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1.
Int Arch Occup Environ Health ; 97(4): 353-363, 2024 May.
Article in English | MEDLINE | ID: mdl-38430240

ABSTRACT

OBJECTIVE: Several studies investigated the link between agricultural occupational exposures and DNA damage, in an attempt to bring elements of biological plausibility to the increased cancer risk associated with them. However, only a few of these studies focused on females. METHODS: The comet assay was performed on PBMC (Peripheral Blood Mononuclear Cells) samples from 245 females working in open field farming and cattle raising, located in the Normandy area of France. Individual questionnaires on tasks performed were administered at the time of sampling to directly assess exposures. Environmental exposures were issued from a questionnaire assessing the farm productions. Linear regression analyses were done using the DNA damage scores. RESULTS: Regarding direct exposures, several tasks associated with exposure to potentially harmful chemicals were not associated with DNA damage, but a longer duration of use of herbicide on meadows (p = 0.05) or of cleaning and upkeep of agricultural equipment (p = 0.06) revealed higher DNA damage levels, although the number of exposed women was low. Several indirect and/or environmental exposures were associated with DNA damage in multivariate analyses: a larger surface of meadows (p = 0.006) or the presence of poultry (p = 0.03) was associated with less DNA damage, while the presence of swine (p = 0.01) was associated with higher DNA damage. Smokers and former smokers had less DNA damage than non-smokers (p = 0.0008 and p = 0.03). CONCLUSIONS: We report modified levels of DNA damage for those environmentally exposed to meadows, poultry and pig farming, underlining the need for a better knowledge of the potential health risks experienced by females in this setting.


Subject(s)
Leukocytes, Mononuclear , Occupational Exposure , Female , Humans , Animals , Cattle , Swine , Comet Assay , Farmers , DNA Damage , Occupational Exposure/adverse effects , Agriculture
2.
Sci Total Environ ; 919: 170816, 2024 Apr 01.
Article in English | MEDLINE | ID: mdl-38346656

ABSTRACT

Since 2014, the Agricultural Operator Exposure Model (AOEM) has been the harmonised European model used for estimating non-dietary operator exposure to pesticide. It is based on studies conducted by the pesticide companies and it features 13 different crops including non-agricultural areas such as amenity grasslands. The objective of this study was to compare the dermal exposure measured during a field study conducted in a non-agricultural area with the corresponding values estimated by the model AOEM. The non-controlled field study was conducted in France in 2011 and included 24 private and public gardeners who apply glyphosate with knapsack sprayers. Dermal exposure was measured using the whole-body method and cotton gloves. Each measured value had an estimated value given by AOEM and we tested their correlation using linear regression. The model overestimated body exposure for all observations and there was no correlation between values. However, it underestimated hand exposure by 42 times and it systematically underestimated the exposure when the operators were wearing gloves, especially during the application. The model failed at being conservative regarding hand exposure and highly overestimated the protection afforded by the gloves. At a time of glyphosate renewed approval in Europe, non-controlled field studies conducted by academics are needed to improve AOEM model, especially in the non-agricultural sector. Indeed, among the 34 studies included in the model, none were conducted on a non-agricultural area and only four assessed the exposure when using a knapsack sprayer. Moreover, knapsack sprayers being the main equipment used worldwide in both agricultural and non-agricultural settings, it is also crucial to integrate new data specific to this equipment in the model. Operator exposure should be estimated with accuracy in the registration process of pesticides to ensure proper safety as well as in epidemiological studies to improve exposure assessment.


