Overexpression of SLC2A1, ALDOC, and PFKFB4 in the glycolysis pathway drives strong drug resistance in 3D HeLa tumor cell spheroids.

The 3D multicellular tumor spheroid (MTS) model exhibits enhanced fidelity in replicating the tumor microenvironment and demonstrates exceptional resistance to clinical drugs compared to the 2D monolayer model. In this study, we used multiomics (transcriptome, proteomics, and metabolomics) tools to explore the molecular mechanisms and metabolic differences of the two culture models. Analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathways revealed that the differentially expressed genes between the two culture models were mainly enriched in cellular components and biological processes associated with extracellular matrix, extracellular structural organization, and mitochondrial function. An integrated analysis of three omics data revealed 11 possible drug resistance targets. Among these targets, seven genes, AKR1B1, ALDOC, GFPT2, GYS1, LAMB2, PFKFB4, and SLC2A1, exhibited significant upregulation. Conversely, four genes, COA7, DLD, IFNGR1, and QRSL1, were significantly downregulated. Clinical prognostic analysis using the TCGA survival database indicated that high-expression groups of SLC2A1, ALDOC, and PFKFB4 exhibited a significant negative correlation with patient survival. We further validated their involvement in chemotherapy drug resistance, indicating their potential significance in improving prognosis and chemotherapy outcomes. These results provide valuable insights into potential therapeutic targets that can potentially enhance treatment efficacy and patient outcomes.

Data-Independent Acquisition Shortens the Analytical Window of Single-Cell Proteomics to Fifteen Minutes in Capillary Electrophoresis Mass Spectrometry.

Separation in single-cell mass spectrometry (MS) improves molecular coverage and quantification; however, it also elongates measurements, thus limiting analytical throughput to study large populations of cells. Here, we advance the speed of bottom-up proteomics by capillary electrophoresis (CE) high-resolution mass spectrometry (MS) for single-cell proteomics. We adjust the applied electrophoresis potential to readily control the duration of electrophoresis. On the HeLa proteome standard, shorter separation times curbed proteome detection using data-dependent acquisition (DDA) but not data-independent acquisition (DIA) on an Orbitrap analyzer. This DIA method identified 1161 proteins vs 401 proteins by the reference DDA within a 15 min effective separation from single HeLa-cell-equivalent (∼200 pg) proteome digests. Label-free quantification found these exclusively DIA-identified proteins in the lower domain of the concentration range, revealing sensitivity improvement. The approach also significantly advanced the reproducibility of quantification, where ∼76% of the DIA-quantified proteins had <20% coefficient of variation vs ∼43% by DDA. As a proof of principle, the method allowed us to quantify 1242 proteins in subcellular niches in a single, neural-tissue fated cell in the live Xenopus laevis (frog) embryo, including many canonical components of organelles. DIA integration enhanced throughput by ∼2-4 fold and sensitivity by a factor of ∼3 in single-cell (subcellular) CE-MS proteomics.

Enzymeless detection and real-time analysis of intracellular hydrogen peroxide released from cancer cells using gold nanoparticles embedded bimetallic metal organic framework.

Abnormal cell growth and proliferation can lead to tumor formation and cancer, one of the most fatal diseases worldwide. Hydrogen peroxide (H2O2) has emerged as a cancer biomarker, with its concentration being crucial for distinguishing cancer cells from normal cells. Herein, a cost-effective and enzymeless electrochemical sensing system for the monitoring of intracellular H2O2 has been constructed. The sensor is fabricated using gold nanoparticles embedded bimetallic copper/nickel metal organic framework (Au-CNMOF) immobilized reduced graphene oxide (RGO) modified screen printed electrode (SPE). The synthesized materials were characterized and confirmed by XRD, FTIR, SEM with EDS, and electrochemical analysis. The fabricated sensor displayed a redox peak at a formal potential (E°) of -0.155 V, corresponding to CuII/I redox couple of CNMOF in 0.1 M phosphate buffer. Electrochemical investigations revealed that the proposed sensor has a large electrochemical active surface area (1.113 cm2) and a higher surface roughness (5.67). Additionally, the sensor demonstrated excellent electrocatalytic activity towards H2O2 at -0.3 V, over a wide linear detection range from 28.5 µM to 4.564 mM with a limit of detection of 4.2 µM (S/N=3). Furthermore, the proposed sensor exhibits excellent stability, repeatability, reproducibility, and good anti-interference activity. Ultimately, the sensor was validated through real-time analysis of H2O2 released from cancer cells, successfully quantifying the released H2O2. The developed sensor holds great promise for real-time H2O2 analysis, with potential applications in clinical diagnostics, biological research and environmental monitoring.

