Search Results (925)

Breast cancer cells exhibit a reversed pH gradient and metabolic plasticity that promote proliferation, invasion, and resistance to therapy. Natural products such as chlorogenic acid (CGA) and cinnamaldehyde (CA) have shown emerging anticancer potential. However, their effects on intracellular pH and metabolic transport systems remain undefined. Therefore, the aim of this study was to characterize these parameters in breast cancer and non-tumorigenic breast cells. This study evaluated the physiochemical properties of CGA and CA using LC–MS, under pH conditions (pH 1.2, 7.4, and 9.0) mimicking the gastrointestinal track (GIT). Additionally, LC–MS-based human liver microsome (HLM) assays with NADPH were used to evaluate susceptibility to CYP-mediated metabolism to evaluate first-pass metabolic stability. Intracellular uptake kinetics were quantified at multiple time points using LC–MS. Following CGA:CA treatment, intracellular pH (pH_i) was measured in cancerous MDA-MB-231 and non-tumorigenic MCF-10A breast cell lines using SNARF-1 targeted ratio-metric fluorescence approach. Expression of OATP1B1, GLUT1, and MCT1 were analyzed by Western and immunofluorescence respectively, to assess potential cellular uptake of CGA:CA through OATP1B1 and their effects on glucose uptake and lactate and proton transport. Physiochemical results demonstrated that the compounds ranged from fully stable (pH 1.2 and 7.4) to completely unstable (pH 9.0). HLM incubation indicated no CYP-mediated hepatic metabolism. Treatment results showed that there was rapid intracellular uptake of CGA and CA in cancer cells and that CGA:CA lowered pH_i in both MDA-MB-231 and MCF-7 cells, while pH_i remained mostly unchanged in MCF-10A cells. Protein analysis revealed that CGA:CA treatment downregulated GLUT1 and MCT1 expression in cancer cells, suggesting impaired glycolytic activity and lactate shuttling. OATP1B1 expression was significantly suppressed in cancer cells, suggesting feedback inhibition of the solute carrier protein. Collectively, these findings indicate that CGA and CA exhibit favorable biochemical stability and disrupt intracellular pH regulation and metabolic transporter expression in breast cancer cells. Importantly, normal cells are not significantly affected. Thus, CGA:CA demonstrates therapeutic potential for breast cancer through pH_i and metabolic modulation. Full article

Vivipary adversely affects the production process of the cucumber seed industry and greatly limits the popularization of cucumber varieties. Identification of the cucumber seed vivipary phenotype and screening of vivipary-associated genes will provide important theoretical value and practical significance for solving this problem in agricultural production. In this study, cucumber near-isogenic lines with significant differences in vivipary traits (viviparous line F and non-viviparous line BF) were successfully screened and used to construct genetic populations. Bulk segregant analysis (BSA) and QTL-seq were performed to fine-map the major-effect quantitative trait locus associated with vivipary variation. Based on QTL and BSA analyses, CsaV3_3G044640 (designated CsCYP82D47), which encodes a cytochrome P450 family protein, was identified as a candidate gene associated with cucumber vivipary. CsCYP82D47 exhibits obvious tissue specificity and is highly expressed in leaves, sprouts, and seeds. However, no significant difference in CsCYP82D47 expression was detected between viviparous and non-viviparous cucumber materials. Further sequence analysis revealed multiple mutation sites in this gene between different genotypes. Specifically, the CYP82D47 protein in viviparous materials harbours one amino acid insertion (L63) and two missense mutations (M71L and S124L). In addition, the altered leucine residue distribution in viviparous cucumber may enlarge the substrate channel and enhance substrate catalytic efficiency, which may contribute to the vivipary phenotype. In summary, this study identifies a promising candidate gene potentially related to cucumber vivipary, which lays a foundation for further exploration of the molecular mechanism underlying cucumber vivipary and provides a potential genetic resource for cucumber molecular breeding. Full article

