Search Results (15,322)

Botrytis cinerea is a highly versatile pathogenic fungus, causing significant damage across a wide range of plant species. A central focus of this review is the recent advances made through proteomics, an advanced molecular tool, in understanding the mechanisms of B. cinerea infection. Recent advances in mass spectrometry-based proteomics—including LC-MS/MS, iTRAQ, MALDI-TOF, and surface shaving—have enabled the in-depth characterization of B. cinerea subproteomes such as the secretome, surfactome, phosphoproteome, and extracellular vesicles, revealing condition-specific pathogenic mechanisms. Notably, in under a decade, the proportion of predicted proteins experimentally identified has increased from 10% to 52%, reflecting the rapid progress in proteomic capabilities. We explore how proteomic studies have significantly enhanced our knowledge of the fungus secretome and the role of extracellular vesicles (EVs), which play key roles in pathogenesis, by identifying secreted proteins—such as pH-responsive elements—that may serve as biomarkers and therapeutic targets. These technologies have also uncovered fine regulatory mechanisms across multiple levels of the fungal proteome, including post-translational modifications (PTMs), the phosphomembranome, and the surfactome, providing a more integrated view of its infection strategy. Moreover, proteomic approaches have contributed to a better understanding of host–pathogen interactions, including aspects of the plant’s defensive responses. Furthermore, this review discusses how proteomic data have helped to identify metabolic pathways affected by novel, more environmentally friendly antifungal compounds. A further update on the advances achieved in the field of proteomics discovery for the organism under consideration is provided in this paper, along with a perspective on emerging tools and future developments expected to accelerate research and improve targeted intervention strategies. Full article

Skin aging is closely related to mitochondrial dysfunction and cell cycle abnormalities, and developing intervention strategies targeting mitochondrial quality control is an important direction for anti-aging research. In this study, we investigated the anti-aging mechanism of Camellia japonica flower (CJF) extract and its active ingredient hyperoside based on a doxorubicin (DOX)-induced endogenous senescence model in human skin fibroblasts (HSFs). LC-MS proteomics analysis revealed that CJF extract and hyperoside specifically activated the FUNDC1-mediated mitochondrial autophagy pathway, significantly ameliorated the DOX-induced decrease in mitochondrial membrane potential and the accumulation of reactive oxygen species (ROS), and alleviated the cellular S-phase blockade and reversed the high expression of senescence-associated β-galactosidase (SA-β-gal). Further studies showed that the two cleared damaged mitochondria by enhancing mitochondrial autophagy and restoring cellular energy metabolism homeostasis while promoting type III collagen and elastin synthesis and repairing the expression of Claudin 1 related to skin barrier function. For the first time, the present study reveals the molecular mechanism of CJF extract in delaying skin aging by regulating the FUNDC1-dependent mitochondrial autophagy pathway, which provides a theoretical basis and a candidate strategy for developing novel anti-aging agents targeting mitochondrial quality control. Full article

Glioblastoma (GBM) is a highly aggressive brain tumor marked by invasive growth and therapy resistance. Tumor cells adapt to hostile conditions, such as hypoxia and nutrient deprivation, by activating survival mechanisms including autophagy and metabolic reprogramming. Among GBM-associated changes, hypersialylation, particularly, the aberrant expression of polysialic acid (PSA), has been linked to increased plasticity, motility, and immune evasion. PSA, a long α2,8-linked sialic acid polymer typically attached to the NCAM, is abundant in the embryonic brain and re-expressed in cancers, correlating with poor prognosis. Here, we investigated how PSA expression was regulated in GBM cells under nutrient-limiting conditions. Serum starvation induced a marked increase in PSA-NCAM, driven by upregulation of the polysialyltransferase ST8SiaIV and an autophagy-dependent recycling of sialic acids from degraded glycoproteins. Inhibition of autophagy or sialidases impaired PSA induction, and PSA regulation appeared dependent on p53 function. Immunohistochemical analysis of GBM tissues revealed co-localization of PSA and LC3, particularly around necrotic regions. In conclusion, we identified a novel mechanism by which GBM cells sustain PSA-NCAM expression via autophagy-mediated sialic acid recycling under nutrient stress. This pathway may enhance cell migration, immune escape, and stem-like properties, offering a potential therapeutic target in GBM. Full article

