Cytokinesis-block Micronucleus Assay Research Articles

Medical personnel occupationally exposed to low-dose ionising radiation in Federation of Bosnia and Herzegovina: A cytogenetic study.

Medical radiation exposures have been reduced significantly with modern equipment and protection measures. Biomonitoring of medical personnel can provide information concerning possible effects of radiation exposure. However, chromosome aberration (CA) analysis is now recommended only when the estimated effective dose is 200 mSv or higher. In this retrospective study in Bosnia and Herzegovina, we have measured the cytogenetic status of medical workers and healthy volunteers (controls). Peripheral blood samples from 66 medical workers exposed to low-dose ionising radiation and 89 non-exposed volunteers were collected for chromosome aberrations (CA) analysis and the cytokinesis-block micronucleus (CBMN) assay. Higher rates of chromatid and chromosome breaks, acentric fragments, double minutes, micronuclei, and micronucleated binuclear cells were observed in the control group, while the rate of nucleoplasmic bridges was higher in the medical workers group.

Associating Air Pollution with Cytokinesis-Block Micronucleus Assay Parameters in Lymphocytes of the General Population in Zagreb (Croatia).

Air pollution is recognized as one of the most serious public health issues worldwide and was declared to be a leading environmental cause of cancer deaths. At the same time, the cytokinesis-block micronucleus (CBMN) assay serves as a cancer predictive method that is extensively used in human biomonitoring for populations exposed to environmental contamination. The objective of this cross-sectional study is two-fold: to evaluate genomic instability in a sample (N = 130) of healthy, general population residents from Zagreb (Croatia), chronically exposed to different levels of air pollution, and to relate them to air pollution levels in the period from 2011 to 2015. Measured frequencies of CBMN assay parameters were in agreement with the baseline data for the general population of Croatia. Air pollution exposure was based on four factors obtained from a factor analysis of all exposure data obtained for the examined period. Based on the statistical results, we did not observe a significant positive association between any of the CBMN assay parameters tested and measured air pollution parameters for designated time windows, except for benzo(a)pyrene (B[a]P) that showed significant negative association. Our results show that measured air pollution parameters are largely below the regulatory limits, except for B[a]P, and as such, they do not affect CBMN assay parameters’ frequency. Nevertheless, as air pollution is identified as a major health threat, it is necessary to conduct prospective studies investigating the effect of air pollution on genome integrity and human health.

P08-19 In vitro cytokinesis block micronucleus (CBMN) assay to evaluate the genotoxicity of multicomponent nanomaterials – a comparison with their individual counterparts

P08-19 In vitro cytokinesis block micronucleus (CBMN) assay to evaluate the genotoxicity of multicomponent nanomaterials – a comparison with their individual counterparts

Cyto-genotoxicity of crystalline and amorphous niobium (V) oxide nanoparticles in CHO-K1 cells.

Niobium (V) oxide nanoparticles (NINPs) have been widely and increasingly applied in various health products and industrial processes. This merits further study of their toxicity. Here, we investigated the potential of NINPs to induce DNA damage, cytotoxicity, and chromosome instability in cultured CHO-K1 cells. NINPs were physico-chemically characterized. As assessed by comet assay, crystalline and amorphous NINPs were genotoxic at the highest concentrations evaluated. The cytokinesis-block micronucleus assay demonstrated that a 24-h treatment with NINPs, for the crystalline and the amorphous samples, significantly reduced the nuclear division cytotoxicity index. In addition, a 4-h treatment period of crystalline NINPs increased micronucleus (MNi) frequencies. MNi, nucleoplasmic bridges and nuclear buds were detected after exposure of the cells for 24h to crystalline NINPs. In the amorphous sample, chromosome instability was restricted to the induction of MNi, in the 24-h treatment, detected at all tested concentrations. The fluorescence and dark field microscopy demonstrated the uptake of NINPs by CHO-K1 cells and an intracellular distribution outlining the nucleus. Our data advance understanding of the cytotoxic and genotoxic effects of NINPs and should be taken into consideration when setting up guidelines for their use in industrial or health products.

