Search Results (5,664)

Proanthocyanidins (PAs) are a significant class of polyphenolic compounds found in grapes, playing important roles in human health and plant stress resistance. Previous studies have shown that the VvMYBPA1/PA2-VvWDR1-VvMYC2 (MBW) complex can regulate the biosynthesis of proanthocyanidins, and some studies have shown that the homologous genes of VvWRKY26 are involved in the biosynthesis of proanthocyanins and anthocyanins in Arabidopsis thaliana and petunias, but the molecular mechanism of VvWRKY26 in regulating the biosynthesis of proanthocyanins in grapes is still unclear. In this study, we found that the content of proanthocyanidins and the expression of related structural genes were significantly increased by salicylic acid (SA) incubation in grapes during the color transition period. Overexpression of VvWRKY26 in grapevine healing tissues revealed that overexpression of VvWRKY26 significantly promoted the accumulation of proanthocyanidins and up-regulation of related structural genes when compared with the empty vector. Further investigation into the interaction mechanisms through yeast two-hybrid and bimolecular fluorescence complementation assays revealed that VvWRKY26 can interact with VvMYBPA1/PA2, VvMYC2, and VvWDR1 to form VvMYBPA1/PA2-VvWDR1-VvMYC2-VvWRKY26 (W-MBW) complex. Through yeast one-hybrid assays and dual-luciferase reporter analysis, it was confirmed that VvWRKY26 could bind to the promoters of VvANR and VvLAR2 and activate their activity. Finally, through the co-overexpression of VvWRKY26 and MBW complex, it was discovered that the promoting activity of VvANR and VvLAR2, as well as the biosynthesis of PAs, were significantly enhanced, which was much higher than the effect of the MBW complex alone, while the opposite occurred after co-interference. In conclusion, this study explored the role of VvWRKY26 in the biosynthesis of proanthocyanidins in grapes after the interaction with the MBW complex to form W-MBW under SA incubation, providing a new regulatory mechanism for the biosynthesis of proanthocyanidins in grapes. Full article

Background/Objectives: Making sustainable choices and transforming guidelines into effective bacterial control practices for viable and safe hatching eggs is a challenge for many researchers. Gradually, scientific findings are strengthening the case for using antibacterial protocols with essential oils (EOs) for hatching eggs, which could lead to changes in traditional egg sanitization management and stimulate new research. The present study aimed to measure the contamination levels and incubation outcomes of hatching eggs sanitized with Zingiber officinale (ZOEO), Cymbopogon flexuosus (CFEO), and Rosmarinus officinalis (ROEO) essential oils. Methods: Hatching eggs from commercial broiler breeders were sanitized with solutions of ZOEO, CFEO, and ROEO prepared in grain alcohol and compared with formaldehyde and non-sanitized eggs. Bacterial contamination, eggshell integrity, incubation parameters, embryonic trachea histology, genotoxicity, and irritation potential were evaluated under commercial conditions. Results: It has been demonstrated that these EOs significantly reduce eggshell and yolk sac contamination, promote hatchability rates above 93% with good-quality chicks, and do not cause alterations in the embryonic trachea or potential genetic damage to the chicks. Conclusions: ZOEO, CFEO, and ROEO can be recommended as sanitizers for hatching eggs. Full article

Phosphorus (P) is a critical factor in enhancing agricultural yield improvement, but the over-application of P fertilizers has led to the widespread accumulation of ineffective P in soils worldwide. Artificial humic acid (AHA) has gained recognition as a new method for enhancing P effectiveness in soils. This study aims to explore the patterns and mechanisms underlying the effect of AHA on P effectiveness. A 60-day indoor incubation experiment was conducted using a soil column system, in which the soil was fractionated into five distinct particle size classes: 2 mm, 4 mm, 6 mm, 8 mm, and 10 mm. Findings revealed that AHA effectively promoted the accumulation of Olsen-P in fine-textured soils. Following the application of AHA, the fraction of particles with a size of 2 mm exhibited the highest increase in Olsen-P, at 15.4%, whereas the fraction with a size of 8 mm showed the lowest increase, at 0.2% relative to the control, at the 60th day. Additionally, AHA promoted the migration of HCl-P while enhancing the immobilization of Olsen-P. During the initial cultivation phase, the concentrations of HCl-P in the topsoil (0 cm) differed little from those in the deeper soil (40 cm). As cultivation progressed, the concentrations of NaOH-P and HCl-P in the 0 cm soil decreased more markedly than those at the 40 cm depth by the later cultivation stage. Finally, the structural equation modeling results indicated that among NaHCO₃-P, NaOH-P, and HCl-P, NaOH-P had the most significant effect on Olsen-P. These findings offer valuable insights into how AHA could be used to improve the effectiveness of P in soils. Full article

