Search Results (11,138)

Exosomes, small extracellular vesicles secreted by various cell types, have gained significant attention in cancer investigations. Isolation and characterization of exosomes derived from DOK (dysplastic oral keratinocyte), SCC (squamous cell carcinoma) and HaCaT (normal skin keratinocyte) cell lines and microRNA profiling were conducted. Magnetic sorting was applied to obtain pure exosomes. Morphology and size were characterized by transmission electron microscopy and nanoparticle tracking analysis. Validation of membrane exosomal markers (CD9, CD63) was performed via Western blotting. MiR-21, miR-31, and miR-133 levels were analyzed in exosomes and parent cells by qPCR. Biological effects of the exosomes were tested by adding them to fibroblast cultures and determining the expression of relevant carcinogenesis markers by qPCR. Exosomes appeared as cup-shaped nano-sized particles, and there was no difference regarding particle diameter and concentration between the three types of exosomes. The oncogenic miR-21 was significantly upregulated both in SCC and SCC-derived exosomes compared to DOK and HaCaT cells and their respective exosomes. However, miR-31 unexpectedly showed the highest expression in normal cells and the lowest in HaCaT exosomes. MiR-133, the tumor suppressor miRNA, was downregulated in both SCC and DOK cells compared to normal (HaCaT) cells, while the opposite situation was observed in exosomes, with HaCaT cells showing the lowest levels of miR-133. The differences in exosome content were reflected in signaling pathway activation in exosome-treated fibroblasts, with SCC exosomes exerting the most potent effect on several cancer-related pathways, notably PIK3/AKT, PTEN, and NOTCH signaling cascades. Full article

With the large-scale closure of coal mines leading to groundwater pollution, in order to systematically identify the sources of major chemical ions in surface water and groundwater. This study comprehensively applied methods such as Piper’s trilinear diagram, linear fitting, and correlation analysis to quantitatively analyze the hydrochemical characteristics of closed coal mining areas in southwest Shandong and to clarify the sources of geochemical components in surface water and groundwater, and the PMF model was used to analyze the sources of chemical components in mine water and karst water. The results show that the concentrations of TDS ( Total Dissolved Solids), SO₄²⁻, Fe, and Mn in the mine water of the closed coal mine area are higher than in the karst water. Both water bodies are above groundwater quality standards. Ca²⁺, SO₄²⁻, and HCO₃⁻ dominate the ionic components in surface water and different types of groundwater. The hydrochemical types of surface, pore, and mine waters are mainly SO₄-HCO₃-Ca, whereas SO₄-HCO₃-Ca and HCO₃-SO₄-Ca dominate karst waters. SO₄²⁻ is the leading ion in the TDS of water bodies. The mineralization process of surface water is mainly controlled by the weathering of silicate minerals, while that of the groundwater is mainly controlled by the dissolution of carbonate minerals. The impact of mining activities on surface water and groundwater is significant, while the impact of agricultural activities on surface water and groundwater is relatively small. The degree of impact of coal mining activities on SO₄²⁻ concentrations in surface water, pore water, and karst water, in descending order, is karst water, surface water, and pore water. The PMF (Positive Matrix Factorization) model analysis results indicate that dissolution of carbonate minerals with sulphate and oxidation dissolution of sulfide minerals are the main sources of chemical constituents in mine waters. Carbonate dissolution, oxidation dissolution of sulfide minerals, domestic sewage, and dissolution of carbonate minerals with sulphate are ranked as the main sources of chemical constituents in karst water from highest to lowest. These findings provide a scientific basis for the assessment and control of groundwater pollution in the areas of closed coal mines. Full article

Lesch–Nyhan disease (LND) is associated with a complete deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT) activity due to mutations in the HPRT1 gene. Although the physiopathology of LND-related neurological manifestations remains unknown, a defective neuronal developmental process is the most widely accepted hypothesis. We generated an HPRT-deficient line from the pluripotent human embryonic cell line NT2/D1 by CRISPR-Cas9 and induced its differentiation along neuroectodermal lineages by retinoic acid treatment. As levels of folic acid in the culture media may affect results in LND models, we employed physiological levels of folate. The effect of HPRT deficiency on neural development-related gene expression was evaluated using two methodological approaches: a directed qPCR array of genes related to neuronal differentiation, and global gene expression by RNAseq. HPRT-deficient pluripotent cells presented altered expression of genes related to pluripotency in human embryonic stem cells, such as DPPA3 and CFAP95, along with genes of the homeobox gene family. HPRT-deficient pluripotent cells were able to differentiate along neuro-ectodermal lineages but presented consistent dysregulation of several genes from the homeobox gene family, including EN1 and LMX1A. GO enrichment analysis of up- and downregulated genes in HPRT-deficient cells showed that the most significant biological processes affected are related to development and nervous system development. Full article