Mechanistic insights into the impact of WIN 55, 212-2, a synthetic cannabinoid, on adhesion molecules PECAM-1 and VE-cadherin in HeLa cells: implications on cancer processes.

The endocannabinoid (eCB) system comprises endogenous ligands, cannabinoid receptors (CBRs), and their regulatory proteins; its alteration leads to many diseases including cancer. Thus, becomes a therapeutic target for synthetic cannabinoids aimed to control cancer cell proliferation, migration, adhesion, and invasion. However, little is known about adhesion molecules regulation through CBRs activation. The aim of this study was to evaluate the effects of a CB1/CB2 agonist, WIN-55, 212-2 (WIN), on the regulation of adhesion molecules platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial cadherin (VE-cadherin) in HeLa cells. CBRs expression was evaluated by immunofluorescence staining in HeLa cells and cell viability (thiazolyl blue tetrazolium bromide), cell adhesion (crystal violet), adhesion molecules expression and location (Western blot and immunofluorescence staining assays) were all assessed on cells treated with different WIN concentrations. Receptors CB1, CB2, and G-protein-coupled receptor 55 were expressed in HeLa cells. Additionally, biphasic effects were observed in their metabolic activity and adhesive properties: low WIN concentrations resulted in significant increases whereas, high ones decreased them compared to controls (p < 0.0001), demonstrating that WIN elicits opposite effects depending on the concentration and exposure time. PECAM-1 was detected in HeLa cell's cytoplasm, membrane, and perinuclear region, whereas VE-cadherin had a nuclear distribution. There were no significant differences in PECAM-1 and VE-cadherin expression and location, suggesting that WIN does not modulate these proteins. These findings support the potential use of WIN due to its anticancer properties without dysregulating adhesion molecules. WIN possible contribution to inhibit cancer progression should be further investigated.

3-Ethoxysalicylaldimine-based symmetrical azine-linked luminogen exhibiting ESIPT and bright orange colour AIE behaviour with live cell bioimaging application.

Herein, we describe the design and development of a new cell-permeable aggregation-induced emission (AIE) active 3-ethoxysalicylaldimine-based symmetrical azine molecule HDBE. The synthesized compound underwent comprehensive investigation of different spectroscopic methods, like NMR, mass and single crystal X-ray diffraction analysis. The fluorophore HDBE exhibited the bright orange colour AIE behaviour in THF-H2O mixture. The drastic enhancement of emission was achieved upon adding the water to the THF solution of HDBE, with a concentration of 90%. Along with the dynamic light scattering (DLS) and quantum yield measurements, the formation of aggregates was also verified by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analysis. Further, HDBE demonstrated excited state intramolecular proton transfer (ESIPT) characteristics in different polarity of solvents, which was corroborated by absorption, emission and lifetime spectroscopical investigations. The detailed scrutiny of X-ray structure of HDBE displayed the two strong intramolecular hydrogen bonding interactions, while solid-state fluorescent spectra showed dual emission that corresponds to enol and keto form confirming the ESIPT feature. Further, the synthesized AIE molecule was non-toxic and cell-permeable, making it easy to label as a biomarker in live HeLa cells via fluorescent bioimaging. These studies offer a quick and easy way to develop both AIE and ESIPT-coupled molecules for live cell bioimaging applications.

Genetically Encoded Trensor Circuits Report HeLa Cell Treatment with Polyplexed Plasmid DNA and Small-Molecule Transfection Modulators.

