Binucleated Cells With Micronuclei Research Articles

Long-term exposure to cypermethrin and piperonyl butoxide cause liver and kidney inflammation and induce genotoxicity in New Zealand white male rabbits

Cypermethrin (CY) is a frequently used class II pyrethroid pesticide, while piperonyl butoxide (PBO) plays a major role in the pesticide formulation of synthetic pyrethroids. Synthetic pyrethroids are metabolized in mammals via oxidation and ester hydrolysis. PBO can prevent the metabolism of CY and enhances its pesticide effect. While this potentiation effect reduces the amount of pesticide required to eliminate insects, it is not clear how this mixture affects mammals. In our in vivo experiment, New Zealand white male rabbits were exposed to low and high doses of CY, PBO, and their combinations, for 4 months. Genotoxicity and cytotoxicity were monitored by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the cytokinesis block proliferation index (CBPI) in lymphocytes. After two months of exposure, a statistically significant increase in the frequency of BNMN was observed for all exposed animals (p < 0.001) in a dose-dependent way. MN were significantly elevated compared to controls (p < 0.001), with high dose groups reaching a 442% increase when co-exposed. BNMN and MN continued to increase after four months. Histopathological examination of lesions showed damage involving inflammation, attaining lymphoplasmatocytic infiltration in the high dose groups. Both CY and PBO cause liver and kidney inflammation and induce genotoxicity.

Study of the genotoxic and cytotoxic effects of the α-, β-, and γ- Hexachlorocyclohexane isomers in human lymphocyte cells using the cytokinesis-block micronucleus assay

The genotoxic potential of hexachlorocyclohexane (HCH) isomers (α-, β-, and γ-) which are organochlorine pesticides was tested in peripheral blood lymphocyte cultures from two donors by using the cytokinesis-block micronucleus assay. Micronucleus (MN) frequency, binucleated cells with micronucleus (BNMN), and cytokinesis-blocked proliferation index (CBPI) were determined as genotoxic and cytotoxic endpoints. At the concentration ranges tested (12.5–100 μg.L −1), all HCH isomers induced dose-dependent cytotoxic effects, γ-HCH being the most toxic. This isomer was also able to induce significant increase in MN frequency and BNMN cells indicating a genotoxic potential at 50 and 100 μg.L −1. The genotoxic test of β-HCH showed a positive induction of MN and BNMN cells at the highest concentration of 100 μg.L −1 and a significant cytotoxicity at 50 μg.L −1. Under the experimental condition used, α-HCH was unable to induce any significant increase in MN frequency confirming that α-HCH is a non-genotoxic agent.

Long-term exposure of rabbits to imidaclorpid as quantified in blood induces genotoxic effect

The present in-vivo study focuses on the genotoxic effect of the neonicotinoid pesticide imidacloprid (IMI) in rabbits. The purpose of the study was to establish a possible relationship between exposure to the pesticide (dose and duration) and genotoxicity. Furthermore, an analytical method for the simultaneous determination of IMI and its major metabolite 6-chloronicotinic acid (6-ClNA) in blood was developed and validated. The isolation of the two analytes from blood was performed by liquid–liquid extraction with dichloromethane. Analysis was performed by Liquid Chromatography – Atmospheric Pressure Chemical Ionization – Mass Spectrometry (LC-APCI-MS). The method was applied on the determination of IMI and 6-ClNA in serum samples obtained from rabbits fed with the insecticide at two low doses. Furthermore, parameters of genotoxicity and cytotoxicity were evaluated by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the Cytokinesis Block Proliferation Index (CBPI), in lymphocytes of exposed rabbits. The results revealed a genotoxic effect of IMI for both exposed groups. There were statistically significant differences in the frequencies of BNMN and MN between control and exposed groups but there was no dose-dependence, neither time-dependence of the genotoxic effect for the administered doses. This is the first time that long term exposure to IMI in rabbits was studied for the determination of its genotoxic effect. The genotoxic effect of IMI as it is depicted by the current study is in accordance with previous studies.

