Several human studies indicate that mobile phone specific electromagnetic fields may cause cancer in humans but the underlying molecular mechanisms are currently not known. Studies concerning chromosomal damage (which is causally related to cancer induction) are controversial and those addressing this issue in mobile phone users are based on the use of questionnaires to assess the exposure. We realized the first human intervention trial in which chromosomal damage and acute toxic effects were studied under controlled conditions. The participants were exposed via headsets at one randomly assigned side of the head to low and high doses of a UMTS signal (n = 20, to 0.1 W/kg and n = 21 to 1.6 W/kg Specific Absorption Rate) for 2 h on 5 consecutive days. Before and three weeks after the exposure, buccal cells were collected from both cheeks and micronuclei (MN, which are formed as a consequence of structural and numerical chromosomal aberrations) and other nuclear anomalies reflecting mitotic disturbance and acute cytotoxic effects were scored. We found no evidence for induction of MN and of nuclear buds which are caused by gene amplifications, but a significant increase of binucleated cells which are formed as a consequence of disturbed cell divisions, and of karyolitic cells, which are indicative for cell death. No such effects were seen in cells from the less exposed side. Our findings indicate that mobile phone specific high frequency electromagnetic fields do not cause acute chromosomal damage in oral mucosa cells under the present experimental conditions. However, we found clear evidence for disturbance of the cell cycle and cytotoxicity. These effects may play a causal role in the induction of adverse long term health effects in humans.

In three water-supply reservoirs in the catchment area of the Odra River (Czech Republic), a special fish stock was monitored for control of health to estimate the mutagenic effect of chemicals. The results contribute to obtaining initial information about the morphology of erythrocyte abnormalities classified in 21 categories in 16 fish species in reservoirs with abundant salmonids (the Morávka Reservoir) or with the prevalence of cyprinids (the Kružberk and Šance Reservoirs), not directly exposed to the adverse environmental effects such as industrial, urban, agricultural and intensive farming activities. The different intensities and prevalence of nuclear abnormalities (NA) and cytoplasmic abnormalities (CA) in fish from the same reservoir habitat show that to be able to obtain an objective view of the genotoxic risk of chemicals, it is necessary to respect the different requirements of the fish for the exploitation of the food available in the biotope and to subject all representatives of piscivorous, omnivorous and benthophagous fishes in the reservoir to cytogenetic analysis. The occurrence of certain categories of erythrocyte abnormalities in diseased fish draws attention to the need to know the state of health of the fish and to employ this knowledge to exclude parasitological, viral and other infectious agents. These results are the first report of the frequencies of erythrocyte abnormalities in native fish. They should serve to check which of the categories examined could be of use in assessing the genotoxic risk in other stagnant and running aquatic ecosystems affected by anthropogenic activities.

We are evaluating the use of metabolically competent HepaRG™ cells combined with CometChip® for DNA damage and the micronucleus (MN) assay as a New Approach Methodology (NAM) alternative to animals for follow up genotoxicity assessment to in vitro positive genotoxic response. Naphthalene is genotoxic in human TK6 cells inducing a nonlinear dose-response for the induction of micronuclei in the presence of rat liver S9. of naphthalene. In HepaRG™ cells, naphthalene genotoxicity was assessed using either 6 (CometChip™) or 12 concentrations of naphthalene (MN assay) with the top dose used for assessment of genotoxicity for the Comet and MN assay was 1.25 and 1.74 mM respectively, corresponding to approximately 45% cell survival. In contrast to human TK6 cell with S9, naphthalene was not genotoxic in either the HepaRG™ MN assay or the Comet assay using CometChip®. The lack of genotoxicity in both the MN and comet assays in HepaRG™ cells is likely due to Phase II enzymes removing phenols preventing further bioactivation to quinones and efficient detoxication of naphthalene quinones or epoxides by glutathione conjugation. In contrast to CYP450 mediated metabolism, these Phase II enzymes are inactive in rat liver S9 due to lack of appropriate cofactors causing a positive genotoxic response. Rat liver S9-derived BMD10 over-predicts naphthalene genotoxicity when compared to the negative genotoxic response observed in HepaRG™ cells. Metabolically competent hepatocyte models like HepaRG™ cells should be considered as human-relevant NAMs for use genotoxicity assessments to reduce reliance on rodents.

