Search Results (7,976)

Limulus amebocyte lysate (LAL) detects bacterial endotoxin through a serine-protease coagulation cascade in which Factor C responds to lipopolysaccharide and Factor G to (1,3)-β-D-glucan. Sustainable LAL production depends on collection buffers that prevent amebocyte degranulation while preserving these clotting factors. We previously showed that caffeine buffer inhibits degranulation, but caffeine-collected pellets aggregated upon resuspension in 5 mM CaCl₂, unlike phosphate-buffered saline (PBS). We therefore developed a PBS-caffeine collection solution and compared it with caffeine buffer. Over one bleeding season, 121 crabs were bled; blood was collected in caffeine, PBS-caffeine, or PBS-caffeine supplemented with EDTA, EGTA, or both, and LAL activity was measured by chromogenic and turbidimetric assays. Both buffers prevented degranulation and gave comparable LAL activity, but PBS-caffeine reduced aggregation and clotting. Treating PBS-caffeine LAL with 10% PEG-8000 selectively abolished endotoxin-sensitive Factor C activity while preserving (1,3)-β-D-glucan–sensitive Factor G activity, and the resulting Factor G lysate, formulated in 20 mM acetate (pH 5.6), remained stable for 27 months. These results define an improved collection buffer and identify conditions that selectively stabilize Factor G zymogen in liquid form. Full article

At present, treated wastewater may still contain residual nutrients and micropollutants, including heavy metals, pharmaceuticals, and dyes, which can negatively affect receiving water bodies. Increasingly stringent environmental regulations, including Directive (EU) 2024/3019, require both enhanced removal of these contaminants and greater integration of renewable energy sources in wastewater treatment plants. This paper presents a review of biomass-based wastewater polishing technologies employing biological agents such as microalgae, fungi, bacteria, co-cultures and duckweed for the removal of residual contaminants from treated effluents. The compiled data indicate that while optimal conditions can drive pollutant removal efficiencies beyond 90%, system performance varies widely depending on species selection, wastewater characteristics, and operational conditions (e.g., pH, temperature, salinity, nutrient availability, and light intensity). In addition to effluent polishing, the produced biomass can be valorized for bioenergy generation, contributing to renewable energy production and supporting circular economy principles in wastewater treatment plants. Despite these benefits, biomass harvesting remains a major technical and economic bottleneck, often representing a significant share of operational costs and limiting large-scale implementation. Overall, biomass-based treatment technologies are a promising approach for improving effluent quality and supporting renewable energy objectives; however, further advances in biomass recovery are required for broader application. Full article

Non-thermal millimeter-wave (MMW) irradiation represents a promising non-invasive strategy for cancer therapy, yet its effects in physiologically relevant 3D systems remain poorly defined. Here, we evaluated the biological impact of MMW exposure in 3D non-small-cell lung cancer (NSCLC) spheroids (NCI-H1299, A549) and normal WI-38 fibroblasts under active cooling to suppress bulk heating. We demonstrate that cellular responses are governed primarily by power density (PD), irradiation geometry, and genotype-dependent susceptibility. High-PD pyramidal horn (PH) irradiation (~4.9 mW/cm²) induced rapid apoptosis, metabolic collapse, and near-complete loss of clonogenic survival, whereas lower-PD waveguide (WG) irradiation (~0.6 mW/cm²) produced depth-limited, cumulative cytotoxicity. Surviving cancer cells exhibited robust senescence-associated growth arrest, particularly in p53-deficient NCI-H1299 cells, indicating a dual apoptotic–senescent anti-proliferative response. In contrast, WI-38 fibroblasts showed minimal apoptosis and only transient stress-associated senescence, confirming selective tumor vulnerability. Mechanistic modeling suggests that MMW energy couples to glycan-rich membrane domains, generating localized electromagnetic hotspots that trigger calcium influx, mitochondrial dysfunction, and depth-dependent apoptosis. These findings establish a mechanistic basis for selective, non-thermal MMW-induced cytotoxicity in 3D NSCLC models and support further preclinical development of MMW-based therapeutic strategies. Full article

