Search Results (1,562)

Background: The Calcineurin B-like (CBL) and CBL-interacting protein kinase (CIPK) system constitute critical signaling modules mediating plant responses to abiotic stress. Although these families have been studied across various species, their evolutionary dynamics across grasses and the functional plasticity of specific isoforms remain elusive. Methods: A genome-wide analysis of CBL and CIPK families was conducted across five major Poaceae species (Oryza sativa, Triticum aestivum, Zea mays, Sorghum bicolor, and Saccharum spontaneum). Phylogenetic and synteny analyses were analyzed to family expansion and evolution. Cis-regulatory elements analysis in gene promoter regions were examined to predict potential stress-responsive features. Expression profiles of OsCBL and OsCIPK gene families were examined by qRT-PCR under conditions involving PEG-induced osmotic stress, pathogen strain P6 inoculation, and exogenous application of the phytohormones abscisic acid (ABA) and methyl jasmonate (MeJA). Protein–protein interactions between selected CBL (OsCBL4) and CIPK pairs were assessed via Yeast Two-Hybrid (Y2H) and Luciferase Complementation Imaging assays (LCI). Results: Phylogenetic and synteny analyses indicated that segmental duplications have contributed substantially to the expansion of these gene families. Promoter analysis revealed that the majority of CBL and CIPK family members, exemplified by OsCBL4, traditionally characterized as a salt sensor, possesses a cis-element architecture (rich in ABREs and MBS) heavily biased towards dehydration responsiveness. Expression profiling showed that OsCBL4 is significantly hyper-induced by direct osmotic stress (PEG) but exhibits almost no response to exogenous ABA. A subset of kinases genes (e.g., OsCIPK2, 9, 18) displayed PEG-induced expression patterns resembling those of OsCBL4, whereas OsCIPK30 remained transcriptionally unresponsive under the same conditions. Protein interaction assays demonstrated that OsCBL4 physically interacts exclusively with PEG-responsive transcriptionally activated kinases such as OsCIPK9, but failed to interact with the non-responsive OsCIPK30. Conclusions: Our study provides a genomic characterization of CBL and CIPK families across five major Poaceae species. The combined expression and interaction data reveal that OsCBL4-assembles with specific CIPKs into signaling modules during osmotic stress responses in rice, pointing to roles that go beyond salt stress responses. The findings establish a foundation for further functional dissection of CBL-CIPK pathway diversification in abiotic stress adaptation. Full article

The wine aromatic profile is influenced by complex interactions between grapevine genotype and enological practices. Thus, the aim of this study was to investigate the combined effects of grapevine clones and yeast strains on the volatile composition and sensory properties of wines produced from the Croatian indigenous variety Moslavac. Wines from five registered Moslavac clones (PUS-017, PUS-026, PUS-030A, PUS-087, and PUS-111) were produced using two commercially available yeast strains (Lalvin QA23 and Zymaflore Xarom). Significant effects of both clone and yeast strain were observed, particularly for yeast-derived compounds, such as isoamyl alcohol, phenylethyl alcohol, and medium-chain fatty acids. Ester production was generally enhanced by the Xarom yeast strain, although clone differences were also observed. Grape-derived volatile compounds differed significantly among clones, with wines from clones PUS-030A and PUS-087 having higher concentrations of norisoprenoids and terpenes, while PUS-017 wines consistently displayed lower concentrations of volatile compounds. Furthermore, PCA and MLF analyses revealed a clear differentiation between clones, with the yeast strain having a secondary modulatory effect. The sensory results were consistent with chemical data, demonstrating that clonal selection plays a key role in defining aromatic expression and typicity of Moslavac wines. Full article

