Evaluation of the Potential Cytoprotective Effect of Melatonin in Comparison with Vitamin E and Trolox against Cd2+-Induced Toxicity in SH-SY5Y, HCT 116, and HepG2 Cell Lines.

Cadmium (Cd) toxicity poses a significant threat to cellular health, leading to oxidative stress and cell damage. Antioxidant agents, particularly those of natural origin, have been studied as a potential alternative for mitigating heavy metal toxicity. This study aimed to evaluate the cytoprotective effects of the antioxidant melatonin (MLT) in comparison with Vitamin E (VitE) and Trolox against Cd2+-induced cellular toxicity. The MTT assay was employed to assess cell viability in neuronal SH-SY5Y, colorectal HCT 116, and hepatic HepG2 cell lines. The results showed that all three antioxidants offered some level of protection against Cd toxicity, with Vitamin E proving to be the most effective. MLT also demonstrated a substantial cytoprotective effect, especially at the highest Cd concentration of 30 µM. These findings suggest that MLT, alongside Vit E and Trolox, could be valuable in mitigating the detrimental effects of Cd exposure by reducing the oxidative stress in these cellular models.

In vitro assessment of cytotoxicity of spent fluid catalytic cracking refinery catalysts on cell lines and identification of critical toxic metals.

The Purpose of the present study was to quantify the responses of ten cell lines (HeLa, HepG2, HEK293, MDA-MB-231, A498, A549, A357, 3 T3, BALB-C3 T3, and NIH-3 T3) to spent fluid catalytic cracking catalysts (SFCCCs) from different petroleum refineries, and relate these responses to metal concentrations of SFCCC leachates (SFCCCLs). Cytotoxicity of SFCCCs were significantly different depending on cell lines. A357 and 3 T3 cell were the most sensitive, and A498 and HeLa cells were the least sensitive. HEK293 cells showed the least fluctuation in toxic response to different SFCCCLs among all cells. Cytotoxic IC50 values of SFCCCs to 7 kinds of cells were the most correlated with vanadium (V) concentration in SFCCCLs. V is the most critical toxic factor of SFCCC. Glutathione synthesis was induced in HepG2 cells exposed to higher concentrations of SFCCCLs. SFCCCLs with low concentration of V can induce the decrease of GSH/GSSG ratio in HepG2 cells, suggesting that high concentration of V inhibits the detoxification of glutathione.

Next-Gen Dual Transcriptomics for Adult Extrapulmonary Tuberculosis Biomarkers and Host-Pathogen Interplay in Human Cells: A Strategic Review.

Tuberculosis (TB) is a major public health concern that results in significant morbidity and mortality, particularly in middle- to low-income countries. Extra-pulmonary tuberculosis (EPTB) in adults is a form of TB that affects organs other than the lungs and is challenging to diagnose and treat due to a lack of accurate early diagnostic markers and inadequate knowledge of host immunity. Next-generation sequencing-based approaches have shown potential for identifying diagnostic biomarkers and host immune responses related to EPTB. This strategic review discusses on the significance using primary human cells and cell lines for in vitro transcriptomic studies on common forms of EPTB, such as lymph node TB, brain TB, bone TB, and endometrial TB to derive potential insights. While organoids have shown promise as a model system, primary cell lines still remain a valuable tool for studying host-pathogen interplay due to their conserved immune system, non-iPSC origin, and lack of heterogeneity in cell population. This review outlines a basic workflow for researchers interested in performing transcriptomics studies in EPTB, and also discusses the potential of cell-line based dual RNA-Seq technology for deciphering comprehensive transcriptomic signatures, host-pathogen interplay, and biomarkers from the host and Mycobacterium tuberculosis. Thus, emphasizing the implementation of this technique which can significantly contribute to the global anti-TB effort and advance our understanding of EPTB.

Characterization of a pangolin SARS-CoV-2-related virus isolate that uses the human ACE2 receptor.