Objectives: Meniscal integrity is crucial for knee joint stability and the prevention of osteoarthritis (OA) development. Recent studies suggested that mechanical overload and interleukin (IL)-17A may be important intertwined players in meniscal degeneration, but a direct impact of IL-17A on the meniscus has not been investigated. Therefore, the aim of this study was to analyze the effect of IL-17A on meniscal tissue with and without combined mechanical injury (MI). Methods: Meniscal explant disks (1 mm height, 3 mm diameter) were isolated from bovine menisci (preserving the native tibial superficial zone) and exposed to IL-17A [0–100 ng/mL] and/or MI (single compression, 50% strain, strain rate 1 mm/sec). After three days of incubation in a serum-free medium, the proteoglycan release (sGAG; DMMB assay), mRNA level of matrix-degrading enzymes (qRT-PCR), aggrecan degradation (NITEGE immunostaining), and cell death (histomorphometry of nuclear blebbing/apoptosis and condensed nuclei/unspecified cell death) were determined. Statistics: one- and two-way ANOVA with Tukey’s multiple comparisons or Kruskal–Wallis with post hoc testing. Results: IL-17A increased sGAG release in a dose-dependent significant manner. MI also induced the release of sGAG significantly, but the combination with IL-17A showed the highest levels. Both IL-17A and MI individually affected the mRNA levels for ADAMTS4 and MMP-13 slightly, but the combination of both particularly induced a significant increase in mRNA levels. Signals for the ADAMTS4-related aggrecan neoepitope NITEGE were elevated by IL-17A in superficial areas of the excised tissue and by MI in superficial and deeper areas. The combination of both stimuli intensified this signal further. MI increased the number of cells with condensed nuclei significantly and induced apoptosis in a small proportion of cells. IL-17A had no significant impact on the amount of condensed or apoptotic nuclei. Conclusions: Our findings emphasize an interaction between inflammatory cytokine IL-17A signaling and mechanical stress since IL-17A induced matrix degeneration in meniscal tissue, which intensified in combination with a trauma. The latter might create a post-traumatic environment that promotes meniscal degeneration and subsequently osteoarthritis progression. Full article

This study aims to investigate the effects of dietary supplementation with AMPs on the growth performance, antioxidant capacity, and intestinal health of squabs. Furthermore, metagenomic and metabolomic approaches were employed to identify key differential bacterial species and metabolites associated with growth performance, and thereby the potential mechanisms underlying the enhancement of squab growth and development by AMPs being elucidated. One hundred and twenty pairs of healthy adult White Carneau pigeons (2 years old) were randomly divided into two groups, the control group (CK, fed with basal diet) and antimicrobial peptide group (AP, fed with basal diet +200 mg/kg antimicrobial peptide), with 10 replicates per group and 6 pairs of breeding pigeons per replicate. The experiment lasted for 53 days, including 7 days of prefeeding, 18 days of incubation and 28 days of feeding. In this study, squabs were weighed at 0 and 28 days of age to evaluate growth performance. At 28 days of age, duodenal contents were collected to assess digestive enzyme activities, while jejunal and liver tissues were harvested to determine antioxidant capacity. Intestinal morphology was examined using tissue samples from the duodenum, jejunum, and ileum. Finally, ileal contents were collected for a comprehensive analysis of microbial composition and metabolite profiles in the two experimental groups, employing high-throughput sequencing and LC-MS/MS techniques. The results showed that body weight, liver total antioxidant capacity (T-AOC), jejunal malondialdehyde (MDA) content, jejunum and ileum villus height-to-crypt depth ratio (VH/CD) were significantly increased, and jejunal crypt depth (CD) was significantly decreased in the AP group at 28 days of age (p < 0.05). In addition, the microbiome data showed that Lactobacillus in the AP group was a biomarker with significant differences (p < 0.05). Metabolomics analysis showed that the steroid hormone biosynthesis pathway was significantly different between the two groups (p < 0.01). In addition, the content of potentially beneficial metabolites (Biotin, beta-Tocotrienol, 7-Chloro-L-tryptophan and Dihydrozeatin) was significantly increased in the AP group (p < 0.05). These results indicate that dietary AMPs can significantly improve the body weights, liver antioxidant capacity and jejunum and ileum VH/CD of squabs. Full article

