is a pro-fibrotic cytokine that affects extracellular matrix (ECM) deposition and fibroblast activity. 17β-Estradiol (E2), the dominant ovarian steroid during the follicular phase (FLP) of the estrous cycle, can also influence ECM remodeling and fibrosis, through prostaglandin (PG) synthesis. PGs have opposing roles in fibrosis, with PGE₂ showing anti-fibrotic effects and PGF₂α promoting fibrosis. Equine endometrosis, whose main pathological feature is fibrosis, is marked by chronic inflammation and ECM accumulation, and may involve mediators like TGF-β1, PGs, and E2. This study aimed to assess how TGF-β1, E2, and their combination affect PG synthase and receptors transcription (qPCR) and PG concentrations (ELISA) in equine endometrial explants during the FLP, after 24 and 48 h. Prostaglandin-endoperoxide synthase 2 (PTGS-2) mRNA was reduced with TGF-β1 and combination treatments at 24 h. Estradiol and combined treatments downregulated microsomal prostaglandin E synthase1 (PGES) mRNA at 24 h, while prostaglandin F synthase (PGFS) mRNA reduced with TGF-β1 at 24 h and with E2 at 48 h. The PGE₂ concentration was lower in TGF-β1 +E2 group than in controls and TGF-β1 alone at 48 h. In contrast, PGF₂α concentration increased with E2 at 24 h and TGF-β1 and TGF-β1 +E2 treatments at 48 h. Prostaglandin E receptor (EP)2 and 4 mRNA upregulated with the combination treatment, while prostaglandin F receptor (FP) mRNA decreased in all treated groups. These findings suggest that TGF-β1 and E2 interact to regulate PG pathways, with potential to drive fibrotic changes in the equine endometrium, by shifting the balance between anti- and pro-fibrotic mediators like PGE₂ and PGF₂α.

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants increasingly linked to human disease. Perfluorohexanesulfonic acid (PFHxS), a widespread PFAS detected in human serum, has an uncharacterized role in gastric cancer (GC), one of the leading causes of cancer mortality worldwide. Here, we employed an integrated multi-omics approach combining network toxicology, large-scale transcriptomic analyses from TCGA and GEO cohorts, single-cell RNA sequencing, and molecular simulations, followed by in vitro and in vivo validation at environmentally relevant concentrations. We identified 667 common targets of PFHxS and GC and developed an 11-gene gradient boosting machine (GBM) prognostic signature that robustly stratified patient survival across independent cohorts and correlated with distinct tumor immune microenvironments. Mechanistically, PFHxS was predicted and experimentally confirmed to directly bind Kelch-like ECH-associated protein 1 (KEAP1), a key regulator of oxidative stress. Chronic low-dose PFHxS exposure downregulated KEAP1 protein expression, disrupted KEAP1/NRF2 antioxidant signaling, and promoted GC cell proliferation, migration, invasion, and tumor growth in vivo. Together, these findings provide the first molecular evidence that PFHxS acts as an environmental driver of GC progression by targeting KEAP1, while also delivering a clinically relevant prognostic tool. This work highlights a previously unrecognized environmental risk factor for gastric cancer and offers new perspectives for risk assessment, prevention, and therapeutic intervention.

Emerging environmental health issues posed by micro- and nanoplastics (M/NPs) have raised significant concerns. Accumulating evidence suggested that M/NPs can bioaccumulate in gonads and impair fertility in animals, yet the underlying cellular mechanisms and tissue-specific responses remain poorly understood. In this study, we employed in vivo and in vitro models to systematically investigate the impact of polystyrene micro- and nanoplastics (PS-M/NPs, 100 nm and 5 µm) on ovarian development and function in pubertal female mice. Following 35-day exposure, we observed size-dependent reproductive toxicity, with 100 nm PS-NPs causing reduced body weight gain and ovarian size, disrupted folliculogenesis, and altered hormone levels. Leveraging single-cell RNA-sequencing (scRNA-seq), we uncovered profound alterations in intracellular communication networks across seven ovarian cell types. Granulosa cells (GCs) were identified as the primary target of PS-NPs, exhibiting marked transcriptional changes, including dysregulation of FSCN1, a critical actin cytoskeleton regulator. In vitro experiments confirmed that only 100 nm PS-NPs were internalized by GCs, leading to cell cycle arrest, necroptosis, and hormonal dysfunction. Mechanistically, PS-NPs triggered F-actin cytoskeleton remodeling, increasing cell stiffness and histone modifications (H3K4me3, H3K27ac) associated with chromatin accessibility. Integrated ATAC-seq and RNA-seq analyses implicated STAT1 as a key transcriptional regulator driving PS-NP-induced epigenetic and transcriptional changes. Overall, our findings establish the first single-cell resolution atlas of PS-NP-mediated ovarian toxicity, revealing that NPs disrupt reproduction through cytoskeletal damage and epigenetic reprogramming. This work provides unprecedented insights into the molecular and epigenetic consequences of M/NPs in mammalian reproduction, emphasizing the potential health risks of environmental M/NP exposure.