Various SARS-CoV-2-related coronaviruses have been increasingly identified in pangolins, showing a potential threat to humans. Here we report the infectivity and pathogenicity of the SARS-CoV-2-related virus, PCoV-GX/P2V, which was isolated from a Malayan pangolin (Manis javanica). PCoV-GX/P2V could grow in human hepatoma, colorectal adenocarcinoma cells, and human primary nasal epithelial cells. It replicated more efficiently in cells expressing human angiotensin-converting enzyme 2 (hACE2) as SARS-CoV-2 did. After intranasal inoculation to the hACE2-transgenic mice, PCoV-GX/P2V not only replicated in nasal turbinate and lungs, but also caused interstitial pneumonia, characterized by infiltration of mixed inflammatory cells and multifocal alveolar hemorrhage. Existing population immunity established by SARS-CoV-2 infection and vaccination may not protect people from PCoV-GX/P2V infection. These findings further verify the hACE2 utility of PCoV-GX/P2V by in vivo experiments using authentic viruses and highlight the importance for intensive surveillance to prevent possible cross-species transmission.

General and Specific Cytotoxicity of Chimeric Antisense Oligonucleotides in Bacterial Cells and Human Cell Lines.

In the last two decades, antisense oligonucleotide technology has emerged as a promising approach to tackling various healthcare issues and diseases, such as antimicrobial resistance, cancer, and neurodegenerative diseases. Despite the numerous improvements in the structure and modifications of the antisense oligonucleotides (ASOs), there are still specific problems with their clinical efficacy and preclinical cytotoxicity results. To better understand the effects of the ASOs in this paper, we conducted many MTT assays to assess the general and specific cytotoxicity of four new chimeric ASOs in bacterial cells and human cell lines. We demonstrate the absence of inhibitory activity in the human pathogenic bacteria Staphylococcus aureus by non-specific ASOs. The pVEC-ASO1 and pVEC-ASO2 are designed to have no specific targets in S. aureus. They have only partial hybridization to the guanylate kinase mRNA. The pVEC-ASO3 targets UBA2 mRNA, a hallmark cancer pathology in MYC-driven cancer, while pVEC-ASO4 has no complementary sequences. We discovered some cytotoxicity of the non-specific ASOs in healthy and cancer human cell lines. The results are compared with two other ASOs, targeting specific mRNA in cancer cells. All ASOs are delivered into the cell via the cell-penetrating oligopeptide pVEC, which is attached to them. We draw a good correlation between the thermodynamic stability of ASO/target RNA and the toxicity effect in human cell lines. The data obtained signify the importance of thorough bioinformatic analysis and high specificity in designing and developing novel ASOs for safer therapeutic agents in clinical practice.

Strategies for single base gene editing in an immortalized human cell line by CRISPR/Cas9 technology.

The use of CRISPR/Cas9 system has rapidly grown in the last years. Here, the optimization of gene editing of a single-nucleotide polymorphism in a human non-malignant somatic cell line of thyrocytes (Nthy-Ori) was described highlighting strategies for overcoming the problems concerning the delivery and off-targets. We employed both lentivirus and chemical lipids as delivery agents and two strategies for creating the double-strand breaks (DSB). The former induced a DSB by a classical Cas9 nuclease (standard strategy), while the second one employed a modified Cas9 creating a single-strand break (SSB). The knock-in was carried out using a single-stranded donor oligonucleotide or the HR410-PA donor vector (HR). The desired cells could be obtained by combining the double nickase system with the HR vector transfected chemically. This result could be due to the type of DSB, likely processed mainly by non-homologous end joining when blunt (standard strategy) and by HR when overhanging (double nickase). Our results showed that the double nickase is suitable for knocking-in the immortalized Nthy-Ori cell line, while the standard CRISPR/Cas9 system is suitable for gene knock-out creating in/del mutations. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03878-4.

Mechanisms related to carbon nanotubes genotoxicity in human cell lines of respiratory origin.