HeLa cell transfection with plasmid DNA (pDNA) is widely used to materialize biologicals and as a preclinical test of nucleic acid-based vaccine efficacy. We sought to genetically encode mammalian transfection sensor (Trensor) circuits and test their utility in HeLa cells for detecting molecules and methods for their propensity to influence transfection. We intended these Trensor circuits to be triggered if their host cell was treated with polyplexed pDNA or certain small-molecule modulators of transfection. We prioritized three promoters, implicated by others in feedback responses as cells import and process foreign material and stably integrated each into the genomes of three different cell lines, each upstream of a green fluorescent protein (GFP) open reading frame within a transgene. All three Trensor circuits showed an increase in their GFP expression when their host HeLa cells were incubated with pDNA and the degraded polyamidoamine dendrimer reagent, SuperFect. We next experimentally demonstrated the modulation of PEI-mediated HeLa cell transient transfection by four different small molecules, with Trichostatin A (TSA) showing the greatest propensity to boost transgene expression. The Trensor circuit based on the TRA2B promoter (Trensor-T) was triggered by incubation with TSA alone and not the other three small molecules. These data suggest that mammalian reporter circuits could enable low-cost, high-throughput screening to identify novel transfection methods and reagents without the need to perform actual transfections requiring costly plasmids or expensive fluorescent labels.

Effects of Arborvitae (Thuja plicata) Essential Oil on Cervical Cancer Cells: Insights into Molecular Mechanisms.

AIMS: This study aimed to assess the effects of AEO in an in vitro model of cell lines derived from cervical cancer-namely, HeLa and SiHa-by screening for AEO's cytotoxic properties and examining its influence on the modulation of gene expression. BACKGROUND: Cervical cancer stands as a prevalent global health concern, affecting millions of women worldwide. The current treatment modalities encompass surgery, radiation, and chemotherapy, but significant limitations and adverse effects constrain their effectiveness. Therefore, exploring novel treatments that offer enhanced efficacy and reduced side effects is imperative. Arborvitae essential oil, extracted from Thuja Plicata, has garnered attention for its antimicrobial, anti-inflammatory, immunomodulatory, and tissue-remodeling properties; however, its potential in treating cervical cancer remains uncharted. OBJECTIVE: The objective of this study was to delve into the molecular mechanisms induced by arborvitae essential oil in order to learn about its anticancer effects on cervical cancer cell lines. METHODS: The methods used in this study were assessments of cell viability using WST-1 and annexin V- propidium iodide, mRNA sequencing, and subsequent bioinformatics analysis. RESULTS: The findings unveiled a dose-dependent cytotoxic effect of arborvitae essential oil on both HeLa and SiHa cell lines. Minor effects were observed only at very low doses in the HaCaT non-tumorigenic human keratinocyte cells. RNA-Seq bioinformatics analysis revealed the regulatory impact of arborvitae essential oil on genes enriched in the following pathways: proteasome, adherens junctions, nucleocytoplasmic transport, cell cycle, proteoglycans in cancer, protein processing in the endoplasmic reticulum, ribosome, spliceosome, mitophagy, cellular senescence, and viral carcinogenesis, among others, in both cell lines. It is worth noting that the ribosome and spliceosome KEGG pathways are the most significantly enriched pathways in HeLa and SiHa cells. CONCLUSION: Arborvitae essential oil shows potential as a cytotoxic and antiproliferative agent against cervical cancer cells, exerting its cytotoxic properties by regulating many KEGG pathways.

Exploring Tryptophan-based Short Peptides: Promising Candidate for Anticancer and Antimicrobial Therapies.