Subject(s)
Occupational Exposure , Pesticides , Pesticides/analysis , Parks, Recreational , Occupational Exposure/analysis , Agriculture , Glyphosate
3.
Environ Res ; 212(Pt C): 113375, 2022 09.
Article in English | MEDLINE | ID: mdl-35533714

ABSTRACT

INTRODUCTION: Knowledge on the health of greenspace workers is scarce, even though they are exposed to many occupational hazards. The aim of this study was to analyze mortality by cause, prevalence of some non-cancer diseases, and incidence of the main cancers among greenspace workers. METHODS: A sub-cohort of greenspace workers was formed within the AGRICAN cohort. Demographic information, health characteristics and self-reported diseases at enrollment were described separately in terms of frequencies (%), median and Interquartile Range (IQR) for greenspace workers, farmers, and other non-agricultural workers. Causes of death and cancer incidence were identified through linkage with cancer registries from enrollment (2005-2007) to 2015. Hazard ratio (HR) and 95% Confidence Intervals [95% CI] were estimated using Cox proportional hazard regression with age as the underlying timescale. RESULTS: The sub-cohort included 6247 greenspace workers who were higher proportion men, younger and more frequently smokers than farmers and non-agricultural workers. Male and female greenspace workers reported more history of allergic diseases; and males, more history of depression. Compared to other workers male greenspace workers showed a non-significant higher mortality from ischemic cardiological diseases (HR = 1.14 [0.81-1.60]). Incidence was higher in male greenspace workers than farmers for overall cancer (HR = 1.15 [1.04-1.27]), cancer of the prostate (HR = 1.21 [1.02-1.44]), thyroid (HR = 2.84 [1.26-6.41]), testis (HR = 3.98 [1.50-10.58]) and skin melanoma (HR = 2.15 [1.33-3.47]). Non-significant increased risks were also found for sarcomas, larynx and breast. In women, risk of breast cancer was higher in greenspace workers than in farmers (HR = 1.71 [1.17-2.50]). CONCLUSIONS: Whereas greenspace workers have often been included with other pesticide applicators in epidemiological studies, our analyses highlighted the differences between these two populations. They demonstrate the need to study them separately and to investigate more thoroughly the role of specific occupational exposures such as pesticides as well as the effect on women.


Subject(s)
Breast Neoplasms , Occupational Exposure , Pesticides , Farmers , Female , Humans , Incidence , Male , Parks, Recreational
4.
Environ Res ; 203: 111822, 2022 01.
Article in English | MEDLINE | ID: mdl-34352232

ABSTRACT

CONTEXT: The greenspace sector includes a broad range of occupations: gardeners, landscapers, municipal workers, maintenance operators of public facilities, golf-course employees and other sports facilities, horticulturists, plant and tree nursery workers etc. The health impact of occupational pesticide exposure has mainly been studied among farmers. Other professionals such as greenspace workers are also extremely exposed, presenting specific exposure features (practices, types of pesticide used). The aim of this review was to summarize epidemiological literature that examine the relationship between pesticide exposure and the risk of cancer and long-term health effects in greenspace workers. METHOD: Six main groups of greenspace workers were identified and examined through a systematic literature review based on PubMed and Scopus. The studies were then grouped according to their design, health outcomes and the type of population studied. RESULTS: Forty-four articles were selected among the 1679 identified. Fifteen studies were conducted exclusively among greenspace workers, while ten also studied these workers with other pesticide applicators. Six were cohorts from the general population in which greenspace workers were identified. Elevated risks were found in several studies for leukaemia, soft-tissue sarcoma, multiple myeloma, non-Hodgkin lymphoma and Parkinson's disease. DISCUSSION: The majority of studies used rough parameters for defining exposure such as job titles which could lead to the misclassification of exposure, with the risk of false or positive negative conclusions. Health outcomes were mainly collected through registries or death certificates, and information regarding potential confounders was often missing. CONCLUSION: The review identified only 15 studies conducted exclusively among greenspace workers. Elevated risk was found for several sites of cancer and Parkinson's diseases. Further epidemiological research is needed, conducted specifically on these workers, to better characterize this population, its exposure to pesticides and the related health effects.