Background: Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and is strongly associated with atrial structural remodeling driven by activated cardiac fibroblasts. Autophagy has been implicated in AF-related atrial remodeling; however, the non-coding RNA mechanisms that govern autophagic activation in atrial fibroblasts under rapid electrical stress remain poorly understood. Methods: Human cardiac fibroblasts from adult atria (HCF-aa) were subjected to rapid electrical stimulation (RES) at 0.5 V/cm and 10 Hz. Expression levels of exosomal metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), cytoplasmic miR-204-5p, and microtubule-associated protein light chain 3B (LC3B) were measured using quantitative real-time PCR and Western blot analyses. Luciferase reporter assays were performed to confirm direct molecular interactions. The functional roles of MALAT1 siRNA, miR-204-5p mimics/antagomirs, rapamycin, and 3-methyladenine (3-MA) on LC3B expression and autophagic activation were assessed by Western blot and immunofluorescence confocal microscopy for LC3B puncta formation. Results: RES significantly induced exosomal MALAT1 expression in a voltage- and time-dependent manner, peaking at 2 h post-stimulation, while cytoplasmic MALAT1 levels remained unchanged. Cytoplasmic miR-204-5p exhibited an initial transient rise followed by a significant decline at 2 h, inversely correlating with peak MALAT1 levels. LC3B mRNA and protein expression subsequently increased, peaking at 6 and 16 h, respectively. Luciferase reporter assays confirmed that miR-204-5p directly binds both the MALAT1 transcript and the 3′-UTR of LC3B mRNA. MALAT1 knockdown augmented miR-204-5p levels and suppressed LC3B expression, while miR-204-5p overexpression attenuated RES-induced LC3B upregulation and LC3B puncta accumulation. Conversely, miR-204-5p inhibition further enhanced autophagic activation, as evidenced by increased LC3B puncta density. Conclusions: In HCF-aa subjected to RES, MALAT1 functions intracellularly as a competing endogenous RNA to putatively sequester miR-204-5p, thereby de-repressing LC3B expression and promoting autophagic activation. Concurrent exosomal secretion of MALAT1 may additionally serve as a paracrine signal to neighboring cells, though this requires future conditioned-media transfer experiments to confirm. Full article

Inorganic carbon availability is an underexplored factor influencing extracellular emulsification-associated responses in haloalkaliphilic bacteria. Here, we show that Vreelandella zhaodongensis BS253 exhibits distinct physiological and transcriptional responses to CO₂ enrichment and bicarbonate supplementation, accompanied by condition-dependent changes in emulsification activity. Both moderate CO₂ enrichment (5–10%) and NaHCO₃ supported high emulsification values (E24 > 60%). However, CO₂ favored higher emulsification activity relative to biomass, whereas NaHCO₃ promoted greater biomass accumulation and elevated absolute activity. Transcriptomic profiling revealed extensive condition-dependent reprogramming, particularly involving membrane transport, envelope-associated functions, and genes annotated as related to exopolysaccharide biosynthesis. Integrative phenotype-guided analyses prioritized candidate genes statistically associated with the emulsification phenotype. The extracellular emulsification-active material remained active across a broad range of salinity, temperature, pH, and pressure, demonstrating pronounced physicochemical robustness. Together, these findings indicate that inorganic carbon availability modulates emulsification activity and associated transcriptional responses in a haloalkaliphile and highlight extremophilic bacteria as promising platforms for sustainable bioprocesses based on inorganic carbon inputs. Full article

Berry skin color CIE parameters, pH and anthocyanidin profiling of 46 grape accessions were investigated using CIE Lab system, pH measurement and anthocyanidin profiling. CIE parameters separated the samples into three groups: yellowish-green, pinkish-red, and purplish-black, and principal component analysis confirmed clear clustering, with the first two components explaining 99.1% of the variance. After anthocyanidin analysis, cyanidin was detected in all samples, whereas trace-level pelargonidin derivatives were identified by UPLC-MS/MS. Total anthocyanidin content was insufficient to evaluate the quality of berry color, but anthocyanidin composition and relative proportions showed a stronger association with color classification. Yellowish-green berries were enriched in cyanidin, while purplish berries contained more malvidin- and cyanidin. Multivariate analysis identified cyanidin, malvidin, and peonidin derivatives as the main drivers of berry skin color variations. Skin homogenate pH ranged from 3.36 to 4.63 and it was lower in wild grape relatives. Correlation analysis indicated that pH was associated with color parameters. Species-related differences in anthocyanidin glycosylation and acylation were evident, and mono/diglucosides may have potential effects on skin color. Overall, skin color appears to depend on anthocyanidin composition, relative proportions, and pH, offering a chemical basis for grape breeding and fruit quality evaluation. Full article