The construction industry is responsible for 39% of global CO₂ emissions related to energy use, with cement responsible for 5–8% of it. Limestone calcined clay cement (LC³), a ternary blended binder system, offers a low-carbon alternative by partially substituting clinker with calcined clay and limestone. This study investigated the use of waste clay brick powder (WBP), a waste material, as a source of calcined clay in LC³ formulations, addressing both environmental concerns and SCM scarcity. Two LC³ mixtures containing 15% limestone, 5% gypsum, and either 15% or 30% WBP, corresponding to clinker contents of 65% (LC³-65) or 50% (LC³-50), were evaluated against general purpose (GP) cement mortar. Tests included setting time, flowability, soundness, compressive and flexural strengths, drying shrinkage, isothermal calorimetry, and scanning electron microscopy (SEM). Isothermal calorimetry showed peak heat flow reductions of 26% and 49% for LC³-65 and LC³-50, respectively, indicating a slower reactivity of LC³. The initial and final setting times of the LC³ mixtures were 10–30 min and 30–60 min longer, respectively, due to the slower hydration kinetics caused by the reduced clinker content. Flowability increased in LC³-50, which is attributed to the lower clinker content and higher water availability. At 7 days, LC³-65 retained 98% of the control’s compressive strength, while LC³-50 showed a 47% reduction. At 28 days, the compressive strengths of mixtures LC³-65 and LC³-50 were 7% and 46% lower than the control, with flexural strength reductions being 8% and 40%, respectively. The porosity calculated from the SEM images was found to be 7%, 11%, and 15% in the control, LC³-65, and LC³-50, respectively. Thus, the reduction in strength is attributed to the slower reaction rate and increased porosity associated with the reduced clinker content in LC³ mixtures. However, the results indicate that the performance of LC³-65 was close to that of the control mix, supporting the viability of WBP as a low-carbon partial replacement of clinker in LC³. Full article

This study evaluates the effectiveness of the B Impact Assessment (BIA) as a catalyst for integrating sustainability into industrial firms through a qualitative case study of LC Paper, the first B Corp-certified tissue manufacturer globally and a pioneer in applying BIA in the pulp and paper sector. Based on semi-structured interviews, organizational documents, and direct observation, this study examines how BIA influences corporate governance, environmental practices, and stakeholder engagement. The findings show that BIA fosters structured goal setting and the implementation of measurable actions aligned with environmental stewardship, social responsibility, and economic resilience. Tangible outcomes include improved stakeholder trust, internal transparency, and employee development, while implementation challenges such as resource allocation and procedural complexity are also reported. Although the single-case design limits generalizability, this study identifies mechanisms transferable to other firms, particularly those in environmentally intensive sectors. The case studied also illustrates how leadership commitment, participatory governance, and data-driven tools facilitate the operationalization of sustainability. By integrating stakeholder and institutional theory, this study contributes conceptually to understanding certification frameworks as tools for embedding sustainability. This research offers both theoretical and practical insights into how firms can align strategy and impact, expanding the application of BIA beyond early adopters and into traditional industrial contexts. Full article

Entomopathogenic fungi (EPF) are one of the most environmentally friendly ways to control a plethora of chewing insects such as T. pityocampa, G. mellonella, and A. grisella. Bioassay of EPF on these highly damaging pests is considered important in the face of climate change in order to research alternative solutions that are capable of limiting chemical control, the overuse of which increases insects’ resistance to chemical compounds. In this study, the insecticidal virulence of Metarhizium robertsii isolates, retrieved from forest ecosystems, was tested on second-instar larvae of T. pityocampa, G. mellonella, and A. grisella. Bioassays were carried out in the laboratory, where experimental larvae were sprayed with 2 mL of a six-conidial suspension from each isolate. Mortality was recorded for 144 h after exposure. Mean mortality, lethal concentrations, sporulation percentage, and sporulation time were estimated for each isolate. Metarhizium isolates resulted in the highest mortality (89.2% for G. mellonella and 90.2% for A. grisella). Based on the LC₅₀ estimates determined by the concentration–mortality relationships for the tested fungal isolates, we demonstrated significant virulence on larvae of G. mellonella, A. grisella, and T. pityocampa. Our results indicate that entomopathogenic fungi have the potential to become a very useful tool in reducing chemical applications. Full article