Protein–protein interactions (PPIs) are fundamental to biological processes, yet experimental identification of PPIs remains time-consuming and costly, particularly for crop species with limited data. Grape (Vitis vinifera) is a globally important fruit crop that would benefit from improved computational tools for PPI prediction to support functional genomics and molecular breeding. Here, we present GrapePPI, a deep learning framework specifically designed for grape PPI prediction that leverages pre-trained ESM (Evolutionary Scale Modeling) protein embeddings. GrapePPI employs a four-component architecture: ESM embedding extraction, sequence encoding, feature combination, and multi-layer interaction prediction. We evaluated GrapePPI on grape-specific datasets with balanced and imbalanced class distributions, as well as benchmark datasets from yeast and Arabidopsis. On grape data, GrapePPI significantly outperformed state-of-the-art methods including DeepFE-PPI, PIPR, and ESMAraPPI, achieving F1 scores of 89.34% and 85.43% on balanced and imbalanced datasets, respectively, with PR AUC values of 95.29% and 90.87%. GrapePPI also demonstrated strong cross-species generalization, outperforming competing methods on yeast datasets and achieving performance comparable to specialized plant models on Arabidopsis data. Our results establish GrapePPI as an effective and robust tool for grape PPI prediction, with practical applications in functional genomics research and crop improvement programs. Full article

Brucella is a neglected foodborne pathogen, which contaminates milk, dairy products, meat, and meat products of infected animals. However, the role of the Brucella putative effector (BPE) protein family, which relies on the type IV secretion system (T4SS) in Brucella abortus, remains unclear. We demonstrated that BPE159 mediates the regulation of host nuclei in autophagy. The host-interacting protein Eci1 was screened using yeast two-hybridization, molecular docking, and immunoprecipitation, and BPE159-deleted (ΔBPE159) and complementary (ΔBPE159-C) strains were constructed by homologous recombination. We evaluated their growth, survival, and replication and measured the expression of autophagy-related cytokine mRNAs in macrophages. BPE159 was localized in the nucleus of host cells and interacted with Eci1 to downregulate the expression of macrophage autophagy factors, thereby inhibiting host autophagy and enabling the persistence of Brucella. These findings highlight the critical role of BPE159 in mediating autophagy through Eci1 in host cells to promote Brucella survival in host cells. Full article

Sustainable alternatives to synthetic polymer-based sanitary napkins are essential to reduce the environmental impact and health concerns. This study presents a method for using water hyacinth (Eichhornia crassipes), an invasive aquatic weed, as biomass to produce biodegradable absorbent material for sanitary pads. Water hyacinth fibers were treated with an alkaline solution and incorporated into the absorbent core. Morphological, chemical, structural, functional, microbiological, and biodegradability evaluations were then conducted systematically. Scanning electron microscopy showed that non-cellulosic components were successfully removed, producing a rougher surface topology and enhanced fiber interactions. Fourier-transform infrared spectroscopy confirmed structural changes in cellulose after treatment. Additionally, X-ray diffraction showed that the crystallinity index increased from 53.21% in untreated fibers to 62.56% in treated fibers, indicating improved order and stability. The developed absorbent sanitary pad showed rapid fluid uptake, absorbing 10 mL within three seconds while maintaining a skin-compatible neutral pH of 6.87, as specified in Indian Standard IS 5405:1980. Microbial contamination remained low, with a total bacterial count of 360 CFU/g, no yeast or mold at ≤1 CFU/g, and no presence of Staphylococcus aureus. Soil burial tests showed 70% biodegradability at 40 days and approximately 95% at 60 days, indicating high biodegradability. These findings demonstrate the potential of water hyacinth as an inexpensive and environmentally friendly material for manufacturing hygienic sanitary pads, highlighting the sustainability benefits of valorizing invasive biomass and reducing reliance on synthetic polymers. Full article

Acidic mine drainage (AMD) poses a severe global environmental threat due to its high acidity and elevated levels of toxic hexavalent chromium (Cr(VI)), for which biogenic iron sulfide (FeS) nanoparticles have emerged as a promising remediation agent; however, their practical application is hindered by aggregation and oxidative deactivation. This research synthesized biogenic FeS nanoparticles via sulfate-reducing bacteria (SRB) and employed humic acid (HA) as a stabilizing agent to enhance Cr(VI) removal performance in simulated AMD conditions. Single-factor experiments combined with response surface methodology identified the optimal biosynthetic conditions for FeS: yeast extract powder dosage of 2.2 g/L, Fe/S molar ratio of 0.8, and NH₄Cl dosage of 3.1 g/L. Under these conditions, the material achieved 84.25% Cr(VI) removal, with the Fe/S molar ratio identified as the most influential parameter governing synthesis and performance. Introducing HA at an optimal dosage of 2 mg/L drove marked improvements in both nanoparticle yield and reactivity: FeS yield increased to 1096.26 mg/L, Cr(VI) removal efficiency reached 99.62%, and residual Cr(VI) dropped from 15.75 mg/L to just 0.38 mg/L. Kinetic and isotherm analyses, paired with SEM/TEM imaging and zeta potential measurements, revealed that HA stabilization improved particle dispersion and reduced lamellar stacking, resulting in a surface-controlled Cr(VI) removal process. FTIR and 2D-COS analyses demonstrated that HA-derived oxygen-containing functional groups, including O–H/N–H, C=O, and C–O moieties, played a central role in interfacial interactions during Cr(VI) sequestration. XRD results confirmed that Cr(VI) was reduced to Cr(III) and primarily immobilized as low-solubility CrOOH and Cr₂S₃, while the formation of Fe–Cr spinel-like phases remains tentative without X-ray Photoelectron Spectroscopy (XPS) validation. Further investigation via surface-sensitive spectroscopy and dynamic leaching tests is needed to fully assess the long-term stability of the reaction products. Full article

