Search Results (1,362)

This study aims to identify microbial strain resources capable of degrading diflufenican and elucidate their degradation characteristics, with the goal of mitigating the phytotoxicity hazards associated with the prolonged use of this persistent herbicide. A degradation strain was isolated, purified, and screened from wheat field soils that had been subjected to diflufenican treatment using an enrichment culture method. The taxonomic classification of the strain was determined through a comprehensive analysis of its morphology, physiology, biochemistry, as well as its 16S rRNA gene sequence. The results demonstrated that the screened bacterial strain 88-1 could utilize diflufenican as its metabolic carbon source and was identified as Enterobacter hormaechei, a facultative anaerobe. The degradation efficiency of strain 88-1 on diflufenican was closely associated with cultivation time, the initial concentration of the herbicide, temperature, pH, and inoculation amount of the strain. Additionally, the degradation rate exhibits a positive correlation with the biomass of the strain. Under optimal conditions (40 mg/L diflufenican, 30 °C, pH 8.0, 10% inoculum), the highest observed degradation efficiency and viable cell density over the 120 h incubation period were 55.11% and 8.05 × 10⁶ CFU/mL, respectively. Furthermore, rapid biotransformation commenced within 24 h, yielding a cascade of metabolites, with 2-(3-(trifluoromethyl)phenoxy)pyridine-3-carboxamide identified as the primary metabolite. These findings suggest that strain 88-1 holds promise for the bioremediation of soils contaminated with diflufenican. Full article

Cardiac fibroblasts (cFbs) are important in vitro models for studying myocardial fibrosis, extracellular matrix homeostasis, and tissue remodeling. However, establishing primary cultures from adult mouse hearts remains technically demanding due to the limited proliferative capacity of freshly isolated cells, their susceptibility to enzymatic and mechanical stress during tissue dissociation, and the resulting difficulty in obtaining sufficient numbers of viable fibroblasts for downstream experiments. Even small variations in digestion and early culture conditions can affect cell recovery, purity, and expansion, highlighting the need for optimized and reproducible isolation protocols. Here, we compared three enzymatic strategies for establishing primary adult mouse cardiac fibroblast cultures: sequential digestion with collagenase I and trypsin, retrograde coronary perfusion with collagenase II, and continuous digestion with Liberase DH. The three protocols were evaluated by assessing post-isolation cell viability, the composition of recovered non-myocyte populations by flow cytometry, and the ability of isolated cells to establish and expand during the first five days of culture. Retrograde coronary perfusion generated viable fibroblasts but yielded relatively few cells and is most advantageous when fibroblasts are isolated simultaneously with cardiomyocytes from the same heart. Liberase DH digestion recovered a broader spectrum of non-myocyte populations, making it suitable for immediate phenotypic analyses, although it produced less efficient fibroblast expansion in culture. Sequential collagenase I/trypsin digestion showed the best overall performance, with low mortality, negligible CD31⁺ endothelial contamination, and robust expansion into confluent spindle-shaped monolayers within five days. Overall, these findings identify sequential collagenase I/trypsin digestion as an effective and reproducible approach for establishing viable adult mouse cardiac fibroblast cultures for downstream functional and molecular studies. Full article

Lentiviral vector (LV) production for CAR-T therapy remains challenging due to the limited scalability of adherent HEK293 cell cultures. To address this, we adapted HEK293FT cells directly to serum-free FreeStyle 293 Expression Medium, generating a novel suspension cell line, HEK293FT-DS. We characterized growth kinetics, stability over 35 passages, and transient transfection parameters (PEI:DNA ratio, cell density). The suspension-adapted cells grew with >93% viability and a specific growth rate of 0.54 day⁻¹, and maintained stable viable cell density (1.51 ± 0.03 × 10⁶ cells/mL) over 35 passages. The optimal PEI:DNA ratio was 2.5:1, and increasing the transfection cell density to 16 × 10⁶ cells/mL boosted functional titers to approximately 9 × 10⁶ TU/mL in clarified supernatant. Scaled production (1 L culture supernatant per batch) yielded up to 5 × 10⁹ TU. The resulting vectors efficiently transduced primary human T cells (57% CAR-positive cells) in a CliniMACS Prodigy-based process. The platform based on suspension-adapted HEK293FT-DS cells enables high-density transfection and provides a cost-effective, scalable alternative to commercial LV production systems, particularly suited for academic CAR-T cell manufacturing. Full article