5-Aminolevulinic acid (ALA) is a plant growth regulator that enhances salt tolerance, yet the underlying mechanisms remain unclear. In this study, we investigated the function and molecular mechanism of the Protein Phosphatase 2A (PP2A) catalytic subunit MdPP2AC in ALA-induced salt tolerance in detached leaves, calli, and rooted plantlets of 'Gala' apple (Malus × domestica). Results showed that pretreatment with ALA before NaCl stress significantly alleviated NaCl-induced damage, enhanced antioxidant enzyme activities, and improved chloride (Cl-) interception in the roots with less transport to the aboveground. Further analysis revealed that the PP2A activity and MdPP2AC expression were induced by NaCl and further by ALA. Functional studies showed that overexpressing (OE)-MdPP2AC enhanced salt tolerance, whereas RNA interference (RNAi)-MdPP2AC or application of cantharidin (CT, a specific inhibitor of PP2A activity) compromised salt tolerance, but exogenous ALA mitigated CT-aggravated salt injury. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), luciferase complementation imaging (LCI), sequence truncation and site-directed mutagenesis assays confirmed that MdPP2AC can interact with the chloride channel protein MdCLC-c2, and the C-terminal Glu684 residue of MdCLC-c2 is indispensable for the molecular interaction and Cl- transport activation. Function identification with Δgef1 mutant yeast showed that MdCLC-c2 transformation promoted intracellular Cl- accumulation as well as cell salt tolerance, and the effects were further promoted by MdPP2AC transformation and exogenous ALA. Collectively, we propose that MdPP2AC mediates ALA-induced salt tolerance in apple by interacting with MdCLC-c2, which promotes Cl- sequestration into root vacuoles with less transport to the aboveground, thus maintaining Cl- homeostasis at the cellular and plant levels. These findings reveal a novel mechanism whereby PP2AC modulates chloride channel function to maintain ion homeostasis within the ALA signaling pathway, providing critical insights into ALA-mediated apple salt tolerance.

Chronic low-grade inflammation underlies many microvascular complications of diabetes, including diabetic kidney disease (DKD). Lipoxins (LXs), an endogenously produced family of lipid mediators, resolve inflammation and protect against renal scarring as occurs in DKD. This study examined the mechanism by which LXs protect against DKD, focusing on the regulation of VCAM-1 and the recruitment of macrophages to the diabetic glomerulus. LXA4 and two fourth-generation mimetics were assessed in diabetic ApoE knockout mice, followed by in vitro studies in the main renal cell populations, including podocytes, proximal tubular, mesangial, and glomerular endothelial cells. LXs attenuated albuminuria, mesangial expansion, and collagen and fibronectin deposition as both a preventive and delayed intervention in experimental DKD. LXs also consistently attenuated the TNF-α-induced expression of VCAM-1 in all the human and mouse renal cell populations examined. Further analysis identified that the renoprotection was in part mediated by an epigenetic modification of the VCAM-1 gene through H3K4 monomethylation, which did not appear to be dependent on NF-κB activation in human glomerular endothelial cells. LXs protect against DKD by modulating glomerular endothelial cell inflammation and via a novel LX-mediated epigenetic mechanism regulating the VCAM-1 promoter in these cells.

The growing global demand for protein, combined with the urgent need for effective strategies to manage conditions such as obesity and diabetes, highlights legume proteins as valuable sources of derived bioactive peptides with health-promoting properties. In this study, we employed the Caenorhabditis elegans model to investigate the effects of supplementation with hydrolysates derived from fava bean and pea protein on healthspan. Supplementation with fava bean and pea protein hydrolysates reduced fat accumulation and age-related lipofuscin pigment in the worms, without impairing their development. The fava bean protein hydrolysate significantly decreased total reactive oxygen species levels and enhanced stress tolerance to juglone exposure, suggesting the modulatory activity of the mitochondrial oxidative stress response. In contrast, pea protein hydrolysate improved the heat stress resistance of C. elegans, and gene expression and mutant analyses revealed the involvement of the endoplasmic reticulum unfolded protein response (ER-UPR) pathway in mediating its health-promoting effects. Together, our data demonstrate that fava bean and pea protein hydrolysates support healthspan in C. elegans by modulating distinct cellular stress response pathways and pave the way for further investigation in more complex animal models.