Accumulating studies have shown that astrocytes are essential for regulating neurons at both synaptic and circuit levels. The main mechanism of brain astrocytic intracellular Ca²⁺ activity is through the release of Ca²⁺ via the inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) from the endoplasmic reticulum (ER). Studies using IP3R2 knockout mouse models (Itpr2^−/−) have shown that eliminating IP3R2 leads to a significant reduction in astrocytic Ca²⁺ activity However, there is ongoing controversy regarding the effect of this IP3R2-dependent reduction in astrocytic Ca²⁺ transients on neuronal activity. In our study, we employed dual-color two-photon Ca²⁺ imaging to study astrocytes and neurons simultaneously in vibrissa somatosensory cortex (vS1) in awake-behaving wild-type and Itpr2^−/− mice. We systematically characterized and compared both recorded astrocytic and neuronal Ca²⁺ activities in wild-type and Itpr2^−/− mice during various animal behaviors, particularly during the transition period from stillness to locomotion. We report that vS1 astrocytic Ca²⁺ elevation in both wild-type and Itpr2^−/− mice was significantly modulated by free whisking and locomotion. However, vS1 neurons were only significantly modulated by locomotion in wild-type mice, but not in Itpr2^−/− mice. Our study suggests a non-synaptic modulatory mechanism on functions of astrocytic IP3R2-dependent Ca²⁺ transients to local neurons. Full article

Background: Assessing pancreatic ductal adenocarcinoma (PDAC) resectability after neoadjuvant therapy (NAT) remains a diagnostic challenge. Traditional computed tomography (CT) criteria often fail to distinguish viable tumor from fibrosis, necessitating a reassessment of imaging-based standards. Methods: A comprehensive literature review was conducted using PubMed, focusing on prospective and retrospective studies over the past 25 years that evaluated the role of CT and complementary imaging modalities (MRI, PET-CT) in predicting resectability post-NAT in non-metastatic PDAC. Studies with small sample sizes or case reports were excluded. Results: Across studies, conventional CT parameters—particularly >180° vascular encasement—showed a limited correlation with histologic invasion or surgical outcomes after NAT. Persistent vessel contact on CT often reflected fibrosis, rather than active tumor. Dynamic changes, such as regression in the tumor–vessel interface and vessel lumen restoration, correlated more accurately with R0 resection. Adjunct markers like CA 19-9 response and patient performance status further improved resectability prediction. Conclusions: CT-based resectability assessment after NAT should transition from static morphologic criteria to response-based interpretation. Multidisciplinary evaluation integrating radiologic, biochemical, and clinical findings is essential to guide surgical decision-making and improve patient outcomes. Full article

This research work focused on the development of an adsorbent biocomposite material based on polyhydroxybutyrate (PHB) and cellulose acetate derived from sugarcane (Saccharum officinarum) fibre, through cellulose acetylation. The resulting material represents both an accessible and effective alternative for the treatment and remediation of water contaminated with heavy metals, such as Ni (II). The biocomposite was prepared by blending cellulose acetate (CA) with the biopolymer PHB using the solvent-casting method. The resulting biocomposite exhibited a point of zero charge (pHpzc) of 5.6. The material was characterised by FTIR, TGA-DSC, and SEM analyses. The results revealed that the interaction between Ni (II) ions and the biocomposite is favoured by the presence of functional groups, such as –OH, C=O, and N–H, which act as active adsorption sites on the material’s surface, enabling efficient interaction with the metal ions. Adsorption kinetics studies revealed that the biocomposite achieved an optimal adsorption capacity of 5.042 mg/g at pH 6 and an initial Ni (II) concentration of 35 mg/L, corresponding to a removal efficiency of 86.44%. Finally, an analysis of the kinetic and isotherm models indicated that the experimental data best fit the pseudo-second-order kinetic model and the Freundlich isotherm. Full article