Small-molecule kinase inhibitors offer a compelling therapeutic strategy for glioblastoma, yet their clinical efficacy remains severely limited by blood–brain barrier penetration and active efflux transporter extrusion. This review evaluates current medicinal chemistry approaches and translational paradigms to overcome these drug delivery and biological constraints. A critical analysis of the literature reveals that direct structural optimization faces a multidimensional balancing act; next-generation design must prioritize macrocyclization, structural rigidification, and bioisosteric capping to lower polar surface area and evade P-glycoprotein and BCRP efflux. Furthermore, carrier-mediated prodrugs targeting the LAT1 transporter provide a viable rescue strategy for highly potent scaffolds. Reviewing recent clinical failures, such as paxalisib and osimertinib, underscores that single-node monotherapies fail due to compensatory pathway hyperactivation and clonal heterogeneity, whereas multi-targeted agents or rational dual-node combinations prevent rapid tumor adaptation. Additionally, combining kinase inhibitors with DNA damage repair inhibitors, immune checkpoint modulation, or MR-guided focused ultrasound could provide powerful synergistic networks. Finally, bridging the translational gap requires complementing conventional serum-cultured cell lines with patient-derived glioma stem cells and orthotopic xenografts to better recapitulate the cellular architecture of the disease. Ultimately, overcoming the therapeutic challenges in glioblastoma demands a fundamental pivot toward rigorous neuro-pharmacological design and multi-lineage network oncology. Full article

Kombucha is a functional beverage produced through the fermentation of Camellia sinensis by a symbiotic culture of bacteria and yeast (SCOBY). Incorporating cashew (Anacardium occidentale L.) and Cajá (Spondias mombin L.) pulps represents a value-addition strategy to mitigate the post-harvest perishability of native Cerrado fruits. This study evaluated physicochemical kinetics, 21-day refrigerated shelf life, consumer acceptance, and biological effects in Caenorhabditis elegans of flavored kombuchas. During a 9-day primary fermentation, the pH decreased to 2.95 while total acidity reached 5.38%. Following flavoring (10% v/v pulp/juice), cashew kombucha exhibited the highest total phenolic content (4.19 mg GAE/mL) and ferric reducing power (FRAP), whereas Cajá kombucha showed superior DPPH scavenging activity (1236 µM TroloxE/mL). Refrigerated storage (4 °C) maintained high counts of viable microorganisms (>10⁵ CFU/mL), though active residual fermentation induced ongoing sugar cleavage and a transient yeast proliferation peak on day 7 (8.15 log CFU/mL in Cajá kombucha). Toxicity assays using C. elegans revealed the initial mortality (31.5–62.5%); however, adjusting the pH to near-neutrality restored high organism viability (<10% mortality), confirming that mortality was acidity-driven. In pH-adjusted media, cashew kombucha promoted a modest extension of nematode lifespan, whereas Cajá kombucha reduced lipid accumulation. Sensory testing revealed moderate consumer acceptance, with cashew kombucha achieving a higher Acceptance Index (72.92%) than Cajá (67.26%). Overall, fruit supplementation enriches the bioactive potential of green tea kombucha, provided that post-fermentation acidity is appropriately managed. Full article

The escalating challenge of managing vector-borne pathologies necessitates the advancement of vector control strategies that are both highly potent and environmentally benign. This research evaluates the insecticidal efficacy and environmental plasticity of three Bacillus thuringiensis subsp. israelensis (Bti) H14 strains 33, 87/1, and 7-1/3 against laboratory and wild mosquito populations. In controlled laboratory assays, cultures with a spore density of 4.0–4.4 × 10⁹ spores/mL demonstrated significant larvicidal activity against larvae of all instars (L₁–L₄) of Aedes aegypti. The median lethal concentration (LC₅₀) was established between 1.15 and 1.48 µL/L, resulting in cumulative mortality rates of 95–96% within a 48 h exposure window. Proteomic profiling via SDS-PAGE and Western blotting validated the robust synthesis of the signature δ-endotoxin complex, specifically verifying the presence of Cry4Aa, Cry4Ba, Cry11Aa, and Cyt1Aa. Field applications in specific forest (Polissya) and steppe (Forest–Steppe) aquatic biotopes of Ukraine—characterized by distinct physicochemical profiles (differing significantly in light exposure, water hardness, salinity, and suspended solids)—substantiated the stability of the liquid formulations, achieving larval suppression rates of 95–100% within 48 h against wild Aedes-dominated assemblages at application rates of 0.25–1.0 mL/m² (equivalent to 2.5–10.0 L/ha). The demonstrated environmental adaptability suggests these novel strains are viable candidates for broad-spectrum ecological engineering applications. Their integration into sustainable vector control strategies offers a mechanism to enhance biosecurity without compromising the ecological balance of local biocenoses. Full article