Aluminium (Al) toxicity in acid soils severely limits forage productivity, and dissecting Al-tolerance mechanisms is crucial for securing forage supply in acid-soil regions. White clover (Trifolium repens L.), an excellent acid-soil-adapted forage with pronounced Al tolerance, serves as an ideal model for studying legume Al-tolerance mechanisms. We exposed white clover seedlings to gradient Al³⁺ concentrations (0, 2, 4, 6, 10 mmol·L⁻¹) to characterize the core physiological and molecular responses underlying its Al adaptation. Al³⁺ stress inhibited growth in a concentration-dependent manner, suppressing both root and shoot development. Roots adopted a prioritised defence strategy, with enhanced antioxidant enzyme activity and soluble sugar accumulation mitigating oxidative damage. Transcriptome analysis revealed coordinated regulation of key pathways: flavonoid biosynthesis showed a core inhibition-branch-specific activation pattern, photosystem-related genes were upregulated to reinforce photosynthetic function, and hormone signalling networks were extensively rewired with divergent responses among auxin, gibberellin, cytokinin, ABA and JA pathways. White clover copes with Al³⁺ stress via an integrated mechanism featuring root-prioritised defence, photosynthetic maintenance and hormone network remodelling. These findings provide new insights into legume Al tolerance and a framework for breeding Al-tolerant forages. Future studies will quantify Al content in shoots and roots, and perform functional validation of upregulated hormone-related genes to clarify their roles in the Al-tolerance regulatory network.

25-Hydroxyvitamin D (25(OH)D), a metabolite of vitamin D, has demonstrated anticancer properties; however, the role of alternative splicing in mediating these effects remains poorly understood. In this study, we reveal for the first time that 25(OH)D exerts antitumor effects by promoting exon 13 skipping of KDM6A (KDM6A Δexon13), which suppresses the proliferation and stemness of breast cancer cells and lacks H3K27 demethylase activity. Mechanistically, CUT&Tag and RNA-seq analyses demonstrated that KDM6A Δexon13 induces the accumulation of H3K27me3 at the promoter region of TRAP1, thereby inhibiting its transcription. Consequently, the downregulation of TRAP1 reduces Smad2/3 phosphorylation. Furthermore, KHDRBS3 was identified as the splicing factor of KDM6A Δexon13 and was regulated by 25(OH)D. Notably, 25(OH)D exhibited a synergistic effect with GSK-J4, a specific inhibitor of KDM6A, in suppressing breast cancer cell growth. Collectively, our findings uncover a novel anticancer mechanism of 25(OH)D, highlight the critical role of KDM6A Δexon13 in breast cancer progression, and provide further evidence supporting the correction of 25(OH)D deficiency in breast cancer patients.

The extracellular matrix (ECM) defines the biomechanical and biochemical microenvironment of tissues, directing cell behaviour and phenotype. In the ovary, ECM must dynamically remodel in each cycle under hormonal regulation to control follicle development and produce fertilizable oocytes. Dysregulation of this process may result in aberrant formation of ECM as seen in polycystic ovary syndrome (PCOS) whose pathology includes fibrosis of the ovary and which is a major cause of infertility. PCOS is characterised by hyperandrogenism and, here, we investigate the impact of androgens on fibrosis, cell-ECM interactions and mechanosensing. We report an altered network of gene expression related to the genesis of fibrosis. Preantral follicles from C57BL/6 mice (14-15 days postpartum) were stimulated with dihydrotestosterone (DHT, 10nM) in 24/72 hours culture. Expression of fibrosis-associated genes (Eln; Ctgf; Acta2; Plod2; Hpse) significantly increased with androgen (72 h), as did TGF-β signalling (Tgfb1; Tgfb3). We show a direct connection between androgen and mechanosensing within the ovary, with androgen upregulating the mechanosensitive Hippo pathway (Yap1; Lats1; Lats2; Stk3; Stk4; Frmd6) and downstream targets (Ctgf; Axl; Cyr61). Our results highlight hyperandrogenism as a probable driver of the fibrosis in the polycystic ovary, and emphasise the importance of ECM regulation in follicle development and fertility.