Potential genotoxicity and carcinogenicity of carbon nanotubes (CNT), as well as the underlying mechanisms, remains a pressing topic. The study aimed to evaluate and compare the genotoxic effect and mechanisms of DNA damage under exposure to different types of CNT. Immortalized human cell lines of respiratory origin BEAS-2B, A549, MRC5-SV40 were exposed to three types of CNT: MWCNT Taunit-M, pristine and purified SWCNT TUBALL™ at concentrations in the range of 0.0006-200 µg/ml. Data on the CNT content in the workplace air were used to calculate the lower concentration limit. The genotoxic potential of CNTs was investigated at non-cytotoxic concentrations using a DNA comet assay. We explored reactive oxygen species (ROS) formation, direct genetic material damage, and expression of a profibrotic factor TGFB1 as mechanisms related to genotoxicity upon CNT exposure. An increase in the number of unstable DNA regions was observed at a subtoxic concentration of CNT (20 µg/ml), with no genotoxic effects at concentrations corresponding to industrial exposures being found. While the three test articles of CNTs exhibited comparable genotoxic potential, their mechanisms appeared to differ. MWCNTs were found to penetrate the nucleus of respiratory cells, potentially interacting directly with genetic material, as well as to enhance ROS production and TGFB1 gene expression. For A549 and MRC5-SV40, genotoxicity depended mainly on MWCNT concentration, while for BEAS-2B - on ROS production. Mechanisms of SWCNT genotoxicity were not so obvious. Oxidative stress and increased expression of profibrotic factors could not fully explain DNA damage under SWCNT exposure, and other mechanisms might be involved.

Studying the Functional Potential of Ground Ivy (Glechoma hederacea L.) Extract Using an In Vitro Methodology.

Glechoma hederacea L., known as ground ivy, has a long history of use in folk medicine. The main bioactive compounds in ground ivy are polyphenolic compounds known for their potent antioxidant and antimicrobial activities and thus have high potential as functional ingredients against bacterial infections and the occurrence of chronic diseases associated with oxidative stress in the human body. The aim of the present study was to determine the biological activity of ground ivy extract on selected human cell lines, including hepatic (HepG2), tongue (CAL 27), gastric (AGS) and colon (Caco-2) cancer cell lines by evaluating cytotoxicity, formation of reactive oxygen species and genotoxicity. The antioxidant capacity of the extract was additionally evaluated using cellular model macromolecules of protein and DNA, bovine serum album and plasmid phiX174 RF1 DNA. The effect of ground ivy extract on representatives of human microflora, including L. plantarum, E. coli and S. aureus, was also studied. The cytotoxicity of the extract depended on the type of cells treated, and the pro-oxidant effect generally decreased with increasing exposure time. The most pronounced genoprotective effect against hydroxyl radical damage was monitored in model plasmid DNA and occurred at the highest tested concentration (0.25 mg mL-1), with 95.89% preservation of the supercoiled form of the plasmid. This concentration also had the most significant antioxidant activity on the model protein-14.01% more than the positive control prepared using Trolox. The ground ivy extract showed high antimicrobial potential against the pathogenic bacteria E. coli and S. aureus.

Rapid and permanent cytotoxic effects of venom from Chiropsella bronzie and Malo maxima on human skeletal and cardiac muscle cells.

Jellyfish envenomation is a global public health risk; Cubozoans (box jellyfish) are a prevalent jellyfish class with some species causing potent and potentially fatal envenomation in tropical Australian waters. Previous studies have explored the mechanism of action of venom from the lethal Cubozoan Chironex fleckeri and from Carukia barnesi (which causes "Irukandji syndrome"), but mechanistic knowledge to develop effective treatment is still limited. This study performed an in-vitro cytotoxic examination of the venoms of Chiropsella bronzie and Malo maxima, two understudied species that are closely related to Chironex fleckeri and Carukia barnesi respectively. Venom was applied to human skeletal muscle cells and human cardiomyocytes while monitoring with the xCELLigence system. Chiropsella bronzie caused rapid cytotoxicity at concentrations as low as 58.8 µg/mL. Malo maxima venom caused a notable increase in cell index, a measure of cell viability, followed by cytotoxicity after 24-h venom exposure at ≥11.2 µg/mL on skeletal muscle cells. In contrast, the cardiomyocytes mostly showed significant increased cell index at the higher M. maxima concentrations tested. These findings show that these venoms can exert cytotoxic effects and Malo maxima venom mainly caused a sustained increase in cell index across both human cell lines, suggesting a different mode of action to Chiropsella bronzie. As these venoms show different real-world envenomation symptoms, the different cellular toxicity profiles provide a first step towards developing improved understanding of mechanistic pathways and novel envenomation treatment.