New Evidences about Adverse Health Effects Based on Chromate Environmental and Biological Exposure Assessment in Chromate Manufacturing Workers

Background Chromium (Cr) compounds are produced and applied widely. Aims On the basis of chromate exposure assessment, Cr related trace element homeostasis, DNA damage and global DNA methylation status, renal damage need further investigation. Method We recruited more than one hundred workers from chromate producing facilities as testing subjects and matched local residents without chromium exposure history served as controls. Personal sampling of airborne Cr (A-Cr) was conducted and Cr content was quantified by Flame Atomic Absorption Spectrometry. At the end of the sampling shift, blood and urine samples were collected and element concentrations were measured by inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry. Adverse health effects including renal impairment, DNA damage, DNA methylation, and multi-trace element homeostasis were investigated comprehensively. Results 90% of the chromate production workers were exposed to A- Cr lower than 50 mu g m(-3).Chromate caused renal impairment with more severity in the tubule than glomerular. After Cr(VI) exposure, a significant increase in blood Cr, Cd, Fe, Mg, Mo, Se and Zn concentrations was observed, as well as a significant decrease in Ca concentration. Blood Cr (B-Cr) was four times higher than that of the controls, and significantly correlated with A-Cr (r = 0.568, P&lt;0.001). Binucleated cells with micronuclei (BNMN ) frequencies were positively correlated with B-Cr and U- Cr; Ln BNMN was regressed on B-Cr, and U-Cr and A-Cr by linear models.We also found that the decreased serum folate was associated with an increased urinary 8-OhdG, DNA strand breaks and global DNA hypomethylation. Conclusions We provided some new evidences about chromate health adverse effects .Based on the above evidences and others, we therefore proposed a modified health surveillance system for chromate workers.

Micronucleus frequency among Iraqi thyroid disorder patients.

Micronucleus (MN) assay has been extensively used in detection of DNA damage, instability in cancer, and genetic disorders. In the current study, MN, binucleated cells, and nuclear division index (NDI) were investigated in Iraqi patients with thyroid disorders. The results indicated significantly (p < 0.05) increased binucleated cells with micronucleus (BNMN) frequencies in thyroid cancer group (37.58 ± 3.07) versus other thyroid disorder groups (6.60 ± 1.29, 14.90 ± 1.69, 15.56 ± 1.76). On the other hand, the frequency of micronucleus per 1,000 and the NDI were significantly (p < 0.05) decreased in hypothyroidism (MN 1.55 ± 0.36) (NDI 0.009 ± 0.001) versus other thyroid disorder groups (MN: 6.05 ± 0.97, 6.09 ± 0.53, 5.34 ± 0.56) (NDI: 0.049 ± 0.003, 0.032 ± 0.002, 0.025 ± 0.002), with no difference versus healthy group (0.0 ± 0.0). The number of BNMN and MN are parallel to the severity of thyroid disorders which were 6.60 ± 1.29, 14.90 ± 1.69, 15.56 ± 1.76, and 37.58 ± 3.07 for hypothyroidism, thyroid toxic goiter, thyroid nontoxic goiter, and thyroid cancer, respectively. The number of BNMN and MN are parallel to the severity of thyroid disorders which were 6.60 ± 1.29, 14.90 ± 1.69, 15.56 ± 1.76, and 37.58 ± 3.07 for hypothyroidism, thyroid toxic goiter, thyroid nontoxic goiter, and thyroid cancer, respectively. The results also indicate that there were no significant differences among age and sex groups as related with BNMN formation within each thyroid disorder groups and healthy control group.

Possible Role of the WDR3 Gene on Genome Stability in Thyroid Cancer Patients

The role of the WDR3 gene on genomic instability has been evaluated in a group of 115 differentiated thyroid cancer (DTC) patients. Genomic instability has been measured according to the response of peripheral blood lymphocytes to ionizing radiation (0.5 Gy). The response has been measured with the micronucleus (MN) test evaluating the frequency of binucleated cells with MN (BNMN), both before and after the irradiation. No differences between genotypes, for the BNMN frequencies previous the irradiation, were observed. Nevertheless significant decreases in DNA damage after irradiation were observed in individuals carrying the variant alleles for each of the three genotyped SNPs: rs3754127 [−8.85 (−15.01 to −2.70), P<0.01]; rs3765501 [−8.98 (−15.61 to −2.36), P<0.01]; rs4658973 [−8.70 (−14.94 to −2.46), P<0.01]. These values correspond to those obtained assuming a dominant model. This study shows for the first time that WDR3 can modulate genome stability.