ESTABLISHMENT OF THE IN VITRO DOSE-RESPONSE CALIBRATION CURVE FOR X-RAY-INDUCED MICRONUCLEI IN HUMAN LYMPHOCYTES.

The cytokinesis-block micronucleus assay has proven to be a reliable technique for biological dosimetry. This study aimed to establish the dose-response curve for X-ray-induced micronucleus. Peripheral blood samples from three healthy donors were irradiated with various doses and scoring criteria by the micronuclei (MN) in binucleated cells. The results showed that the frequency of MN increased with the elevation of radiation dose. CABAS and Dose Estimate software were used to fit the MN and dose into a linear quadratic model, and the results were compared. The linear and quadratic coefficients obtained by the two software were basically the same and were comparable with published curves of similar radiation quality and dose rates by other studies. The dose-response curve established in this study can be used as an alternative method for in vitro dose reconstruction and provides a reliable tool for biological dosimetry in accidental or occupational radiation exposures.

Plastic additive di(2-ethylhexyl)phthalate (DEHP) causes cell death and micronucleus induction on a bottlenose dolphin’s (Tursiops truncatus) in vitro-exposed skin cell line

Marine plastic pollution is one of the most concerning worldwide environmental issues, and research is day by day demonstrating its adverse effects on marine ecosystems. Nevertheless, little is still known about the toxic potential on marine fauna of chemical additives released by plastic debris. Here we investigated the cyto- and genotoxicity of the most used plasticizer in plastic production, di(2-ethylhexyl)phthalate (DEHP), on a skin cell line (TT) derived from the bottlenose dolphin (Tursiops truncatus), a species particularly exposed to the accumulation of this lipophilic pollutant, being a coastal top predator rich in fatty subcutaneous tissues. Dolphin cell cultures were exposed to increasing DEHP doses (0.01–5 mM) to evaluate effects on cell viability, cell death, and induction of DNA damage. On the hypothesis that bottlenose dolphin cells show greater resistance to DEHP toxicity than terrestrial mammals, as already shown for other pollutants, the same parameters were analyzed on exposed Chinese hamster ovary (CHO) cell lines. Both MTT and Trypan Blue assays showed no significant decrease in dolphin’s cell viability after 24-h DEHP exposure. No induction of primary DNA damage was detected by the comet assay, whereas the cytokinesis-block micronucleus assay revealed significant micronuclei induction and inhibition of cell proliferation starting from the lowest DEHP doses. DEHP had similar but sharper and significant effects on cell viability in CHO cells, also causing a much greater induction of necrosis than that recorded on dolphin cells. For both cell lines, the lack of induction of primary DNA damage (i.e., strand breaks) together with the increase of micronuclei yield after DEHP treatment suggests an aneugenic effect of the phthalate, that is, the loss of entire chromosomes during cell division. Overall, the potential chromosome loss detected could constitute a threat for species of marine mammals constantly exposed to plastic marine litter.

Hydrochar and hydrochar co-compost from OFMSW digestate for soil application: 3. Toxicological evaluation

Modern societies produce ever-increasing amounts of waste, e.g. organic fraction of municipal solid waste (OFMSW). According to the best available techniques, OFMSW should be treated through anaerobic digestion to recover biogas and subsequently composted. An innovative scheme is under investigation, where anaerobic digestion is combined with hydrothermal carbonization (HTC) and composting. The final product, referred to as hydrochar co-compost (HCO), is under study to be used as an unconventional soil improver/fertilizer. Recent studies showed that HCO is not phytotoxic. However, nothing is known about the toxicity of HCO on cells as part and organisms as a whole. This study aims to investigate in vitro genotoxicity and cytotoxicity of the HCO and its precursors in the production process. In particular, we tested water and methanolic extracts of HCO (WEHCO and MEHCO) from one side and methanolic extracts of hydrochar (MEH) and OFMSW digestate (MED) as well as liquor produced downstream HTC (HTCL) from the other side. Genotoxicity was investigated using cytokinesis-block micronucleus assay in Chinese Hamster Ovarian K1 (CHO-K1) cells. Cytotoxicity was tested in vitro against a panel of human cells line. Zebrafish embryo toxicity upon MEH treatment was also investigated. Results show that incubation of CHO-K1 cells with all the tested samples at different concentrations did not cause any induction of micronucleus formation compared to the vehicle-treated control. Treatment of cells with MEH, MED, HTCL and MEHCO, but not WEHCO, induced some degree of cytotoxicity and MEH showed to be more cytotoxic against tested cells compared to the MEHCO. Toxicity effect at the highest tested concentrations of MEH on zebrafish embryos resulted in coagulation, induction of pericardial edema and death. In conclusion, the hydrochar co-compost cytotoxicity is similar to standard compost cytotoxicity. Hence composting the hydrochar from OFMSW digestate is a good step to eliminate the cytotoxicity of hydrochar.