Ceramic production generates large quantities of fired waste that is durable, non-biodegradable, and increasingly difficult to manage through conventional waste disposal practices. This study analyzes how ceramic waste is transformed into sustainable value in Jingdezhen, China, a city where ceramic production and cultural heritage have developed over more than a millennium. Using a qualitative multiple-case design, this study examines representative cases from three ceramic waste reutilization pathways: industrial reuse, environment-oriented reuse, and craft-based artistic reuse. The analysis shows that ceramic waste creates sustainable value through three interconnected processes: material transformation, economic activation, and cultural re-signification. Industrial cases primarily promote resource recovery and product innovation; public and environmental projects improve environmental awareness by integrating ceramic waste into urban spaces; and artistic practices reinterpret discarded ceramics as a medium for historical reflection, cultural expression, and public engagement. Based on these findings, the study proposes a material economic cultural analytical framework that explains how these value dimensions interact to transform ceramic waste from an environmental burden into a strategic resource. This study goes beyond documenting feasible ceramic waste recycling models by demonstrating that effective circular resource management in heritage-based industrial regions depends not only on technical recycling practices but also on cultural connotations and public recognition, which, together, generate value across environmental, economic, and cultural significance. These findings extend the scope of circular economy research by demonstrating a viable pathway for heritage-based industrial regions to leverage their own cultural heritage in transforming ceramic waste into environmental, economic, and cultural value. They also provide a practical model for other heritage-based regions seeking to align waste management with sustainable development. Full article

Objectives: Antiviral vaccines are usually created by inactivating viruses using physical or chemical methods. Inactivation of poliovirus with β-propiolactone (BPL) has advantages, including the absence of a requirement for long-term incubation with the inactivating agent, which minimizes the risk of negative effects of the potentially dangerous substance on the virus and reduces the duration of the inactivation process. Methods: BPL was applied at 0.2% (w/v) concentration under two conditions: 4 °C for 24 h and 37 °C for 3 h. Inactivation completeness was confirmed on Vero cell culture, while immunogenicity was assessed in guinea pigs via neutralizing antibody test. Conclusions: Two variants of virus inactivation made it possible to obtain inactivated samples that retained their immunogenic properties. BPL-inactivated samples retained sufficient D-antigen levels and elicited neutralizing antibodies comparable to or exceeding those from formaldehyde-inactivated controls. The inactivation method at 37 °C provided faster inactivation, while at 4 °C it provided a smooth decrease in virus titer. These results confirm that β-propiolactone is a viable alternative to formaldehyde for the production of inactivated polio vaccine (IPV), providing rapid inactivation of the virus compared to inactivation with formaldehyde while maintaining immunogenicity, as confirmed by guinea pig control. Full article

The growing demand for plant-based functional foods has encouraged the development of non-dairy fermented beverages able to deliver viable microorganisms and improve the technological and nutritional value of plant matrices. This study aimed to develop a prototype fermented peanut beverage using Lactiplantibacillus plantarum strains and to evaluate their technological performance, stability, and functional potential. Two plant derived strains, L. plantarum F1B and LpAv, and the commercial probiotic L. plantarum 299v were assessed in animal-free culture media and used to ferment peanut extract supplemented with sucrose and yeast extract. Fermented beverages were characterized through microbiological, physicochemical, rheological, peptide profile, refrigerated storage, and simulated gastrointestinal digestion. The selected formulation allowed all strains to reach pH ≤ 4.5 after 6.5 h, with LAB counts of 8.6–9.0 log CFU/mL. Lactic acid was the main fermentation product, and oxalic acid levels were significantly reduced. Fermentation increased peptide signals and improved rheological behavior, generating pseudoplastic beverages with higher viscosity than the control. LAB counts remained ≥8 log CFU/mL after 28 days at 4 °C, although gastrointestinal resistance after storage was strain-dependent. Taken together, L. plantarum strains showed promising technological and functional properties for developing fermented peanut-based beverages. The nutritional properties of peanut, together with the fact that it supports the growth and survival of LAB in a fermented beverage, positions peanuts as a candidate which deserves further studies in the space of fermented functional foods. Full article