Hub genes identification and association of key pathways with hypoxia in cancer cells: A bioinformatics analysis.

Three human cancer cell lines (A549, HCT116, and HeLa) were used to investigate the molecular mechanisms and potential prognostic biomarkers associated with hypoxia. We obtained gene expression data from Gene Expression Omnibus (GEO) datasets GSE11704, GSE147384, and GSE38061, which included 5 hypoxic and 8 control samples. Using the GEO2R tool and Venn diagram software, we identified common differentially expressed genes (cDEGs). The cDEGs were then subjected to Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis by employing DAVID. The hub genes were identified from critical PPI subnetworks through CytoHuba plugin and these genes' prognostic significance and expression were verified using Kaplan-Meier analysis and Gene Expression Profiling Interactive Analysis (GEPIA), respectively. The research showed 676 common DEGs (cDEGs), with 207 upregulated and 469 downregulated genes. The STRING analysis showed 673 nodes and 1446 edges in the PPI network. We identified 4 significant modules and 19 downregulated hub genes. GO analysis revealed all of them were majorly involved in ribosomal large subunit assembly and biogenesis, rRNA processing, ribosome biogenesis, translation, RNA & protein binding frequently at the sites of nucleolus and nucleoplasm while 11 were significantly associated with a better prognosis of hypoxic tumors. Our research sheds light on the molecular mechanisms that underpin hypoxia in human cancer cell lines and identifies potential prognostic biomarkers for hypoxic tumors.

Efficient tagging of endogenous proteins in human cell lines for structural studies by single-particle cryo-EM.

CRISPR/Cas9-based genome engineering has revolutionized our ability to manipulate biological systems, particularly in higher organisms. Here, we designed a set of homology-directed repair donor templates that enable efficient tagging of endogenous proteins with affinity tags by transient transfection and selection of genome-edited cells in various human cell lines. Combined with technological advancements in single-particle cryogenic electron microscopy, this strategy allows efficient structural studies of endogenous proteins captured in their native cellular environment and during different cellular processes. We demonstrated this strategy by tagging six different human proteins in both HEK293T and Jurkat cells. Moreover, analysis of endogenous glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in HEK293T cells allowed us to follow its behavior spatially and temporally in response to prolonged oxidative stress, correlating the increased number of oxidation-induced inactive catalytic sites in GAPDH with its translocation from cytosol to nucleus.

Anti-Inflammatory and Antioxidant Effects of Leaves and Sheath from Bamboo (Phyllostacys edulis J. Houz).

Bamboo (Phyllostacys edulis J. Houz) has become an emerging forest resource of economic and ecological significance with health benefits. Since the beneficial effects of the non-edible parts of bamboo have not been thoroughly explored, we characterized in this study bamboo leaf (BL) and sheath (BS) extracts. The total phenol and flavonoid content (TPC and TFC), antioxidant activity (ABTS, DPPH, FRAP and ß-carotene bleaching test) and anti-inflammatory properties were determined. Leaves exhibited a TPC value of 73.92 mg equivalent (eq) gallic acid/g fresh weight (FW) and a TFC value of 56.75 mg eq quercetin/g FW. Ultra-High-Performance Liquid Chromatography (UHPLC) coupled with photo diode array detector (PDA) analysis revealed evidence for the presence of protocatechuic acid, isoorientin, orientin and isovitexin in BL, whereas BS was rich in phenolic acids. Both samples demonstrated a significant ability to scavenge radicals against ABTS·+, with an inhibitory concentration of 50% of 3.07 µg/mL for BL and 6.78 µg/mL for BS. At a concentration of 0.1 and 0.2 mg/mL, BS decreased reactive oxygen species production without hampering cell viability in HepG2 liver cells, while at the same concentrations, BL exhibited cytotoxicity in HepG2 cells. In addition, 0.1 and 0.2 mg/mL BS and BL reduced Interleukin-6 and Monocyte Chemoattractant Protein-1 production in human lipopolysaccharide-stimulated THP-1 macrophages, without affecting cell viability. These findings highlight the anti-inflammatory and antioxidant properties of BL and BS, corroborating their different potential applications in the nutraceutical, cosmetic and pharmaceutical industries.