In order to elucidate the high-temperature reaction process of solid waste-based high belite sulphoaluminate cement containing residual gypsum in clinker (NHBSAC) and obtain the formation laws of each mineral in clinker, this article studied its high-temperature reaction kinetics. Through QXRD analysis and numerical fitting methods, the formation of C₄A₃$\overline{\mathrm{S}}$, β-C₂S, and CaSO₄ in clinker under different calcination systems was quantitatively characterized, the corresponding high-temperature reaction kinetics models were established, and the reaction activation energies of each mineral were obtained. The results indicate that the content of C₄A₃$\overline{\mathrm{S}}$ and β-C₂S increases with the prolongation of holding time and the increase in calcination temperature, while CaSO₄ is continuously consumed. Under the control mechanism of solid-state reaction, the formation and consumption of minerals follow the kinetic equation. C₄A₃$\overline{\mathrm{S}}$ and β-C₂S satisfy the D₄ equation under diffusion mechanism control, and CaSO₄ satisfies the R₃ equation under interface chemical reaction mechanism control. The activation energy required for mineral formation varies with different temperature ranges. The activation energies required to form C₄A₃$\overline{\mathrm{S}}$ at 1200–1225 °C, 1225–1275 °C, and 1275–1300 °C are 166.28 kJ/mol, 83.14 kJ/mol, and 36.58 kJ/mol, respectively. The activation energies required to form β-C₂S at 1200–1225 °C and 1225–1300 °C are 374.13 kJ/mol and 66.51 kJ/mol, respectively. This study is beneficial for achieving flexible control of the mineral composition of NHBSAC clinker, providing a theoretical basis and practical experience for the preparation of low-carbon cement and the optimization design of its mineral composition. Full article

We inspected a sector of the Apennines (central–southern Italy) in geographic and structural continuity with the Quaternary-active extensional belt but where clear geomorphic and seismological signatures of normal faulting are unexpectedly missing. The evidence of active tectonics in this area, between Abruzzo and Molise, does not align with geodetic deformation data and the seismotectonic setting of the central Apennines. To investigate the apparent disconnection between active deformation and the absence of surface faulting in a sector where high lithologic erodibility and landslide susceptibility may hide its structural evidence, we combined multi-scale and multi-source data analyses encompassing morphometric analysis and remote sensing techniques. We utilised high-resolution topographic data to analyse the topographic pattern and investigate potential imbalances between tectonics and erosion. Additionally, we employed aerial-photo interpretation to examine the spatial distribution of morphological features and slope instabilities which are often linked to active faulting. To discern potential biases arising from non-tectonic (slope-related) signals, we analysed InSAR data in key sectors across the study area, including carbonate ridges and foredeep-derived Molise Units for comparison. The topographic analysis highlighted topographic disequilibrium conditions across the study area, and aerial-image interpretation revealed morphologic features offset by structural lineaments. The interferometric analysis confirmed a significant role of gravitational movements in denudating some fault planes while highlighting a clustered spatial pattern of hillslope instabilities. In this context, these instabilities can be considered a proxy for the control exerted by tectonic structures. All findings converge on the identification of an ~20 km long corridor, the Castel di Sangro–Rionero Sannitico alignment (CaS-RS), which exhibits varied evidence of deformation attributable to active normal faulting. The latter manifests through subtle and diffuse deformation controlled by a thick tectonic nappe made up of poorly cohesive lithologies. Overall, our findings suggest that the CaS-RS bridges the structural gap between the Mt Porrara–Mt Pizzalto–Mt Rotella and North Matese fault systems, potentially accounting for some of the deformation recorded in the sector. Our approach contributes to bridging the information gap in this complex sector of the Apennines, offering original insights for future investigations and seismic hazard assessment in the region. Full article

Background: Huang Qin Decoction (HQD) is a well-established Traditional Chinese Medicine (TCM) formulation recognized for its application in the treatment of colorectal cancer (CRC). However, the precise therapeutic mechanisms remain inadequately defined. Methods: This study integrates metabolomics from a mouse model and network pharmacology to screen potential targets and bio-active ingredients of HQD. The pharmacological activity of HQD for CRC was evidenced via single-cell RNA sequencing (scRNA-seq), molecular docking, and molecular dynamics simulations. Atomic force microscopy (AFM) assays and cellular experimental validation were used to confirm the relative mechanisms. Results: The metabolite profile undergoes significant alterations, with metabolic reprogramming evident during the malignant progression of CRC liver metastasis. Network pharmacology analysis identified that HQD regulates several metabolic pathways, including arginine biosynthesis, alanine, aspartate, and glutamate metabolism, nitrogen metabolism, phenylalanine metabolism, and linoleic acid metabolism, by targeting key proteins such as aspartate aminotransferase (GOT1), cytochrome P450 1A2 (CYP1A2), and carbonic anhydrase 2 (CA2). ScRNA-seq analysis indicated that HQD may enhance the functionality of cytotoxic T cells, thereby reversing the immunosuppressive microenvironment. Virtual verification revealed a strong binding affinity between the identified hub targets and active constituents of HQD, a finding subsequently corroborated by AFM assays. Cellular experiments confirmed that naringenin treatment inhibits the proliferation, migration, and invasion of CRC cells by downregulating GOT1 expression and disrupting glutamine metabolism. Conclusions: Computational prediction and in vitro validation reveal the active ingredients, potential targets, and molecular mechanisms of HQD against CRC liver metastasis, thereby providing a scientific foundation for the application of TCM in CRC treatment. Full article