Intergeneric and interspecific hybridization has been employed for the breeding of Phalaenopsis to transfer desirable traits between species, producing novel phenotypes with improved size, color, form, and flower-bearing ability. These characteristics are often enhanced; however, many of these hybrids are triploids and have reduced or complete sterility, for example, Phalaenopsis Queen Beer \'Mantefon\', an important novelty-type cultivar in Asia, particularly in China, Japan, and Republic of Korea. Despite the increasing demand for the crop for ornamental purposes, little is known about its cytogenetics, which is essential for breeding and, consequently, crop improvement. In this study, karyotyping using fluorescence in situ hybridization, meiotic chromosome behavior analysis, pollen staining, and in vitro viability germination tests were performed to understand the cause of hybrid sterility and pollen abnormality in Phalaenopsis Queen Beer \'Mantefon\' from a cytogenetic perspective. Viability tests revealed pollen infertility at all flower developmental stages, confirmed by the absence of pollen tube growth. Aberrant chromosomal behavior was observed in pollen mother cells (PMCs), frequently forming univalents, chromosomal bridges, and laggards during the entire meiotic process. PMCs were also divided irregularly into sporads with varying numbers of micronuclei, which may be responsible for pollen sterility in this cultivar. Altogether, the cytogenetic analyses provided insights into the pollen development of Phalaenopsis Queen Beer \'Mantefon\' and the conceivable causes of its infertility.

Amygdalin (AMY), a plant secondary metabolite containing nitrile, is a major component of the seeds of Rosaceae family plants. It is known that this compound has many pharmacological activities such as cancer prevention, antipyretic, and cough suppressant. In this study, the genotoxic and modulatory effects of amygdalin were assessed by chromosomal aberration (CA), sister chromatid exchange (SCE), and cytokinesis-block micronucleus assay (CBMN) assays using human peripheral lymphocytes (HPLs) in the absence and presence of metabolic activator (S9 mix). Lymphocytes were exposed to various concentrations of amygdalin (0.86, 1.72, 3.43, 6.86, and 13.75 μg/mL) alone and in combination with mitomycin-C (MMC, 0.20 μg/mL) or cyclophosphamide (CP, 12 μg/mL). The mitotic index (MI), replication index (RI), cytokinesis-block proliferation index (CBPI), and cytostasis were also evaluated to determine cytotoxicity. Amygdalin alone did not exhibit genotoxic and cytotoxic effects at all the tested concentrations both in the absence and presence of the S9 mix. In contrast, amygdalin significantly reduced the frequencies of CA (especially at 48 h treatments), SCE, and MN (except 0.86 μg/mL in pre- and simultaneous treatment) induced by MMC in all the tested concentrations and treatment protocols. It has also considerably decreased CP-induced CA and SCE frequencies at all the concentrations (except 0.86 μg/mL) in simultaneous treatment. This study demonstrated that amygdalin alone was not genotoxic, on the contrary, it has revealed modulatory effects against chemotherapy agents that induced genomic damage in human lymphocytes, suggesting its chemopreventive potential.

The use of pesticides to prevent and control pests also increases food production. Pesticides are widely used by contemporary farmers, especially in Brazil, where the economy is based on agriculture. The objective of this study was to evaluate the genotoxic potential of pesticide use in rural workers in Maringá, Paraná, Brazil. DNA damage in whole blood cells was measured by the comet assay, while the frequency of cell types, abnormalities, and nuclear damage was estimated using the buccal micronucleus cytome assay. Samples of buccal mucosa were collected from 50 male volunteers (27 not exposed to pesticides and 23 occupationally exposed to pesticides). Among them, 44 volunteered for blood sampling (24 unexposed and 20 exposed). In the comet assay, the exposed farmers had a higher damage index than non-exposed ones. There were also statistically significant differences between the groups in the buccal micronucleus cytome assay. Farmers exhibited an increase in basal cell numbers, and cytogenetic alterations, represented by condensed chromatin and karyolitic cells. Comparisons between cell morphologies and epidemiological factors indicated an increased number of condensed chromatin and karyolitic cells in individuals who were responsible for preparation and transportation of pesticides to agricultural machines. Thus, the participants in this study who were exposed to pesticides were more sensitive to genetic damage, and thereby, more susceptible to diseases resulting from such damage. These results demonstrated that health policies should be developed for pesticide-exposed farmers to better mitigate risks and damage to their health.

