Search Results (4,790)

Background: Gandouling (GDL) is a compound prepared in Chinese medicine and demonstrates favorable clinical efficacy. Studies have shown that sinusoid capillarization promoted hepatic fibrosis and was a potential target for preventing and treating liver fibrosis in Wilson’s disease (WD). This study aimed to explore whether GDL inhibited the sinusoid capillarization in WD by blocking the communication between hepatic stellate cells (HSCs) and liver sinusoidal endothelial cells (LSECs). Methods: In this study, Atp7b-H1071Q (TX) mice were used as the WD model mice, and CuSO₄⋅5H₂O treated LX-2 cells were used as the HSC activation model. We used scanning electron microscopy, vascular tube formation assay, Western blot, cell transfection, and co-culture system to study how GDL blocked the communication between HSCs and LSECs, as well as its inhibitory effect on the sinusoid capillarization. Results: We found that GDL alleviated liver fibrosis in TX mice, inhibited HSC activation, and sinusoid capillarization in TX mice. Excessive secreted VEGFA by LX-2 cells promoted the sinusoid capillarization, played the role of a messenger molecule, and GDL blocked the VEGFA-mediated HSCs-LSECs communication. Furthermore, bioinformatics analysis, molecular docking, and molecular dynamics suggested that GDL may exert its effect by modulating the PDGFRβ/ERK/VEGFA signaling axis. We validated the above observation through experiments, that GDL reduced PDGFRβ/ERK signal pathway in LX-2 cells, inhibited the expression of messenger molecule VEGFA, blocked HSCs-LSECs communication, inhibited sinusoid capillarization, and improved WD. Conclusions: GDL blocked the communication between HSCs and LSECs and inhibited the sinusoid capillarization associated with liver fibrosis in WD by the PDGFRβ/ERK/VEGFA signaling axis. Full article

People living with HIV-1 (PLWH) are part of the so-called “fragile” populations to which COVID-19 vaccines were/are strongly recommended. The fact that most widely used COVID-19 vaccines rely on the production of a biologically active SARS-CoV-2 Spike protein expressed by synthetic mRNA poses the relevant question of whether and how this vaccination influences the fate of the HIV-1 reservoir. This report presents a detailed analysis of the literature data on the effects of SARS-CoV-2 Spike and COVID-19 vaccines on HIV-1 latently infected cells. Despite being limited in number, the experimental evidences consistently indicate that vaccine mRNA and/or SARS-CoV-2 Spike can effectively reactivate latent HIV-1. This conclusion has been drawn after “in vitro”, “ex vivo”, and “in vivo” assays, and with virus-associated Spike, soluble Spike, or its intracellular expression, as well as with COVID-19 mRNA vaccines. On the other hand, real-world observations on vaccinated PLWH under antiretroviral therapy (ART) provided evidence of HIV-1 reactivation almost exclusively in PLWH with unsuppressed viremia, as measured in terms of size of the HIV-1 reservoir. Although several issues still need to be clarified through urgent additional investigations, these data suggest the possibility that the Spike protein and/or the vaccine mRNA molecules affect the HIV-1 latency in PLWH. Full article

Cancer patients are highly vulnerable to severe COVID-19, requiring models that capture tumor–virus interactions. We investigated tumor- and variant-specific effects of SARS-CoV-2 Gamma and Delta infections using patient-derived organoids (PDOs) from metastatic breast, lung, and colorectal cancers. Viral infection was quantified by Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) 24 h post-infection, and morphological changes and immune mediators were profiled. Genomic analysis using whole-exome sequencing was performed to identify contributing host-related gene alterations. The Delta variant produced consistently higher viral loads in lung and breast PDOs, while colorectal PDOs showed variable susceptibility. Infection led to reduced area and perimeter and increased circularity across all tumor types. Immune profiling revealed distinct responses: Gamma decreased Interferon alpha (IFNα) in lung PDOs and increased E-selectin in colorectal PDOs. Delta broadly reduced inflammatory mediators in lung [10 kDa interferon gamma-induced protein (IP-10) and Intercellular adhesion molecule 1 (ICAM-1)] and breast [Interleukin-6 (IL-6), Interleukin-13 (IL-13), and Interleukin-17A (IL-17A)] PDOs, while increasing Macrophage inflammatory protein 1-beta (MIP-1β) in colorectal PDOs. Host gene variants involved in trafficking (FYCO1 and RAB7A) and immune signaling (FOXA2, SFTPD, STAT3, and TET2) were associated with differential infection profiles. These findings show that SARS-CoV-2 induces variant- and tumor-specific morphological and immunological changes in cancer PDOs, highlighting the potential of this model to unravel host–virus interactions and identify genetic factors that shape infection outcomes in cancer. Full article