Subject(s)
Agricultural Workers' Diseases , Neoplasms , Nervous System Diseases , Occupational Exposure , Pesticides , Epidemiologic Studies , Humans , Neoplasms/chemically induced , Neoplasms/epidemiology , Occupational Exposure/analysis , Parks, Recreational , Pesticides/analysis , Pesticides/toxicity
5.
Sci Total Environ ; 660: 1513-1521, 2019 Apr 10.
Article in English | MEDLINE | ID: mdl-30743943

ABSTRACT

Wastewater reuse provides valuable solutions to solve the societal challenges of decreasing availability and limiting access to secure water resources. The present study quantifies the environmental performance of nectarine orchards irrigation using treated municipal wastewater (TMW) and surface water using a unique dataset based on field experimental data. Climate change, toxicity (for human and freshwater), eutrophication (marine and freshwater) and acidification impacts were analysed using the impact assessment method suggested by the International Reference Life Cycle Data System (ILCD). The water footprint associated to the life cycles of each system has been estimated using the Available WAter REmaining (AWARE) method. Monte Carlo simulation was used to assess data uncertainty. The irrigation of nectarine orchards using TMW performs better than the irrigation using surface water for eutrophication impact categories. Compared with surface water resources, the potential impacts of TMW reuse in agriculture on climate change and toxicity are affected by the wastewater treatment phase (WWT). Only eutrophication and acidification burdens are generated by in-field substitution of surface water with TMW. Considering human and ecosystem water demand, the irrigation with TMW increases water consumption of 19.12 m3 per kg of nectarine produced. Whereas, it shows a positive contribution to water stress (-0.19 m3) if only human water demand is considered. This study provides important results that allow for a better understanding of the potential environmental consequences of TMW reuse in agriculture. It suggests that embracing the type of WWTs, the replacement of fertilizers, the effects on water scarcity and ecosystem quality might be useful to redefine water reuse regulations and increase public acceptance for the reuse of TMW in agriculture. Moreover, this study reveals the need for developing consensus and standardized guidance for life cycle analysis of water reuse applications.


Subject(s)
Agricultural Irrigation/methods , Environment , Wastewater/chemistry , Climate Change , Eutrophication , Mediterranean Region , Prunus , Water Supply
6.
Environ Res ; 132: 360-9, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24837246

ABSTRACT

Physical contact with branches, leaves, fruit or vegetables in previously treated crops is responsible for the transfer of pesticides to the worker's skin in agricultural tasks such as harvesting, pruning, thinning, cutting or sorting. Few studies have documented workers' exposure during re-entry in vineyards. In the PESTEXPO study, we described levels of exposure and analyzed their determinants during re-entry and harvesting in vineyards in the Bordeaux area, France. Between 2002 and 2007, volunteers performing re-entry tasks (N=46 days) or harvesting (N=48 days) after dithiocarbamate or folpet treatment were observed. Detailed information on the tasks was collected and dermal contamination was assessed using patches placed on the skin and hand-washing at the end of each working phase. Daily median contamination was 1 967.7 µl of mixture during re-entry (90(e) percentile: 5 045.3 µl) and 18.7 µl during harvesting (90(e) percentile: 911.4 µl). The type of task was the parameter found to be the most strongly associated with contamination. For re-entry, the highest contaminations were observed during raising of wires and cutting of branches. During the harvest, the contamination was maximal for grape-picking. The delay since the last treatment and the rate of active ingredient per hectare played a role, together with other factors such as meteorological factors, crop and farm characteristics, gloves and clothes. Our results underline the necessity to take into account exposures during re-entry and harvest when considering pesticide exposure, both for epidemiological research and preventive action.