Aqueous precipitation and crystallization are central to impurity removal, product formation, and resource recovery in mineral and chemical processing, but robust inline monitoring remains challenging because supersaturation is not measured directly and conductivity signals are affected by temperature, composition drift, bubbles, solids, polarization, and fouling. Electrical conductivity (EC) is attractive as a low-cost, rugged process analytical tool, yet its usefulness depends on mechanistic interpretation: EC reflects charge-carrier concentration and mobility rather than supersaturation itself. This review organizes the literature into a layered framework covering (i) measurement integrity and deployment, (ii) bulk-signal extraction in multiphase media, (iii) estimation of latent variables such as dissolved concentration or supersaturation proxies, and (iv) control readiness based on conductivity-derived targets. Frequency-aware conductivity extraction, event-anchored verification, and observer-based estimation are treated as optional, complementary modules. A Ca-carbonate/CaCO₃ system is used as an illustrative case because its coupling among conductivity, pH/speciation, supersaturation, and precipitation is especially transparent, although the framework is intended for broader processing systems, including complex liquors and slurries. Opportunities are also highlighted for nanomaterials to improve both precipitation control and EC information content. Full article

Biochar can enhance microbial-mediated organic phosphorus mineralization, but the underlying mechanisms remain unknown in forest soils with varying pH values. An incubation experiment was conducted using karst (alkaline) and non-karst (acidic) forest soils. Four amounts of bagasse biochar were applied (0, 5, 10, and 15 t·hm⁻²) to assess their effects on soil phosphorus availability and microbial community structure. Olsen-P content of alkaline and acidic forest soils increased with increasing bagasse biochar addition and incubation time, especially in non-karst forest soil. The structure and diversity of phoD-harboring bacterial community of acidic forest soil were significantly altered by the amount of bagasse biochar added and the incubation time, whereas those in alkaline karst forest soil were not significantly affected. The relative abundance of the dominant order Burkholderiales reached (43%) in acidic forest soil, significantly exceeding the (9%) recorded in alkaline karst forest soil. The phoD bacteria in acidic forest soil had more complex microbial networks and were more closely related to phosphorus fractions than those in alkaline forest soil. Structural equation modeling indicated that soil phosphorus availability was directly controlled by bagasse biochar input in acidic forest soil, with an indirect pathway linked to phoD bacterial community structure. The contribution of phoD bacteria to the variation in phosphorus availability was higher in acidic forest soil than in alkaline forest soil based on variance partitioning, indicating that enhancing soil phosphorus availability with bagasse biochar depends on the amount added, soil type, and its regulation of phoD bacterial communities. Full article

Background/Objectives: DNA-targeting small molecules that induce replication stress represent a promising strategy in anticancer drug development. 1,8-Naphthalimide (NI) derivatives are well-established DNA-intercalating agents, and heterocyclic annulation offers a rational approach to enhancing their potency and tumor selectivity. Here, we report the synthesis and biological evaluation of a novel series of benzofuran-containing naphthalimide derivatives, with particular focus on the lead dinitro-substituted compound 5d. Methods: Cytotoxic activity was assessed using the MTT assay in A549 (p53 wild-type), H1299 (p53-null), and MRC-5 cells. Long-term antiproliferative effects were evaluated by clonogenic survival assay. Cell cycle distribution was analyzed by propidium iodide staining and flow cytometry. Replication stress and DNA damage were quantified by EdU incorporation and γH2AX immunofluorescence, respectively. Apoptosis was assessed by Annexin V/PI staining and caspase-3/7 activation assay. p53 nuclear accumulation and autophagy induction were evaluated by immunofluorescence and Western blot, using LC3 as an autophagic marker. Results: All compounds exhibited cytotoxic activity in the nanomolar range, with 5d emerging as the most potent and selective. Clonogenic survival was significantly reduced, indicating durable suppression of proliferative capacity. Treatment with 5d induced G₁ arrest in A549 cells and the accumulation of H1299 cells in G₂/M, consistent with p53-dependent and p53-independent checkpoint activation, respectively. EdU incorporation was markedly reduced, while γH2AX intensity increased, collectively supporting a replication stress-driven mechanism of DNA damage. Apoptosis was confirmed by increased Annexin V-positive populations and caspase-3/7 activation. LC3 puncta formation and LC3-I/LC3-II conversion were increased, indicating LC3 processing and autophagosome accumulation consistent with the activation of autophagy-related processes. Conclusions: 5d induces a cellular phenotype consistent with replication stress, including reduced EdU incorporation, γH2AX accumulation, cell cycle arrest, and apoptotic cell death in a p53 status-dependent manner. These findings establish benzofuran-annulated naphthalimides as a promising scaffold for the development of anticancer agents that exploit replication stress vulnerabilities in tumor cells. Full article