Flammulina filiformis is a widely cultivated edible mushroom valued for its taste and nutrition. However, its stipe often develops a fibrous and stringy texture that unpleasantly lodges between teeth during chewing. Texture analysis confirmed a distinct toughness gradient, with the upper stipe being more brittle and less tough than the lower part. UHPLC-MS/MS-based metabolomics of these regions identified 953 metabolites, predominantly spanning lipids and lipid-like molecules, organic acids and derivatives, and nucleosides, nucleotides, and analogues. Comparative analysis revealed that the tender upper stipe was characterized by a widespread downregulation of primary metabolites, including severe depletion of key signaling molecules (cAMP, cGMP) and amino acids such as L-tryptophan. In contrast, the tough lower stipe was enriched with metabolites indicative of an oxidative environment, notably a broad spectrum of oxidized lipids and phenolic compounds. KEGG pathway analysis attributed this dichotomy to distinct metabolic programs. While the upper stipe exhibited downregulation in tryptophan and purine metabolism, the lower stipe was enriched for pathways associated with redox homeostasis and lipid peroxidation, including glutathione metabolism and lipid peroxidation. The co-accumulation of oxidized lipids and phenolics suggests a potential mechanism for oxidation-driven tissue fortification. This study reveals a spatially programmed metabolic basis for the textural differentiation in F. filiformis stipes, providing a framework for understanding tissue development and highlighting potential regulatory targets for breeding varieties with improved eating quality. Full article

The creation of a specific culture medium for colorectal organoids in 2011 heralded a new era in human primary cultures by enabling the indefinite expansion of normal and pathological epithelial organoids. The original formula has been used ever since, with only minor, lab-specific modifications. The goal of culturing organoids from different tissues has relied on saving and propagating the pluripotent stem cell. The “magic bullet” and all its subsequent derivatives have pursued this goal. Consequently, agonist and antagonist signals are chronically activated in the organoid medium, forcing organoid cells (as well as any other co-cultured cellular model) into constrained signaling pathways. This extremely artificial condition is often overlooked in experimental approaches and may bias the results. Furthermore, some molecules in the organoid medium have unpredictable off-target effects that significantly impact the behavior and maturation of certain cell populations. Nicotinamide, gastrin and PGE2 inhibit immune responses. SB202190, A83-01 and vanadate (from advanced DMEM-F12) modify intracellular signaling. N-AcetylCysteine and Primocin modify the redox response and mitochondrial metabolism, respectively. Thus, the unintentional addition of these molecules to the organoid medium introduces biases under specific experimental settings. While the original organoid medium formula is the gold standard for propagating organoids in vitro, more focused, reliable conditions are necessary for specific organoid-based tests. Full article

A library of thirty-one quinazoline derivatives was assessed as potential inhibitors of epidermal growth factor receptor kinase (EGFR) using 3D-QSAR methods, namely Comparative Molecular Field Analysis (CoMFA) and Comparative Molecular Similarity Indices Analysis (CoMSIA). Training and test sets were generated by aligning the molecules to the lowest-energy conformer of the most active compound. The optimized models exhibited strong statistical performance, with R² values of 0.981 (CoMFA) and 0.978 (CoMSIA), and cross-validation coefficients (Q²) of 0.645 and 0.729, respectively. External validation confirmed their predictive power, yielding R² values of 0.929 and 0.909. Guided by these models, eighteen new quinazoline candidates were designed and evaluated for drug likeness and ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) properties using in silico approaches. Molecular docking and molecular dynamics simulations highlighted the binding features and stability of these derivatives, with compound Pred65 demonstrating superior affinity and stability compared to Erlotinib. Collectively, the study provides valuable insights for the optimization of quinazoline scaffolds as EGFR inhibitors, supporting the development of promising anticancer leads. Full article