To compare the micronucleus (MN) score in all the major diagnostic categories as per \"The Bethesda System for Reporting Cervical Cytology\" 2014 including negative for intraepithelial lesions and malignancy (NILM), inflammatory, abnormal squamous cells of undetermined significance (ASC-US), abnormal squamous cells cannot exclude high-grade squamous intraepithelial lesion (HSIL) (ASC-H), low-grade squamous intraepithelial lesion (LSIL), HSIL, and invasive carcinoma (IC) and to assess the role of MN scoring as a biomarker for predicting risk of carcinoma.
A total of 1000 conventional cervical smears stained with Papanicolaou (Pap) stain, comprising unsatisfactory for evaluation (86), NILM (140), inflammatory (696), ASC-US (23), ASC-H (16), LSIL (18), HSIL (15), and IC (6) were studied independently by two pathologists, and the number of MN cells per 1000 epithelial cells in high-power (×400) and oil immersion (×1000) was counted and expressed as MN score per 1000 cells.
The mean MN score ± standard deviation was found to be 0.99 ± 0.744 in NILM cases, 0.67 ± 0.782 in inflammatory cases, 1.57 ± 0.507 in ASC-US cases, 1.63 ± 0.50 in ASC-H cases, 1.56 ± 0.511 in LSIL cases, 2.47 ± 0.516 in HSIL cases, and 3.0 ± 0.00 in IC cases. A step-wise increase was observed in MN score from inflammatory to IC categories.
MN score is a reliable and easy test that can be used in conjunction with routine cervical PAP to assess the risk of malignant transformation in the uterine cervix as a biomarker for predicting the risk of carcinoma.
Résumé Objectifs et objectifs: comparer le score du micronucléus (MN) dans toutes les principales catégories de diagnostic selon “le système Bethesda pour signaler la cytologie cervicale” 2014, y compris négatif pour les lésions intraépithéliales et la malignité (Nilm), inflammatoire et anormal des cellules squameuses de signification indéterminées (Nilm), inflammatoire et anormale des cellules squameuses de signification indéterminées (Nilm), inflammatoire et anormale des cellules pure ASC - US), les cellules squameuses anormales ne peuvent pas exclure la lésion intraépithéliale épidermoïde de haute qualité (HSIL) (ASC - H), la lésion intraépithéliale squameuse à faible teneur (LSIL), le carcinome invasif (IC) et pour évaluer le rôle de MN La notation en tant que biomarqueur pour prédire le risque de carcinome. Matériaux et méthodes: un total de 1000 frottis cervicaux conventionnels colorés avec une tache de papanicolaou (PAP), comprenant insatisfaisant l\'évaluation (86), nilm (140), inflammatoire (696), ASC - US (23), ASC - H (16), LSIL (18), HSIL (15) et IC (6) ont été étudiés indépendamment par deux pathologistes, et le nombre de cellules Mn pour 1000 cellules épithéliales dans la puissance (× 400) et l\'immersion à l\'huile (× 1000) ont été comptées et exprimé en score MN par 1000 cellules. Résultats: Le score MN moyen ± l\'écart type s\'est révélé être de 0,99 ± 0,744 dans des cas nilms, 0,67 ± 0,782 dans des cas inflammatoires, 1,57 ± 0,507 dans les cas ASC - US, 1,63 ± 0,50 dans les cas ASC - H, 1,56 ± 0,511 dans LSIL cas, 2,47 ± 0,516 dans les cas HSIL et 3,0 ± 0,00 dans les cas IC. Une augmentation de pas de pas a été observée dans le score MN des catégories inflammatoires vers IC. Conclusions: Le score MN est un test fiable et facile qui peut être utilisé en conjonction avec le PAP cervical de routine pour évaluer le risque de transformation maligne dans le col utérine en tant que biomarqueur pour prédire le risque de carcinome. Mots-clés: Frottis cervical, micronucleus, dépistage.