Poor aqueous solubility and consequently low bioavailability of various NSAIDs (non-steroidal anti-inflammatory drugs) usually result in high and multiple dosing with potentially serious side effects. Therefore, systems for the effective transport of NSAIDs through the GIT (gastrointestinal tract), ensuring enhanced bioavailability, remain in high demand. In the present work, we studied chitosan (CS) hydrogel lyophilizates as carrier systems for a model NSAID, namely ibuprofen (IBU). The CS-IBU lyophilizates were prepared from homogeneous or heterogeneous CS-IBU hydrogels to assess their influence on the resulting lyophilizate microstructure and IBU dissolution profiles. To gain a complex view of the CS-IBU behavior and its practical consequences, dissolution profiles of free IBU (reference) and CS-associated IBU (CS-IBU) were examined and compared to each other at variable pH (1.2 and 6.5) in two separate dissolution systems and in one discontinuous dissolution system mimicking GIT conditions. The results of dissolution experiments were supported by kinetic model data. This study demonstrated that the dissolution of IBU from the CS-IBU lyophilizates is affected by two main pH-dependent competitive effects; i.e., dissolved CS acts as an IBU solubilizer and the undissolved CS matrix serves as an IBU trap, which could be used in the rational design of innovative stimuli (pH)-responsive oral dosage forms of IBU. Full article

The redox behavior of nickel complexes with sulfur-containing ligands remains of considerable interest due to their significant value in coordination chemistry, catalysis, and bioorganic modeling. In this context, it is important to investigate how aqueous media and acid–base equilibria influence the stability and transformation pathways of such complexes. In this work, the electrochemical behavior of nickel complexes with phosphorylated dithiocarbamate was studied using cyclic voltammetry at various scan rates and pH values. Compared to similar systems in organic solvents, the complexes exhibited additional oxidation and reduction signals, indicating coupled chemical steps. The pH dependence of these peaks confirmed the role of hydroxo groups in the oxidation processes. Varying the scan rate revealed competition between ligand exchange pathways. At low and moderate scan rates, tris-dithiocarbamate nickel(III/IV) complexes are formed, whereas at higher scan rates, hydroxo-containing compounds make a greater contribution. Based on the experimental results and standard redox potentials derived from quantum chemical calculation data, a general scheme for the resulting electrochemical processes was proposed. The results demonstrate the key role of aqueous media and pH in regulating the redox process of nickel complexes with phosphorylated dithiocarbamate. Full article

The Asian corn borer, Ostrinia furnacalis, is a major pest in China and across East and Southeast Asia, serving as the primary target of Bt maize expressing Cry proteins. Evolution of resistance to Bt toxins represents a critical challenge in plant protection. The high-dose/refuge strategy is more effective when resistance is recessively inherited and fitness costs are present. Here, we characterize the inheritance pattern and fitness costs of Cry1Ab resistance in O. furnacalis using a resistant strain exhibiting a resistance ratio of >1400-fold. The LC₅₀ values of F₁ hybrids from reciprocal crosses between resistant and susceptible strains were 2.44 (1.90–3.12) μg/g and 2.01 (1.53–2.61) μg/g, respectively, with no significant difference, indicating autosomal inheritance. The effective dominance (h) of F₁ offspring decreased with increasing concentration, suggesting that resistance was concentration-dependent. Analysis of observed versus expected mortality in backcross progeny (F₁ × resistant strain) indicated that Cry1Ab resistance is likely governed by more than one genetic locus. Compared with the susceptible strain, resistant individuals exhibited prolonged larval development (18.6 d vs. 17.2 d, p < 0.001), reduced pupation (42.5% vs. 60.8%, p < 0.001) and adult emergence rates (60.3% vs. 87.8%, p < 0.001), while fecundity was not significantly affected. These results verify the existence of fitness costs associated with Bt resistance. Our findings provide important insights into the mechanistic basis of Cry1Ab resistance and will assist in designing proactive management strategies to delay resistance evolution in field populations of O. furnacalis. Full article