The growing global demand for ethical, resource-efficient protein sources has renewed serious interest in cultured meat as a viable alternative to conventional livestock production. Two revolutionary biotechnological systems, induced pluripotent stem cell (iPSC) reprogramming and CRISPR-Cas9-mediated genome editing, when combined, offer unparalleled accuracy and scalability for the biofabrication of avian flesh. In this review, we present a comprehensive pipeline that involves the ectopic expression of Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc) to reprogram primary somatic cells derived from Gallus gallus into induced pluripotent stem cells (iPSCs). This process is subsequently followed by targeted genome editing to enhance myogenic potential, growth efficiency, nutritional composition, and disease resistance. iPSCs are cultivated in a xeno-free bioreactor following genome editing, and subsequently directed to develop into myoblasts and mature myotubes. Three-dimensional tissue biofabrication is realized by combining biomaterial scaffolds and perfusion bioreactor systems, structuring an authentic muscle tissue matrix. These engineering platforms enable precise control over microenvironmental parameters, including oxygenation and nutrient perfusion. The resulting biofabricated poultry product is compositionally optimized, free of antibiotic residues, and exhibits a significantly reduced environmental footprint than poultry that is grown in the traditional way. This all-in-one solution solves important problems in food security, animal welfare, land use efficiency, and greenhouse gas emissions while also setting up a scalable biomanufacturing framework for making proteins for the next generation. Full article

Background: Incorporating probiotic bacteria such as BC, a spore-forming probiotic bacterium known for its resistance, into extruded snacks has emerged as an innovative approach to developing functional foods. Little is known about how the addition influences metabolomic changes during storage. Methods: The mung bean snacks were produced by extrusion, followed by BC fortification. The control snack without seasoning and the two BC treatments (0.1% and 0.5% w/w) were stored at room temperature for 3 months. Culture-based methods and qPCR were performed for microbial analysis. Metabolites were extracted, and metabolite profiles were evaluated using an LC-MS-based approach. Results: 0.5% BC was the most stable in the snack, with viable counts of 7.10 log CFU/g. The PCA plot demonstrated distinct clustering patterns among groups, despite no significant difference by PERMANOVA. In negative mode, L-tyrosine showed higher accumulation in the 0.1 BC group, whereas Threonine, L-histidine, and fatty acid metabolites were highly abundant in the 0.5 BC treatment. The positive mode showed discriminant metabolites, with L-glutamic acid, Isoleucine, and L-phenylalanine enriched in the 0.5 BC treatment. Overall, BC altered citrate and phenylalanine metabolism, and each treatment affected aminoacyl-tRNA biosynthesis. Conclusions: These findings demonstrate the maintenance of culturable B. coagulans in fortified extruded mung bean snacks during storage and provide exploratory insights into metabolite changes associated with amino acid metabolism, lipid oxidation, and flavor development that may be relevant to product quality and storage stability. Full article

The intensification of the urban heat island effect poses a critical threat to the preservation and habitability of compact historic districts. The Alameda de Hércules in Seville exemplifies this vulnerability, where the intersection of heritage protection and extreme Mediterranean summers limits conventional climate adaptation. This study conducts a multi-temporal evaluation of the square’s climate resilience, spanning from its configuration prior to major 21st-century renovations to its current state and future projections, proposing future interventions. By integrating advanced microclimatic simulation and high-fidelity energy modeling, the research assesses a dual-function strategy: the improvement of the thermal environment while implementing non-intrusive photovoltaic pavements (PVPs) for energy generation. Environmental parameters, including air temperature, mean radiant temperature (MRT), and the universal thermal climate index (UTCI), were analyzed alongside the renewable energy potential of the site’s mobility infrastructure. Four heritage-sensitive interventions were tested: PV-integrated bicycle lanes, shading canopies, reflective pavement, and permeable paved grass. The results demonstrate that the canopies and paved grass zones can lower surface temperature up to 3.7–4.3 °C, reduce UTCI stress up to 2.3–3.0 °C, and decline MRT up to 10.6 °C. These values correspond to the maximum reductions achieved in specific zones. However, the PVP can locally increase surface temperature by about 4.7 °C and the reflective pavements increase MRT by around 10.4 °C, while generating an estimated annual energy yield of 174.19 MWh. The analysis under future climate projections suggests that these strategies remain equally effective under future scenarios. These findings confirm that PV-integrated urban surfaces offer a viable, reversible, and replicable approach to retrofitting historic public spaces, harmonizing climate-adaptive cooling with decentralized energy production without compromising the site’s cultural significance. Full article

