Exploring In-Silico, In-Vitro Antioxidant, and Cytotoxic Potential of Valerian wallichii by On Cervical Epithelial Carcinoma (HeLa) Cell Lines.

Traditional medicinal practices often utilize herbal remedies for treating various diseases. This study focuses on exploring the phytochemical constituents, in-silico, in-vitro antioxidant, and anticancer activities of Valerian wallichii root extracts on human cervical epithelial carcinoma (HeLa) cell lines. The molecular docking approach was employed to predict the ligand molecule's orientation within the receptor like Epidermal growth factor receptor tyrosine kinase domain (PDB ID: 1M17) using Schrodinger's GLIDE model. Among the selected phytocompounds, hesperidin exhibited promising inhibitory activity against EGFR (Epidermal Growth Factor Receptor) domain with -8.701 kcal/mol docking score and interactions with MET 769, ASP 831, ASP776, LEU694 and ASN818 residues as compared to standard doxorubicin with -7.6 kcal/mol docking score and interactions with ASP 831, ASN818 and ASP776 residues and further, various antioxidant activity was assessed and In-vitro anticancer activity against HeLa cell lines was evaluated by hydroalcoholic root extracts demonstrated antioxidant capacities, and selective cytotoxicity was observed, with IC50 : 45.759±0.42 µg/mL for the overall extract and IC50 : 30.245±0.58 µg/mL for the EAF fraction as compared to standard doxorubicin with IC50 : 25.9891±0.25 µg/mL, respectively. The present study concluded that Valerian wallichii L possesses potential human cervical epithelial carcinoma cell line inhibition properties based on the computer aided drug design models and in-vitro activity.

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.

Fungi fabrication, characterization, and anticancer activity of silver nanoparticles using metals resistant Aspergillus niger.

The purpose of this study was to assess the bio-fabrication possibilities of pre-isolated (from bauxite mine tailings) metal-tolerant Aspergillus niger biomass filtrate and the anticancer potential of synthesized silver nanoparticles (AgNPs) tested with a Human Cervical cancer cell line (HeLa cells: Henrietta Lacks cells). The nitrate reduction test demonstrated that A. niger has the ability to reduce nitrate, and filtrate derived from A. niger biomass efficiently fabricated AgNPs from AgNO3, as demonstrated by a visible color change from pale greenish to brownish. The UV-visible spectroscopy analysis revealed an absorbance peak at 435 nm, which corresponded to the AgNPs. These AgNPs have been capped and stabilized with several functional groups related to various bioactive molecules such as aldehyde, benzene rings, aldehydic, amines, alcohols, and carbonyl stretch protein molecules. Fourier-Transform Infrared Spectroscopy (FTIR) analysis confirmed the capping and stabilizing chemical bonding pattern. Scanning Electron Microscopy (SEM) revealed that the synthesized AgNPs were spherical, with an average size of 21.38 nm. This bio-fabricated AgNPs has in-vitro anticancer potential when tested against the HeLa cell line due to its potential size and shape. At 100 g mL-1 concentrations of this bio-fabricated AgNPs, the anticancer activity percentage was found to be 70.2%, and the IC50 value was found to be 66.32 g m-1. These findings demonstrated that the metal-tolerant A. niger cell filtrate could produce AgNPs with anticancer potential.

Synthesis and characterization of TiO2 NPs by aqueous leaf extract of Coleus aromaticus and assess their antibacterial, larvicidal, and anticancer potential.

The frequent applications of synthetic chemical insecticides and drugs create resistance among insects and microbes, creating a new threat to human and environmental welfare. This investigation focused on evaluating the possibilities of fabricating and characterizing the titanium dioxide nanoparticles (TiO2 NPs) from titanium dioxide (TiO2) through the aqueous leaf extract of Coleus aromaticus. Their biological applications were studied against the larvae of Aedes aegypti human pathogenic bacteria, and cancer cell line. The results revealed that the aqueous leaf extract had the metal reducing proficiency to produce nanoparticles from TiO2. The synthesized TiO2 NPs were initially confirmed by visible color changes and Ultraviolet-Visible Spectrophotometer analysis that showed a predominant peak at 332 nm. Furthermore, the nanocrystals, structural alignment, functional groups and elemental compositions were studied by following standard operating protocol in XRD (X-ray Powder Diffraction), FTIR (Fourier Transform Infrared Spectroscopy), TEM (Transmission Electron Microscopy), and EDX (Energy-Dispersive X-ray Spectroscopy) techniques, respectively. The results attained from these techniques confirmed that the plant mediated and fabricated particles were in the nanoscale range (12-33 nm) with a hexagonal shape. The synthesized TiO2 NPs had an outstanding (1000 µg mL-1) larvicidal activity against the four stages of instars larvae of Ae. aegypti at 1000 µg mL-1. It also had an excellent antibacterial potential against E. faecalis (33 mm), followed by S. boydii (30 mm) at 30 mg L-1 concentration. The green fabricated TiO2 NPs had a fabulous (92.37%) cytotoxic activity on the HeLa cell line at 100 µg mL-1 dosage within one day of exposure. The entire results concluded that the C. aromaticus mediated TiO2 NPs have excellent biological applications and thus, could be considered for the welfare of human beings.