Intracerebral hemorrhage (ICH) is a type of stroke with high mortality and disability rates. The hemoglobin and iron ions released by ruptured red blood cells after ICH can induce programmed cell death characterized by lipid peroxide accumulation—a defining feature of ferroptosis—which is one of the key mechanisms for the occurrence and progression of secondary brain injury after ICH. Betaine (BET), a natural amino acid derivative, is known to be an antioxidant, but its protective effect and molecular mechanisms in ICH-induced ferroptosis have not been studied yet. In this study, we investigated the effect of BET intervention on ICH-induced ferroptosis and possible mechanisms in vitro and in vivo, and we evaluated the expression of ferroptosis and oxidative stress molecules through in vivo and in vitro experiments. We analyzed the distribution of nuclear factor E2-related factor 2 (Nrf2) and assessed neurobehavioral function, hematoma volume, and iron content in the brain tissue of mice with ICH. BET upregulates nuclear factor E2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) signaling, reducing long-chain acyl-CoA synthetase 4 (ACSL4), reactive oxygen species (ROS), and malondialdehyde (MDA) while increasing glutathione (GSH) and glutathione peroxidase 4 (GPX4) levels. It also decreases brain iron accumulation, aids hematoma clearance, and protects against ferroptosis and oxidative damage post ICH. Inhibition of Nrf2 with ML385 diminishes BET’s neuroprotective effects, highlighting the pathway’s importance in BET’s mechanism of action. BET boosts antioxidant capacity via the Nrf2/HO-1 pathway; inhibits ferroptosis; reduces oxidative stress, brain edema, and iron accumulation post ICH; and aids hematoma clearance, offering neuroprotection. Full article

Vehicle exhaust gases remain one of the key sources of atmospheric air pollution and pose a serious threat to ecosystems and public health. This study presents an experimental investigation into reducing the toxicity of gasoline internal combustion engine exhaust using ultrasonic waves and infrared (IR) laser exposure. An original hybrid system integrating an ultrasonic emitter and an IR laser module was developed. Four operating modes were examined: no treatment, ultrasound only, laser only, and combined ultrasound–laser treatment. The concentrations of CH, CO, CO₂, and O₂, as well as exhaust gas temperature, were measured at idle and under operating engine speeds. The experimental results show that ultrasound provides a substantial reduction in CO concentration (up to 40%), while IR laser exposure effectively decreases unburned hydrocarbons CH (by 35–40%). The combined treatment produces a synergistic effect, reducing CH and CO by 38% and 43%, respectively, while increasing the CO₂ fraction and decreasing O₂ content, indicating more complete post-oxidation of combustion products. The underlying physical mechanisms responsible for the purification were identified as acoustic coagulation of particulates, oxidation, and photodissociation of harmful molecules. The findings support the hypothesis that combined ultrasonic and laser treatment can enhance real-time exhaust gas purification efficiency. It is demonstrated that physical treatment of the gas phase not only lowers the persistence of by-products but also promotes more complete oxidation processes within the flow. Full article

ORF8 is the second most mutated protein in SARS-CoV-2. It can form oligomers such as trimers and can bind to the IL-17RA/RC receptor. To understand the possible role of ORF8 in SARS-CoV-2, the first step of this study involved predicting the ORF8 trimer structure and the complex structure of the ORF8 monomer bound to the IL-17RA receptor using docking and molecular dynamics simulation methods. It was found that ORF8 molecules bound to the central ORF8 molecule through covalent and noncovalent interactions exhibit similar RMSD and RMSF values as the central ORF8 molecule and form a similar buried surface area, but display different numbers of hydrogen bonds and varying dynamic correlations. Additionally, trimer formation increases the dynamic correlation of the noncovalently bound ORF8 unit. ORF8 can bind with the IL-17RA receptor stably. Regions on ORF8, including C25–I47, L60–S67, T80–C90, and S103–E110, and regions on IL-17RA, including L1–H63 and D122–M165, are involved in the binding interface of the complex. ORF8 becomes less rigid when bound to IL-17RA than in its monomer, dimer, and trimer forms. Based on dihedral angle correlation predictions, binding of ORF8 to IL-17RA reduces internal correlations within ORF8 while strengthening correlations within IL-17RA. The G50–T80 region of ORF8 appears to be critical for interaction with IL-17RA, and the L1–V150 region of IL-17RA should be critical for its dynamics once bound to ORF8. These results help elucidate the structure and dynamics of ORF8 in SARS-CoV-2. Full article