This study investigated lactic fermentation of green buckwheat beverages formulated at 8% (A, AA) and 10% (B, BB) solids using single- and multiple-strain cultures of Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lacticaseibacillus paracasei and Lacticaseibacillus rhamnosus. Fermentation at 37 °C rapidly reduced pH from slightly alkaline values (7.44–7.57) to approximately 4.2–4.5 within 3–8 h, while viable counts increased from near-zero to 8–9 log₁₀CFU mL⁻¹, confirming efficient lactic acid bacteria (LAB) proliferation in all substrates. A general trend was observed in the sugar consumption strategy of studied LAB: sucrose (after hydrolysis) and glucose were almost completely depleted within 6–8 h, fructose was consumed more slowly, and raffinose remained largely unchanged, with the 10% substrate mainly accelerating early sugar turnover without altering final cell densities or the qualitative utilisation pattern. Dry matter changed little during fermentation, whereas total phenolic content (TPC) and total tannin content (TTC) were strongly affected in a matrix- and strain-dependent manner. At 8% solids, fermentation promoted substantial TTC reduction and, for several cultures, a net decrease in extractable phenolics. In contrast, 10% formulations, particularly those inoculated with L. acidophilus and L. paracasei (alone or in combination), partially preserved or increased TPC while achieving more moderate tannin losses. Overall, green buckwheat proved to be a promising substrate for developing fermented beverages in which solids level and starter composition can be tuned to combine rapid acidification and high LAB viability with tailored sugar depletion and favourable modulation of phenolic and tannin fractions. Full article

Botrytis cinerea, the causal agent of gray mold disease, is a major phytopathogen responsible for substantial losses in horticultural crops. In this study, cultivation conditions for Bacillus velezensis R22 were optimized to maximize overall antifungal activity against B. cinerea. A Plackett–Burman design was used to identify medium components affecting antifungal activity in flask cultures, followed by response surface methodology based on a central composite design (CCD) to optimize sucrose concentration, temperature, and agitation speed in a stirred bioreactor. Maximum antifungal activity was obtained at 17.45 g/L initial sucrose, 31.8 °C, and 293 rpm. The biological relevance of the optimized culture was confirmed in a tomato infection model, in which gray mold severity was reduced by 85.3% relative to the untreated control and by 59.9% relative to the non-optimized R22 culture. The same CCD approach was subsequently applied to determine cultivation conditions that maximize the concentration of R22 viable cells. The optimal parameters for 24-h growth (35.46 g/L sucrose, 36.5 °C, and 454 rpm) differed markedly from those identified for maximal antifungal activity. When evaluated on uninfected tomato plants, cultures produced under conditions favoring higher cell density showed enhanced plant growth-promoting activity compared to the non-optimized culture. Mass spectrometric analysis of lipopeptide extracts revealed that the enhanced antifungal activity was accompanied by an increased abundance of long-chain homologs across all major lipopeptide families, particularly surfactins. Thus, our results indicate that maximizing overall antifungal activity may be of greater practical significance than optimization of the individual fungicidal agent. Full article

Elite swimmers are exposed to sustained high training loads, early-morning sessions, and restricted recovery opportunities, all of which may increase psychological strain and compromise sleep. This study examined the effects of an 8-week Mindfulness–Acceptance–Insight–Commitment (MAIC) program, embedded within routine high-load training, on athlete-specific psychological stress, subjective sleep quality, and mindfulness in elite swimmers. A randomized controlled mixed-methods design was used. Thirty elite swimmers from a provincial high-performance program in China were randomly assigned to an MAIC group or a usual-practice control group (n = 15 per group). Quantitative outcomes were assessed at baseline, post-intervention, and three-month follow-up using the Athlete Psychological Strain Questionnaire, salivary cortisol, the Pittsburgh Sleep Quality Index, and the Athlete Mindfulness Questionnaire. Semi-structured interviews were conducted with all athletes in the MAIC group after the intervention. Mixed-design ANOVAs revealed significant Group × Time interactions for athlete-specific psychological stress, salivary cortisol, sleep quality, and mindfulness. Compared with the control group, the MAIC group showed lower psychological strain and cortisol, better subjective sleep quality, and higher mindfulness at post-intervention. At follow-up, improvements in psychological stress and mindfulness remained evident relative to baseline, whereas lower salivary cortisol and more favorable self-reported sleep quality remained evident relative to the control group. Qualitative findings further showed that MAIC was experienced as feasible, low-burden, and readily integrated into the training context. Athletes described attentional resets, acceptance-based responses to discomfort, and brief post-session or pre-sleep practices as helpful for regulating cognitive reactivity and arousal. Overall, MAIC appears to be a culturally grounded and practically viable adjunct strategy for supporting psychological regulation and self-reported sleep quality in elite swimmers during demanding training periods. Full article