Ubiquitin-specific protease 22 (USP22) regulates epigenetic gene expression by deubiquitinating histone H2B (H2Bub1) and upregulating oncogenic proteins and pathways, while antagonizing p53-mediated tumor suppression. USP22 is frequently overexpressed in cancers and associated with therapy resistance and poor prognosis yet remains largely untargeted pharmacologically. Here, using a fluorescence-based USP22 deubiquitinase assay to screen the LOPAC^®1280 library, we identified β-Lapachone, a natural ortho-naphthoquinone with strong anticancer activities, as a potent USP22 inhibitor. β-Lapachone potently inhibited USP22 enzymatic activity, with a half-maximal inhibitory concentration (IC₅₀) of ~0.75 μM, and molecular docking revealed its occupation of the catalytic pocket adjacent to the USP22 active-site triad, supporting a potential binding mode. Functionally, β-Lapachone suppressed proliferation and induced apoptosis in A549 and H1299 RAS-mutant lung adenocarcinoma (LUAD) cells, while USP22 knockout conferred marked resistance, indicating partial USP22 dependence. In patient-derived LUAD models, β-Lapachone inhibited sphere formation and reduced CD133⁺ cancer stem cell populations. Notably, it synergized with cisplatin to enhance DNA damage and apoptosis. In vivo, β-Lapachone significantly suppressed tumor growth in a syngeneic KRAS-mutant/p53-Null mouse lung cancer model and further potentiated cisplatin-induced antitumor effects. Collectively, these findings identify β-Lapachone as a potent inhibitor of USP22 and validate USP22 inhibition as a key mechanism underlying its anticancer activity in LUAD cells, both in vitro and in vivo. Full article

The migrating myoelectric complex (MMC) is the electrical basis of fasting small intestinal motility. Although oxytocin (OT) regulates gastrointestinal functions through oxytocin receptors (OTRs), its effect on jejunal MMC activity during fasting remains unclear. This study investigated the effects of OT on jejunal MMC activity in fasted rats and evaluated the involvement of OTRs, glucagon-like peptide-1 receptors (GLP-1Rs), and nitric oxide (NO) pathways. Bipolar electrodes were implanted at three jejunal sites in adult male Sprague Dawley rats for MMC recordings. After recovery and 18 h fasting, OT was administered intraperitoneally (4–32 µg/kg) following one hour of basal recording. To assess mechanisms, rats were pretreated with the OTR antagonist atosiban (2 mg/kg), the GLP-1R antagonist exendin (9–39) (200 µg/kg), or the nitric oxide synthase inhibitor NG-nitro-L-arginine (L-NNA; 5 mg/kg) before OT (16 µg/kg). Oxytocin dose-dependently reduced spike frequency and MMC cycle number (p < 0.05–0.001 vs. vehicle). Atosiban completely reversed these effects (p < 0.001 vs. OT), while exendin (9–39) partially attenuated them (p < 0.01–0.001 vs. OT). L-NNA showed no significant effect. These findings indicate that OT inhibits jejunal MMC activity via OTR-dependent mechanisms with partial involvement of GLP-1R signaling but not NO pathways. Full article

This study reports the preparation of a nanocomposite using a black cumin surface as a carbon framework on which zinc oxide-doped manganese ferrite nanoparticles were deposited and grown. A simple precipitation method was used to prepare the nanocomposite. The resulting composite was characterized using various characterization analyses such as FTIR, XRD, EDX, SEM, TEM, and TGA. The composite surface was highly conformed with functional groups, and the nanocomposite was formed due to electrostatic and non-electrostatic interactions between the carbon framework and the nanoparticles. X-ray analysis revealed a crystalline structure with crystal sizes up to 45 nm. Microscopic images revealed the surface morphology, confirming the irregular distribution of particles within the composite. The resulting composite material was used for adsorption application. The composite material was tested for the removal of Congo red dye from water. It was found that under optimal conditions, a dose of 2 g per liter of absorbent removed nearly 100% of dye from a 10 mL volume of 10 mg per liter Congo red solution within 90 min and 7 pH. A monolayer adsorption was confirmed by the isotherm analysis. The monolayer adsorption capacity for the present study was ~13.0 mg per gram. The adsorption kinetics suggested the fitting of pseudo-second order. Based on the findings, it was concluded that the chemical mechanism was responsible for the present adsorption process. The regeneration study demonstrates the stability of current adsorbent up to two cycles only. This nanocomposite is the first of its kind which promotes the creation of nanocomposites in the future by using natural materials and reduces the dependency on activated carbon. Full article