Relationship between chromatin bridges in anaphase and chromosomal aberrations induced by TMP+UVA (365 nm) in CHO cells

In a recent paper, the hypothesis of 'conservative pairing' between complementary DNA strands belonging to both sister chromatids has been proposed as a phenomenon that could account for, at least in part, sister chromatid pairing in late G(2)/mitosis. The hypothesis was verified through a cytogenetic approach, studying the so-called 'sister chromatid chromatin bridges' (SCCBs), induced in the previous G(2)/mitosis by a crosslinking (TMP + UVA 365 nm) treatment in CHO cells (Rizzoni, M., E. Cundari, P. Perticone and B. Gustavino (1993) Chromatin bridges between sister chromatids induced in late G(2) mitosis in CHO cells by trimethylpsoralen + UVA, Experimental Cell Res., 209, 149-155; [1]). The purpose of the present paper is the study of the relationship between chromatin bridges without fragments in ana-telophase, which were demonstrated to be SCCBs, and chromosomal aberrations, in order to investigate their mechanism of induction. The evolution along the time of the two classes of mitotic anomalies was studied and a comparison was carried out to verify whether the bridges rise as a direct and immediate effect of the treatment or represent the misrepair-mediated effect of it. The present data show that single bridges without fragments come from a direct effect of photoinduced crosslinks in late G(2)/mitosis. Moreover TMP + 365 nm UVA treatment shows an S-dependent clastogenic effect. The proposed hypothesis of 'conservative pairing' between DNA strands of sister chromatids is further supported

Micronucleus induction by low doses of X-rays in Vicia faba root tips

Studies on the micronucleus test in Vicia faba root tips (VM test) were carried out in order to estimate the effects at low doses of X-rays (1, 2, 4, 8 and 12 R). The control value of micronucleus frequency is about 0.44/1000 cells. The dose where the micronucleus frequency is twice that of the control was estimated at 1.384 R. There was a linear kinetic dose response for the low-dose range studied here

Effect of cytochalasin B on the induction of chromosome missegregation by colchicine at low concentrations in human lymphocytes

The aim of the present work was to investigate the possible interference of cytochalasin B (cyt B) with low concentration treatment with colchicine in the induction of chromosome/chromatid loss and micronuclei in human lymphocytes mitotically activated in vitro. Thus, cells from a single female donor were treated with colchicine (10 or 25 nM, from 24 h after PHA addition to fixation at 66 h) either in the presence or absence of cyt B. Single lagging chromosomes/chromatids were scored in bipolar ana-telophases and greater damage (disrupted and c-anaphases) was scored in cells at anaphase. Micronuclei were scored in the first 4000 nuclei observed in both cyt B-treated (in mononucleate and binucleate cells) and untreated cultures. With the same criterion, FISH analysis was performed on 2000 nuclei where chromosome 7 and 11 centromeric DNA probes were used in pairs. Our results showed that: (i) the frequency of laggards and of micronuclei increased with colchicine concentration but in the presence of cyt B there was a lower frequency of both (with a mean reduction of ~ 49%); (ii) FISH analysis showed a colchicine concentration-dependent increase in nuclei with three spots for chromosome 7; (iii) a colchicine concentration-dependent increase in tetraploid cells was observed. This increase was particularly remarkable (5-fold) in cells grown in the presence of cyt B compared with cyt B-untreated cells. The observed 'cyt B effects' can be explained if it is assumed that in cytokinesis-blocked cells there is a shorter distance between the poles. As a consequence: (i) laggards would be engulfed in the nearest daughter nucleus with a consequent lower induction of micronuclei; (ii) segregating sister chromatids in heavily impaired anaphases would not travel a sufficient distance to give rise to two daughter nuclei, leading to an increased frequency of polyploid nuclei

A glass spark counter for high rate environments

The performance of a glass spark counter prototype, built with glass electrodes of about 1010 Ω cm volume resistivity, is described. The measure