Investigation of the Genotoxic, Antigenotoxic and Antioxidant Profile of Different Extracts from Equisetum arvense L.

The present study investigated the cyto-genotoxic and antigenotoxic effects of four different extracts of Equisetum arvense L. (common name: field horsetail) on human lymphocytes. Specifically, Soxhlet’s prepared extracts from E. arvense L., using different solvents (S1: methanol (MeOH)-, S2: ethanol (EtOH)-, S3: water-, and S4: ethanol/water (EtOH-W)-) were analyzed for (a) their total phenolic and flavonoid content (TPC and TFC, respectively), (b) their antioxidant activity (AA), via the DPPH, FRAP and ABTS assays, and (c) their cyto-genotoxic and/or protective efficiency against the mutagenic agent mitomycin C, via the Cytokinesis Block MicroNucleus assay. All extracts showed increased TPC, TFC, and AA values in almost all cases. S1, S3 and S4 demonstrated no cytotoxic potential, whereas S2 was cytotoxic only at the highest concentrations. Genotoxicity was not observed in the tested extracts. The highest antigenotoxic activity was observed for EtOH-W (S4) extract, which was found to be rich in flavonoids, flavonoid-O-glycosides, phytosterols, phenolic and fatty acids as well as in minerals and mainly in K, Ca, Mg, Si and P, as assessed by using various mass spectrometry techniques. Those findings confirm that E. arvense L. extracts could be valuable candidates for medicinal applications and pharmaceutical products, thus alleviating the effects of more conventional drugs.

Quality and Purity of L929 Cell Cultures: Isoenzyme Profiles As a Sensitive Marker of Cross-Contamination.

Objective: Inappropriate handling of cells can generate modifications in the genomic DNA. The additional risk is cross-contamination. Isoenzyme analysis with gel agarose electrophoresis is a known, fast, and cheap technique for detection of species-specific isoforms of intracellular enzymes. The aim of the experimental work was to check if variations in the cell growth conditions can affect morphology and/or nuclear anomalies including micronuclei (MN) in the L929 cells; and to define how sensitive and selective is the classic gel agarose electrophoresis for analysis of isoforms of the selected enzymes to detect cross-contamination of L929 cultures with HeLa cells or with the related species, such as CHO-K1 cells, in the case of unavailability of the commercial kits. Methods: The experiments were done with use of the National Collection of Type Cultures clone 929 (L929)-mouse fibroblasts from subcutaneous connective tissue; HeLa-human cervix adenocarcinoma; and CHO-K1-epithelial-like hamster ovary cells. Cell morphology was evaluated with a light/fluorescence microscope. MN were determined with the cytokinesis-block micronucleus assay, and the isoenzyme analysis was performed using gel agarose electrophoresis. Results: As shown, the overgrown cultures result in a significant increase of the MN in the L929 cells. The band patterns for lactate dehydrogenate, glucose-6-phosphate dehydrogenase, or malate dehydrogenase allow identification of the single L929, HeLa, or CHO-K1 cell line and to detect the cross-contamination, even up to 0.4%. Conclusions: There can be no exceptions from the recommended cell culture conditions in the passage scheme. The sensitivity of the gel agarose separation depends on the cells and on the type of enzyme tested and seems to be sufficient in a quick screening of the CHO-K1, L929, or HeLa cell cultures through the possible mutual contamination.

