Objectives and achievements of the HUMN project on its 26th anniversary.

Micronuclei (MN) are a nuclear abnormality that occurs when chromosome fragments or whole chromosomes are not properly segregated during mitosis and consequently are excluded from the main nuclei and wrapped within nuclear membrane to form small nuclei. This maldistribution of genetic material leads to abnormal cellular genomes which may increase risk of developmental defects, cancers, and accelerated aging. Despite the potential importance of MN as biomarkers of genotoxicity, very little was known about the optimal way to measure MN in humans, the normal ranges of values of MN in healthy humans and the prospective association of MN with developmental and degenerative diseases prior to the 1980's.

Entinostat Enhances the Efficacy of Chemotherapy in Small Cell Lung Cancer Through S-phase Arrest and Decreased Base Excision Repair.

Purpose: Acquired chemoresistance is a frequent event in small cell lung cancer (SCLC), one of the deadliest human malignancies. Histone deacetylase inhibitors (HDACi) have been shown to synergize with different chemotherapeutic agents including cisplatin. Accordingly, we aimed to investigate the dual targeting of HDAC inhibition and chemotherapy in SCLC.

Use of the single cell gel electrophoresis assay for the detection of DNA-protective dietary factors: Results of human intervention studies.

The single cell gel electrophoresis technique is based on the measurement of DNA migration in an electric field and enables to investigate via determination of DNA-damage the impact of foods and their constituents on the genetic stability. DNA-damage leads to adverse effects including cancer, neurodegenerative disorders and infertility. In the last 25 years approximately 90 human intervention trials have been published in which DNA-damage, formation of oxidized bases, alterations of the sensitivity towards reactive oxygen species and chemicals and of repair functions were investigated with this technique.

Search for the optimal genotoxicity assay for routine testing of chemicals: Sensitivity and specificity of conventional and new test systems.

Many conventional in vitro tests that are currently widely used for routine screening of chemicals have a sensitivity/specificity in the range between 60 % and 80 % for the detection of carcinogens. Most procedures were developed 30-40 years ago. In the last decades several assays became available which are based on the use of metabolically competent cell lines, improvement of the cultivation conditions and development of new endpoints.

Recommendations and quality criteria for micronucleus studies with humans.

Micronucleus (MN) analyses in peripheral blood lymphocytes and exfoliated cells from different organs (mouth, nose, bladder and cervix) are at present the most widely used approaches to detect damage of genetic material in humans. MN are extranuclear DNA-containing bodies, which can be identified microscopically. They reflect structural and numerical chromosomal aberrations and are formed as a consequence of exposure to occupational, environmental and lifestyle genotoxins.

"Micronuclei and Disease" special issue: Aims, scope, and synthesis of outcomes.

The purpose of the "Micronuclei and Disease" special issue (SI) is to: (i) Determine the level of evidence for association of micronuclei (MN), a biomarker of numerical and structural chromosomal aberrations, with risk of specific diseases in humans; (ii) Define plausible mechanisms that explain association of MN with each disease; (iii) Identify knowledge gaps and research needed to translate MN assays into clinical practice. The "MN and Disease" SI includes 14 papers. The first is a review of mechanisms of MN formation and their consequences in humans.

Micronucleus assays with the human derived liver cell line (Huh6): A promising approach to reduce the use of laboratory animals in genetic toxicology.

The inadequate representation of enzymes which catalyze the activation/detoxification of xenobiotics in cells that are currently used in genotoxicity testing of chemicals leads to a high number of false positive results and the number of follow up studies with rodents could be reduced by use of more reliable in vitro models. We found earlier that several xenobiotic drug metabolizing enzymes are represented in the human derived liver cell line Huh6 and developed a protocol for micronucleus (MN) experiments which is in agreement with the current OECD guideline. This protocol was used to test 23 genotoxic and non-genotoxic reference chemicals; based on these results and of earlier findings (with 9 chemicals) we calculated the predictive value of the assay for the detection of genotoxic carcinogens.

Impact of infections, preneoplasia and cancer on micronucleus formation in urothelial and cervical cells: A systematic review.

Approximately 165,000 and 311,000 individuals die annually from urothelial (UC) and cervical (CC) cancer. The therapeutic success of these cancers depends strongly on their early detection and could be improved by use of additional diagnostic tools. We evaluated the current knowledge of the use of micronucleus (MN) assays (which detect structural and numerical chromosomal aberrations) with urine- (UDC) and cervix-derived (CDC) cells for the identification of humans with increased risks and for the diagnosis of UC and CC.

Micronuclei as biomarkers of DNA damage, aneuploidy, inducers of chromosomal hypermutation and as sources of pro-inflammatory DNA in humans.

Micronuclei (MNi) are among the most widely studied biomarkers of DNA damage and chromosomal instability in humans. They originate from chromosome fragments or intact chromosomes that are not included in daughter nuclei during mitosis. The main reasons for their formation are a lack of functional centromere in the chromosome fragments or whole chromosomes or defects in one or more of the proteins of the mitotic system that, consequently, fails to segregate chromosomes properly.