Soybean is a major legume crop in Togo. However, there is no scientific basis for guiding planting density recommendations to enhance yield. This study evaluated six soybean varieties across multiple planting densities in two major soybean-growing areas of Togo over two years. Data were collected on morphological traits and yield and were subjected to variance analysis, heritability estimate and mean comparison using R software version 4.4.1. Heritability was high (>0.8) for most traits except for number of branches per plant (0.54) and pod weight per plant (0.50). The local check variety (TGX1910) consistently outperformed introduced lines at all planting densities, except for hundred-seed weight. Plant height increased with density, whereas branch number, pods per plant, and grains per plant decreased, reflecting soybean’s self-regulation. Grain yield ranged from 1.17 t/ha at 200,000 plants/ha to 3.83 t/ha at 333,000 plants/ha. Despite reduced individual plant productivity at higher densities, plot yield increased up to an optimal density of ~333,000 plants/ha, before declining at the highest density of 450,000 plants/ha. Wider row spacing (60 cm) slightly enhanced yield compared to narrower spacing (40 cm). These results demonstrate that optimal planting density maximizes population-level yield, and planting soybean at 333,000 plants/ha with 60 cm row spacing is recommended for Togo. Full article

For large-scale floating offshore wind turbines, existing sensitivity studies have not fully addressed the combined effects of multiple uncertain input parameters on multiple output responses. Meanwhile, conventional Sobol indices quantify contribution magnitude but do not indicate effect direction. Based on the IEA 15 MW semi-submersible benchmark model, this study investigates the sensitivity of mooring tension and platform motion dynamic responses at a normal operating condition under power production. Integrated dynamic simulations were performed to generate response data. Eight uncertain parameters were considered, including the key mechanical and hydrodynamic coefficients of mooring lines as well as mass distribution and hydrodynamics-related key parameters for the platform. A polynomial chaos expansion surrogate model was used for the global sensitivity analysis, based on the Sobol index, Spearman coefficient, and a newly proposed modified Sobol index. The results indicate weak parameter interactions, with first-order Sobol indices dominating. The platform mass makes the largest contribution, with first-order Sobol indices approaching 1.0 for the mean tensions of all three mooring lines and 0.995 and 0.999 for the mean surge and heave displacements, respectively. The mooring line normal drag coefficient reaches a first-order Sobol index of 0.805 for the standard deviation of the upwind mooring line tension. The pitch response is influenced by multiple parameters. The Spearman coefficients confirmed the dominant parameters and identified their effect directions. By integrating variance contribution with effect direction, the modified Sobol index provides a more interpretable assessment of parameter effects. These findings can support parameter prioritization, mooring system design, and digital-twin model updating for floating offshore wind turbines. Full article

Adenoviruses are detected in wastewater at significantly higher frequencies than most other enteric viruses. The aim of this study was to determine the abundance of human adenoviruses in samples collected from wastewater treatment plants in Sofia, Plovdiv, Burgas, and Varna using quantitative PCR (qPCR). During the study period in 2024, one or two peaks in adenovirus levels were detected in each city, with viral loads ranging from 10 to 30,000 genome copies/mL. Selected wastewater samples from Sofia were used to inoculate three different cell lines. Productive adenovirus replication was observed only in the Vero E6 cell line. Shotgun sequencing of DNA extracted from infected cells enabled the assembly of six complete adenovirus genomes from three qPCR-positive samples. Taxonomic analysis identified viruses belonging to the Mastadenovirus adami, Mastadenovirus blackbeardi, Mastadenovirus caesari, and Mastadenovirus dominans species, with the predominant genotype (three of six genomes) showing the highest similarity to HAdV-A31. Comparative genomic analysis provided evidence of molecular evolution driven by multiple recombination events. The recovered genomes contained hypervariable regions composed of genetic elements closely related to genotypes previously reported from different geographical regions. Overall, the integrated approach employed in this study demonstrates the value of combining wastewater surveillance, virus isolation, and whole-genome sequencing to monitor adenovirus circulation, characterize dominant variants, and assess their potential clinical significance. Full article