In vitro fertilization (IVF) exposes embryos to environmental stressors that can disrupt early development and confer long-term health risks, though the mechanisms remain poorly understood. Here, we tested the hypothesis that reducing incubation temperature during the first zygotic cleavage would promote long-term developmental stability in IVF-conceived offspring. Using a mouse model, we compared the long-term effects of standard (37 °C) versus reduced (35 °C) IVF culture temperature on energy balance, circadian rhythms, sleep architecture, and brain histone modifications. Although offspring from both IVF groups exhibited increased body mass without notable effects on glucose metabolism, significant disruptions in circadian rhythms and sleep–wake patterns were detected. The 37 °C group exhibited altered amplitudes in oxygen consumption rhythms and respiratory exchange ratios, as well as pronounced alterations in sleep–wake patterns, including reduced sleep duration and increased nighttime activity. The 35 °C group displayed intermediate phenotypes, substantiating the importance of optimizing embryo incubation parameters. These metabolic and behavioral changes were paralleled by altered histone modifications in the cerebral cortex of IVF offspring, suggesting an epigenetic basis for circadian misalignment. Our results identify disrupted circadian rhythm and sleep architecture as a novel mechanism contributing to metabolic dysfunction in IVF-conceived offspring. The partial mitigation of these effects through reduced culture temperature underscores the importance of optimizing IVF protocols to minimize long-term epigenetic and metabolic risks. Full article

Background/Objectives: Carvacrol and thymol are monoterpenes present in phenolic-rich essential oils extracted from aromatic plants that exhibit antimicrobial activity. This study evaluates the antiprotozoal effect of carvacrol, thymol and their precursor, p-Cymene, against Giardia lamblia and investigates their mechanism of action and cytotoxicity profile. Methods: G. lamblia susceptibility, cell viability, swelling and adhesion abilities following application of carvacrol, thymol and p-Cymene were assessed. Ultrastructural changes were evaluated using electron microscopy. Cytotoxicity was determined in mammalian cell lines (murine macrophages RAW 264.7 and bovine aortic endothelial cells) exposed to the same IC₅₀ concentrations effective against G. lamblia. Results: Carvacrol and thymol led to significant inhibition of G. lamblia trophozoite proliferation (IC₅₀ ≅ 50 µg/mL). After 7 h of incubation, total cell number decreased by 30% (p < 0.01) with carvacrol and by 50% (p < 0.001) with thymol, accompanied by reduced motility and adhesion (<20% attached cells). At IC₅₀ concentrations, G. lamblia trophozoites exposed to carvacrol and thymol underwent considerable ultrastructural alterations (e.g., aberrant-shaped cells, mitochondrial swelling and autophagosomal structures). Reduced trophozoite motility and adhesion capacity were also observed. In mammalian cells, thymol showed no significant cytotoxicity, whereas carvacrol significantly reduced viability in both cell lines. In contrast, p-Cymene showed no antigiardial activity. Conclusions: Our data suggests that carvacrol and thymol disrupt G. lamblia trophozoite integrity, possibly through alterations in membrane permeability and osmoregulatory processes. In conclusion, these compounds reveal in vitro antigiardial activity, supporting their potential as antigiardial drugs. Full article