Subject(s)
Agriculture/statistics & numerical data , Occupational Exposure/analysis , Pesticides/analysis , Adult , Algorithms , Female , Humans , Male , Multivariate Analysis , Skin , Vitis
7.
Neuro Oncol ; 13(12): 1370-8, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21980160

ABSTRACT

An increase in the incidence of CNS tumors has been observed in many countries in the last decades. The reality of this trend has been much debated, as it has happened during a period when computer-assisted tomography and MRI have dramatically improved the detection of these tumors. The Gironde CNS Tumor Registry provides here the first data on CNS tumor incidence and trends in France for all histological types, including benign and malignant tumors, for the period 2000-2007. Incidence rates were calculated globally and for each histological subtype. For trends, a piecewise log-linear model was used. The overall annual incidence rate was found to be 17.6/100 000. Of this rate, 7.9/100 000 were neuroepithelial tumors and 6.0/100 000 were meningiomas. An overall increase in CNS tumor incidence was observed from 2000 to 2007, with an annual percent change (APC) of +2.33%, which was explained mainly by an increase in the incidence of meningiomas over the 8-year period (APC = +5.4%), and also more recently by an increase in neuroepithelial tumors (APC = +7.45% from 2003). The overall increase was more pronounced in women and in the elderly, with an APC peaking at +24.65% in subjects 85 and over. The increase in the incidence rates we observed may have several explanations: not only improvements in registration, diagnosis, and clinical practice, but also changes in potential risk factors.


Subject(s)
Central Nervous System Neoplasms/epidemiology , Meningeal Neoplasms/epidemiology , Neoplasms, Neuroepithelial/epidemiology , Adolescent , Adult , Aged , Aged, 80 and over , Central Nervous System Neoplasms/mortality , Child , Child, Preschool , Female , France/epidemiology , Humans , Incidence , Infant , Infant, Newborn , Male , Meningeal Neoplasms/mortality , Middle Aged , Neoplasms, Neuroepithelial/mortality , Registries , Risk Factors , Survival Rate , Time Factors , Young Adult
9.
Food Chem Toxicol ; 45(12): 2420-5, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17655998

ABSTRACT

To investigate the exposure of livestock and farm workers to mycotoxins during the last months of silage use, the mycoflora and the mycotoxins in a mature silage (11-months-old) were studied. A multimycotoxin method was developed to evaluate the toxigenic in vitro ability of fungal strains. The screening of potentially toxigenic fungi isolated from the mature silage showed that six Fusaria (Fusarium culmorum, Fusarium equiseti, Fusarium graminearum, Fusarium oxysporum, Fusarium solani and Fusarium verticillioides) and one Aspergillus (Aspergillus fumigatus) were able to produce mycotoxins on nutrient agar. Seven major mycotoxins (aflatoxin B(1), citrinin, deoxynivalenol, fumonisin B(1), gliotoxin, ochratoxin A and zearalenone) were also searched in the corn silage by high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Among the three mycotoxins (citrinin, gliotoxin and deoxynivalenol) detected in the silage, gliotoxin, a strongly immunosuppressive mycotoxin, occurred in the mature silage at level up to 877 ppb, which was associated with the presence of A. fumigatus in the silage.


Subject(s)
Aspergillus/metabolism , Food Microbiology , Fusarium/metabolism , Mycotoxins/biosynthesis , Zea mays/microbiology , Chromatography, High Pressure Liquid , Humans , Mass Spectrometry , Mycotoxins/chemistry
10.
Aquat Toxicol ; 79(1): 65-77, 2006 Aug 12.
Article in English | MEDLINE | ID: mdl-16834997