Background: Manufacturing supply chains must increasingly coordinate cost, environmental impact, and service continuity under demand uncertainty and limited capacity. Methods: This study develops a stochastic mixed-integer linear programming framework for carbon-aware production–distribution planning in an automotive supply chain. The model jointly optimizes production quantities, inventory levels, shipments, truck usage, and lost sales over a multi-period horizon. Demand uncertainty is represented through scenarios, while production- and transportation-related emissions are monetized using an internal carbon price. Lost-sales penalties capture service degradation when demand cannot be fulfilled by the focal plant, and a rolling-horizon analysis evaluates planning responsiveness as demand information is updated. The framework is applied to an industrially inspired, capacity-constrained automotive case with multiple products, production lines, destinations, and demand scenarios. Computational experiments assess carbon pricing, lost-sales penalties, demand volatility, deterministic versus stochastic planning, and rolling-horizon replanning. Results: Results show that carbon pricing mainly acts as an economic valuation mechanism under the studied fixed-structure configuration, whereas lost-sales penalties strongly influence service performance. Demand volatility increases unmet demand, and lower emissions may reflect lower fulfilled demand rather than improved efficiency. Conclusions: The study provides a decision-support framework for evaluating cost–carbon–service trade-offs under stochastic demand while acknowledging single-plant and fixed-routing limitations. Full article

Background: Circular RNAs (circRNAs) constitute an emerging research field, as these RNA molecules play a crucial role in cellular functions and the progression of various human pathologies. Little is known about alternative circularization leading to the formation of distinct circRNAs from the same primary transcript, the role of circRNAs with slightly different back-splice junctions (BSJs) resulting in very similar circRNA sequences—called circRNA isoforms—and the extent to which the same primary transcripts produce alternative circRNAs. In this study, we discovered alternative circRNAs produced by two apoptosis-related genes, BAX and BCL2L12, expressed in established human cell lines originating from myelodysplastic syndrome (MDS) and different types of acute myeloid leukemia (AML). Methods: After total RNA extraction from one MDS cell line and five AML cell lines, first-strand cDNA synthesis, and multiple nested PCRs with distinct sets of divergent primers (10 and 16 primer pairs for BAX and BCL2L12 circRNAs, respectively) annealing in each exon of BAX and BCL2L12 genes, amplicon libraries were prepared and sequenced by both nanopore sequencing and NGS. Detailed bioinformatic analysis was then performed, based on existing bioinformatic tools and our own algorithms. Results: Our approach led to the identification of 72 BAX circRNAs and 52 BCL2L12 circRNAs with distinct expression patterns in MDS and AML cell lines. Most of these circRNAs—either merely exonic or exonic–intronic—were detected for the very first time. Furthermore, several BAX circRNA isoforms were detected in a unique cell line. Moreover, the back-splice sites joined together to form the BSJ of each circRNA were non-canonical, in many cases. The identified circRNAs are predicted to sponge distinct sets of miRNAs, some of which are known to regulate the activity of pivotal pathways. Conclusions: Overall, our findings support the notion that alternative splicing and back-splicing lead to the production of tens of distinct circRNAs from the same human gene. Full article