BACKGROUND: Ultra-short peptides are essential therapeutic agents due to their heightened selectivity and reduced toxicity. Scientific literature documents the utilization of dipeptides, tripeptides, and tetrapeptides as promising agents for combating cancer. We have created a range of tryptophan-based peptides derived from literature sources in order to assess their potential as anticancer drugs. METHODS: We present the results of our study on the antibacterial and anticancer effectiveness of 10 ultra-short peptides that were produced utilizing microwave-assisted solid phase peptide synthesis. The synthesized peptides underwent screening for in vitro antibacterial activity using the agar dilution method. RESULTS: HPLC, LC-MS, 1H NMR, and 13C NMR spectroscopy were used to analyze the synthesized peptides. In tests using the HeLa and MCF-7 cell lines, the synthesized peptides' anticancer efficacy was assessed. The study found that two peptides showed potential median inhibitory concentration (IC50) values of 3.9±0.13 µM and 1.8±0.09 µM, respectively, and showed more activity than the reference medication doxorubicin. CONCLUSION: The antibacterial activity of synthesized peptides 3b and 4b was found to be better than the other synthetic peptides. MIC value of roughly 5-50 µg/mL for peptides 3a, 4c, and 4d showed strong antifungal activity against Candida albicans. The synthesized peptides were also evaluated for their anticancer activity against HeLa and MCF-7 cell lines, and found that peptides 3e and 4e were more potent than other peptides against doxorubicin.

Data-Driven Modelling of Substituted Pyrimidine and Uracil-Based Derivatives Validated with Newly Synthesized and Antiproliferative Evaluated Compounds.

The pyrimidine heterocycle plays an important role in anticancer research. In particular, the pyrimidine derivative families of uracil show promise as structural scaffolds relevant to cervical cancer. This group of chemicals lacks data-driven machine learning quantitative structure-activity relationships (QSARs) that allow for generalization and predictive capabilities in the search for new active compounds. To achieve this, a dataset of pyrimidine and uracil compounds from ChEMBL were collected and curated. A workflow was developed for data-driven machine learning QSAR using an intuitive dataset design and forwards selection of molecular descriptors. The model was thoroughly externally validated against available data. Blind validation was also performed by synthesis and antiproliferative evaluation of new synthesized uracil-based and pyrimidine derivatives. The most active compound among new synthesized derivatives, 2,4,5-trisubstituted pyrimidine was predicted with the QSAR model with differences of 0.02 compared to experimentally tested activity.

Palladium catalysed cross coupling reactions on 2,3-isoxazol-17α-ethynyltestosterone, their anti-cancer activity, molecular docking studies and ADMET analysis.

In the current study, the Sonogashira coupling reaction of danazol with aryl halides was carried out, yielding new aryl substituted danazol derivatives. The synthetic compounds were examined for anti-cancer potential on the HeLa human cervical cancer cell line, and they showed promising cytotoxic action. Synthesized compounds 2, 4 and 5 inhibited the growth of HeLa cervical cancer cells, potentially making them effective anti-cancer drugs in the future. Furthermore, molecular docking studies were performed to evaluate the inhibitory impact of danazol derivatives on the Human Papillomavirus (HPV) target protein (1F9F). The docking results showed a significant inhibitory action against the cervical cancer protein (1F9F). The binding energy (ΔG) values of 1, 2, 3, 4 and 5 against the protein 1F9F were -8.01, -8.70, -9.43, -9.58 and -9.75 kcal/mol, indicating a high affinity of the synthesized compounds to bind with the HPV target proteins compared to their parent compound danazol (1). ADMET analyses of all derivatives have also been carried out.

Antibacterial and Antitumoral Potentials of Phytosynthesized Silver/Silver Oxide Nanoparticles Using Tomato Flower Waste.

This study presents the phytosynthesis of silver-based nanoparticles using tomato flower waste extracts for the first time in the literature. The determination of total polyphenolic and flavonoid contents in the extracts showed high gallic acid equivalents (6436-8802 mg GAE/kg dm) and high quercetin equivalents (378-633 mg QE/kg dm), respectively, dependent on the extraction method. By the Ultra Performance Liquid Chromatography technique, 14 polyphenolic compounds were identified and quantified in the tomato flower waste extracts. The abundant phenolic compounds were caffeic acid (36,902-32,217 mg/kg) and chlorogenic acid (1640-1728 mg/kg), and the abundant flavonoid compounds were catechin (292-251 mg/kg) and luteolin (246-108 mg/kg). Transmission electron microscopy of the nanoparticles revealed a particle size range of 14-40 nm. Fourier Transform infrared spectroscopy and X-ray diffraction studies confirmed the phytosynthesis of the silver/silver oxide nanoparticles. These findings hold significant results for the antibacterial and antitumoral potential applications of the obtained nanoparticles, opening new areas for research and development and inspiring further exploration. The impact of this research on the field of metallic nanoparticle phytosynthesis is substantial, as it introduces a novel approach and could lead to significant advancements in the field.