Serine metabolism is a critical vulnerability in cancer; however, its role in mediating therapeutic resistance in non-small cell lung cancer (NSCLC) remains incompletely understood. In this study, we identify key enzymes in the serine synthesis pathway (SSP), namely PHGDH, PSAT1 and PSPH, as well as the serine transporter SLC1A4, which are significantly overexpressed in lung cancer and correlate with poor patient prognosis. We show that serine contributes to carboplatin resistance in NSCLC, particularly in lung squamous cell carcinoma (LUSC). Notably, the LUSC lineage-specific oncogene ΔNp63α serves as a master transcriptional regulator of serine biosynthesis, directly transactivating the expression of PHGDH, PSAT1, PSPH, and SLC1A4. ΔNp63α-driven serine biosynthesis supports nucleotide synthesis and enhances antioxidant defense, enabling cancer cells to survive carboplatin-induced DNA damage and oxidative stress, thereby promoting therapeutic resistance. The combined inhibition of endogenous serine synthesis and restriction of exogenous serine/glycine significantly overcomes ΔNp63α-mediated carboplatin resistance. Our findings establish the ΔNp63α-SSP axis as a critical mechanism driving carboplatin resistance in LUSC. These results highlight dual-targeted disruption of serine availability as a promising therapeutic strategy to overcome chemotherapy resistance in ΔNp63α-driven LUSC. This study underscores the importance of lineage-specific metabolic dependencies as essential targets for precision oncology in NSCLC.

Interferon tau (IFNT), a pregnancy recognition factor in ruminants, maintains the corpus luteum and induces the expression of many genes in the uterus; however, the functions of these genes remain unclear. In this study, we investigated the effect of IFNT on endothelial cell-specific molecule 1 (ESM1) expression, along with the subsequent roles of ESM1 in bovine vascular endothelial cells in an in vitro cell culture environment. Quantitative polymerase chain reaction (qPCR) analysis showed that IFNT treatment significantly increased the expression levels of ESM1 in endometrial epithelial cells compared with those in non-treated control cells. qPCR and Western blot analyses showed an increasing trend in vascular endothelial growth factor (VEGF) expression following ESM1 treatment in bovine vascular endothelial cells. In addition, ESM1 treatment tended to increase cell proliferation and significantly increased the formation of tube-like structures of endothelial cells compared to those in non-treated control cells. These results suggest that IFNT induces ESM1 expression in endometrial epithelial cells. Moreover, ESM1 may contribute to angiogenesis via the expression of angiogenic factors, such as VEGF, in bovine endothelial cells.

Brassica napus, a major oilseed crop in southern China, faces dual challenges of cadmium (Cd(II)) contamination and potassium (K) deficiency in soils. However, the mechanisms by which K alleviates Cd(II) toxicity in B. napus remain poorly understood. This study, therefore, aimed to elucidate the physiological, biochemical, and molecular mechanisms underlying K-mediated Cd(II) detoxification in B. napus using a hydroponic experimental system. The results demonstrated that elevated K application significantly alleviated Cd(II)-induced toxicity. A major finding was the substantial recovery of photosynthetic activity, with K supplementation increasing the photosynthetic rate by 40.7 % under Cd(II) stress, which corresponded with a significant reduction in biomass loss and decreased concentrations of malondialdehyde and H2O2. Mechanistically, this alleviation was linked to a fortified antioxidant defense system, evidenced by enhanced activities of catalase (CAT) and peroxidase (POD) by 54.2 % and 15.7 %, respectively. Transcriptomic analysis provided deeper insights, revealing that K-mediated tolerance involves enhanced Cd(II) sequestration. K supplementation significantly up-regulated genes encoding tonoplast-localized transporters (CAX and HMA) responsible for vacuolar Cd(II) import, while simultaneously down-regulating the vacuolar Cd(II) efflux gene NRAMP4. This was complemented by an increase in ion-bound pectin and cellulose in the cell wall, further restricting Cd(II) mobility. Furthermore, K appeared to modulate key signaling pathways, as the expression of PYL genes, encoding abscisic acid (ABA) receptors, was significantly induced. Collectively, our findings demonstrate that K alleviates Cd(II) toxicity in B. napus through a multi-pronged strategy encompassing the enhancement of antioxidant capacity and the promotion of Cd(II) sequestration in both vacuoles and cell walls, potentially regulated via an ABA-dependent pathway. This research provides a theoretical basis for optimizing K fertilization to improve phytoremediation efficiency in Cd-contaminated agricultural soils.