Background: Cancer cells live in a dynamic and favorable ecosystem conducive to their growth and survival, known as the tumor microenvironment (TME). The TME is replete with various proinflammatory cell types that mediate crosstalk via the secretion of cytokines and chemokines to facilitate tumor growth and development. One cardinal proinflammatory cytokine that is present in the TME is interleukin-1 (IL-1). IL-1 promotes tumor angiogenesis and cancer cell metastasis; thus IL-1 receptor antagonist (IL-1Ra) is of clinical interest. Our lab previously reported that chronic exposure to exogenous IL-1 can select for cancer cells that evolve insensitivity to IL-1 signaling, thus rendering IL-1-targeting therapies, like IL-1Ra, irrelevant. While immune cells are the primary source of TME IL-1, cancer cells can also produce and secrete IL-1 to engage in autocrine and/or paracrine signaling. In this context, cancer cell exposure to multiple other sources of exogenous IL-1 beyond autocrine production might also amplify extrinsic IL-1 signaling in these cells, but the consequences of sustained amplified IL-1 signaling on the regulation, function, and therapeutic response of these cancer cells need to be explored. Methods: Using the IL-1-secreting 5637 bladder cancer (BlCa) cell line, we generated chronic IL-1 sublines by spiking the growth medium with additional IL-1α or IL-1β chronically for 6 months. Once established, we assessed subline acute IL-1 sensitivity, paracrine signaling and response to IL-1Ra. Results: Following chronic exposure to elevated exogenous IL-1 levels, the 5637 BlCa cell line retains sensitivity to acute IL-1 and IL-1Ra and maintains the ability to induce IL-1-dependent endothelial cell activation, which is reversed with IL-1Ra. These data suggest that for cancer cells that engage in cell autonomous IL-1 signaling, sustained extrinsic IL-1 exposure does not dampen IL-1 or IL-1Ra sensitivity, supporting the context-dependent use of IL-1 antagonists as rational therapeutics in both acute and chronic inflammatory TMEs. Full article

Lycopene liquid beverages are functional products with strong antioxidant activity, but their quality control is hindered by the lack of rapid and simultaneous detection methods for multiple indicators. This study investigated a near-infrared spectroscopy (NIRS)—based approach for the simultaneous quantification of pH, moisture, and lycopene content in lycopene liquid beverages. A combined preprocessing strategy integrating Savitzky-Golay smoothing and standard normal variate (SG-SNV) was optimized via grid search to reduce spectral noise and scattering effects. Model interpretability was further enhanced by SHapley Additive exPlanations (SHAP) analysis. Furthermore, four variable selection algorithms (MWPLS, UVE-SPA, ICO, and CARS) were compared to improve model performance and interpretability. The results showed that SG-SNV preprocessing notably improved the prediction performance for pH (R²_CV = 0.9398, R²_PRE = 0.9624) and lycopene (R²_CV = 0.9860, R²_PRE = 0.9576), while moisture prediction remained at a comparable level (R²cv = 0.9617, R²_PRE = 0.9727). SHAP analysis identified spectral regions that may be associated with each component, providing chemically plausible explanations. For variable selection, the optimal algorithm differed across the three quality indicators. For moisture, CARS achieved the highest prediction performance (R²_PRE = 0.9946), while ICO provided comparable accuracy with slightly better model stability. For pH, UVE–SPA produced the strongest generalization ability (R²_PRE = 0.9705) with only 5 selected variables. For lycopene, none of the variable selection methods improved upon the preprocessing-only model (R²_PRE = 0.9576); MWPLS retained a parsimonious 21-variable model but with reduced predictive accuracy (R²_PRE = 0.8888). This study explores a preliminary rapid and non-destructive NIR analytical approach for the simultaneous detection of multiple quality indicators in lycopene liquid beverages, suggesting the feasibility of chemometric modeling for this application. The results provide a preliminary foundation for future development of rapid multi-indicator analytical methods. However, this work was conducted entirely under controlled laboratory conditions using a small, gradient-designed sample set with no production-line or online validation; it should therefore be regarded as a proof-of-concept feasibility study. Substantial further work—including validation with larger, naturally variable sample cohorts—would be needed before the approach could be considered for practical deployment. Full article