In vitro cancer cell line luminescence-based validation of anticancer phytocompounds obtained from Leucas biflora against HELA cervical and A549 lung cancer cells.

Current research aims to screen the anticancer prospective of Leucas biflora phytocompounds against apoptotic regulator target protein essential for cancer progression. In gas chromatography-mass spectrometry analysis major phytocompounds such as tetracosahexaene, squalene, phytol, 22-stigmasten-3-one, stigmasterol, fluorene, and 1,4-dihydro were identified in ethanolic leaf extract of Leucas biflora. In vitro, the free radical scavenging potential of ethanolic leaf extract of Leucas biflora was examined through its DPPH and ABTS radical scavenging potential IC50 value 15.35 and 13.20 µg/ml, respectively. Dose-dependent cytotoxicity was monitored against both A549 lung cancer and HELA cervical cancer cells. Leucas biflora ethanolic leaf extract highly reduces the cell viability of both HELA and A549 cells in in vitro cytotoxicity assays. Leucas biflora ethanolic extract produces 23.76% and 29.76% viability rates against A549 lung and HELA cervical cancer cell lines, and their IC50 values differ slightly at 95.80 and 90.40 µg/ml, respectively. In molecular docking analysis lung cancer target protein-ligand complex 5Y9T-16132746 showed a maximum score of -14 kcal/mol by exhibiting stable binding affinity and interactions among all screened complexes. Based on docking score nine phytocompounds from Leucas biflora and two reference standard drugs were chosen for further analysis. Further validation reveals that the fluorene, 1,4-dihydro possess good ADMET, Bioactivity and density functional theory indices.

[Effect of total saponins from Panacis Majoris Rhizoma on proliferation, apoptosis and autophagy of human cervical carcinoma HeLa cells].

This paper investigated the effect of total saponins from Rhizoma Panacis Majoris on the proliferation, apoptosis, and autophagy of human cervical carcinoma HeLa cells. The saponin content was detected by ultraviolet-visible spectrophotometry. Cell coun-ting kit-8(CCK-8) assay, 4,6-diamidino-2-phenylindole(DAPI) staining, and flow cytometry were used to detect the effects of total saponins of Panacis Majoris Rhizoma on cell viability, morphology, cell cycle and apoptosis of HeLa cells. Western blot was used to detect the expression of apoptosis-related proteins B cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cleaved caspase-9, and cleaved caspase-3, autophagy-related proteins Beclin-1 and SQSTM1(p62), and the proteins related to the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) and mitogen-activated protein kinase(MAPK) signaling pathways. It was found that the yield and saponin content of total saponins from Rhizoma Panacis Majoris were 6.3% and 78.3%, respectively. Total saponins from Rhizoma Panacis Majoris could significantly inhibit the proliferation(P<0.001), effect the nuclear morphology, block the G_0/G_1 cycle, and induce cell apoptosis in HeLa cells with a concentration-dependent manner. In addition, total saponins from Rhizoma Panacis Majoris up-regulated the expression of pro-apoptotic proteins Bax, cleaved caspase-9, and cleaved caspase-3, and autophagy-related protein p62(P<0.05), while down-regulated the expression of anti-apoptotic protein Bcl-2 and autophagy-related protein Beclin-1(P<0.01). Total saponins from Rhizoma Panacis Majoris could promote the expression of p-p38/p38, p-Jun N-terminal kinase(JNK)/JNK, p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR proteins in PI3K/Akt/mTOR and MAPK signaling pathways(P<0.05). In contrast, the effect on p-ERK/ERK expression was not obvious. Therefore, total saponins from Rhizoma Panacis Majoris may inhibit autophagy and promote apoptosis of HeLa cells through the activation of the PI3K/Akt/mTOR, c-JNK, and p38 MAPK signaling pathways, which indicates that total saponins from Rhizoma Panacis Majoris may have a potential role in cervical cancer treatment.