Biothiols, including cysteine (Cys), homocysteine (Hcy), and glutathione (GSH), are crucial for physiological regulation and their imbalance poses severe health risks. Herein, we developed a pH-responsive liquid crystal (LC)-based sensing platform for detection of biothiols by doping 4-n-pentylbiphenyl-4-carboxylic acid (PBA) into 4-n-pentyl-4-cyanobiphenyl (5CB). Urease catalyzed urea hydrolysis to produce OH⁻, triggering the deprotonation of PBA, thereby inducing a vertical alignment of LC molecules at the interface corresponding to dark optical appearances. Heavy metal ions (e.g., Hg²⁺) could inhibit urease activity, under which condition LC presents bright optical images and LC molecules maintain a state of tilted arrangement. However, biothiols competitively bind to Hg²⁺, the activity of urease is maintained which enables the occurrence of urea hydrolysis. This case triggers LC molecules to align in a vertical orientation, resulting in bright optical images. This pH-driven reorientation of LCs provides a visual readout (bright-to-dark transition) correlated with biothiol concentration. The detection limits of Cys/Hcy and GSH for the PBA-doped LC platform are 0.1 μM and 0.5 μM, respectively. Overall, this study provides a simple, label-free and low-cost strategy that has a broad application prospect for the detection of biothiols. Full article

Brown algae such as Ascophyllum nodosum (AN) and Fucus vesiculosus (FV) are gaining considerable attention as functional feed additives due to their health-beneficial properties. This study evaluated the antioxidant potential of AN and FV extracts in intestinal epithelial cells and the in vivo model Caenorhabditis elegans (C. elegans). Aqueous AN and FV extracts were characterized for total phenolic content (TPC), antioxidant capacity (TEAC, FRAP), and phlorotannin composition using LC-HRMS/MS. Antioxidant effects were assessed in vitro, measuring AAPH-induced ROS production in Caco-2 and IPEC-J2 cells via H₂DCF-DA, and in vivo, evaluating the effects of paraquat-induced oxidative stress and AN or FV treatment on worm motility, GST-4::GFP reporter expression, and gene expression in C. elegans. FV exhibited higher total phenolic content, antioxidant capacity (TEAC, FRAP), and a broader phlorotannin profile (degree of polymerization [DP] 2–9) than AN (DP 2–7), as determined by LC-HRMS/MS. Both extracts attenuated AAPH-induced oxidative stress in epithelial cells, with FV showing greater efficacy. In C. elegans, pre-treatment with AN and FV significantly mitigated a paraquat-induced motility decline by 22% and 11%, respectively, compared to PQ-stressed controls. Under unstressed conditions, both extracts enhanced nematode healthspan, with significant effects observed at 400 µg/g for AN and starting at 100 µg/g for FV. Gene expression analysis indicated that both extracts modulated antioxidant pathways in unstressed worms. Under oxidative stress, pre-treatment with AN and FV significantly reduced GST-4::GFP expression. In the nematode, AN was more protective under acute stress, whereas FV better supported physiological function in the absence of stressors. These findings demonstrate that AN and FV counteract oxidative stress in intestinal epithelial cells and in C. elegans, highlighting their potential as stress-reducing agents in animal feed. Full article

This study was designed to systematically identify novel umami peptides in lager beer, clarify their molecular interactions with the T1R1/T1R3 receptor, and determine their specific effects on multidimensional sensory attributes. The peptides were characterized by LC-MS/MS combined with de novo sequencing, and 906 valid sequences were obtained. Machine-learning models (UMPred-FRL, Tastepeptides-Meta, and Umami-MRNN) predicted 76 potential umami peptides. These candidates were docked to T1R1/T1R3 with the CDOCKER protocol, producing 57 successful complexes. Six representative peptides—KSTEL, DELIK, DIGISSK, IEKYSGA, DEVR, and PVPL—were selected for 100 ns molecular-dynamics simulations and MM/GBSA binding-energy calculations. All six peptides stably occupied the narrow cleft at the T1R1/T1R3 interface. Their binding free energies ranked as DEVR (−44.09 ± 5.47 kcal mol⁻¹) < KSTEL (−43.21 ± 3.45) < IEKYSGA (−39.60 ± 4.37) ≈ PVPL (−39.53 ± 2.52) < DELIK (−36.14 ± 3.11) < DIGISSK (−26.45 ± 4.52). Corresponding taste thresholds were 0.121, 0.217, 0.326, 0.406, 0.589, and 0.696 mmol L⁻¹ (DEVR < KSTEL < IEKYSGA < DELIK < PVPL < DIGISSK). TDA-based sensory validation with single-factor additions showed that KSTEL, DELIK, DEVR, and PVPL increased umami scores by ≈21%, ≈22%, ≈17%, and ≈11%, respectively, while DIGISSK and IEKYSGA produced marginal changes (≤2%). The short-chain peptides thus bound with high affinity to T1R1/T1R3 and improved core taste and mouthfeel but tended to amplify certain off-flavors, and the long-chain peptides caused detrimental impacts. Future formulation optimization should balance flavor enhancement and off-flavor suppression, providing a theoretical basis for targeted brewing of umami-oriented lager beer. Full article