Acute myocardial hypoxia/ischemia is associated with abnormal accumulation of myocardial lipids, including dihydroceramides. Here, we characterized how dihydroceramides are remodeled in response to hypoxia and assessed how dihydroceramide remodeling correlates to human cardiac pathophysiology. Hypoxia resulted in a marked accumulation of very-long-chain (VLC)-dihydroceramides in cultured HL-1 cardiomyocytes. In humans, we identified a correlation between the abundance of VLC-dihydroceramides in myocardial biopsies and arrhythmias and heart failure and showed that cardiac expression of CERS2, coding for an enzyme that promotes synthesis of VLC-dihydroceramides, was associated with signaling pathways linked to cardiac arrhythmia and cardiomyopathy. In cultured HL-1 cardiomyocytes, we showed that CerS2 knockdown reduced accumulation of VLC dihydroceramides and altered the expression of mediators regulating Ca²⁺ cycling and electrical conduction. In conclusion, our findings indicate that increased abundance of VLC-dihydroceramides, promoted by increased activity of CerS2 in response to hypoxia, could play a role in cardiac arrhythmias and heart failure. Full article

Rapid urbanization and industrial development have significantly altered land use and cover across the globe, intensifying urban thermal environments and exacerbating the urban heat island (UHI) effect. Gujranwala, Pakistan, represents an industrial growth that has driven substantial land use/land cover (LULC) changes and temperature increases; however, the directional and distance-based patterns of these changes remain unquantified. Therefore, this study is conducted to examine spatiotemporal changes in LULC and variations in the Urban Thermal Field Variation Index (UTFVI) between 2001 and 2021 and to project future scenarios for 2031 and 2041 using (1) Earth Observation Indices (EOIs) with machine learning (ML) classifiers (Random Forest) for precise LULC mapping through the Google Earth Engine (GEE) platform, (2) Cellular Automata–Artificial Neural Networks (CA-ANNs) for future scenario projection, and (3) Gradient Directional Analysis (GDA) to quantify directional (16-axis) and distance-based (concentric zones) patterns of urban expansion and thermal variation from 2001–2021. The study revealed significant LULC changes, with built-up areas expanding by 7.5% from 2001 to 2021, especially in the east, northeast, and southeast directions within a 20 km radius. Due to urban encroachment, vegetation and cropland decreased by 1.47% and 1.83%, respectively. The urban thermal environment worsened, with the highest land surface temperature (LST) rising from 41 °C in 2001 to 55 °C in 2021. Additionally, the UTFVI showed expanding areas under the ‘strong’ and ‘strongest’ categories, increasing from 30.58% in 2001 to 33.42% in 2041. Directional analysis highlighted severe thermal stress in the southern and southwestern areas linked to industrial activities and urban sprawl. This integrated approach provides a template for analyzing urban thermal environments in developing cities, supporting targeted mitigation strategies through direction- and distance-specific planning interventions to mitigate UHI impacts. Full article