Development and characterization of hyaluronic acid surface scaffolds Encorafenib loaded polymeric nanoparticles for colorectal cancer targeting

For treatment of colorectal cancer and to improve intravascular bioavailability, Encorafenib (ECF) encapsulated Hyaluronic Acid (HA) decorated polycaprolactone (PCL), and polyhydroxybutyrate (PHB) polymeric nanoparticles were prepared. The DLS, SEM, and TEM data show an increase in particle size (<200 nm) after hyaluronic acid conjugation over polymeric nanoparticles. AFM confirms that HA-linked nanoparticles have a rougher surface. FTIR, DSC, XRD, and TGA confirm that HA conjugates form amorphous nanoparticles. The in-vitro drug release study indicated initial burst release of ECF followed by sustain release profile. In vitro cytotoxicity testing revealed that HA-ECF-PHB-NPs significantly inhibited HCT116 cell lines compared to the other three nanoparticles and ECF drug solutions. The genotoxicity of polymeric nanoparticles was assessed by cytokinesis-block micronucleus (CBMN) assay, indicating DNA damage in HCT116 cells and micronuclei and nuclear buds’ formation. The produced nanoparticles can lower ROS, possibly due to their good cellular internalisation and apoptotic arrest. Histopathology of colon tissue following nanoparticle injection showed no injury. Haemolysis, LDH assay showed nanoparticles' biocompatibility. According to fluorescence research, epithelial colon cells accumulate more fluorescent nanoparticles after i.v. administration. The ECF polymeric nanoparticles fabricated with HA can be considered a promising drug delivery system for colorectal cancer therapy.

The overall assessment of simultaneous photocatalytic degradation of Cimetidine and Amisulpride by using chemical and genotoxicological approaches

Pharmaceutical Active Compounds (PhACs) are of particular interest among the emerging contaminants detected in the aquatic environment. Commonly, PhACs exist as complex mixtures in aquatic systems, causing potential adverse effects to the environment and human health than those of individual compounds. Based on the increasing interest in the contamination of water resources by PhACs, the photocatalytic degradation of Cimetidine and Amisulpride as a mixture in combination with their toxic and genotoxic effects before and after the treatment were evaluated for the first time. The toxic, genotoxic and cytotoxic effects were investigated using the Trypan Blue Exclusion Test and the Cytokinesis Block MicroNucleus (CBMN) assay in cultured human lymphocytes.The photocatalytic degradation of the PhACs was studied in ultrapure water and environmentally relevant matrices using UV-A and visible (Vis) irradiation and C–TiO2 (TiO2 Kronos vlp 7000) as photocatalyst. High removal percentages were observed for both compounds under UV-A and Vis irradiation in ultrapure water. In lake and drinking water a slower degradation rate was shown that could be attributed to the complex composition of these matrices. Scavenging experiments highlighted the significant role of h+ and O2●- in the degradation mechanisms under both irradiation sources. Oxidation, dealkylation and deamination were the main degradation pathways. Regarding the individual compounds, Amisulpride was found to be more cytotoxic than Cimetidine. No significant differences of the genotoxic effects during the treatment were observed. However, a slight increase in cytotoxicity was observed at the first stages of the process. At the end of the process under both UV-A and Vis light, non-significant cytotoxic/toxic effects were observed. Based on the results, heterogeneous photocatalysis can be considered as an effective process for the treatment of complex mixtures without the formation of harmful transformation products.

Selected fungi of the genus Lactarius - screening of antioxidant capacity, antimicrobial activity, and genotoxicity