Anzer honey is well known in Turkey and used for its medicinal properties, especially for pharyngitis, tonsillitis, ulcers and cancer. In this study, we investigated whether Anzer honey, which is shown to have antioxidant, anti-tumoral, and anti-inflammatory properties, has a protective effect against X-ray induced genotoxic damage by cytogenetic methods. Peripheral blood lymphocytes isolated from 20 healthy volunteers were divided into two groups and cultivated by conventional methods. Study group lymphocytes were treated with 10% diluted honey while those in the control group were not. Both groups were exposed to a high dose (2 Gy) X-ray at the 48th hour of culture. Conventional cytogenetic staining and Giemsa banding methods were applied to evaluate chromosomal breakage and ring formation. Micronucleus frequencies were determined by the cytokinesis-block micronucleus (CBMN) assay. Paired sample t test was used to compare groups. Anzer honey, which was analyzed melissopalynologically, was used. Micronucleus frequency was significantly decreased in the study group (CI = 348.75 ± 31, median 326, min. 98, max. 704) compared to the control group (CI = 489.10 ± 27, median 500, min. 216, max. 645) (p = .001). Chromosomal breakage was also significantly decreased in the study group (CI = 118.70 ± 16, median 109, min. 12, max. 316) compared to the control group (CI = 233.60 ± 25, median 225, min. 65, max. 492) (p < .0001). This is the first study indicating that genotoxic damage in the peripheral blood lymphocytes of healthy volunteers induced by X-radiation may be prevented or alleviated by adding Anzer honey in vitro. These results encourage further research about the protective effects of honey. RESEARCH HIGHLIGHTS: Anzer honey has a genoprotective effect against radiation-induced genotoxicity, probably by preventing oxidation damage.

Vitex megapotamica (Spreng) Moldenke is commonly known as tarumã, it is an important medicinal and edible fruit plant. It is native to regions of tropical and subtropical climate in greater proportion than temperate zones and widely distributed in Central America, South America, Asia, and Africa. In Brazil, it is present in the Atlantic Forest and Cerrado biomes. Despite its widespread use, there are no minimum standards for quality control or information on genotoxicity. Therefore, the aim of this study was to present a detailed description of the short-term genotoxicity assays of V. megapotamica and to provide parameters of a preparation routinely used in traditional folk medicine. For genotoxicity assays, five groups were used with eight wistar rats in each group. For this, three doses of the V. megapotamica extract in doses (100, 300, and 900 mg/kg) or negative control (filtered water) were administered orally and positive control cyclophosphamide monohydrate (20 mg/kg; Sigma-Aldrich®) was applied by the intraperitoneal route after 24 h. At the end, whole blood was collected in a tube containing EDTA for the comet test and later the animals were euthanized. For the micronucleus test, femurs were removed, and bone marrow was collected. In the comet assay, V. megapotamica crude extract did not show significant DNA damage at all doses tested. The micronucleus assay showed no significant increase in the frequency of inducing micronuclei at any dose examined. It can be concluded that the safety parameters in genotoxicity studies reveal that V. megapotamica has no toxicity, which characterizes the important quality control of this plant species.

The contamination of freshwaters by heavy metals represents a great problem, posing a threat for human and environmental health. Cadmium is classified as carcinogen to humans and its mechanism of carcinogenicity includes genotoxic events. In this study a recently developed eco-friendly cellulose-based nanosponge (CNS) was investigated as a candidate in freshwater nano-remediation process. For this purpose, CdCl2 (0.05 mg L-1) contaminated artificial freshwater (AFW) was treated with CNS (1.25 g L-1 for 2 h), and cellular responses were analyzed before and after CNS treatment in Dreissena polymorpha hemocytes. A control group (AFW) and a negative control group (CNS in AFW) were also tested. DNA primary damage was evaluated by Comet assay while chromosomal damage and cell proliferation were assessed by Cytome assay. AFW exposed to CNS did not cause any genotoxic effect in zebra mussel hemocytes. Moreover, DNA damage and cell proliferation induced by Cd(II) turned down to control level after 2 days when CNS were used. A reduction of Cd(II)-induced micronuclei and nuclear abnormalities was also observed. CNS was thus found to be a safe and effective candidate in cadmium remediation process being efficient in metal sequestering, restoring cellular damage exerted by Cd(II) exposure, without altering cellular physiological activity.