Background: Women with systemic autoimmune rheumatic disease (SARD) are at higher risk of developing infection-related complications, anxiety, and depression. Using the example of the COVID-19 pandemic, we aimed to explore the impact of this external stressor on symptoms of depression, anxiety, and stress in a sample of women with SARD in a cross-sectional study design. Methods: Females aged 18–50 with SARD were enrolled from 04/2021 to 04/2022 at the Medical University of Vienna or through an online self-help group, while snowball sampling was used to recruit an age-matched healthy control group. Participants completed questionnaires including: (1) demographic information, medical history, and access to healthcare; (2) the Depression, Anxiety, and Stress Scale (DASS-21); and (3) the Coronavirus Anxiety Scale (CAS). Parameters were compared between groups using Chi-squared, Fisher’s exact, and Mann–Whitney U tests. Linear regression analysis was used to investigate which individual factors predicted the DASS-21 in women with SARD. Results: The study sample consisted of 226 women (n = 99 with SARD and n = 127 healthy controls). Women with SARD reported lower DASS-21 stress (p = 0.008) and CAS scores (p = 0.057) than the control group. There were no significant differences in DASS-21 anxiety or depression scores. Among women with SARD, a linear regression model identified the most important predictors of DASS-21 as access to rheumatological care (p = 0.002) and recent disease activity (p = 0.028). Conclusions: Despite the pandemic, women with SARD reported mental health outcomes equal to or better than those of the healthy control group. Continued access to rheumatological care may serve as an important protective factor for their mental health during large-scale crises like pandemics. Full article

Background: In light of increasing breast cancer survival rates, it is essential to address cancer-related cognitive impairment (CRCI), a common yet often underestimated symptom. Methods: A randomised controlled trial is proposed involving 50 newly diagnosed participants, divided into a control group (CG) and an intervention group (IG). Both groups will receive an educational leaflet, while the IG will also take part in an individualised cognitive training programme based on everyday cognition (80 sessions distributed across four periods, compiled in a training dossier). Cognitive, emotional, and functional variables will be assessed before and after the intervention: cognitive function (MoCA test), everyday cognition (PECC), anxiety (Hamilton), functionality (LB), sleep quality (PSQI), quality of life (ECOG), and subjective memory complaints (FACT-COG). Expected results: Findings may guide future interventions and tailored protocols to alleviate CRCI in breast cancer patients undergoing active treatment. Ethics and dissemination: This study was approved by the Ethics Committee of the University of Salamanca (PI 2023 12 1478-TD). Full article

Platelets play critical roles in haemostasis and thrombosis. The platelet activation process is driven by agonist-induced rises in cytosolic [Ca²⁺]_i, where the patterns of Ca²⁺ responses are still incompletely understood. In this study, we developed a number of techniques to model the [Ca²⁺]_i curves of platelets from a single blood donor. Fura-2-loaded platelets were quasi-simultaneously stimulated with various agonists, i.e., thrombin, collagen, or CRP, in the presence or absence of extracellular Ca²⁺ entry, secondary mediator effects, or Ca²⁺ reuptake into intracellular stores. To understand the calibrated time curves of [Ca²⁺]_i rises, we developed two non-linear models, a multilayer perceptron (MLP) network and an autoregressive network with exogenous inputs (NARX). The trained networks accurately predicted the [Ca²⁺]_i curves for combinations of agonists and inhibitors, with the NARX model achieving an R² of 0.64 for the trend prediction of unforeseen data. In addition, we used the same dataset for the construction of a partial least square (PLS) linear regression model, which estimated the explained variance of each input. The NARX model demonstrated that good fits could be obtained for the nanomolar [Ca²⁺]_i curves modelled, whereas the PLS model gave useful interpretable information on the importance of each variable. These modelling results can be used for the development of novel platelet [Ca²⁺]_i-inhibiting drugs, such as the drug 2-aminomethyl diphenylborinate, blocking Ca²⁺ entry in platelets, or for the evaluation of general platelet signalling defects in patients with a bleeding disorder. Full article

Thiophene derivatives are particularly attractive for application in drug development for their versatile pharmacological properties. We synthesized a series of four compounds with thiophene carboxamide as a scaffold. The structures were established based on HR-MS and 1D- and 2D-NMR. The purity of the compounds was established to be greater than 92% by thin-layer chromatography and NMR. The cytotoxic effects of the newly synthesized compounds were evaluated against the normal HaCaT cell line and A375, HT-29, and MCF-7 cancer cell lines. The cytotoxic assessment revealed that two compounds exhibit a significant cytotoxic effect on all cancer cell lines. To investigate their potential underlying mechanisms of action, several tests were performed: immunofluorescence imaging, caspase-3/7 assay, mitochondrial membrane potential (JC-1) assay, and 2′,7′–dichlorofluorescein diacetate (DCFDA) assay. MB-D2 proved to be the most cytotoxic and effective in terms of caspase 3/7 activation, mitochondrial depolarization and decrease in ROS production; these effects did not occur in normal HaCaT cells, revealing that MB-D2 has a high selectivity against A375 cancer cells. Full article