ABSTRACT

The aim of the present work was to investigate genotoxicant accumulation and biological responses of zebra mussels and blue mussels collected along a pollution gradient in the Seine estuary and in the Seine Bay. The sampling area included three contaminated and one reference sites for each species. The study focused on polyaromatic hydrocarbons (PAH), lindane, polychlorobiphenyls (PCB) and metals known to be potential genotoxicants and/or reactive oxygen species (ROS) inducers. Enzymatic activities related to cellular defence systems including the phase II enzyme glutathione S-transferase (GST) and three antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were measured in gills. DNA adducts and DNA strand breaks (Comet assay) were measured in digestive gland and hemocytes, respectively. Species differences were observed in metal accumulation (As and Pb), GPx activity and DNA adduct formation. A marked upstream-downstream gradient was reported for PAH body burden and to a lesser extent for PCB and metals with the highest values measured just downstream the industrialized area of Rouen. GST and SOD activities in gills of bivalves were positively related to PAH and metals body burden, respectively. Activation of those cellular defences may prevent accumulation of electrophilic metabolites and free radicals and thus may protect DNA and others macromolecules against oxidation and adduction. Although DNA strand breaks and bulky adducts were detected in both species, levels were relatively low and no significant site differences were observed in June 2003. Our results indicate a clear relationship between genotoxicant accumulation and positive activation of detoxification and antioxidant systems but poor consequences in term of DNA damage for wild population of mussels inhabiting the Seine estuary.


Subject(s)
Dreissena/drug effects , Enzymes/drug effects , Mytilus edulis/drug effects , Water Pollutants, Chemical/toxicity , Animals , Comet Assay/methods , DNA Adducts/drug effects , DNA Damage , Dreissena/chemistry , Environmental Exposure , France , Hexachlorocyclohexane/analysis , Hexachlorocyclohexane/toxicity , Metals/analysis , Metals/toxicity , Mytilus edulis/chemistry , Polychlorinated Biphenyls/analysis , Polychlorinated Biphenyls/toxicity , Polycyclic Aromatic Hydrocarbons/analysis , Polycyclic Aromatic Hydrocarbons/toxicity , Rivers , Water Pollutants, Chemical/analysis
12.
Mutat Res ; 587(1-2): 90-102, 2005 Nov 10.
Article in English | MEDLINE | ID: mdl-16140568

ABSTRACT

Seasonal variations of DNA-adduct levels in peripheral blood cells were evaluated in open field farmers (n=26) by use of the 32P-postlabelling assay. Samples were collected before (sample S0) and during (sample S4) the period of intensive pesticide use. A similar sampling procedure was applied to a referent group (n=29). Exposure to pesticides was estimated via a detailed questionnaire. For the group of farmers, an increase in mean adduct level was observed during the season (mean RALS0=3.9+/-3.4 x 10(-10), mean RALS4=13.3+/-15.7 x 10(-10); p=0.008; RAL=relative adduct level). The mean adduct levels were significantly different between farmers and referents only in the S4 samples, with higher levels for farmers (p=0.02). The number of different adducts per individual was higher for farmers at S4 when compared with S0 (p=0.02) and compared with the referents at S4 (p=0.03). However, the increase of the adduct level in farmers did not seem to be attributable to the occurrence of specific new adducts in S4 as compared with S0, but was supposedly due to intensification of pre-existing spots and/or appearance of new unspecific ones. This would be in agreement with indirect genotoxic (epigenetic) effects known for several pesticides, even though a direct mechanism cannot be ruled out definitively. The implication of the pesticides used by the farmers in the modulation of DNA-adduct patterns was explored by analysis of exposure data obtained from the questionnaire.


Subject(s)
DNA Adducts , Occupational Exposure , Pesticides/poisoning , Adult , Agriculture , DNA Adducts/blood , Humans , Male , Seasons
13.
Mutagenesis ; 20(5): 365-73, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16105905

