Antifibrotic effect of silymarin on arecoline-induced fibrosis in primary human buccal fibroblasts: an in silico and in vitro analysis.

BACKGROUND: This study aimed to assess silymarin's anticancer and antifibrotic potential through in silico analysis and investigate its impact on in vitro arecoline-induced fibrosis in primary human buccal fibroblasts (HBF). METHODS & RESULTS: The study utilized iGEMDOCK for molecular docking, evaluating nine bioflavonoids, and identified silymarin and baicalein as the top two compounds with the highest target affinity, followed by subsequent validation through a 100ns Molecular Dynamic Simulation demonstrating silymarin's stable behavior with Transforming Growth Factor Beta. HBF cell lines were developed from tissue samples obtained from patients undergoing third molar extraction. Arecoline, a known etiological factor in oral submucous fibrosis (OSMF), was employed to induce fibrogenesis in these HBFs. The inhibitory concentration (IC50) of arecoline was determined using the MTT assay, revealing dose-dependent cytotoxicity of HBFs to arecoline, with notable cytotoxicity observed at concentrations exceeding 50µM. Subsequently, the cytotoxicity of silymarin was assessed at 24 and 72 h, spanning concentrations from 5µM to 200µM, and an IC50 value of 143µM was determined. Real-time polymerase chain reaction (qPCR) was used to analyze the significant downregulation of key markers including collagen, epithelial-mesenchymal transition (EMT), stem cell, hypoxia, angiogenesis and stress markers in silymarin-treated arecoline-induced primary buccal fibroblast cells. CONCLUSION: Silymarin effectively inhibited fibroblast proliferation and downregulated genes associated with cancer progression and EMT pathway, both of which are implicated in malignant transformation. To our knowledge, this study represents the first exploration of silymarin's potential as a novel therapeutic agent in an in vitro model of OSMF.

Immunomodulatory Gene-Splicing Dysregulation in Tumorigenesis: Unmasking the Complexity.

Cancer is a global health concern with rising incidence, morbidity, and mortality. The interaction between the tumor and immune cells within the tumor microenvironment is facilitated by signaling pathways driven by immunomodulatory proteins. Alternative splicing regulates the production of multiple immunomodulatory proteins with diverse functionality from a single mRNA transcript. Splicing factors are pivotal in modulating alternative splicing processes but are also subject to regulation. The dysregulation of alternative splicing may result from splicing factor (SF) abnormal expression levels and mutations in the cis and trans-acting elements and small nuclear RNA (snRNA) molecules. Aberrant splicing may generate abnormal mRNA transcripts encoding isoforms with altered functions that contribute to tumorigenesis or cancer progression. This review uncovers the complexity of immunomodulatory genes splicing dysregulation in oncogenesis. Identifying specific immunomodulatory splicing isoforms that contribute to cancer could be utilized to improve current immunotherapeutic drugs or develop novel therapeutic interventions for cancer.

Clinicopathological Significance of Cancer Stem Cell Markers (OCT-3/4 and SOX-2) in Oral Submucous Fibrosis and Oral Squamous Cell Carcinoma.

Oral submucous fibrosis (OSMF) is highly prevalent in South East Asia with higher rates of malignant transformation in Indian subcontinent. Numerous biomarkers are now being studied to predict disease prognosis and detect malignant alterations at an early stage. Patients with clinically and biopsy-proven oral submucous fibrosis and oral squamous cell carcinoma were included in the study as the experimental group, while patients without a tobacco or betel nut habit who had their third molars surgically removed were included as the healthy control group. For the immunohistochemistry (IHC) investigation, 5-µm slices from formalin-fixed, paraffin-embedded tissue blocks (FFPE) were obtained. Fresh tissues (n = 45) from all three groups were collected and gene expression was studied using relative quantitation-based qPCR. The protein expression of octamer-binding transcription factor 3/4 (OCT 3/4) and sex-determining region Y-box 2 (SOX 2) was evaluated in the experimental group and compared with healthy controls. The IHC results showed a significant correlation with the expression of OCT 3/4 (p value = 0.000; χ2 = 20.244) and SOX 2 (p value = 0.006; χ2 = 10.101) among OSCC and OSMF patients in comparison to healthy controls. Both OCT 3/4 and SOX 2 showed overexpression of four-fold and three-fold in OSMF when compared to OSCC and healthy controls, respectively. This study shows the significant importance of cancer stem cell markers OCT 3/4 and SOX 2 to assess the disease prognosis in OSMF.

Unravelling the emerging carcinogenic contaminants from industrial waste water for prospective remediation by electrocoagulation - A review.