ABSTRACT It is well-known that mushrooms of the genus Lactarius constitute a natural food resource providing health benefits as a nutritient. This genus contains 4 mushrooms identified as L. deliciosus, L. volemus, L. sanguifluus, L. semisanguifluus and L. piperatus which were collected in Serbia. The aim of this study aimed was to identify and characterize the content of phenolic compounds and examine the antioxidant potential of 5 wild edible mushrooms. L. sanguifluus contained the highest content of total phenol compounds (75.25 mg gallic acid equivalents (GAE) per g dry extract weight (GAE/g DE) and exhibited the greatest antioxidant activity through the ability to remove radicals as evidenced by ABTS assay (8.99 mg of trolox equivalents (TE) per g dry extract weight (mg TE/g DE); total reducing power (TRP) assay mg ascorbic acid equivalents per mg of dry extract weight (0.42 mg AAE/g DE) and CUPRAC (14.23 mg TE /g DE). L. deliciosus methanolic extract produced greatest scavenging of the DPPH radical (46%). The methanol mushroom extracts were screened for in vitro antimicrobial activity against a panel of pathogenic bacterial strains using the microdilution method. Of all the extracts tested, L. sanguifluus extract showed the best antibacterial properties. The cytokinesis block micronucleus assay results for the examined mushrooms demonstrated that extracts at a concentration of 3 μg/ml decreased the number of micronuclei (MN) in the range of 19–49% which is significant bearing in mind that radioprotectant amifostine reduced the frequency of MN by only 16.3%. Data thus demonstrate that the 5 wild edible mushrooms of genus Lactarius contain constituents that are beneficial not only as nutrients but also have the potential as antioxidants, antibacterial and antigenotoxic properties.

Automatic reagent handling and assay processing of human biospecimens inside a transportation container for a medical disaster response against radiation.

Biological materials can be shipped off-site for diagnostic, therapeutic and research purposes. They usually are kept in certain environments for their final application during transportation. However, active reagent handling during transportation from a collection site to a laboratory or biorepository has not been reported yet. In this paper, we show the application of a micro-controlled centrifugal microfluidic system inside a shipping container that can add reagent to an actively cultured human blood sample during transportation to ensure a rapid biodosimetry of cytokinesis-block micronucleus (CBMN) assay. The newly demonstrated concept could have a significant impact on rapid biodosimetry triage for medical countermeasure in a radiological disaster. It also opens a new capability in accelerated sample processing during transportation for biomedical and healthcare applications.

The Biological Effects of Polystyrene Nanoplastics on Human Peripheral Blood Lymphocytes.

Environmental exposure to microplastics (MPs) and nanoplastics (NPs) is an increasing concern from human health perspectives. Little information on the genotoxic and cytotoxic potential of NP particles in human cells is available. We aimed to assess the cytotoxic and genotoxic potential of polystyrene nanoplastics (PSNPs) at different concentrations (2000μg/mL, 1000μg/mL, and 500μg/mL) by using chromosomal aberration (CA) and cytokinesis-block micronucleus assays (CBMN) on human peripheral lymphocytes. Dose-dependent hemolytic activity and cell viability were observed against the PSNPs exposure. Increased chromosomal aberrations, such as chromosomal breaks and dicentric chromosomes, and an increase in nucleoplasmic bridge (NBP) formation and nuclear budding (NBUD) were observed. The frequency of mitotic index (MI) decreased significantly in the PSNP-exposed groups from lower to higher concentrations. A significant increase in micronuclei (MN) formation and cytostasis% and a dose-dependent reduction in nuclear division index (NDI) in PSNP-exposed groups indicated oxidative stress-mediated cytotoxicity, DNA damage, and genomic instabilities due to PSNP exposure in human lymphocyte cells. This study highlights the importance of understanding the toxic mechanisms and associated chronic and acute health effects on humans due to exposure to this pervasive environmental pollutant.

Analysis of the In Vitro Toxicity of Nanocelluloses in Human Lung Cells as Compared to Multi-Walled Carbon Nanotubes.

Cellulose micro/nanomaterials (CMNM), comprising cellulose microfibrils (CMF), nanofibrils (CNF), and nanocrystals (CNC), are being recognized as promising bio-nanomaterials due to their natural and renewable source, attractive properties, and potential for applications with industrial and economical value. Thus, it is crucial to investigate their potential toxicity before starting their production at a larger scale. The present study aimed at evaluating the cell internalization and in vitro cytotoxicity and genotoxicity of CMNM as compared to two multi-walled carbon nanotubes (MWCNT), NM-401 and NM-402, in A549 cells. The exposure to all studied NM, with the exception of CNC, resulted in evident cellular uptake, as analyzed by transmission electron microscopy. However, none of the CMNM induced cytotoxic effects, in contrast to the cytotoxicity observed for the MWCNT. Furthermore, no genotoxicity was observed for CNF, CNC, and NM-402 (cytokinesis-block micronucleus assay), while CMF and NM-401 were able to significantly raise micronucleus frequency. Only NM-402 was able to induce ROS formation, although it did not induce micronuclei. Thus, it is unlikely that the observed CMF and NM-401 genotoxicity is mediated by oxidative DNA damage. More studies targeting other genotoxicity endpoints and cellular and molecular events are underway to allow for a more comprehensive safety assessment of these nanocelluloses.