ABSTRACT

The aetiology of brain tumours remains unclear. Occupational exposures to pesticides and organic solvents are suspected risk factors. The case-control study CEREPHY (221 cases, 442 controls) carried in the Departement de la Gironde in France revealed a significantly increased risk of brain tumours for subjects most exposed to pesticides. In some cancers, TP53 mutations could reflect exposure to specific carcinogens. These mutations are present in approximately 30% of astrocytic brain tumours. In a pilot study, we explored the hypothesis that pesticide or solvent exposure could raise the frequency of TP53 mutations in brain tumour cells. We investigated TP53 mutations in exons 2-11 by denaturing high performance liquid chromatography (DHPLC) and sequencing, and p53 accumulation by immunohistochemistry in brain tumour of the 30 patients from CEREPHY study with a history of occupational exposure to pesticides (n = 21) and/or organic solvents (n = 14) for whom tumoral tissue was available. Included cases concerned 27% of CEREPHY cases exposed to pesticides and, based on the cumulative index of occupational exposure, they were more exposed to pesticides. There were 12 gliomas, 6 meningiomas, 7 neurinomas, 2 central nervous system lymphomas and 3 tumours of other histological types. We detected TP53 mutations in three tumours, which is similar to the expected number (3.3) calculated from 46 published studies referenced in the IARC TP53 mutations database, taking into account histological types. Considering TP53 mutations previously detected in the laboratory by DHPLC and the frequency of TP53 polymorphisms detected in this sample (similar to published data), the TP53 mutations rate is probably not underestimated. These preliminary results, even if it was on a limited number of tumours, are not in favour of the role of pesticide or organic solvent exposure in the occurrence of TP53 mutations in brain tumours.


Subject(s)
Brain Neoplasms/chemically induced , Mutagens/toxicity , Occupational Exposure , Pesticides/toxicity , Solvents/toxicity , Tumor Suppressor Protein p53/genetics , Brain Neoplasms/chemistry , Brain Neoplasms/genetics , Case-Control Studies , Chromatography, High Pressure Liquid , DNA Mutational Analysis , DNA, Neoplasm/analysis , Exons/genetics , Female , Humans , Male , Mutation , Nucleic Acid Denaturation
14.
Occup Environ Med ; 60(12): 910-7, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14634181

ABSTRACT

AIMS: To determine haematological parameters, urine mutagenicity (on three Salmonella typhimurium strains), and DNA damage (using the comet assay) in mononuclear leucocytes of farmers before and after a one-day spraying period of pear and apple trees with the fungicide captan in usual conditions. METHODS: Fruit growers were exposed to captan during the 1998 (n = 12) and/or the 2000 spraying seasons (n = 17). Biological samples were collected on the morning of the day of spraying (S1), the evening after spraying (S2), and the morning of the day after (S3). The UK Predictive Operator Exposure Model (UK-POEM) was used to quantify pesticide exposure intensity. RESULTS: No effect was observed on haematological parameters for these two spraying seasons. Proportions of mutagenic urine samples did not significantly differ between S1 and S2/S3 sampling points. In contrast with strains TA97a and YG1041 mainly sensitive to frameshift mutations, a positive trend was observed between the difference (S3-S1) of mutagenic power on strain TA102 detecting base-pair mutations and the exposure predicted value given by UK-POEM, mainly due to parameters related to protective clothing. No significant variations in DNA damage levels were observed between S1 and S3, nor were correlations observed with parameters of pesticide exposure. CONCLUSIONS: A one-day spraying period with captan and other pesticides does not significantly induce DNA damages in mononuclear leucocytes. In contrast, an inefficient protective clothing could correlate with an increase in urine mutagenicity as assessed by the TA102 tester strain.


Subject(s)
Agriculture , Captan/toxicity , DNA Damage , Fungicides, Industrial/toxicity , Occupational Exposure/adverse effects , Adult , Captan/administration & dosage , Fruit , Fungicides, Industrial/administration & dosage , Humans , Life Style , Lymphocytes/drug effects , Male , Middle Aged , Mutagenicity Tests , Protective Clothing/statistics & numerical data , Urine/chemistry
15.
Br J Cancer ; 82(2): 263-9, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10646875