Melatonin is recognised as a multifunctional regulatory molecule that enhances plant tolerance to abiotic stresses, but its effectiveness is often strongly genotype-dependent. This study aimed to elucidate how exogenous melatonin (200 µM) modulates the physiological and biochemical responses of wheat during drought and subsequent recovery in two genotypes with contrasting grain pigmentation: the standard cv. Bohemia (red grain) and an experimental purple-pericarp (PP) line. Plants were exposed to drought at the early vegetative stage (BBCH 15), and gas exchange, leaf water potential, and biochemical markers (proline, malondialdehyde, phenolics, and flavonoids) were assessed during drought and after rehydration. In cv. Bohemia, water deficit led to a pronounced decrease in CO₂ assimilation, stomatal conductance, and leaf water potential, accompanied by strong increases in proline (Pro) and malondialdehyde (MDA). Melatonin application in this genotype markedly reduced the accumulation of Pro and MDA and accelerated the recovery of gas exchange, indicating a significant protective effect. The lower Pro levels in melatonin-treated Bohemia plants suggest that melatonin mitigated the perceived stress intensity, thereby reducing the physiological demand for osmotic adjustment. In contrast, the PP line exhibited higher inherent stability of the photosynthetic apparatus and more moderate biochemical shifts; its recovery was almost complete and independent of melatonin. Overall, these results indicate that the functional benefit of exogenous melatonin is greater in genotypes with a lower intrinsic stress-buffering capacity. This study highlights the importance of considering constitutive genotype traits and the recovery phase when using physiological regulators to improve wheat drought resilience. Full article

SARS-CoV-2, the virus responsible for coronavirus disease COVID-19, is a highly transmissible pathogen that has caused substantial global morbidity and mortality. The ongoing COVID-19 pandemic caused by this virus has had a significant impact on public health and the global economy. One approach to combating COVID-19 is the development of broadly neutralizing antibodies for prevention and treatment. In this work, we performed an in silico ligand-based screening of the PDB database to search for unique anti-SARS-CoV-2 motifs. The collected data were organized and presented in a classified SARS-CoV-2 Ligands Database, categorized based on the number of ligands and structural components of the spike glycoprotein. The database contains 1797 entries related to the structures of the spike glycoprotein (UniProt ID: P0DTC2), including both full-length molecules and their fragments (individual domains and their combinations) with various ligands, such as angiotensin-converting enzyme II and antibodies. The database’s capabilities allow users to explore various datasets according to the research objectives. To search for motifs in the receptor-binding domain (RBD) most frequently involved in antibody binding sites, antibodies were classified into four classes according to their location on the RBD; for each class, special binding motifs are revealed. In the RBD binding sites, specific tyrosine-containing motifs were found. Data obtained may help speed up the creation of new antibody-based therapies, and guide the rational design of next-generation vaccines. Full article

Background and Objectives: SARS-CoV-2 produces potentially pathogenic molecules, such as single-stranded RNA and spike proteins, which can potentially activate microglial cells. In this study, we aimed to investigate whether SARS-CoV-2 spike protein S1 and mRNA vaccines can cause neurotoxicity directly or through microglial involvement. Materials and Methods: Primary cerebellar granule cell cultures isolated from Wistar rats and organotypic hippocampal slice cultures from transgenic C57BL/6J mice were used in the experiments. Imaging and quantitative analysis of cell viability, proliferation, and phagocytic activity were performed using light and fluorescence microscopy. Results: The exogenous SARS-CoV-2 S1 protein at 50 µg/mL concentration induced neuronal cell death in neuronal–glial co-cultures and stimulated microglial proliferation during the first 3 days of exposure without an effect on inflammatory cytokine secretion. Single application of Tozinameran/Riltozinameran and Original/Omicron BA. 4–5 vaccines did not affect neuronal viability and total neuronal number in cell co-cultures after 7 days of exposure. In contrast, three repeated treatments with mRNA vaccines at 6 ng/mL caused microglial proliferation without affecting microglial phagocytosis and TNF-α release. In organotypic brain slice cultures, only Tozinameran/Riltozinameran stimulated microglial cell proliferation in female brain slices, while male brain slices remained unaffected by both vaccines, indicating sex-dependent effects. Conclusions: The findings suggest that mRNA vaccines do not exert neurotoxic effects in primary neuronal–glial co-cultures, but induce microglial proliferation, particularly in female brains in the absence of inflammatory cytokine release. SARS-CoV-2 S1 protein at high concentrations directly induces neuronal death. Full article