The micronucleus assay as a biological dosimeter in hospital workers exposed to low doses of ionizing radiation

The health risk associated with low levels of ionizing radiation is still a matter of debate. A number of factors, such as non-target effects, adaptive responses and low-dose hypersensitivity, affect the long-term outcome of low-dose exposures. Cytogenetic bio-dosimetry provides a measure of the absorbed dose, taking into account the individual radiation sensitivity. The aim of the present study is to evaluate the value of the micronucleus (MN) test as a bio-dosimeter in hospital workers exposed to low doses of ionizing radiation. Blood samples were obtained from 30 subjects selected among workers exposed to X- and gamma-radiation, and 30 controls matched for sex, age and smoking from the same hospital. Micronucleus frequencies were analyzed by use of the cytokinesis-block method. The MN frequency was compared among the groups considering the confounding factors and the length of employment. No increase in the number of bi-nucleated cells with MN (BNMN), but a significant increase in the number of mono-nucleated cells with micronuclei (MOMN) was observed in exposed subjects compared with the controls. The relationship between MN frequency and accumulated dose (mSv) was evaluated. The length of employment did not affect the extent of MN frequency, but an increase of BNMN and MOMN cells was observed based on the accumulated radiation dose. Our study shows the sensitivity of the MN test in the detection of cytogenetic effects of cumulative exposure levels, suggesting the potential usefulness of this assay in providing a biological index in medical surveillance programs.

Evaluation of the Correlation Between Genetic Damage and Occupational Chromate Exposure Through BNMN Frequencies

To evaluate the correlation between chromate exposure and binucleated cells with micronuclei (BNMN) frequencies in chromate production workers and explore the ideal biomarker to detect early DNA damage. There are 79 workers and 112 peasants in this study. Chromium concentrations in the air (CrA), blood (CrB), and urine (CrU) were detected. BNMN frequencies were also counted. The CrA, CrB, CrU, and BNMN frequencies of the exposure group were higher than those of the control group; BNMN frequencies were positively correlated with CrB and CrU; natural logarithm (Ln) BNMN was regressed on CrB, CrU, and CrA by linear models. The increases in CrB, CrU, and BNMN frequencies caused by lower CrA level suggest BNMN frequencies could be used to detect early DNA damage in the chromate production health surveillance.

Are thyroid cancer patients sensitive to ionising radiation?

Purpose: To determine the ionising radiation sensitivity of peripheral blood lymphocytes in a group of differentiated thyroid cancer (DTC) patients.Material and methods: A total of 53 thyroid cancer patients (26 women and 27 men) and 50 donors (23 women and 27 men) were included in the study. The cytokinesis-block micronucleus assay (CBMN) in G0 peripheral blood lymphocytes was carried out using the cytochalasin B technique. Four cultures were established per each donor, two were irradiated with 0.5 Gy 137Cs g-rays, while the other two remained untreated.Results: No significant differences were observed in the frequency of binucleated cells with micronuclei (BNMN) between patients and controls, for both spontaneous and after the irradiation frequencies. Nevertheless, a positive and significant correlation was found between the frequencies of both spontaneous and after irradiation DNA damage, for control and patient groups.Conclusions: We have found that DTC patients do not present particular sensitivity to ionising radiation when an in vitro treatment is performed in G0 stage of the cell cycle, but this result does not discard the hypothesis about an increased sensitivity in other stages of the cell cycle in DTC patients.

Biomonitoring of Genotoxic Risk in Agricultural Workers from Five Colombian Regions: Association to Occupational Exposure to Glyphosate