Genotoxicity and in vitro investigation of Gefitinib-loaded polycaprolactone fabricated nanoparticles for anticancer activity against NCI-H460 cell lines

For non-small cell lung cancer (NSCLC) treatment, a BCS class II drug, Gefitinib, was widely used. Due to poor bioavailability, uncontrollable drug release, Gefitinib witnessed side effects. To circumvent such associated problems, optimized Gefitinib encapsulated polycaprolactone (PCL) nanoparticles with three different molecular weights of PCL (average Mn∼10,000, Mn∼45,000 & Mn∼80,000) were developed using Box–Behnken design while understanding the influence of critical process parameters of the nanoparticles. For morphological characterizations, SEM, TEM, AFM were used. Hemocompatibility, platelet aggregation, and erythrocyte membrane integrity tests were used to test nanoparticles for biocompatibility; excellent biocompatibility was reported during these tests. The in-vitro drug release studies confirmed that Gefitinib-PCL10,000NPs, Gefitinib-PCL45,000NPs, and Gefitinib-PCL80,000 NPs, show significant initial burst effects, and later nanoparticles possessed zero-order kinetics. The genotoxicity of PCL nanoparticles was assessed by cytokinesis-block micronucleus (CBMN) assay, indicating DNA damage in NCI-H460 cell and micronuclei and nuclear buds’ formation. Further, reactive oxygen species studies, MTT cytotoxicity assays at 24 & 48 h, stability, in-vitro cellular uptake of optimized fluorescent Gefitinib PCL80,000NPs, and apoptosis studies were also carried out. As a result, investigating stable Gefitinib-loaded poly-caprolactone (PCL) nanoparticles could open up new research avenues, potentially lowering side effects and improving Gefitinib's profile in the treatment of NSCLC.

In vitro genotoxic and mutagenic effects of water samples from Sapucaia and Esteio streams (Brazil) under the influence of different anthropogenic activities

Pollution of aquatic ecosystems is associated with the discharge of mainly industrial and urban effluents, which may cause damage to public health. This study aims to evaluate the cytotoxic, genotoxic, and mutagenic potential of surface water samples under the influence of different anthropogenic effluents in a human-derived liver cell line (HepG2). Samples were collected in Esteio and Sapucaia streams (Rio Grande do Sul; Brazil), which flow into the Sinos River, a source of water supply for more than one million people. Physicochemical and microbiological analyses were performed as well as an analysis of inorganic elements using the PIXE technique (Particle-Induced X-Ray Emission). The presence of pharmaceutical compounds and caffeine was evaluated by gas chromatography coupled to mass spectrometry. The cytotoxicity, genotoxicity, and mutagenicity of the samples were evaluated in HepG2 cells by cell viability assays, alkaline Comet Assay and Cytokinesis-block micronucleus (CBMN) assay. We verified alterations in the physicochemical and microbiological parameters and detected caffeine, diethyltoluamide, and different inorganic elements that corresponded to elements from domestic and industrial effluents and agricultural runoff. Although the samples in the concentration used were not cytotoxic, water samples from all sites induced DNA damage. However, it is difficult to attribute these damages to a specific substance since the factors are a complex mixture of different compounds. Despite this, it is observed that both urban and industrial contributions had a similar effect in the cells evaluated. Such results demonstrate the need to perform biomonitoring of surface waters under anthropogenic influence, especially those that flow into rivers that are a source of public supply water. We also highlight the need for research into emerging pollutants in these aquatic environments.