Breast cancer is a heterogeneous disease that exists in multiple subtypes, some of which still lack targeted and effective therapy. A major challenge is to unravel their underlying molecular mechanisms and bring to light novel therapeutic targets. In this study, we investigated the role of WNT-inducible signaling pathway protein 1 (WISP1) matricellular protein in the acquirement of an invasive phenotype by breast cancer cells. To this aim, we treated non-invasive MCF7 cells with WISP1 and assessed the expression levels of macrophage migration inhibitory factor (MIF) and its cellular receptor CD74. Next, we examined the expression of epithelial-to-mesenchymal transition (EMT) markers as well as molecular effectors of the tumor microenvironment, such as CD44, the main hyaluronan receptor that also acts as a co-receptor for MIF, the hyaluronan oncogenic network, and specific matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs). The results showed that WISP1 potently induces the expression of MIF cytokine and affects the expression of specific extracellular matrix molecules with established roles in the promotion of malignant properties. Notably, Src kinases and MIF are critically involved in these processes. Collectively, the present study demonstrates for first time a WISP1/Src/MIF axis as well as its ability to induce an invasive phenotype in MCF7 cells and highlights novel cellular and molecular processes involved in the epithelial-to-mesenchymal transition and the development of invasive breast cancer. This suggests that specific cues from the tumor microenvironment can activate a migratory/invasive phenotype in a subpopulation of cells residing within the heterogeneous breast tumor. Full article

Background/Objectives: Post-COVID-19 pulmonary fibrosis (PCPF) and idiopathic pulmonary fibrosis (IPF) exhibit significant clinical and pathophysiological overlap, suggesting convergent molecular pathways driving fibrosis. This prospective longitudinal study investigates the sustained dysregulation of matrix metalloproteinases (MMP)-2 and MMP-9 and its relationship with evolving systemic inflammation and endothelial dysfunction in convalescent COVID-19 patients, with comparative analysis to IPF. Methods: We conducted a prospective observational study of 86 patients at 6 and 12 months post-SARS-CoV-2 infection, stratified by high-resolution CT evidence of PCPF (FB+ group, n = 32) or absence of fibrosis (FB− group, n = 54). Gene expression of MMP-2 and MMP-9 in peripheral blood leukocytes and circulating levels of MMP-2, MMP-9, pro-inflammatory cytokines (TNF-α, IL-6), and endothelial dysfunction markers (Endothelin-1 [ET-1], adhesion molecules) were quantified via qRT-PCR and ELISA. A pre-pandemic healthy control group (HD, n = 20) and an IPF patient group (n = 10) served as comparators. Results: A significant, sustained elevation of MMP-2 and MMP-9 was observed in all post-COVID-19 patients versus HDs, most pronounced in the FB+ group and qualitatively similar to IPF. A critical divergence emerged: FB− patients showed resolution of systemic inflammation (reduced TNF-α, IL-6), whereas FB+ patients exhibited persistent cytokine elevation. Critically, a delayed, severe endothelial dysfunction, characterized by a profound surge in ET-1 and elevated adhesion molecules, manifested exclusively in the FB+ cohort at 12 months. Positive correlations linked plasma MMP-2/9 levels with ET-1 (r_s = 0.65, p = 0.004; r_s = 0.49, p = 0.009) and ET-1 with sICAM-1 (r_s = 0.68, p = 0.01). Conclusions: The development of PCPF is associated with a distinct pathogenic triad: sustained MMP dysregulation, failure to resolve inflammation, and severe late-phase endothelial dysfunction. The correlative links between these components suggest a self-reinforcing loop. This systemic signature mirrors patterns in IPF, underscoring shared final pathways in fibrotic lung disease and identifying the MMP–inflammation–endothelial axis as a promising target for biomarker development and therapeutic intervention. Full article

Volatile organic compounds (VOCs) produced by Medicinal Aromatic Plants (MAPs) are bioactive signaling molecules that play key roles in plant defense, acting against pathogens and triggering resistance responses. Intercropping with VOC-emitting MAPs can therefore enhance disease resistance. This study investigated VOCs emitted by sage (Salvia officinalis) as potential resistance inducers in grapevine (Vitis vinifera) against Plasmopara viticola, the causal agent of downy mildew, under consociated growth conditions. Sage and grapevine plants were co-grown in an airtight box system for 24 or 48 h, after which grape leaves were inoculated with P. viticola. Disease assessments were integrated with grapevine leaf metabolic profiling to evaluate responses to VOC exposure and pathogen infection. Untargeted and targeted metabolomic analysis revealed that sage VOCs consistently reprogrammed grapevine secondary metabolism, without substantial differences between 24 and 48 h exposures. Lipids, phenylpropanoids, and terpenoids were markedly accumulated following VOC exposure and persisted following inoculation. Correspondingly, leaves pre-exposed to sage VOCs exhibited a significant reduction in disease susceptibility. Overall, our results suggest that exposure to sage VOCs induces signaling and metabolic reprogramming in grapevine. Further research should elucidate how grapevines perceive and integrate these signals, as well as the broader processes underlying MAP VOC-induced defense, and evaluate their translation into sustainable viticultural practices. Full article