In order to assess possible human effects associated with glyphosate formulations used in the Colombian aerial spray program for control of illicit crops, a cytogenetic biomonitoring study was carried out in subjects from five Colombian regions, characterized by different exposure to glyphosate and other pesticides. Women of reproductive age (137 persons 15–49 yr old) and their spouses (137 persons) were interviewed to obtain data on current health status, history, lifestyle, including past and current occupational exposure to pesticides, and factors including those known to be associated with increased frequency of micronuclei (MN). In regions where glyphosate was being sprayed, blood samples were taken prior to spraying (indicative of baseline exposure), 5 d after spraying, and 4 mo after spraying. Lymphocytes were cultured and a cytokinesis-block micronucleus cytome assay was applied to evaluate chromosomal damage and cytotoxicity. Compared with Santa Marta, where organic coffee is grown without pesticides, the baseline frequency of binucleated cells with micronuclei (BNMN) was significantly greater in subjects from the other four regions. The highest frequency of BNMN was in Boyacá, where no aerial eradication spraying of glyphosate was conducted, and in Valle del Cauca, where glyphosate was used for maturation of sugar cane. Region, gender, and older age (≥35 yr) were the only variables associated with the frequency of BNMN measured before spraying. A significant increase in frequency of BNMN between first and second sampling was observed in Nariño, Putumayo, and Valle immediately (<5 d) after spraying. In the post-spray sample, those who reported direct contact with the eradication spray showed a higher quantitative frequency of BNMN compared to those without glyphosate exposure. The increase in frequency of BNMN observed immediately after the glyphosate spraying was not consistent with the rates of application used in the regions and there was no association between self-reported direct contact with eradication sprays and frequency of BNMN. Four months after spraying, a statistically significant decrease in the mean frequency of BNMN compared with the second sampling was observed in Nariño, but not in Putumayo and Valle del Cauca. Overall, data suggest that genotoxic damage associated with glyphosate spraying for control of illicit crops as evidenced by MN test is small and appears to be transient. Evidence indicates that the genotoxic risk potentially associated with exposure to glyphosate in the areas where the herbicide is applied for coca and poppy eradication is low.

Genotoxic damage in Maras powder consumers from Kahramanmaras province of Turkey

In the present study, chromosomal aberrations (CAs) and micronucleus (MN) levels in the lymphocytes from 60 male individuals consisting of 40 habitues of Maras powder (a kind of smokeless tobacco) and 20 unexposed subjects were determined to investigate the possible inducing effect of Maras powder. The consumers of Maras powder had no exposure to any other known mutagens or toxicants. The mean exposure period to Maras powder was 12.25 + 0.93 years (range 3-22). Data obtained from microscopic examination of the slides was analyzed by SPSS (10.0) package programme. Mean frequency of CA and MN was found to be significantly higher in Maras powder consumers as compared to controls. Similarly, there was a significant elevation in the level of aberrant cells (Ab.Cells) with CAs and binucleated cells with MN (BNMN) in habitues. Spearman's rho correlation analysis indicated a significant increase in the frequency of CA and MN with increase in both age and years of exposure in consumers. Our finding of a significant elevation of CA and MN frequencies in peripheral lymphocytes from smokeless tobacco consumers demonstrated a potential cytogenetic hazard associated with Maras powder exposure.

Cytogenetic damage in female Chilean agricultural workers exposed to mixtures of pesticides

The VIII Region of Bio-Bio is a major fruit-growing area of Chile that makes intensive use of agricultural pesticides. The cytogenetic damage associated with exposure to mixtures of pesticides was evaluated by comparing peripheral blood lymphocyte micronucleus (MN) frequencies in a group of 64 female agricultural workers and 30 female controls. The exposed subjects worked during the spring and summer in thinning and pruning fruit trees and in harvesting and packing different fruits, such as raspberries, grapes, apples, and kiwis. They did not use any protective measures during their work activities. A significant increase in the frequency of binucleated cells with micronuclei (BNMN) was found in the exposed women as compared with the controls (36.94 +/- 14.47 vs. 9.93 +/- 6.17 BNMN/1000 BN cells; P < 0.001). The frequency of BNMN varied as a function of age in both the exposed and control groups, but no correlation was found between BNMN frequency and the duration of exposure. Also, smoking and other habits had no effect on MN frequency. Our study confirms that occupational exposure to pesticide mixtures results in cytogenetic damage.

Cytogenetic biomonitoring of workers from laboratories of clinical analyses occupationally exposed to chemicals

A cytogenetic monitoring study was carried out on a group of workers in clinical analysis laboratories to investigate the risk of occupational exposure to chronic low levels of chemicals. Thirty-four clinical laboratories have been involved in the study. In these laboratories, toxicants and analytical procedures utilized have been characterized. The individual occupational exposure of workers was assessed by use of a questionnaire concerning the chemical substances utilized. About 300 different chemicals have been identified. Cytogenetic analyses (chromosomal aberration and micronucleus tests) were carried out on a strictly selected group of 50 workers enrolled from these laboratories and compared to 53 controls (healthy blood donors) matched for gender and age. The exposed group shows a significantly higher frequency of genetic damage than the control group. Both chromatid and chromosome aberration frequencies in workers appear significantly higher than in controls. Similarly, comparison between micronucleated cells rates of exposed and unexposed groups show significantly higher frequencies of binucleated cells with micronucleus (BNMN) and of total micronuclei (MN tot) in workers than in controls.