ABSTRACT

A prospective study was conducted to evaluate the impact of home enteral tube feeding on quality of life in 39 consecutive patients treated for head and neck or oesophageal cancer at the Centre François Baclesse in Caen, France. Patients were taken as their own controls. Quality of life was evaluated using the EORTC QLQ-C30 core questionnaire, and the EORTC H&N35 and OES24 specific questionnaires. The feeding technique tolerance was evaluated using a questionnaire specifically developed for this study. Two evaluations were made, the first a week after hospital discharge (n = 39) and the second 3 weeks later (n = 30). Overall, the global health status/quality of life scale score slightly improved; among symptoms, scale scores that significantly improved (P < 0.05) concerned constipation, coughing, social functioning and body image/sexuality. The physical feeding technique tolerance was acceptable while the technique was psychologically less tolerated with two-thirds of the patients longing to have the tube removed. One third of the patients was also uncomfortable about their body image. Home enteral tube feeding was responsible for not visiting family or close relations in 15% of patients, and not going out in public in 23%. We conclude that home enteral tube feeding is a physically well accepted technique although a substantial proportion of patients may experience psychosocial distress.


Subject(s)
Enteral Nutrition/psychology , Esophageal Neoplasms , Head and Neck Neoplasms , Quality of Life , Stress, Psychological , Adult , Aged , Body Image , Esophageal Neoplasms/therapy , Family Relations , Female , Head and Neck Neoplasms/therapy , Health Status , Home Care Services , Humans , Male , Middle Aged , Patient Satisfaction , Prospective Studies , Sexual Behavior
16.
Cytometry ; 36(2): 117-22, 1999 Jun 01.
Article in English | MEDLINE | ID: mdl-10554159

ABSTRACT

BACKGROUND: The ability of the comet assay to quantify DNA strand breaks and alkali labile sites has been widely demonstrated. In this study, this assay was tested for its ability to identify DNA fragmentation occurring during apoptosis in comparison with standard DNA flow cytometry analysis. METHODS: Staurosporine-induced apoptosis in CHO cells is an adequate model to study a rapid time- and dose-dependent appearance of this process. RESULTS: Nuclear staining with DAPI confirmed the induction of apoptosis with a typical chromatin condensation and fragmentation. Analysis of propidium-iodide- (PI) stained DNA by flow cytometry showed the presence of a pre-G1 peak, characteristic of apoptotic cells, 6 h after drug treatment. The detection of highly damaged cells (HDC) by the comet assay after 3 h treatment occurred earlier than the detection of apoptotic cells by flow cytometry. However, HDC were missed when the DNA fragmentation was too high, preventing accurate quantification of late apoptotic cells. CONCLUSIONS: The comet assay is more sensitive than standard DNA flow cytometry to detect early DNA fragmentation events occurring during apoptosis. However, the comet assay modified by omitting electrophoresis was necessary to quantify apoptotic fraction at later stages.


Subject(s)
Apoptosis/drug effects , Comet Assay/methods , Flow Cytometry/methods , Staurosporine/pharmacology , Animals , CHO Cells , Cell Survival/drug effects , Cricetinae , DNA Damage , Fluorescent Dyes , Indoles , Kinetics , Staining and Labeling
17.
Histochem Cell Biol ; 112(2): 155-61, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10460469

ABSTRACT

Comet, TUNEL, and annexin V assays were used to identify DNA fragmentation and plasma membrane alterations occurring during staurosporine-induced apoptosis in Chinese hamster ovary cells. TUNEL assay detected apoptotic cells after 6 h treatment. The occurrence of annexin V immunofluorescence staining after 1 h treatment confirms that exposure of phosphatidylserine (PS) residues is an early biochemical feature of apoptosis. According to intensity, three annexin staining patterns were distinguished, related to different steps in the apoptotic process. The detection of highly damaged cells by the comet assay after 3 h treatment occurred earlier than the detection of DNA modifications by the TUNEL assay, but later than the exposure of PS residues. However, late apoptotic cells, otherwise characterized by plasma membrane disruption and high annexin V staining, were not detected by the comet assay. In this case, comet assay modified by omitting electrophoresis (halo assay) was more sensitive for an accurate quantification of the apoptotic fraction.