Indirect mitotic nondisjunction in Vicia faba and Chinese hamster cells

The hypothesis of indirect mitotic nondisjunction was tested in plant and mammalian cells. This hypothesis states that micronuclei derived from lagging chromosomes or chromatids are able to perform DNA synthesis and undergo mitotic condensation synchronously with main nuclei. Hence, as chromosomes, they can be moved to spindle poles together with the chromosomes of the main nuclei during mitosis. In that way chromosomes "lost" as micronuclei can be reincorporated in the main nuclei. In order to test this, both Vicia faba meristematic cells and cells of a Chinese hamster line (Cl-1) were treated with low doses of colchicine. Mitotic anomalies, micronuclei and cells with a polyploid or aneuploid karyotype were scored at different fixation times. A detailed analysis was performed on single chromosome misdistributions, as well as on micronuclei and cells with aneuploid karyotypes derived from single chromosome misdistributions. Indirect mitotic nondisjunction was shown to play a primary role in the origin of aneuploid karyotypes in Vicia faba, but not in Cl-1 cells

Genotoxicity testing for radon exposure: Dolichopoda (Orthoptera, Rhaphidophoridae) as potential bio-indicator of confined environments

Abstract Radon represents the major source of natural radioactivity in confined environments. Despite the clear evidence of a direct association between residential exposure and human lung cancer provided by case-control studies, results relating indoor exposure and genotoxic/mutagenic effect induction are still contradictory. The present study attempts to estimate the genotoxic effects induced by exposure to radioactive radon in wild cricket populations sampled from caves where varying concentrations of radon are present. Cave crickets are also tested as possible bio-indicator organisms of genotoxic potential of contaminated residential and confined environments. Six caves in Central Italy are considered covering a broad spectrum of radon radioactivity concentration (221–26, 000 Bq/m3). Dolichopoda specimens were sampled from each cave; both haemocytes and brain cells taken from individuals were tested for responsiveness to DNA damage induced by radon through the Comet assay. Specimens from the least radioactive cave, housed in controlled conditions for 60 days before analysis, were used as control group. Statistically significant increase of DNA damage was found in all groups of individuals from each cave, for both cell types. Very low values of all Comet parameters were found in control group individuals, which gave indications of a good responsiveness of the organism to the variable environmental levels of radioactive contamination. Results indicate that cave crickets represent a reliable tool for the detection of genotoxic potential induced by radioactive contamination of confined environments and can be proposed as a possible bio-indicator system for air (-radioactive) pollution related to indoor exposure

Modulating effects of humic acids on genotoxicity induced by water disinfectants in Cyprinus carpio

The use of chlorinated disinfectants during drinking-water production has been shown to generate halogenated compounds as a result of interactions of humic acids with chlorine. Such chlorinated by-products have been shown to induce genotoxic effects and consumption of chlorinated drinking-water has been correlated with increased risk for cancer induction in human populations. The aim of this work was to test the potential genotoxic effects on circulating erythrocytes of the fish Cyprinus carpio exposed in vivo to well-waters disinfected with sodium hypochlorite (NaClO), chlorine dioxide (ClO2) or peracetic acid (CH3COO2H, PAA), in the absence or presence of standard humic acids (HA). The effects were measured by use of the micronucleus (MN) and the single-cell gel electrophoresis (Comet) assays at different sampling times after a 3-day exposure period. The exposure to chlorine disinfectants without the addition of HA produced a clear toxic effect. Significant cytogenetic damage (i.e. MN induction) was detected in fish populations exposed to both NaClO and ClO2 with humic acids. In the Comet assay, a significant decrease of DNA migration was observed in erythrocytes of specimens after exposure to NaClO-disinfected water without HA. No effects were observed in any other experimental condition. (c) 2005 Elsevier B.V. All rights reserved

Influence of Temperature on Mutagenicity in Plants Exposed to Surface Disinfected Drinking Water.