Sucrose phosphate synthase (SPS) is a key rate-limiting enzyme that regulates carbon partitioning and stress tolerance in plants. In this study, we systematically characterized the SPS gene family in maize (Zea mays L.) and identified key members and their interaction networks involved in drought responses. A total of seven ZmSPS genes were identified through genome-wide bioinformatics analyses. Motif composition, gene structure, phylogenetic relationships, and synteny analyses indicated that the ZmSPS gene family is highly conserved among monocot species. Promoter analysis revealed that the upstream regions of ZmSPS genes are enriched with multiple stress responsive cis-acting elements. Drought stress treatments combined with quantitative real-time PCR (RT-qPCR) analyses showed that the expression of ZmSPS3 was significantly upregulated with increasing stress duration. Furthermore, yeast two-hybrid assays demonstrated that ZmSPS3 physically interacts with protein kinases and F-box proteins. These interactions suggest a potential involvement of ZmSPS3 in post-translational modification and protein stability regulation during osmotic stress. As a potential candidate gene responsive to drought, ZmSPS3 provides a preliminary basis for understanding the complex drought-response networks in maize. Full article

Heat stress, intensified by global warming, poses a great threat to plant growth and crop production. However, the molecular mechanisms underlying heat stress response (HSR) remain largely unclear. In this study, we identified and characterized SlFBX38, an F-box gene in tomato. SlFBX38 was predominantly expressed in leaves and fruits, and its expression levels were induced by heat stress and various phytohormones, including ABA, JA and SA. Subcellular location analysis revealed that SlFBX38 resides in both the nucleus and cytoplasm in N. benthamiana leaf cells, but it displays no transcriptional activity. Overexpression of SlFBX38 (OE) lines conferred enhanced heat stress tolerance, as evidenced by improved photosynthetic efficiency, elevated accumulation of ascorbic acid (AsA), stronger protective enzyme activities, and upregulation of HSR-related genes in SlFBX38-OE lines under heat stress condition. To identify potential interacting proteins, yeast two-hybrid (Y2H) library screening and further Y2H verification indicate that SlFBX38 may interact with SlbHLH058. Collectively, these findings establish SlFBX38 as a positive regulator of thermotolerance in tomato and provide a basis for further mechanistic studies of its role in HSR. Full article

The growing demand for clean-label, plant-based foods is accelerating the development of vegan emulsified products that avoid synthetic additives while delivering appealing sensory and health-related attributes. We formulated naturally colored, mayonnaise-like oil-in-water emulsions using 55% canola oil and yeast protein extracts (YPEs) as emulsifiers and polyphenol-rich ingredients derived from red cabbage and butterfly pea flower. The resulting systems were characterized for rheological behavior, texture, droplet-size distribution, lipid oxidation (peroxide value) and microbiological stability. Two distinct YPEs produced emulsions with different microstructural and mechanical properties, highlighting the role of protein composition on emulsion architecture. Incorporation of anthocyanin-rich polyphenol matrices (red cabbage extracts characterized by predominantly simple acylations and butterfly pea flower extracts containing complex acylations, both at similar purities) modulated emulsion structuring and stability during storage, beyond color delivery. Overall, polyphenol addition strengthened emulsion structure, as evidenced by a significant increase in plateau modulus from 621 Pa to 1428 Pa in emulsions with complete YPE and butterfly pea extract and mitigated lipid oxidation, supporting their use as partial replacement options for additives such as EDTA in clean-label formulations. These findings provide a practical basis for designing functional, and visually attractive vegan emulsions that align with consumer demand for additive-reduced products. Full article