Design, synthesis and evaluation of novel norfloxacin analogs as potent anticancer and antioxidant agents.

Aim: To synthesize a novel series of norfloxacin analogs and to evaluate biological activity. Methodology: Novel norfloxacin analogs were synthesized and characterized by NMR and mass spectrometry. Antiproliferative and antioxidant properties were studied. Results: Compound 2f was the most potent against HeLa cell-line with 100% inhibition of cell viability IC50 = 3.1 ± 0.2 µM. All compounds exhibit moderate to excellent antioxidant properties. Docking study demonstrates higher binding affinity of compounds with respective anticancer (B-cell lymphoma-2) and (tyrosinase) antioxidant targets. In silico absorption, distribution, metabolism and excretion profile of compounds proves all synthesized compounds follow Lipinski's rule of drug likeness, non toxic and possess passive gastrointestinal absorption. Conclusion: The biological profile suggest that the synthesized norfloxacin analogs can be a novel scaffold for future anticancer drug development.

[Box: see text].

Phytomediated Copper Oxide Nanoparticles Derived from the Fronds of Adiantum venustum D.Don: Evaluation of their Biomedical Potential.

The green synthesis of copper oxide nanoparticles (CuO) mediated by crude ethanolic extract and the n-butanol fraction of Adiantum venustum represents a groundbreaking approach in nanotechnology, combining ecological sustainability with advanced functionality. This innovative method leverages the natural bioactive compounds present in A. venustum to produce CuO nanoparticles, which exhibit remarkable antioxidant, anti-inflammatory, antimicrobial, and anti-proliferative properties. The green synthesized nanoparticles were characterized using a variety of techniques, as XRD confirmed the crystalline nature of the CuO nanoparticles, with a crystallite size of 14.65 nm for CuO-C and 18.73 nm for CuO-B. The grain sizes of CuO-C (14.09 ± 0.17 nm) and CuO-B (67.88 ± 2.08 nm) were determined using transmission electron microscopy micrographs. Furthermore, the synthesized nanomaterial and the crude ethanolic extract, n-butanol fraction, were examined for their biological potentials namely antioxidant, anti-inflammatory, antimicrobial, and anti-proliferative activity against HeLa cancer cells. Among the synthesized nanomaterials, copper oxide nanoparticles synthesized by utilizing the n-butanol fraction have appeared as a potential biomedical agent. CuO-B has arisen as an antioxidant agent with IC50 values of 44.63 ± 0.49 µg/mL, 48.49 ± 0.17 µg/mL, and 35.39 ± 0.61 µg/mL for DPPH, FRAP, and reducing power assay, respectively. Furthermore, the significant antibacterial potential of CuO-B against gram-positive (S. aureus MIC 46.88 µg/mL) and gram-negative (K. pneumonia MIC 23.48 µg/mL) bacterial strains cannot be neglected either. Along with this, the IC50 value (138.07 µg/mL) of CuO-B against HeLa cells proved it to be a potential anticancerous agent. Hence, this novel approach emphasized that these synthesized nanoparticles have tremendous biological potential and can be applied to various fields of agriculture and biomedicine.

Biogenic silver nanomaterials synthesized from Ocimum sanctum leaf extract exhibiting robust antimicrobial and anticancer activities: Exploring the therapeutic potential.