Identify essential genes based on clustering based synthetic minority oversampling technique.

Prediction of essential genes in a life organism is one of the central tasks in synthetic biology. Computational predictors are desired because experimental data is often unavailable. Recently, some sequence-based predictors have been constructed to identify essential genes. However, their predictive performance should be further improved. One key problem is how to effectively extract the sequence-based features, which are able to discriminate the essential genes. Another problem is the imbalanced training set. The amount of essential genes in human cell lines is lower than that of non-essential genes. Therefore, predictors trained with such imbalanced training set tend to identify an unseen sequence as a non-essential gene. Here, a new over-sampling strategy was proposed called Clustering based Synthetic Minority Oversampling Technique (CSMOTE) to overcome the imbalanced data issue. Combining CSMOTE with the Z curve, the global features, and Support Vector Machines, a new protocol called iEsGene-CSMOTE was proposed to identify essential genes. The rigorous jackknife cross validation results indicated that iEsGene-CSMOTE is better than the other competing methods. The proposed method outperformed λ-interval Z curve by 35.48% and 11.25% in terms of Sn and BACC, respectively.

Preparation of 6-Mercaptopurine Loaded Liposomal Formulation for Enhanced Cytotoxic Response in Cancer Cells.

6-Mercaptopurine (6-MP) is a well-known immunosuppressive medication with proven anti-proliferative activities. 6-MP possesses incomplete and highly variable oral absorption due to its poor water solubility, which might reduce its anti-cancer properties. To overcome these negative effects, we developed neutral and positively charged drug-loaded liposomal formulations utilizing the thin-film hydration technique. The prepared liposomal formulations were characterized for their size, polydispersity index (PDI), zeta potential, and entrapment efficiency. The average size of the prepared liposomes was between 574.67 ± 37.29 and 660.47 ± 44.32 nm. Positively charged liposomes (F1 and F3) exhibited a lower PDI than the corresponding neutrally charged ones (F2 and F4). Entrapment efficiency was higher in the neutral liposomes when compared to the charged formulation. F1 showed the lowest IC50 against HepG2, HCT116, and MCF-7 cancer cells. HepG2 cells treated with F1 showed the highest level of inhibition of cell proliferation with no evidence of apoptosis. Cell cycle analysis showed an increase in the G1/G0 and S phases, along with a decrease in the G2/M phases in the cell lines treated with drug loaded positively charged liposomes when compared to free positive liposomes, indicating arrest of cells in the S phase due to the stoppage of priming and DNA synthesis outside the mitotic phase. As a result, liposomes could be considered as an effective drug delivery system for treatment of a variety of cancers; they provide a chance that a nanoformulation of 6-MP will boost the cytotoxicity of the drug in a small pharmacological dose which provides a dosage advantage.

Antioxidant and Anti-Inflammatory Effects of Extracts from Pulsed Electric Field-Treated Artichoke By-Products in Lipopolysaccharide-Stimulated Human THP-1 Macrophages.