To reduce the environmental impact of plastic packaging in the fruit supply chain, this study developed an edible natural CH-CS-EGCG coating (collagen hydrolysate-chitosan-epigallocatechin gallate) for mango preservation. The coating was prepared using an optimized CH:CS mass ratio of 1:4 with 3 wt% EGCG, exhibiting enhanced mechanical properties and low water vapor permeability. SEM and FT-IR analyses confirmed the successful incorporation of EGCG into the CH-CS matrix through hydrogen bonding, hydrophobic interactions, and electrostatic forces. The CH-CS-EGCG coating demonstrated superior antioxidant activity: its ABTS radical scavenging capacity and DPPH scavenging capacity were 234% and 422% higher, respectively, than those of the CH-CS coating. It also effectively inhibited bacterial growth, achieving a 92% inhibition rate against Staphylococcus aureus after 24 h of incubation. When applied to mango preservation, the CH-CS-EGCG coating significantly slowed down fruit decay and deterioration, extending the shelf life by 6 days. The CH-CS-EGCG coating offers a promising eco-friendly alternative for fruit preservation, broadening the applications of EGCG and advancing research in edible coatings. Full article

Background/Aim: Photodynamic therapy uses a photosensitizer and light to generate reactive oxygen species that kill tumor cells and can shift inflammatory signaling. Hypericin is a potent photosensitizer, but its immunomodulatory impact in breast cancer needs clarification. We evaluated the phototoxic and cytokine-modulating effects of hypericin-mediated photodynamic therapy in MCF-7 human breast adenocarcinoma cells. This study examines how HYP-PDT affects MCF-7 breast cancer cells by assessing viability and cytokine secretion to guide the development of targeted, immune-enhancing PDT protocols. Methods: MCF-7 cells were incubated with hypericin at 0, 0.125, 0.25, 0.5, or 1 μM, then exposed to light doses of 0, 1, 2, or 5 J/cm². Viability was measured 24 h later by MTT; selected conditions were also assessed by Trypan Blue. Cell supernatants collected after sublethal treatment were analyzed for IL-6, IL-8, IL-10, and TNF-α using a multiplex immunoassay. Experiments were repeated four times. Statistical analyses followed the study’s plan for group comparisons. Results: At 1 J/cm², MTT values did not differ from matched dark controls across hypericin concentrations. At 2 and 5 J/cm², some conditions showed increased MTT signal relative to controls, indicating higher metabolic activity; Trypan Blue performed at 0 J/cm² showed a concentration-dependent reduction in viability with hypericin. Hypericin-PDT decreased IL-6 and IL-8 concentrations and increased TNF-α in MCF-7 supernatants. No statistically significant changes were detected for IL-10. Conclusions: Hypericin-PDT altered inflammatory readouts in MCF-7 cells, with reductions in IL-6 and IL-8 and an increase in TNF-α, consistent with a pro-inflammatory shift. Viability results suggest condition-dependent changes in metabolic activity or survival effects that warrant confirmation with matched cell counts across all light doses. These findings support further standardized dosimetry and multi-line validation of hypericin-PDT in breast cancer models. Full article

Global warming is expected to increase the frequency and intensity of extreme precipitation, yet its effects on soil greenhouse gas (GHG) emissions and functional stability remain uncertain. This study explored the impact of extreme soil moisture conditions on farmland and forest soil under three scenarios: 60% field water capacity (W1), soil saturation (W2), and 10 cm of standing water (W3). We used a laboratory incubation to evaluate how three extreme soil moisture regimes—60% of field water capacity (W1), soil saturation (W2), and 10 cm of standing water (W3)—affect GHG emissions and the functional stability of farmland and forest soils. Forest soils exhibited significantly higher global warming potential (GWP) than farmland across all regimes (p < 0.05). Relative to W1, farmland GWP increased by 0.14% under W3, whereas forest GWP increased by 13.7% under W2 (p < 0.05). Extreme soil moisture conditions markedly elevated total organic C (TOC) and ammonium N (NH₄⁺–N) contents in soil solutions from both farmland and forest, with increases of 25.0% and 6.0% for TOC and 78.6% and 69.6% for NH₄⁺–N, respectively. Conversely, nitrate N (NO₃⁻–N) content in farmland soil decreased by 3.54% and 6.96% under W2 and W3 treatments, while forest soil NO₃⁻–N increased by 39.68% under W2 and decreased by 39.13% under W3. Functional stability declined under extreme precipitation and was positively correlated with total CO₂ emissions, GWP, and TOC (p < 0.001), as well as with total N₂O emissions and soil total C (p < 0.05). Overall, forest soils maintained greater functional stability than farmland under extreme moisture. These findings clarify how extreme soil-moisture events influence soil functional stability in a warming climate and highlight the potential for post-event recovery of soil functions. Full article