Low temperature stress is a major abiotic constraint on agricultural productivity, especially in temperature-sensitive crops like lettuce. Nano-selenium has demonstrated considerable potential in improving plant stress resilience. In this study, lettuce plants exposed to low-temperature stress were treated with five concentrations (N1: 1 mg L-1; N2: 3 mg L-1; N3: 9 mg L-1; N4: 27 mg L-1) of nano-selenium. The optimal concentration of nano-selenium was determined to be N3 (9 mg L-1). Integrated transcriptomic and metabolomic analyses revealed that nano-selenium application significantly enhanced photosynthetic efficiency, antioxidant defenses, and metabolic adaptation under cold stress. A total of 25,593 differentially expressed genes (DEGs) and 20 key metabolites were identified. Enriched metabolic pathways included arginine and proline metabolism, amino sugar and nucleotide sugar metabolism, and glycerophospholipid metabolism. Under low-temperature conditions, nano-selenium treatment markedly improved cold tolerance by modulating proline metabolism-promoting its biosynthesis while inhibiting its catabolism-resulting in substantial proline accumulation. Furthermore, nano-selenhanced cellular structural integrity through two distinct mechanisms: (1) reinforcing cell wall architecture via enhanced amino sugar metabolism, thereby mitigating low-temperature-induced membrane damage; and (2) optimizing glycerophospholipid composition, particularly by regulating phosphatidylcholine and phosphatidylethanolamine biosynthesis through key enzyme modulation, which helped maintain membrane fluidity and stability under cold stress. These findings advance our understanding of nano-selenium-mediated stress tolerance and underscore its potential application in sustainable agriculture.

Methotrexate (MTX) is used in treating several malignancies. However, MTX neurotoxicity remains a significant clinical side effect, leading to cell division malformation, and neurogenesis impairment. Chrysin, a flavonoid compound found in natural products, demonstrates various biological characteristics, including neuroprotective and antioxidant properties. The purpose of this study was to investigate the ameliorative effect of chrysin on oxidative damage and neurogenesis impairment caused by MTX. Male Sprague-Dawley rats were randomly divided into four groups, including the vehicle, MTX (75 mg/kg), chrysin (10 mg/kg), and chrysin+MTX groups. Chrysin was orally administered for 15 days. MTX was administered intravenously on days 8 and 15. The hippocampal neural stem cells were evaluated using sex determining region Y-box 2 (sox2) and nestin immunofluorescence staining. Antioxidant enzyme expression and the levels of oxidative stress marker were assessed. Additionally, the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), brain-derived neurotrophic factor (BDNF), cAMP-response element binding (CREB), and phosphorylated CREB (pCREB) were evaluated using Western blotting. Results showed that MTX significantly decreased the activity of antioxidant enzymes and produced oxidative stress. MTX also impaired neurogenesis, evidenced by decreased sox2 and nestin-positive cells and decreased expression of Nrf2, BDNF, CREB, and pCREB in the hippocampus and prefrontal cortex. However, chrysin significantly reversed the effects of MTX on these parameters. In conclusion, chrysin exhibits neuroprotective effects against MTX-induced neurogenesis impairment by upregulating antioxidant enzyme activity, reducing oxidative stress, and improving protein expression related to neurogenesis.

Grapevine Trunk Diseases (GTDs) are caused by phytopathogenic fungi that compromise grapevine productivity and wine quality. Most GTDs preventive treatments are chemical-based and environmentally harmful. One goal of the European Green Deal is to develop sustainable agriculture which does not harm the environment and reduces pesticide use and an alternative to those treatments may be the use of elicitors such as oligosaccharides from fungi. Many studies confirm that oligosaccharides activate the defence response. The experiment was carried out in vineyards of Tempranillo and Airén cvs. Asymptomatic and symptomatic vines were treated with mannans. Leaves and grapes were taken and pigments and phenols content, polyphenol oxidase (PPO) and superoxide dismutase (SOD) activities and gene expression of several defence enzymes were determined. The mannan addition to symptomatic vines was more positive for the leaves than for the grapes, palliating the damage caused by the disease, especially in the cv. Tempranillo. On the one hand, in the leaves, mannans caused an increase in phenols and PPO activity and expression; on the other hand, in grapes, although phenols increased, the other parameters did not. Mannans increased the expression levels of chalcone synthase (CHS1, CHS3), phenylalanine ammonia lyase (PAL), SOD, and PPO in asymptomatic leaves of both cultivars. In symptomatic leaves, CHS3 and PAL expression decreased in both cultivars, while CHS1 and PPO increased only in Tempranillo. In grapes, the expression of the genes varied due to the development of the disease. The mannan treatment seemed to reduce the oxidative stress caused by GTDs, but, above all, mannans would act as a biostimulant activaing the defence system of asymptomatic vines that would help them respond more successfully to a possible pathogenic fungi infection, that although this response depended on the cultivar.