Disinfection of surface drinking water, particularly water chlorination, produces by-products with potential genotoxic and/or carcinogenic activity. A study carried out at a pilot plant for drinking water disinfection of lake water revealed mutagenic activity of three different disinfectants (sodium hypochlorite, chlorine dioxide and peracetic acid) in different seasons using in situ mutagenicity assays, both in animal (micronucleus test) and in plant organisms (anaphase chro- mosomal aberration and micronucleus tests). The effects of the disinfectants appeared to be modulated by the season of exposure. In this study, we tried to understand if (and to what extent) the temperature parameter could actually play an independent role in the registered seasonal variation of mutagenic effects, neglecting the variation of other parameters, e.g. physical conditions and chemical composition of the lake water. Therefore plants (Allium cepa for chromosomal aberration test and Vicia faba for micronucleus test) were exposed to the same disinfected lake-water samples at differ- ent temperatures (10°C, 20°C and 30°C), according the ones registered during the in situ experiment. Long-term expo- sure at the temperatures of 20°C (both Vicia faba and Allium cepa) and 30°C (Vicia faba only) to disinfected waters in- duced clear mutagenic effects. These results show that temperature is an important variable which should be taken into account when in situ exposure of plants is planned for mutagenicity testing. Also, different plant systems clearly show specific temperature ranges suitable for their growth, thereby indicating the need for an accurate selection of the test organism for a specific experimental plan

The OPERA magnetic spectrometer

The OPERA neutrino oscillation experiment foresees the construction of two magnetized iron spectrometers located after the lead-nuclear emulsion targets. The magnet is made up of two vertical walls of rectangular cross section connected by return yokes. The particle trajectories are measured by high precision drift tubes located before and after the arms of the magnet. Moreover, the magnet steel is instrumented with Resistive Plate Chambers that ease pattern recognition and allow a calorimetric measurement of the hadronic showers. In this paper we review the construction of the spectrometers. In particular, we describe the results obtained from the magnet and RPC prototypes and the installation of the final apparatus at the Gran Sasso laboratories. We discuss the mechanical and magnetic properties of the steel and the techniques employed to calibrate the field in the bulk of the magnet. Moreover, results of the tests and issues concerning the mass production of the Resistive Plate Chambers are reported. Finally, the expected physics performance of the detector is described; estimates rely on numerical simulations and the outcome of the tests described above.Comment: 6 pages, 10 figures, presented at the 2003 IEEE-NSS conference, Portland, OR, USA, October 20-24, 200

Comet assay and micronucleus test in circulating erythrocytes of Cyprinus carpio specimens exposed in situ to lake waters treated with disinfectants for potabilization

The detection of a possible genotoxic effect of surface water treated with disinfectants for potabilization is the aim of the present work. The Comet assay and the micronucleus test were applied in circulating erythrocytes of Cyprinus carpio. Young specimens (20-30g) were exposed in experimental basins, built within the potabilization plant of Castiglione del Lago (Perugia, Italy). In this plant the water of the Trasimeno Lake is treated and disinfected for potabilization before it is distributed to the people in the net of drinkable water. A continuous flow of water at a constant rate was supplied to basins; the water was continuously treated at a constant concentration with one of the three tested disinfectants (sodium hypochlorite, peracetic acid and chloride dioxide), one control basin being supplied with untreated water. Three sampling campaigns were performed: October 2000, February 2001 and June 2001. Repeated blood samplings through intracardiac punctures allowed to follow the same fish populations after different exposure times: before introduction of the disinfectant, and 10 or 20 days afterwards. An additional blood sampling was performed 3h after addition of the disinfectant in other, simultaneously exposed, fish populations. Genotoxic damage was shown in fish exposed to water disinfected with sodium hypochlorite and chloride dioxide. The Comet assay showed an immediate response, i.e. DNA damage that was induced directly in circulating erythrocytes, whereas micronuclei reached their highest frequencies at later sampling times, when a genotoxic damage in stem cells of the cephalic kidney is expressed in circulating erythrocytes. The quality of the untreated surface water seems to be the most important parameter for the long-term DNA damage in circulating erythrocytes. © 2003 Elsevier B.V. All rights reserved