Chemical disinfection of removable acrylic dental prostheses minimizes the risk of denture stomatitis caused by the opportunistic fungal pathogen Candida albicans. We previously reported that neutral-pH electrolysed oxidising water (EOW), a hypochlorous acid (HOCl)-based biocide, is effective at inhibiting C. albicans biofilm formation on denture resins. Knowledge about the mechanism of action of EOW on C. albicans is lacking. This study investigated the molecular mechanism of action of neutral-pH EOW against C. albicans cells that were incubated with sub-inhibitory concentrations of EOW-HOCl (treatment with 0.125× MIC₉₀ EOW-HOCl (15 µM; T0.125) or treatment with 0.5× MIC₉₀ EOW-HOCl (59 µM; T0.5)). RNA-sequencing (RNA-seq) was used to identify differentially expressed genes (DEGs) which were validated by qRT-PCR. Ninety-five DEGs were identified between the treated and untreated cells after a 60 min exposure. A moderate sub-inhibitory EOW-HOCl concentration (T0.125) caused significant upregulation (log₂ fold change > +2) of genes responsive to oxidative stress (EBP1, GAP6, PRN1, HSP21), weak organic acid stress (PRN1), and heat-shock (HSP21). A higher sub-inhibitory concentration (T0.5) caused a significant downregulation of most DEGs (notably, −1.9 to −3 log₂ fold reduction in SUT1, HNM3, STP4 expression), cessation of growth, and an upregulation of genes involved in ammonia transport, carbohydrate metabolism, and the unfolded protein and apoptotic response pathways (ATO2, IRE1). Our findings reveal HSP21 and PRN1 to be possible key players in protecting C. albicans cells against HOCl, a natural biocide of the innate immune system. Full article

The increasing demand for sustainable substrates in agriculture and urban greening calls for alternatives to peat, whose extraction poses significant environmental risks. This study assesses the potential of olive pomace biochar (OB), wood biochar (WB), and green compost (GC), alone or in combination, to partially replace peat in growing media and improve substrate properties and plant development. Ten different substrates were formulated by substituting 10–20% of a commercial peat-based substrate with these organic amendments, using the commercial substrate alone as a control. The effects of such replacements were evaluated in the following experiments: a germination test conducted in Petri dishes using four forage species (Medicago polymorpha, Lolium perenne, Festuca arundinacea, and Lolium rigidum); and two parallel pot experiments lasting 100 days each (one with M. polymorpha and L. perenne, and another with young Olea Europaea var. Arbequina saplings). This study evaluated the impact on plant development, as well as the physical properties and composition of the substrates during the incubation process. Germination and survival of forage species were comparable or improved in most treatments, except those including 20% OB, which consistently reduced germination—likely due to high electrical conductivity (>10dS/m). In the pot experiments, substrate pH and total carbon content increased significantly with biochar addition, particularly with 20% WB, which doubled total C relative to control. Both forage species (Medicago polymorpha and Lolium perenne) and the olive saplings (Olea Europaea) exhibited normal growth, with no significant differences in biomass, water content, or physiological stress indicators when compared to the control group. Nutrient uptake was found to be stable across treatments, although magnesium levels were below sufficiency thresholds without triggering visible deficiency symptoms. Overall, combining compost and biochar—particularly WB and GC—proved to be a viable strategy to reduce peat use while maintaining substrate quality and supporting robust plant growth. This approach proved effective across the different plant varieties tested, including Medicago polymorpha, Lolium perenne, and young olive plants, which together encompass a wide spectrum of agronomic and horticultural applications as well as contrasting growth and nutrient requirements. Adverse effects on early plant development can be avoided by carefully selecting and characterizing biochars, with specific attention to salinity and C/N ratio. This finding is crucial for the successful large-scale implementation of sustainable alternatives to peat. Full article