The use of oenological additives is an emerging trend in winemaking aimed at improving technological properties. Recent studies suggest that these additives may also influence aroma persistence after wine consumption by modulating the retention of aroma compounds in the oral cavity. The aim of this study was to evaluate the effect of three commercial oenological additives, hydrolysable tannins (gallotannin and ellagitannin) and yeast mannoproteins, on the oral aroma retention of selected aroma compounds in red and white wines. Eight aromatised wines were prepared, including three red and three white wines with additives and two control wines without additives. Thirty-eight volunteers rinsed with each wine following the Spit-Off Odorant Measurement (SOOM) procedure. Oral aroma retention was calculated by comparing aroma levels in expectorated samples with those in wines prior to oral processing. Results showed that additive type significantly affected oral aroma retention (p < 0.05), depending on both the aroma compound and the wine matrix. In red wines, tannins increased the oral retention of most aroma compounds (5–20%), whereas in white wines, tannins reduced aroma retention. Mannoproteins enhanced oral aroma retention (5–40%) in both wine types. These results highlight the role of interactions between oenological additives, aroma compounds, and the wine matrix in modulating oral aroma retention. Full article

Wine alcoholic fermentation occurs in a dynamic biochemical environment where interactions between the vessel and the product can cause inorganic and organic species to migrate into the fermenting must or wine. At low pH and with rising ethanol levels, fermentation tanks made of stainless steel, concrete or cementitious materials, ceramics, or polymers exhibit material-specific behaviors that may promote the release of toxic trace elements or alter technologically important ions. These changes can affect yeast physiology, fermentation kinetics, and matrix stability, directly impacting wine safety and quality. They may also influence the evolution of key fermentation metabolites and phenolic constituents, thereby affecting process performance, color development, oxidative stability, and other quality-related attributes. This review synthesizes current evidence on migration mechanisms and examines how vessel composition shapes the chemical and microbiological profile of fermentation. It also critically evaluates biosensor technologies—covering both biorecognition elements and signal-transduction strategies—and assesses the transition from laboratory prototypes to in situ or at-line implementations capable of detecting both migration-related events and process-relevant compositional changes with operational value for HACCP-based control. Electrochemical, optical, bienzymatic, and nanozyme-enabled platforms are discussed in terms of selectivity, matrix compatibility, and long-term functional stability under polyphenol and protein interference, CO₂ variability, fouling and biofouling, and calibration drift. Particular attention is given to analytes associated with vessel-derived migrants and to biosensor targets related to fermentation metabolites and phenolic indicators, which support dynamic process monitoring and quality-focused decision making. Considering regulatory compliance requirements across the EU, US, and Asia, we propose a practical pathway for integrating biosensors into HACCP monitoring by treating vessel–product interactions as critical control points, while laboratory reference methods remain essential for verification and compliance documentation. Full article

Titanium and its alloys are widely used in endoprostheses. The naturally formed titanium dioxide film on titanium surfaces improves chemical stability and enhances implant biocompatibility. However, oxidised titanium surfaces may also promote bacterial adhesion and biofilm formation, contributing to implant-associated infections. Therefore, surface modification represents a key strategy for controlling microbial–implant interactions. This article focuses widely used titanium alloy Ti-6Al-4V treated with a laser beam, which induces surface colour changes as a result of oxide formation. Laser processing enables controlled formation of micro- and nanoscale features, structural reconstructions, and defects that may influence the surface electrical charge and, consequently, cell immobilisation. Thus, the surface colour, electrical potential, and cell immobilisation capacity are likely interrelated. From a manufacturing perspective, titanium oxide colouring facilitates quality control and process reproducibility, as surface colour provides a rapid, non-destructive visual indicator of oxide thickness and treatment consistency. This study aims to identify correlations among surface colour, electrical potential, and cell immobilisation capacity on laser-treated titanium alloys. A relationship between the optical properties, electronic structure, and biological response of laser-processed titanium oxide films is established. Specifically, the blue colour saturation of the oxide film is inversely correlated with the electron work function. A more saturated blue corresponds to a lower work function, indicating a higher positive surface charge density. This shift is attributed to changes in electron affinity, likely resulting from laser-induced structural reconstruction and defect formation within the oxide layer. The proposed changes in electronic structure are supported by modifications in the electronic density of states, analysed using near-threshold photoelectron spectroscopy. The biological response is directly linked to these physical changes: enhanced immobilisation of yeast (Saccharomyces cerevisiae) cells on the treated alloy surface correlates with the electron work function. These results may assist in the development of controlled titanium oxide surfaces with enhanced biocompatibility. Full article