Genetic Load and Potential Mutational Meltdown in Cancer Cell Populations.

Large genomes with elevated mutation rates are prone to accumulating deleterious mutations more rapidly than natural selection can purge (Muller's ratchet). As a consequence, it may lead to the extinction of small populations. Relative to most unicellular organisms, cancer cells, with large and nonrecombining genome and high mutation rate, could be particularly susceptible to such "mutational meltdown." However, the most common type of mutation in organismal evolution, namely, deleterious mutation, has received relatively little attention in the cancer biology literature. Here, by monitoring single-cell clones from HeLa cell lines, we characterize deleterious mutations that retard the rate of cell proliferation. The main mutation events are copy number variations (CNVs), which, estimated from fitness data, happen at a rate of 0.29 event per cell division on average. The mean fitness reduction, estimated reaching 18% per mutation, is very high. HeLa cell populations therefore have very substantial genetic load and, at this level, natural population would likely face mutational meltdown. We suspect that HeLa cell populations may avoid extinction only after the population size becomes large enough. Because CNVs are common in most cell lines and tumor tissues, the observations hint at cancer cells' vulnerability, which could be exploited by therapeutic strategies.

The synthesis and biological evaluation of nucleobases/tetrazole hybrid compounds: A new class of phosphodiesterase type 3 (PDE3) inhibitors.

Spired by the chemical structure of Cilostazol, a selective phosphodiesterase 3A (PDE3A) inhibitor, several novel hybrid compounds of nucleobases (uracil, 6-azauracil, 2-thiuracil, adenine, guanine, theophylline and theobromine) and tetrazole were designed and successfully synthesized and their inhibitory effects on PDE3A as well as their cytotoxicity on HeLa and MCF-7 cancerous cell lines were studied. Obtained results show the linear correlation between the inhibitory effect of synthesized compounds and their cytotoxicity. In some cases, the PDE3A inhibitory effects of synthesized compounds are higher than the Cilostazol. Besides, compared to a standard anticancer drug methotrexate, some of the synthesized compounds showed the higher cytotoxicity against the HeLa and MCF-7 cancerous cell lines.

Green synthesis of silver nanoparticles using Indian Belladonna extract and their potential antioxidant, anti-inflammatory, anticancer and larvicidal activities.

KEY MESSAGE: Atropa acuminata aqueous leaf extract biosynthesized silver nanoparticles showed strong antioxidant, anticancerous (HeLa cells) and anti-inflammatory activities. Besides, this bio syn-AgNP also proved effective against mosquito vectors causing malaria, dengue and filariasis. Present study highlights eco-friendly and sustainable approach for the synthesis of silver nanoparticles (AgNP) using aqueous leaf extract of A. acuminata, a critically endangered medicinal herb. The addition of 1 mM silver nitrate to aqueous leaf extract resulted in the synthesis of AgNP when solution was heated at 60 °C for 30 min at pH 7. Absorption band at 428 nm, as shown by UV-Vis spectroscopy confirmed the synthesis of AgNP. XRD patterns revealed the crystalline nature of AgNP and TEM analysis showed that most of the nanoparticles were spherical in shape. Zeta potential of AgNP was found to be - 33.5 mV which confirmed their high stability. FT-IR investigations confirmed the presence of different functional groups involved in the reduction and capping of AgNP. The synthesized AgNP showed effective DPPH (IC50-16.08 µg/mL), H2O2 (IC50-25.40 µg/mL), and superoxide (IC50-21.12 µg/mL) radical scavenging activities. These plant-AgNP showed significant inhibition of albumin denaturation (IC50-12.98 µg/mL) and antiproteinase activity (IC50-18.401 µg/mL). Besides, biosynthesized AgNP were found to have strong inhibitory effect against a cervical cancer (HeLa) cell line (IC50-5.418 µg/mL) as well as larvicidal activity against 3rd instar larvae of Anopheles stephensi (LC50-18.9 ppm, LC90-40.18 ppm), Aedes aegypti (LC50-12.395 ppm, LC90-36.34 ppm) and Culex quinquefasciatus (LC50-17.76 ppm, LC90-30.82 ppm) and were found to be non-toxic against normal cell line (HEK 293), and a non-target organism (Mesocyclops thermocyclopoides). This is the first report on the synthesis of AgNP using aqueous leaf extract of A. acuminata, validating their strong therapeutic potential.

Acinetobacter baumannii outer membrane protein A induces HeLa cell autophagy via MAPK/JNK signaling pathway.