A follow-up study on micronucleus frequency in Spanish agricultural workers exposed to pesticides.

To determine whether occupational exposure to a complex mixture of pesticides results in a significant increase in the level of cytogenetic damage, a follow-up study was planned on 39 greenhouse workers from Almería (southeastern Spain). Taking into account that pesticide exposure can be season-related, two blood samples were taken from each individual at different times: one in a period of high exposure (sample A, spring-summer) and the other in a period of lower exposure (sample B, autumn-winter). Using the cytokinesis block micronucleus technique the frequency of binucleated cells with micronuclei (BNMN) and the cytokinesis blocked proliferation index (CBPI) were determined in peripheral blood lymphocytes. The results obtained indicate that there were no statistically significant differences in BNMN frequencies between the two sampling periods nor between exposed and controls. ANCOVA analysis of repeated measures revealed that the age of the individuals showed a direct relation with BNMN in the first study period. With regard to CBPI, a significant and season-related effect was found.

Occupational Exposure to Lead and Induction of Genetic Damage

To investigate whether occupational exposure to lead is genotoxic, we evaluated data from 103 lead-exposed workers and 78 matched controls. These data correspond to three different sampling periods, and we measured genetic damage as increases in the frequency of binucleated cells with micronuclei (BNMN) in peripheral blood lymphocytes. The levels of exposure were determined according to the lead levels in blood. Clearly significant increases in BNMN were observed in the exposed groups when compared to the control group. In addition, for the overall population (n = 181), we observed a clear relationship between lead levels in blood and BNMN (r = 0.497; p &lt; 0.001). When we examined four exposure levels--very low exposure (&lt; 1.20 microM/L), low exposure (1.20-1.91 microM/L), high exposure (1.92-2.88 microM/L), and very high exposure (&gt; 2.88 microM/L)--we found significant differences in the genetic damage induction. We conclude that exposure to levels of lead higher than 1.20 microM/L may pose an increase in genetic risk. In addition, our data show that blood lead level is a good indicator of genetic damage induction.

Occupational exposure to lead and induction of genetic damage.

To investigate whether occupational exposure to lead is genotoxic, we evaluated data from 103 lead-exposed workers and 78 matched controls. These data correspond to three different sampling periods, and we measured genetic damage as increases in the frequency of binucleated cells with micronuclei (BNMN) in peripheral blood lymphocytes. The levels of exposure were determined according to the lead levels in blood. Clearly significant increases in BNMN were observed in the exposed groups when compared to the control group. In addition, for the overall population (n = 181), we observed a clear relationship between lead levels in blood and BNMN (r = 0.497; p < 0.001). When we examined four exposure levels--very low exposure (< 1.20 microM/L), low exposure (1.20-1.91 microM/L), high exposure (1.92-2.88 microM/L), and very high exposure (> 2.88 microM/L)--we found significant differences in the genetic damage induction. We conclude that exposure to levels of lead higher than 1.20 microM/L may pose an increase in genetic risk. In addition, our data show that blood lead level is a good indicator of genetic damage induction.

Micronuclei induction by 131I exposure: Study in hyperthyroidism patients

To evaluate the eventual genetic damage induced by therapeutic exposure to 131I, we have studied the presence of micronuclei (MN) in binucleated peripheral blood lymphocytes from a group of 28 hyperthyroidism patients who received 131I sodium iodide, via oral administration. The study was conducted over time and blood samples were obtained before the treatment, and 1 week, 1 month and 3 months after it. The results obtained indicate a positive relationship between dose and BNMN frequency as calculated by the linear regression coefficient, showing significant increases in the frequency of MN and BNMN (binucleated cells with MN) in the subgroup of patients that received more than 500 MBq. Taking into account that the patients studied were treated with relatively low doses of 131I, our positive results support the view that the MN assay is sensitive enough to monitor the chromosome damage resulting from the exposure.