There is a surge in antibiotic consumption because of the emergence of resistance among microbial pathogens. In the escalating challenge of antibiotic resistance in microbial pathogens, silver nanoparticles (AgNPs)-mediated therapy has proven to be the most effective and alternative therapeutic strategy for bacterial infections and cancer treatment. This study aims to explore the potential of OsAgNPs derived from Ocimum sanctum's aqueous leaf extract as antimicrobial agents and anticancer drug delivery modalities. This study utilized a plant extract derived from Ocimum sanctum (Tulsi) leaves to synthesize silver nanoparticles (OsAgNPs), that were characterized by FTIR, TEM, SEM, and EDX. OsAgNPs were assessed for their antibacterial and anticancer potential. TEM analysis unveiled predominantly spherical or oval-shaped OsAgNPs, ranging in size from 4 to 98 nm. The (MICs) of OsAgNPs demonstrated a range from 0.350 to 19.53 µg/ml against clinical, multidrug-resistant (MDR), and standard bacterial isolates. Dual labelling with ethidium bromide and acridine orange demonstrated that OsAgNPs induced apoptosis in HeLa cells. The OsAgNPs-treated cells showed yellow-green fluorescence in early-stage apoptotic cells and orange fluorescence in late-stage cells. Furthermore, OsAgNPs exhibited a concentration-dependent decrease in HeLa cancer cell viability, with an IC50 value of 90 µg/ml noted. The study highlights the remarkable antibacterial efficacy of OsAgNPs against clinically significant bacterial isolates, including antibiotic-resistant strains. These results position the OsAgNPs as prospective therapeutic agents with the potential to address the growing challenges posed by antibiotic resistance and cervical cancer.

Fluorescent sensor/tracker for biocompatible and real-time monitoring of ultra-trace arsenic toxicants in living cells.

Real-time monitoring and tracking of extreme toxins that penetrate into living cells by using biocompatible, low-cost visual detection via fluorescent monitors are vitally essential to reduce health hazards. Herein, we report a simple engineering design of biocompatible and fluorescent sensors/trackers for real-time monitoring and ultra-trace tracking (up to ppb) of extremely toxic substances (such as arsenic species) in living cells. The biocompatible As(V) sensor (BAS) design is fabricated via successful dressing/decoration process of 2-hydroxy 5-methyl isophthalaldehyde fluorescent receptor into hierarchical organic-inorganic carriers that have micro-hollow geodes, swirled caves and nest-shaped cages, and uniform cubic structures. The BAS monitors show evidence for the selective trapping/detecting/tracking of As(V) species in biological cells (i.e., HeLa cells) despite the coexistence of highly competitive and interfered species. Our simple batch-contact sensing assays shows real-space evidence of the continuous monitoring of As(V) species in HeLa cells with ultra-sensitive detection (i.e., with a low detection limit of 0.149 ppb) and rapid recognition (i.e., in the order of seconds). Significantly, the BAS monitors did not affect the cell population and achieved low cytotoxicity and high cell viability during the monitoring/tracking process inside HeLa cells. The high biocompatibility of BAS remarkably allows precise quantification and real-time monitoring/tracking of toxicant targets in living cells.

Kuwanon C Inhibits Tumor Cell Proliferation and Induces Apoptosis by Targeting Mitochondria and Endoplasmic Reticulum.

Kuwanon C is a unique flavonoid found in the mulberry family, characterized by two isopentenyl groups. While previous research has focused on various properties of kuwanon C, such as antioxidant, hypoglycemic, antimicrobial, food preservation, skin whitening, and nematode lifespan extension, little attention has been given to its potential role in oncological diseases. In this study, we investigate the antitumor effect of kuwanon C in cervical cancer cells and elucidate its specific mechanism of action. We assessed the antitumor effects of kuwanon C using various experimental techniques, including cell proliferation assay, wound healing assays, EdU 488 proliferation assay, mitochondrial membrane potential assay, ROS level assay, cell cycle, apoptosis analysis, and studies on kuwanon C target sites and molecular docking. The results revealed that kuwanon C significantly impacted the cell cycle progression of HeLa cells, disrupted their mitochondrial membrane potential, and induced a substantial increase in intracellular ROS levels. Moreover, kuwanon C exhibited notable anti-proliferative and pro-apoptotic effects on HeLa cells, surpassing the performance of commonly used antitumor drugs such as paclitaxel and cisplatin. Notably, kuwanon C demonstrated superior efficacy while also being more easily accessible compared to paclitaxel. Our study demonstrates that kuwanon C exerts potent antitumor effects by its interaction with the mitochondrial and endoplasmic reticulum membranes, induces a significant production of ROS, disrupts their normal structure, inhibits cell cycle progression, and stimulates apoptotic signaling pathways, ultimately resulting in the death of HeLa tumor cells. As an isopentenyl compound derived from Morus alba, kuwanon C holds great promise as a potential candidate for the development of effective antitumor drugs.