Genotoxic properties of materials used for endoprostheses: Experimental and human data.

Approximately 2 million endoprostheses are implanted annually and metal ions as well as particles are released into the body from the materials which are used. This review describes the results of studies concerning genotoxic damage caused by artificial joints. DNA damage leads to various adverse long-term health effects in humans including cancer.

Genotoxic activities of wastewater after ozonation and activated carbon filtration: Different effects in liver-derived cells and bacterial indicators.

Aim of this study was to investigate the impact of advanced wastewater treatment techniques (combining ozonation with activated carbon filtration) on acute and genotoxic activities of tertiary treated wastewater. Concentrated samples were tested in Salmonella/microsome assays. Furthermore, induction of DNA damage was measured in liver-derived cells (human hepatoma and primary rat hepatocytes) in single cell gel electrophoresis experiments, which are based on the measurement of DNA migration in an electric field.

Micronuclei and disease - Report of HUMN project workshop at Rennes 2019 EEMGS conference.

The "Micronuclei and Disease" workshop was organized by the HUMN Project consortium and hosted by the European Environmental Mutagen and Genomics Society at their annual meeting in Rennes, France, on 23 May 2019. The program of the workshop focused on addressing the emerging evidence linking micronucleus (MN) frequency to human disease. The first objective was to review what has been published and evaluate the level and quality of evidence for the connection between MN frequency and various diseases through all life stages.

Smoking causes induction of micronuclei and other nuclear anomalies in cervical cells.

Introduction: Smoking is an independent cause of cervical cancer, which is the 4th most common malignancy in women. It is currently not known if tobacco consumption causes chromosomal damage (which is a hallmark of human cancer) in cervical cells and if age and the hormonal status have an impact on tobacco induced genetic instability in the cervix.

Methods: We conducted a study with pre- and post-menopausal women smokers and never-smokers (25/group).

Use of human derived liver cells for the detection of genotoxins in comet assays.

One of the problems of in vitro genotoxicity testing is the inadequate representation of drug metabolizing enzymes in indicator cells which are currently used. An alternative are human derived liver cell lines which retained the activities of enzymes that catalyze the activation and detoxification of genotoxins. Several cell lines were identified which were used in comet experiments.

Micronucleus Assay with Tetrad Cells of Tradescantia.

The Tradescantia micronucleus assay has been used since 50 years for the detection of genotoxins (including carcinogens) in the environment. A large database concerning the effects of individual chemicals and complex environmental mixtures (soil, air and waters) has accumulated. In contrast to other mutagenicity test systems, the effects of low concentrations of heavy metals, radionuclides, certain herbicides, pesticides and gaseous mutagens can be detected and it is also possible to conduct in situ biomonitoring studies with plant.

Environmental risk assessment of widely used anticancer drugs (5-fluorouracil, cisplatin, etoposide, imatinib mesylate).

Anticancer drugs are among the most toxic chemicals, which are commercially produced; therefore, their release in aquatic ecosystems raised concerns in regard to potential adverse effects. This article describes the results of risk assessments concerning their environmental safety, which are based on data generated in the frame of a coordinated EU project ("Cytothreat"). Eight research institutions participated in the project and four widely used anticancer drugs with different mechanisms of therapeutic action (5-fluorouracil 5FU, cisplatin CDDP, imatinib mesylate IM and etoposide ET) were tested in a variety of indicator organisms (cyanobacteria, algae, higher plants, rotifers, crustacea, fish and also in human and fish derived cell lines) in acute/subacute/chronic toxicity assays.

Cytome micronucleus assays with a metabolically competent human derived liver cell line (Huh6): A promising approach for routine testing of chemicals?

One of the main problems of in vitro genotoxicity tests is the inadequate representation of drug metabolizing enzymes in most indicator cell lines which are currently used. We identified recently a human derived liver cell line (Huh6) which detected induction of DNA damage by representatives of different groups of promutagens without enzyme mix and showed that these cells are more suitable in terms of reproducibility and sensitivity as other currently used liver derived lines. We developed a protocol for micronucleus (MN) cytome assays with these cells and validated the procedure in experiments with representatives of different groups of directly and indirectly acting genotoxic carcinogens (MMS, cisplatin, PhIP, IQ, NDMA, B(a)P, AFB1, etoposide, and H O ).

Counteraction of Oxidative Stress by Vitamin E Affects Epigenetic Regulation by Increasing Global Methylation and Gene Expression of and Dose Dependently in Caco-2 Cells.

Obesity- or diabetes-induced oxidative stress is discussed as a major risk factor for DNA damage. Vitamin E and many polyphenols exhibit antioxidative activities with consequences on epigenetic regulation of inflammation and DNA repair. The present study investigated the counteraction of oxidative stress by vitamin E in the colorectal cancer cell line Caco-2 under normal (1 g/l) and high (4.