Retromer is an evolutionarily conserved protein complex first identified in budding yeast. It was originally described for its essential role in endosome-to-Golgi retrieval of lysosomal hydrolase receptors. Retromer is now known to mediate trafficking of many endosomal cargoes. The mammalian retromer is constituted by a core heterotrimer encoded by the vacuolar protein sorting (VPS) gene products VPS26, VPS35, and VPS29. To mediate cargo recognition and endosomal sorting into various pathways, this trimer can cooperate with phosphoinositide-binding sorting nexin family members. Defective retromer functioning has been associated with alterations in cellular homeostasis, leading to disease. To gain insights into how it may mediate these broad processes, a proteomic strategy in polarized Madin-Darby canine kidney cells was devised to identify retromer-interacting proteins. Subsequent validation of one of the candidates, i.e., cytokeratin 19, led to the unexpected finding that retromer localizes to the pericentriolar region in dividing cells and subsequently translocates to the midbody during cytokinesis. Retromer was found interacting with CK19, and its antisense depletion led to delocalization from CK19. Subcellular fractionation and live cell monitoring of depleted cells provided evidence of a role by retromer in post-metaphase progression and in epithelial cell migration, thereby connecting retromer with key processes of cellular dynamics. Full article

In recent years, passive cell manipulation in microfluidic devices has emerged as a crucial tool for biomedical and biotechnological applications, allowing control over cell positioning and behavior without the need for chemical labels or complex external forces. However, achieving precise and tunable modulation of cell dynamics remains a challenge, particularly with low-cost and non-invasive methods. In this work, we present a novel approach that leverages controlled acousto-mechanical perturbations (AMPs) to modulate cell arrangement and behavior in microchannels. By coupling a smartphone-driven audio speaker with a microfluidic device, acoustic signals are converted into mechanical vibrations of the tubing, generating AMPs that interact with hydrodynamically driven flows. Experiments with yeast cells and silica beads under different flow conditions revealed that acoustic stimulation induced periodic flow dynamics, with yeast cells showing tunable, flow-dependent responses while inert particles exhibited weak and stable modulation. Frequency-domain analysis highlighted a dominant response synchronized with the applied acoustic protocol, accompanied by higher-frequency components characteristic of acoustic actuation. These results demonstrate that simple, low-cost acoustic actuation revealed distinct dynamical responses between rigid inert particles and deformable biological cells and enable label-free cellular manipulation. The proposed platform offers a versatile, non-invasive, and accessible approach for controlled cell manipulation in microfluidics. Full article

This study focused on optimizing endoglucanase production using a peculiar fungal co-culture comprising Rhizopus arrhizus and Aspergillus fumigatus, identified through morphological and 18S rDNA analyses. The co-culture achieved the highest enzyme production after 72 h of fermentation with alkaline-treated substrates. Scanning Electron Microscopy (SEM) revealed substantial structural disruption in pretreated biomass, enhancing enzyme accessibility. Among the tested substrates, pea hulls proved to be the most effective for enzyme production. Optimization of physical and nutritional parameters was performed using Design of Experiments (DOE) approaches, specifically Plackett–Burman Design (PBD) for screening and Central Composite Design (CCD) for fine optimization. The maximum endoglucanase activity of 119.58 U/mL/min was obtained under the optimized conditions of 27.5 °C, pH 5.5, inoculum age 3.5 days, and supplementation with 1.5% fructose, 1.25% yeast extract, 1.25% sodium nitrate, and 1.25% Tween 80. Analysis of Variance (ANOVA) confirmed the significance of these parameters and their interactions at a 95% confidence level, with a strong model fit (R² = 0.9052). This study demonstrates the potential of waste pea hulls as a cost-effective substrate for enzyme production, supporting waste valorization and contributing to a circular bioeconomy through sustainable biomass utilization. Full article