Saproxylic beetles are key bioindicators of forest ecosystem quality and play essential roles in deadwood decomposition and nutrient cycling. However, their populations are increasingly threatened by habitat fragmentation, deadwood removal, and climate-driven environmental changes. For this reason, an integrated sampling method can increase the detection of species with varying ecological traits. We evaluated the effectiveness of integrative sampling methodologies to assess saproxylic beetle diversity within Sila National Park, a Mediterranean forest ecosystem of high conservation value, specifically in two beech forests and four pine forests. The sampling methods tested included Pan Traps (PaTs), Malaise Traps (MTs), Pitfall Traps (PTs), Bait Bottle Traps (BBTs), and Visual Census (VC). All specimens were identified to the species level whenever possible, using specialized dichotomous keys and preserved in the Entomological Collection TB, Unical. Various trap types captured a different number of species: the PaT collected 32 species, followed by the PT with 24, the MT with 16, the VC with 7, and the BBT with 5 species. Interestingly, biodiversity analyses conducted using PAST software version 4.17 revealed that PaTs and MTs recorded the highest biodiversity indices. The GLMM analysis, performed using SPSS software 29.0.1.0, demonstrated that various traps attracted different species with different abundances. By combining multiple trapping techniques, we documented a more comprehensive community composition compared to single-method approaches. Moreover, PaTs, MTs, and PTs recorded 20%, 40%, and 33% of the Near Threatened species, respectively. We report new records for Sila National Park, including the LC species Pteryngium crenulatum (Curculionidae) and the NT species Grynocharis oblonga (Trogossitidae). For the first time in Calabria, the LC species Triplax rufipes (Erotylidae) and the NT species Oxypleurus nodieri (Cerambycidae) and Glischrochilus quadrisignatus (Nitidulidae) were collected. Our results emphasize the importance of method diversity in capturing species with distinct ecological requirements and highlight the relevance of saproxylic beetles as indicators of forest health. These findings support the adoption of multi-method sampling protocols in forest biodiversity monitoring and management programs, especially in biodiversity-rich and structurally heterogeneous landscapes. Full article

Heavy metal contamination in freshwater ecosystems poses persistent threats to aquatic organisms and public health. This study evaluates the transgenerational toxicity of cadmium chloride and copper sulfate on Culex pipiens, focusing on development, reproduction, and midgut histopathology over two successive generations. Larval bioassays showed cadmium chloride to be more toxic than copper sulfate, with early instars exhibiting higher sensitivity (LC₅₀ = 8.66 μg/L for Cd; 175.63 μg/L for Cu). Both metals significantly delayed larval and pupal development, reduced fecundity, and decreased egg hatchability in a dose-dependent manner. Histopathological examination revealed midgut epithelial degeneration, vacuolation, and brush border loss, with copper sulfate inducing more severe cytotoxicity. These findings confirm that sublethal, chronic metal exposure can impair physiological and reproductive traits across generations. Moreover, this study highlights the utility of mosquitoes as sensitive bioindicators of aquatic pollution, and underscores the long-term ecological implications of heavy metal contamination on vector dynamics and disease transmission. Full article

The two-spotted spider mite, Tetranychus urticae, is one of the polyphagous pests of several crops and forestry, resistant to numerous conventional chemicals. Due to the negative side effects of harmful chemical pesticides, such as environmental pollution, and risks to human health, the introduction of effective and low-risk alternatives is essential. The promising pesticidal effects of essential oils (EOs) isolated from Artemisia annua have been documented in recent studies. In the present study, the acaricidal effects of an A. annua EO, along with its two dominant monoterpenoids, 1,8-cineole and camphor, were investigated against adults of T. urticae. Artemisia annua EO, 1,8-cineole, and camphor, with 24 h-LC₅₀ values of 0.289, 0.533, and 0.64 µL/L air, respectively, had significant toxicity by fumigation against T. urticae adults. Along with lethality, A. annua EO and monoterpenoids had significant inhibitory effects on the activity of detoxifying enzymes, including α- and β-esterases, glutathione S-transferases, and cytochrome P-450 monooxygenase. According to the findings of the present study, A. annua EO and its two dominant monoterpenoids, 1,8-cineole and camphor, with significant toxicity and inhibitory effects on detoxifying enzymes, can be introduced as available, effective, and eco-friendly acaricides in the management of T. urticae. Full article