In this study, pulsed electric field (PEF-3 kV/cm; 5 kJ/kg) pretreatment was used to intensify the extractability of valuable intracellular compounds from artichoke by-products during a subsequent aqueous extraction (solid-liquid ratio = 1:10 g/mL, T = 20 °C; t = 120 min). Total phenolic content (TPC), antioxidant activity (DPPH, ABTS) and HPLC-PDA analysis of the artichoke extract (AE) and the biological effects on human cell lines were determined. Chlorogenic acid was found to be the most abundant phenolic compound (53% of the TPC) in the AE. The extract showed good antioxidant properties in a concentration-dependent manner. The potential biological effects of AE were investigated using THP-1 macrophages stimulated by lipopolysaccharides (LPS) as an in vitro model system of oxidative stress. Reduced reactive oxygen species production upon treatment with AE was found. Moreover, AE was able to reduce the secretion of the pro-inflammatory mediators Interleukin-6 and Monocyte Chemoattractant Protein-1 in LPS-stimulated macrophages, as determined by qRT-PCR and ELISA assays. These results highlighted the anti-inflammatory and antioxidant properties of the extracts from PEF-treated artichoke by-products, corroborating their potential application as a source of functional ingredients obtained through a feasible and sustainable process.

Cytotoxic activity of strawberry tree (Arbutus unedo L.) honey, its extract, and homogentisic acid on CAL 27, HepG2, and Caco-2 cell lines.

Strawberry tree (Arbutus unedo L.) honey (STH), also known as "bitter honey", is a traditional medicine widely used in the Mediterranean area. Regardless of geographical origin, it usually has a very high content of phenolic compounds and strong antioxidant capacity. Yet, little is still known about the effects of STH, its phenolic extract (STHE), and its main bioactive compound - homogentisic acid (HGA) - at the cell level. The aim of this study was to estimate total phenolic content, DPPH radical scavenging activity, and ferric reducing antioxidant power of STH made in Croatia and investigate cytotoxic and pro-oxidative effects of STH, STHE and HGA on three human cell lines: tongue squamous cell carcinoma (CAL 27), hepatocellular carcinoma (HepG2), and epithelial colorectal adenocarcinoma cells (Caco-2) cells. These substances were tested at four concentrations (0.5-5× average human daily intake of STH) and over 30 min and 1 and 2 h. Croatian STH had a total phenolic content of 1.67 g gallic acid equivalents (GAE) per kg of honey, DPPH radical scavenging activity of 2.96 mmol Trolox equivalents (TE) per kg of honey, and ferric reducing antioxidant power (FRAP) of 13.5 mmol Fe2+ per kg of honey. Our results show no clear and consistent time- or concentration-dependent cytotoxicity in any of the cell lines. ROS levels in all the three cell types at almost all exposure times were not significantly higher than control. The most important observation is that the tested substances have low cytotoxicity and high biocompatibility, regardless of concentration, which is a good starting point for further research of their biological effects in other models.

Wheat Germ Spermidine and Clove Eugenol in Combination Stimulate Autophagy In Vitro Showing Potential in Supporting the Immune System against Viral Infections.

Impaired autophagy, responsible for increased inflammation, constitutes a risk factor for the more severe COVID-19 outcomes. Spermidine (SPD) is a known autophagy modulator and supplementation for COVID-19 risk groups (including the elderly) is recommended. However, information on the modulatory effects of eugenol (EUG) is scarce. Therefore, the effects of SPD and EUG, both singularly and in combination, on autophagy were investigated using different cell lines (HBEpiC, SHSY5Y, HUVEC, Caco-2, L929 and U937). SPD (0.3 mM), EUG (0.2 mM) and 0.3 mM SPD + 0.2 mM EUG, significantly increased autophagy using the hallmark measure of LC3-II protein accumulation in the cell lines without cytotoxic effects. Using Caco-2 cells as a model, several crucial autophagy proteins were upregulated at all stages of autophagic flux in response to the treatments. This effect was verified by the activation/differentiation and migration of U937 monocytes in a three-dimensional reconstituted intestinal model (Caco-2, L929 and U937 cells). Comparable benefits of SPD, EUG and SPD + EUG in inducing autophagy were shown by the protection of Caco-2 and L929 cells against lipopolysaccharide-induced inflammation. SPD + EUG is an innovative dual therapy capable of stimulating autophagy and reducing inflammation in vitro and could show promise for COVID-19 risk groups.