Molecular Actions of Enicostemma hyssopifolium Whole Plant Extract on HPV18-Infected Human Cervical Cancer (HeLa) Cells.

OBJECTIVE: Enicostemma hyssopifolium (E. hyssopifolium) contains several bioactive compounds with anti-cancer activities. This study was performed to investigate the molecular effects of E. hyssopifolium on HPV18-containing HeLa cells. METHODS: The methanol extract of E. hyssopifolium whole plant was tested for cytotoxicity by MTT assay. A lower and higher dose (80 and 160 µg/mL) to IC50 were analyzed for colonization inhibition (Clonogenic assay), cell cycle arrest (FACS analysis), and induction of apoptosis (AO/EtBr staining fluorescent microscopy and FACS analysis) and DNA fragmentation (comet assay). The HPV 18 E6 gene expression in treated cells was analyzed using RT-PCR and qPCR. RESULTS: A significant dose-dependent anti-proliferative activity (IC50 - 108.25±2 µg/mL) and inhibition of colony formation cell line were observed using both treatments. Treatment with 80 µg/mL of extract was found to result in a higher percent of cell cycle arrest at G0/G1 and G2M phases with more early apoptosis, while 160 µg/mL resulted in more cell cycle arrest at SUBG0 and S phases with late apoptosis for control. The comet assay also demonstrated a highly significant increase in DNA fragmentation after treatment with 160 µg/mL of extract (tail moments-19.536 ± 17.8), while 80 µg/mL of extract treatment showed non-significant tail moment (8.152 ± 13.0) compared to control (8.038 ± 12.0). The RT-PCR and qPCR results showed a significant reduction in the expression of the HPV18 E6 gene in HeLa cells treated with 160 µg/mL of extract, while 80 µg/mL did not show a significant reduction. CONCLUSION: The 160 µg/mL methanol extract of E. hyssopifolium demonstrated highly significant anti-cancer molecular effects in HeLa cells.

Antimicrobial, Antioxidant and Cytotoxic Effects of Essential Oil, Fatty Acids and Bioactive Compounds of Beta vulgaris var. crassa (Fodder Beet).

Beta vulgaris var. crassa is undoubtedly a very important plant that is not used enough in the world. In this study, it was aimed to determine the cytotoxic activities of the components (essential oils, fatty acids, total phenol and flavonoid) found in the leaf parts of Beta vulgaris var. crassa against PC-3, MCF-7 and HeLa cancer cell lines. In addition, the effectiveness of these ingredients against bacteria and fungi that can cause serious health problems in humans was tested. In experiments, three tumor cell lines were exposed to various plant extract concentrations (31.25, 62.5, 125, 250, 500 and 1000 µg/mL) for 72 h. It was found that plant extracts showed high (SI: 2.14 > 2) cytotoxicity to PC-3 cells, moderate (SI: 1.62 < 2) to HeLa cells, and low (SI: 0.93 < 2) cytotoxicity to MCF-7 cells. Also, different plant extract concentrations were found to cause an inhibition rate of 16.3-22.3% in Staphylococcus aureus, 16.8-23.5% in Streptococcus pyogenes and 12-16.2% in Cutibacterium acnes. Similarly, inhibition rates were determined between 9.5-20.7% for Candida albicans, 3.5-7.7% for Candida auris, and 5.5-15.1% for Candida glabrata. The results showed that the plant extract exhibited a concentration-dependent cytotoxic and antimicrobial effect against both cancer cell lines and microbial pathogens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01269-8.