Bioabsorbable scaffolds from synthetic polyesters are widely used in the field of tissue engineering. However, their hydrophobic surface and lack of suitable functional groups are the main limitations related to cell attachment. The aim of this research was to modify the surface of polycaprolactone (PCL) scaffolds using a bioactive coating containing heparin bound via albumin spacer and platelet lysate over heparin. Porous scaffolds were produced by electrospinning from 10% PCL (w/w) solution in methylene chloride (25 kV voltage, 100 mm distance between electrodes and 4 mL/h feedrate), which demonstrated 5.5 ± 1.1 MPa Young’s modulus, 2.5 ± 0.4 MPa tensile strength and 321 ± 29% elongation at break. Bioactive coating does not change the structure and mechanical properties of the scaffolds. Treated scaffolds are biocompatible and have no cytotoxic effect in direct contact with cells. Functionalization also promotes the in vitro adhesion and proliferation of human adipose mesenchymal stromal cells. After 7 days of incubation, the PCL scaffold modified with the heparin–platelet lysate complex had a cell density of 185.6 ± 15.7 cells/mm² compared to 79.5 ± 7.8 cells/mm² for nontreated control. The intramuscular implantation of scaffolds revealed that immobilization of heparin alone prolongs the acute phase of the inflammatory reaction. However, subsequent treatment with platelet lysate minimizes the inflammatory reaction, slows the rate of implant absorption, and accelerates vascularization. The results obtained show that the developed bioactive coating improves the cellular properties of PCL electrospun scaffolds and can be used to form in vivo tissue-engineered constructs. Full article

Infertility is a growing global health concern, with male infertility contributing to nearly half of all cases. While conventional semen analysis often overlooks functional impairments, oxidative stress has emerged as a key factor affecting sperm quality. Notably, oxidative stress is elevated in obesity, a rising epidemic affecting more than 1 in 8 people worldwide. This study examines the role of deoxycholic acid (DCA), a secondary bile acid that is elevated in obesity, and its potential to induce oxidative stress and impair sperm function. Semen samples from healthy donors were incubated with DCA, and its effects on sperm motility, viability, capacitation, and oxidative stress markers were assessed. Sperm motility and viability were evaluated using computer-assisted semen analysis (CASA) and hypo-osmotic swelling (HOS) tests, while sperm capacitation was measured via tyrosine phosphorylation (P-Tyr) and acrosome reaction (AR). Oxidative stress markers were quantified using flow cytometry. While progressive motility and viability remained unchanged, DCA reduced hyperactive motility, P-Tyr, and acrosome reaction and increased oxidative stress markers in spermatozoa. These findings suggest that secondary bile acids can disrupt sperm function through oxidative mechanisms, affecting non-conventional semen parameters that may go undetected in standard analyses. This underscores the gut-testis axis’s role in male infertility and highlights the need for more comprehensive diagnostics and targeted therapies. Full article

Background/Objectives: Nanotherapies are a strategy to combat Candida resistance. This study analyzed the impacts of iron oxide nanoparticles (IONPs) functionalized with a chitosan (CS) layer acting as carriers of chlorhexidine (CHX) on an oral candidiasis microcosm biofilm. Methods: Saliva samples from three healthy donors were used to form biofilms, to which Candida species were added to reproduce an oral candidiasis microcosm. Biofilms were cultivated for 72 h on glass coverslips using an active adhesion model. Biofilms without Candida served as a control model. The nanocarrier loaded with CHX at 78 (IONPs-CS-CHX78) or 156 µg/mL (IONPs-CS-CHX156) was co-incubated with the biofilms for 24 h. Controls included isolated IONPs, CS, and CHX, in addition to an untreated group (NC). Assays for biomass production, metabolism, microbial load, and lactic acid production were conducted to assess antibiofilm effects. Biofilm structure, viability, and thickness were also examined by confocal microscopy. Statistical analysis was performed using one-way ANOVA or Kruskal–Wallis, subsequently accompanied by the Student–Newman–Keuls post hoc test (p < 0.05). Results: CHX and IONPs-CS-CHX156 were the most effective agents against all tested biofilm models, significantly reducing metabolism, microbial load (bacterial and fungal), and viability. For the oral candidiasis biofilm, the nanocarrier did not affect biomass or biofilm thickness but led to a significant increase in lactic acid levels compared to NC. Conclusions: It is concluded that the nanocarrier of CHX exhibits a significant reducing effect on oral candidiasis microcosm biofilms at half the concentration required for non-carried CHX. This nanostructure can be explored in the development of antiseptic or disinfectant solutions for managing oral candidiasis. Full article