The need of the hour relies on finding new but sustainable ways to curb rising pollution levels. The accelerated levels of urbanization and increase in population deplete the finite resources essential for human sustenance. In this aspect, water is one of the non-renewable sources that is running out very fast and is polluted drastically day by day. One way of tackling the problem is to reduce the pollution levels by decreasing the usage of chemicals in the process, and the other is to find ways to reuse or reduce the contaminants in the effluent by treatment methods. Most of the available water recycling or treatment methods are not sustainable. Some of them even use toxic chemicals in the processing steps. Treatment of organic wastes from industries is a challenging task as they are hard to remove. Electrocoagulation is one of the emerging water treatment technologies that is highly sustainable and has a comparatively cheaper operating cost. Being a broad-spectrum treatment process, it is suitable for treating the most common water pollutants ranging from oils, bacteria, heavy metals, and others. The process is also straightforward, where electrical current is used to coagulate the contaminates. The presence of carcinogens in these waste water increases the need for its treatment towards further use. The present investigation is made as an extensive analysis of the emerging carcinogens and their various sources from process industries, especially in the form of organic waste and their removal by electrocoagulation and its coupled techniques. The paper also aims to ascertain why the electrocoagulation technique may be a better alternative compared with other methods for the removal of carcinogens in organic wastewater, an analysis which has not been explored before.

Evaluation of antimicrobial, anticancer potential and Flippase induced leakage in model membrane of Centella asiatica fabricated MgONPs.

The use of chemically synthesized nanoparticles and crude plant extracts as antimicrobial -anticancer agents have many limitations. In this study, we have used Centella asiatica extract (CaE) having relatively less explored but tremendous medicinal properties, as reducing and stabilizing agents to green synthesize magnesium oxide nanoparticles (MgONPs) using magnesium nitrate. In comparison to the bulk material, capabilities of Ca-MgONPs as an improved antibacterial, antifungal, and anticancer agent in human prostatic carcinoma cells (PC3), as well as membranolytic capability in model cell membrane, were studied. The phyto-functionalized Ca-MgONPs were characterized using UV-Visible spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Energy Dispersive X-Ray Spectroscopy (EDX), X-ray Diffraction (XRD), Fourier Transform Infra-Red Spectroscopy (FT-IR) and Atomic Force Microscopy (AFM). Observation of characteristic peaks by spectroscopic and microscopic analysis confirmed the synthesis of Ca-MgONPs. The Ca-MgONPs showed broad spectrum of bactericidal activity against both gram-positive and gram-negative bacteria and fungicidal activity against two species of the Candida fungus. The Ca-MgONPs also exhibited dose-dependent and selective inhibition of proliferating PC3 cells with IC50 of 123.65 ± 4.82 µg/mL at 24 h, however, without having any cytotoxicity toward non-cancerous HEK293 cells. Further studies aimed at understanding the probable mechanism of toxicity of Ca-MgONPs in PC3 cells, the results indicated a significant reduction in cell migration capacities, increment in cytosolic ROS, loss of mitochondrial transmembrane potential, DNA damage and S-phase cell cycle arrest. Ca-MgONPs also induced pore formation in a synthetic large unilamellar vesicle. Thus, Ca-MgONPs might be useful in the effective management of several human pathogens of concern and some more cancer types.

Efficiency of microbial fuel cells in the treatment and energy recovery from food wastes: Trends and applications - A review.

The rising global population and their food habits result in food wastage and cause an obstacle in its treatment and disposal. Due to the rapid shift in the lifestyle of the human population and urbanization, almost one-third of the food produced is wasted from various sectors like domestic sources, agricultural sectors, and industrial sectors. These food resources squandered are rich in organic biomolecules which can cause complications upon direct disposal in the environment. Conventional disposal methods like composting, landfills and incineration demand high costs besides causing severe environmental and health issues. To overcome these demerits of the conventional methods and to avoid the loss of rich organic food resources, there is an immediate need for a sustainable and eco-friendly solution for the valorization of the food wastes. Microbial fuel cells (MFCs) are gaining attention, due to their ideal approach in the production of electricity and parallel treatment of organic food wastes. The MFCs are significant as an innovative approach using microorganisms and oxidizing the organic food wastes into bio-electricity. In this review, the recent advancements and practices of the MFCs in the field of food waste treatment and management along with electricity production are discussed. The major outcome of this work highlights the setting up of MFC for the treatment of higher volumes of food waste residues and enhancing the bioelectricity production in an optimal condition. For further improvements in the food waste treatments using MFCs, greater understanding and more research needs are to be focused on the commercialization, different operational modes, operational types, and low-cost fabrication coupled with careful examination of scale-up factors.