Autophagy is an evolutionary conserved self-balancing process that plays an important role in maintaining cellular homeostasis via the clearance of damaged organelles and misfolded proteins. Infection-triggered autophagy specifically inhibits the invasion of intracellular bacterial replication and hence protects the cells from microbial infections. It has been reported that Acinetobacter baumannii trigger cell autophagy. However, the role of its virulence protein OmpA remains unclear. Therefore, this study aimed to explore the effects of Acinetobacter baumannii OmpA on cell autophagy and its underlying molecular mechanisms. The results showed that OmpA induced autophagy in HeLa and RAW264.7 cells, increased LC3BII expression, and hindered p62 degradation. Moreover, OmpA triggered incomplete autophagy by interfering the fusion of autophagosomes with lysosomes. Besides, OmpA activated MAPK/JNK signaling pathway and enhanced the phosphorylation levels of JNK, p38, and ERK, c-Jun. Inhibition of JNK signaling pathway suppressed OmpA-induced autophagy in HeLa cells. Ab wild-type strains carrying OmpA triggered incomplete autophagy and resulted in a large number of IL-1ß production. Ab-△OmpA strain (OmpA gene mutation) restored autophagic flux and reduced the accumulation of p62 and the release of IL-1ß in HeLa cells. Rapamycin activated autophagy to inhibit OmpA-induced IL-1ß secretion and protect HeLa cells from inflammatory damage. Collectively, these results suggest that OmpA can induce autophagy in HeLa cells through MAPK/JNK signaling pathway. Pre-treatment with Rapamycin activates autophagy and protects against cell death.

New thiazol-hydrazono-coumarin hybrids targeting human cervical cancer cells: Synthesis, CDK2 inhibition, QSAR and molecular docking studies.

Motivated by the potential anticancer activity of both coumarin and 2-aminothiazole nuclei, a new set of thiazol-2-yl hydrazono-chromen-2-one analogs were efficiently synthesized aiming to obtain novel hybrids with potential cytotoxic activity. MTT assay investigated the significant potency of all the target compounds against the human cervical cancer cell lines (HeLa cells). Cell cycle analysis showed that the representative compound 8a led to cell cycle cessation at G0/G1 phase indicating that CDK2/E1complex could be the plausible biological target for these newly synthesized compounds. Thus, the most active compounds (7c and 8a-c) were tested for their CDK2 inhibitory activity. The biological results revealed their significant CDK2 inhibitory activity with IC50 range of 0.022-1.629 nM. Moreover, RT-PCR gene expression assay showed that compound 8a increased the levels of the nuclear CDK2 regulators P21 and P27 by 2.30 and 5.7 folds, respectively. ELISA tequnique showed also that compound 8a led to remarkable activation of caspases-9 and -3 inducing cell apoptosis. QSAR study showed that the charge distribution and molecular hydrophobicity are the structural features affecting cytotoxic activity in this series. Molecular docking study for the most potent cytotoxic compounds (7c and 8a-c) rationalized their superior CDK2 inhibitory activity through their hydrogen bonding and hydrophobic interactions with the key amino acids in the CDK2 binding site. Pharmacokinetic properties prediction of the most potent compounds showed that the newly synthesized compounds are not only with promising antitumor activity but also possess promising pharmacokinetic properties.

Large-Scale Reanalysis of Publicly Available HeLa Cell Proteomics Data in the Context of the Human Proteome Project.

The practice of data sharing in the proteomics field took off and quickly spread in recent years as a result of collective effort. Nowadays, most journal editors mandate the submission of the original raw mass spectra to one of the databases of the ProteomeXchange consortium. With the exception of large institutional initiatives such as PeptideAtlas or the GPMDB, few new studies are however based on the reanalysis of mass spectrometry data. A wealth of information is thus left unexploited in public databases and repositories. Here, we present the large-scale reanalysis of 41 publicly available data sets corresponding to experiments carried out on the HeLa cancer cell line using a custom workflow. In addition to the search of new post-translational modification sites and "missing proteins", our main goal is to identify single amino acid variants and evaluate their impact on protein expression and stability through the spectral counting quantification approach. The X!Tandem software was selected to perform the search of a total of 56 363 701 tandem mass spectra against a customized variant protein database, compiled by the application of the in-house MzVar tool on HeLa-specific somatic and genomic variants retrieved from the COSMIC cell line project. After filtering the resulting identifications with a 1% FDR threshold computed at the protein level, 49 466 unique peptides were identified in 7266 protein entries, allowing the validation of 5576 protein entries in accordance with the HPP guidelines version 2.1. A new "missing protein" was observed (FRAT2, NX_O75474, chromosome 10), and 189 new phosphorylation and 392 new protein N-terminal acetylation sites could be identified. Twenty-four variant peptides were also identified, corresponding to 21 variants in 21 proteins. For three of the nine heterozygous cases where both the variant peptide and its wild-type counterpart were detected, the application of a two-tailed sign test showed a significant difference in the abundance of the two peptide versions.