Genotoxic effects of the ochratoxin A (OTA), its main metabolite (OTα) per se and in combination with fumonisin B1 in HepG2 cells and human lymphocytes

Ochratoxin A (OTA) and fumonisin B1 (FB1) are mycotoxins distributed in a wide variety of foods for human or animal consumption and are classified as possible carcinogens for humans. This study aimed to evaluate the cytotoxic, cytostatic and genotoxic effects of OTA and its main metabolite, ochratoxin α (OTα), FB1 and three combinations of OTA and FB1 at moderate and environmental doses. Cell viability was evaluated through MTT assay and the trypan blue exclusion method. The cytostatic and genotoxic effects were evaluated through the cytokinesis-block micronucleus assay. The results showed synergistic time- and concentration-dependent cytotoxic effects of one of the combinations of OTA and FB1. In contrast, significant differences were observed in the micronuclei (MN) frequency from OTA, OTα and coexposure of OTA + FB1. Some of these combinations increased the frequency of nuclear buds, nucleoplasmic bridges, donut-shaped nuclei, necrotic and apoptotic cells and MN in mononucleated cells. In conclusion, OTA and its main metabolite OTα, as well as the co-exposure of OTA and FB1, cause stable DNA damage at environmentally relevant concentrations, which was greater in metabolically competent cells. More studies are needed to understand the chemical interactions that occur due to the joint presence of mycotoxins, which occurs commonly.

Rapid and automatic detection of micronuclei in binucleated lymphocytes image

Cytokinesis block micronucleus (CBMN) assay is a widely used radiation biological dose estimation method. However, the subjectivity and the time-consuming nature of manual detection limits CBMN for rapid standard assay. The CBMN analysis is combined with a convolutional neural network to create a software for rapid standard automated detection of micronuclei in Giemsa stained binucleated lymphocytes images in this study. Cell acquisition, adhesive cell mass segmentation, cell type identification, and micronucleus counting are the four steps of the software's analysis workflow. Even when the cytoplasm is hazy, several micronuclei are joined to each other, or micronuclei are attached to the nucleus, this algorithm can swiftly and efficiently detect binucleated cells and micronuclei in a verification of 2000 images. In a test of 20 slides, the software reached a detection rate of 99.4% of manual detection in terms of binucleated cells, with a false positive rate of 14.7%. In terms of micronuclei detection, the software reached a detection rate of 115.1% of manual detection, with a 26.2% false positive rate. Each image analysis takes roughly 0.3 s, which is an order of magnitude faster than manual detection.

Zinc Prevents DNA Damage in Normal Cells but Shows Genotoxic and Cytotoxic Effects in Acute Myeloid Leukemia Cells.

Genomic instability is prevented by the DNA damage response (DDR). Micronutrients, like zinc (Zn), are cofactors of DDR proteins, and micronutrient deficiencies have been related to increased cancer risk. Acute myeloid leukemia (AML) patients commonly present Zn deficiency. Moreover, reports point to DDR defects in AML. We studied the effects of Zn in DDR modulation in AML. Cell lines of AML (HEL) and normal human lymphocytes (IMC) were cultured in standard culture, Zn depletion, and supplementation (40 μM ZnSO4) conditions and exposed to hydrogen peroxide (H2O2) or ultraviolet (UV) radiation. Chromosomal damage, cell death, and nuclear division indexes (NDI) were assessed through cytokinesis-block micronucleus assay. The phosphorylated histone H2AX (yH2AX) expression was monitored at 0 h, 1 h, and 24 h after exposure. Expression of DDR genes was evaluated by quantitative real time polymerase chain reaction (qPCR). Zn supplementation increased the genotoxicity of H2O2 and UV radiation in AML cells, induced cytotoxic and antiproliferative effects, and led to persistent yH2AX activation. In contrast, in normal lymphocytes, supplementation decreased damage rates, while Zn depletion favored damage accumulation and impaired repair kinetics. Gene expression was not affected by Zn depletion or supplementation. Zn presented a dual role in the modulation of genome damage, preventing damage accumulation in normal cells and increasing genotoxicity and cytotoxicity in AML cells.