Background: Among natural products, vanillin (Van), a major component of Vanilla planifolia, exhibits multiple bioactivities, including antimicrobial effects. Methods: In this study, Van, its analogues o-vanillin (oVan), iso-vanillin (iVan), ethylvanillin (eVan), and a library of newly synthesized derivatives were evaluated against Helicobacter pylori strains with distinct antibiotic susceptibilities. Time-kill kinetics, antibacterial spectrum, and viability in a normal gastric cell line GES-1, were also assessed. Results: Van showed minimal or no activity (MIC and MBC > 128 µg/mL), whereas structural modifications markedly improved anti-H. pylori activity, with MIC values as low as 4 µg/mL. Compounds 16V, 20oV, and 29eV were among the most potent (MIC₉₀ = 4–16 µg/mL). Activity depended on both the vanilloid core and substituent type. The compounds were inactive against representative Gram-negative and Gram-positive bacteria (MIC > 128 µg/mL). Selected compounds preserved viability in GES-1 cells. Hierarchical clustering, artificial neural clustering, and principal component analysis identified potency-related architectural motifs and strain-specific activity. Docking against H. pylori urease suggested that several compounds, particularly 16V, may interact with the enzyme, providing preliminary support for a possible involvement of this target. Conclusions: Systematic modification of the vanilloid scaffold generated selective and relatively non-cytotoxic anti-H. pylori hit compounds and confirmed the value of natural metabolites in antibacterial drug discovery. Full article

Adenylate cyclase 5 (Adcy5) generates cyclic adenosine monophosphate (cAMP) downstream of G protein-coupled receptor signaling, yet its role in vertebrate pigmentation remains incompletely understood. Here, we generated a CRISPR/Cas9-mediated adcy5 knockout line in Nile tilapia (Oreochromis niloticus) to investigate its function in chromatophore biology. Loss of adcy5 resulted in a pronounced disruption of body coloration, characterized by the absence of vertical black bars and a global reduction in pigmentation. Despite this, chromatophore number was largely unaffected, indicating that Adcy5 is not required for pigment cell specification but is essential for functional pigmentation. At the cellular level, pigment granules in melanophores and erythrophores failed to undergo dispersion and instead remained constitutively aggregated, revealing a primary defect in intracellular pigment granule transport. Consistently, adcy5 mutants exhibited reduced expression of key melanogenesis-associated genes, including mitfa, tyrb, tyrp1a, and tyrp1b, accompanied by decreased melanin content across multiple tissues. Pharmacological activation of cAMP signaling partially rescued the pigment dispersion defect, whereas stimulation of upstream α-MSH signaling produced only limited effects, placing Adcy5 upstream of intracellular cAMP production within the pigment regulatory cascade. Importantly, we further demonstrate that Adcy5 is required for erythrophore pigment granule dispersion, extending its functional role beyond melanophore biology. Together, these findings identify Adcy5 as a conserved regulator integrating cAMP-dependent pigment synthesis and granule transport across multiple chromatophore types in teleost fish. Full article

Edge chamfering is a critical process in glass manufacturing because it suppresses crack propagation and improves both edge strength and operational safety. As such, it has become an essential step in modern glass production. However, in current automated production lines, chamfer quality inspection still relies heavily on manual visual inspection and contact-based measuring instruments, which are limited by low efficiency, strong subjectivity, and poor compatibility with production-line takt time. To overcome these limitations, this paper proposes a vision-based inspection method for transparent glass chamfers using corner analysis, and a dedicated glass chamfer inspection system is developed for experimental validation. First, machine-vision image processing is applied to enhance chamfer-region features and improve detection robustness. Candidate endpoint generation is then combined with the construction of an endpoint-screening coordinate system to achieve stable localization of chamfer endpoints. In addition, quadrant-based spatial constraints are introduced to further enhance endpoint localization stability, enabling automatic measurement of key geometric parameters, including chamfer length, width, and height. On this basis, multiple defect-discrimination criteria are established by integrating geometric tolerance violations with spatial abnormalities in endpoint distribution, thereby enabling automatic identification of chamfer defects. Experimental results show that the proposed method achieves a mean absolute measurement error of 0.087 mm and an average inspection time of 0.505 s per workpiece. The method can also effectively identify chamfer defects, demonstrating its suitability for online inspection while maintaining both dimensional measurement accuracy and defect detection capability. Full article