In vitro Susceptibility of Human Cell Lines Infection by Bovine Leukemia Virus.

Evidence of the presence of bovine leukemia virus (BLV) in human beings and its association with breast cancer has been published in the literature, proposing it as a zoonotic infection. However, not enough evidence exists about transmission pathways nor biological mechanisms in human beings. This study was aimed at gathering experimental evidence about susceptibility of human cell lines to BLV infection. Malignant and non-malignant human cell lines were co-cultured with BLV-infected FLK cells using a cell-to-cell model of infection. Infected human cell lines were harvested and cultured for 3 to 6 months to determine stability of infection. BLV detection was performed through liquid-phase PCR and visualized through in situ PCR. Seven out of nine cell lines were susceptible to BLV infection as determined by at least one positive liquid-phase PCR result in the 3-month culture period. iSLK and MCF7 cell lines were able to produce a stable infection throughout the 3-month period, with both cytoplasmic and/or nuclear BLV-DNA visualized by IS-PCR. Our results support experimental evidence of BLV infection in humans by demonstrating the susceptibility of human cells to BLV infection, supporting the hypothesis of a natural transmission from cattle to humans.

Differential comparison of genotoxic effects of aristolochic acid I and II in human cells by the mass spectroscopic quantification of γ-H2AX.

Aristolochic acids (AAs) are known to be the potent genotoxic carcinogens, of which aristolochic acid I (AAI) and aristolochic acid II (AAII) are the two representative compounds. As the carcinogenic risk of herbs containing AAs is a global health issue, quantitative evaluation of genotoxicity is needed for the risk assessment of AAs. γ-H2AX, which is an acknowledged attractive bifunctional biomarker for simultaneously reflect the DNA damage response and repair, was used to quantitatively determine the DNA damage and repair properties of AAI and AAII in human cell lines, based on our previously developed mass spectrometry method. Results indicated that both AAI and AAII could increase the level of γ-H2AX in cells in a concentration-dependent manner, and the increased level of γ-H2AX induced by AAI was relatively higher than that induced by AAII. Time-effect curves showed that the change tendency of the proportion of γ-H2AX was obviously different in the later period, particularly afterwards 8 h post exposure. Additionally, AAI and AAII induced an opposite change of expression levels of DNA damage repair-associated proteins (ERCC1 and p53) in HepG2 cells, revealing their distinct molecular mechanisms. Findings of the present study are helpful for understanding the genotoxicity mechanism of AAI and AAII.

Pharmacokinetics of PEGylated Gold Nanoparticles: In Vitro-In Vivo Correlation.

Data suitable for assembling a physiologically-based pharmacokinetic (PBPK) model for nanoparticles (NPs) remain relatively scarce. Therefore, there is a trend in extrapolating the results of in vitro and in silico studies to in vivo nanoparticle hazard and risk assessment. To evaluate the reliability of such approach, a pharmacokinetic study was performed using the same polyethylene glycol-coated gold nanoparticles (PEG-AuNPs) in vitro and in vivo. As in vitro models, human cell lines TH1, A549, Hep G2, and 16HBE were employed. The in vivo PEG-AuNP biodistribution was assessed in rats. The internalization and exclusion of PEG-AuNPs in vitro were modeled as first-order rate processes with the partition coefficient describing the equilibrium distribution. The pharmacokinetic parameters were obtained by fitting the model to the in vitro data and subsequently used for PBPK simulation in vivo. Notable differences were observed in the internalized amount of Au in individual cell lines compared to the corresponding tissues in vivo, with the highest found for renal TH1 cells and kidneys. The main reason for these discrepancies is the absence of natural barriers in the in vitro conditions. Therefore, caution should be exercised when extrapolating in vitro data to predict the in vivo NP burden and response to exposure.