Background Plasma membrane proteins (PMPs) play key roles in cell signalling, adhesion, and trafficking, and are attractive therapeutic targets in cancer due to their surface accessibility. However, their typically low abundance limits detection by conventional proteomic approaches. Methods: To improve PMP detection, we employed a surface proteomics workflow combining cell surface biotinylation and affinity purification prior to LC-MS/MS analysis in cervical (SiHa) and bladder (UMUC3) cancer cell lines cultured under normoxic (21% O₂) or hypoxic (0.1% O₂) conditions. Results: In SiHa cells, 43 hypoxia-upregulated proteins were identified exclusively in the biotin-enriched fraction, including ITGB2, ITGA7, AXL, MET, JAG2, and CAV1/CAV2. In UMUC3 cells, 32 unique upregulated PMPs were detected, including CD55, ADGRB1, SLC9A1, NECTIN3, and ACTG1. These proteins were not observed in corresponding whole-cell lysates and are associated with extracellular matrix remodelling, immune modulation, and ion transport. Biotinylation enhanced the detection of membrane-associated pathways such as ECM organisation, integrin signalling, and PI3K–Akt activation. Protein–protein interaction analysis revealed links between membrane receptors and intracellular stress regulators, including mitochondrial proteins. Conclusions: These findings demonstrate that surface biotinylation improves the sensitivity and selectivity of plasma membrane proteomics under hypoxia, revealing hypoxia-responsive proteins and pathways not captured by standard whole-cell analysis. Full article

Background/Objectives: Prior research suggest that poor working memory significantly contributes to the growth of overweight and obesity. This study investigated the behavioral and neural aspects of general and food-specific working memory in females with overweight or obesity (OW/OB). Method: A total of 54 female participants, with 26 in the OW/OB group and 28 in the normal-weight (NW) group, completed a general and a food-related two-back task while an EEG was recorded. Results: In the general task, the OW/OB group showed significantly poorer performance (higher IES) than the NW group (p = 0.018, η² = 0.10), with reduced theta power during non-target trials (p = 0.040, η² = 0.08). No group differences were found for P2, N2, or P3 amplitudes. In the food-related task, significant group × stimulus interactions were observed. The OW/OB group showed significantly higher P2 amplitudes in high-calorie (HC) versus low-calorie (LC) food conditions (p = 0.005, η² = 0.15). LPC amplitudes were greater in the OW/OB group for HC targets (p = 0.036, η² = 0.09). Alpha power was significantly lower in OW/OB compared to NW in HC non-targets (p = 0.030, η² = 0.09), suggesting a greater cognitive effort. Conclusions: These findings indicate that individuals with OW/OB exhibit deficits in general working memory and heightened neural responses to high-calorie food cues, particularly during non-target inhibition. The results suggest an interaction between reward salience and cognitive control mechanisms in obesity. Full article

Background/Objective: Pharmacological interventions often exert systemic effects beyond their primary targets, underscoring the need for a comprehensive evaluation of their metabolic impact. Etodolac is a nonsteroidal anti-inflammatory drug (NSAID) that alleviates pain, fever, and inflammation by inhibiting cyclooxygenase-2 (COX-2), thereby reducing prostaglandin synthesis. While its pharmacological effects are well known, the broader metabolic impact and potential mechanisms underlying improved clinical outcomes remain underexplored. Untargeted metabolomics, which profiles the metabolome without prior selection, is an emerging tool in clinical pharmacology for elucidating drug-induced metabolic changes. In this study, untargeted metabolomics was applied to investigate metabolic changes following a single oral dose of etodolac in healthy male volunteers. By analyzing serial blood samples over time, we identified endogenous metabolites whose concentrations were positively or inversely associated with the drug’s plasma levels. This approach provides a window into both therapeutic pathways and potential off-target effects, offering a promising strategy for early-stage drug evaluation and multi-target discovery using minimal human exposure. Methods: Thirty healthy participants received a 400 mg dose of Etodolac. Plasma samples were collected at five time points: pre-dose, before Cmax, at Cmax, after Cmax, and 36 h post-dose (n = 150). Samples underwent LC/MS-based untargeted metabolomics profiling and pharmacokinetic analysis. A total of 997 metabolites were significantly dysregulated between the pre-dose and Cmax time points, with 875 upregulated and 122 downregulated. Among these, 80 human endogenous metabolites were identified as being influenced by Etodolac. Results: A total of 17 metabolites exhibited time-dependent changes closely aligned with Etodolac’s pharmacokinetic profile, while 27 displayed inverse trends. Conclusions: Etodolac influences various metabolic pathways, including arachidonic acid metabolism, sphingolipid metabolism, and the biosynthesis of unsaturated fatty acids. These selective metabolic alterations complement its COX-2 inhibition and may contribute to its anti-inflammatory effects. This study provides new insights into Etodolac’s metabolic impact under healthy conditions and may inform future therapeutic strategies targeting inflammation. Full article