Geochemical and isotopic analysis of bitumen-lined transport container vessels, so called ‘torpedo jars’, from the Sasanian to Early Islamic period fort of Fulayj in Oman confirms the presence of two distinct compositional categories that can be matched to contemporary sources in different areas of southwest Iran. It appears that the bitumen used to line jars was extracted from different geographic areas, hinting at the existence of multiple production locations for this vessel class. In terms of the function of the jars, organic residue analysis provides a positive identification of biomarkers associated with the storage and transportation of red wine. These results provide the first published confirmation of the long-suspected association between torpedo jars and wine transportation from the core urbanised zone of the Sasanian and Early Islamic imperial heartlands towards the Persian Gulf and Indian Ocean maritime sphere. The results have added significance as the samples are derived from relatively accurately dated archaeological contexts spanning the period before and after the Islamic conquest. Full article

Alkaline soil limits soybean production. This study elucidates the molecular mechanism by which the Gshdz4-GmU2AFb-GmCML27 module regulates soybean alkaline tolerance. Transcriptome analysis, yeast one-hybrid and dual-luciferase assays confirm that the HD-Zip transcription factor Gshdz4 binds to the CAATAA motif in the GmU2AFb promoter and activates its transcription. Subcellular localization verifies the nuclear distribution of GmU2AFb. Overexpression of GmU2AFb improves alkaline tolerance by increasing antioxidant enzyme activities and proline levels, reducing MDA accumulation, facilitating root development, and upregulating alkaline-responsive genes, including GmSOD1, while gene knockout impairs stress resistance. Combined Y2H, BiFC and LCI assays validate the nuclear protein interaction between GmU2AFb and the calcium-binding protein GmCML27, and overexpression of GmCML27 also enhances antioxidant capacity and root growth in soybean. Co-overexpression of the two genes generates obvious synergistic effects; compared with single-gene overexpression lines, co-transgenic plants possess higher antioxidant levels and elevated transcription of downstream alkaline-tolerant genes, accompanied by alleviated growth inhibition under alkaline stress. In summary, Gshdz4 transcriptionally activates GmU2AFb, and the interaction between GmU2AFb and GmCML27 connects RNA splicing with calcium signaling pathways to synergistically trigger downstream defense responses and promote root development, thereby enhancing soybean alkaline tolerance at multiple layers. This work provides candidate genes for molecular breeding of alkali-resistant soybean. Full article

Background: Multiple myeloma (MM) is a mature B-cell disorder characterized by the excessive production of monoclonal immunoglobulins. It is the second most common hematological cancer and predominantly affects older adults. In 2022, there were approximately 188,000 MM new cases worldwide. The disease is associated with a relapsing–refractory course. First-line therapies are often insufficient, making additional treatment options necessary. For patients refractory to lenalidomide and proteasome inhibitors (bortezomib and/or carfilzomib), pomalidomide combined with dexamethasone and an additional active agent can be used as a therapeutic strategy. Objective: This study was conducted to evaluate the bioequivalence and tolerability of two pomalidomide hard capsule formulations to support regulatory approval of a branded generic product in Latin American countries. Methods: An open-label, randomized, single-dose, two-treatment, two-sequence, two-period crossover study was conducted in healthy male subjects. Participants received a single oral dose of the test product, Xetrane^® 4 mg hard capsule (Laboratório LKM S.A., Argentina), and the reference product, Imnovid^® 4 mg hard capsule (Celgene International Sàrl), with a 7-day washout period. Blood samples were collected over 48 h post-dose. Plasma pomalidomide concentrations were determined using a validated LC-MS/MS method, and pharmacokinetic parameters were estimated using non-compartmental analysis. Findings: Thirty-four subjects were enrolled, and 29 completed the study. Geometric mean ratios (90% confidence intervals) for Cmax and AUC_0–t were 106.24% (100.77–112.00) and 94.67% (91.61–97.82), respectively. Both formulations were well tolerated. Bioequivalence between Xetrane^® and Imnovid^® was demonstrated in accordance with regulatory criteria. NCT07694011. Full article