Subject(s)
Annexin A5/analysis , Apoptosis , Staurosporine/pharmacology , Animals , CHO Cells , Cricetinae , DNA Fragmentation , In Situ Nick-End Labeling , Phosphatidylserines/metabolism , Time Factors
18.
Mutat Res ; 419(1-3): 79-90, 1998 Nov 09.
Article in English | MEDLINE | ID: mdl-9804902

ABSTRACT

One herbicide (isoproturon), two fungicides (carbendazim and chlorothalonil) and etoposide (an effective antitumor agent used as a positive control), were tested for their ability to induce cytotoxic and genotoxic effects in Chinese Hamster Ovary (CHOK1) cells. Etoposide induced DNA damage detectable both by the alkaline Single Cell Gel Electrophoresis (SCGE) assay and the chromosomal aberration (CA) test in absence of noticeable cytotoxicity. With the SCGE assay, a clear induction of DNA damage was observed for chlorothalonil within a 0.2 to 1 microM concentration range. In the CA test, chlorothalonil gave also positive results, inducing mainly chromosome breaks. In contrast, no DNA damage was observed with the SCGE assay for carbendazim and isoproturon. In the CA test, carbendazim induced only numerical aberrations in the concentration range of 25 microM to 100 microM, and isoproturon did not induce any significant increase in CA. In conclusion, chlorothalonil appears genotoxic in proliferative CHOK1 cells, and as expected, the aneugenic compound, carbendazim, did not induce DNA strand breaks in the SCGE assay.


Subject(s)
Carbamates , Chromosome Aberrations , DNA Damage , Mutagenicity Tests/methods , Pesticides/toxicity , Phenylurea Compounds , Animals , Benzimidazoles/toxicity , CHO Cells , Cricetinae , Electrophoresis, Agar Gel , Fungicides, Industrial/toxicity , Herbicides/toxicity , Methylurea Compounds/toxicity , Nitriles/toxicity
19.
Cancer Epidemiol Biomarkers Prev ; 7(10): 917-27, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9796638

ABSTRACT

The alkaline comet assay was used to quantify, using visual and image analyses, the level of DNA damage in mononuclear leukocytes of farmers who were occupationally exposed to pesticides. Hematological parameters were also measured on the same samples. Enrollment of farmers was based on handling of heavily used pesticides at particular periods during one spraying season. Forty-one blood samples from 29 different farmers were collected at the beginning of the season (n = 11) and at the intermediate (n = 14) and final (n = 16) periods of intense spraying activity. The mean numbers of lymphocytes and eosinophils were nonsignificantly higher in groups 3, 1, and 4 than they were in group 2. No individual characteristics significantly influenced the mean number of lymphocytes or eosinophils, and no correlation was observed between pesticide exposure-related parameters and hematological parameters. The level of DNA damage was significantly (P < 0.01) higher in groups 3, 1, and 4 than it was in group 2. In addition, DNA damage quantification was not significantly different among investigators or among slides. Prescription medicine, alcohol consumption, and age had no statistically significant effect on DNA damage level. Conversely, smoking (smokers versus non- and ex-smokers) significantly influenced DNA damage level (P < 0.0001). A significant (P < 0.05) negative correlation was detected between the number of days without pesticide spraying and DNA damage level, particularly among non- and ex-smokers. DNA damage detected by the alkaline comet assay seems to reflect ongoing exposure to genotoxic agents but not an accumulation of damage.


Subject(s)
Agrochemicals/adverse effects , DNA Damage/genetics , Environmental Monitoring/methods , Leukocytes, Mononuclear/drug effects , Occupational Exposure/adverse effects , Pesticides/adverse effects , Seasons , Adult , Analysis of Variance , Eosinophils/drug effects , France , Humans , Leukocyte Count/drug effects , Lymphocyte Count/drug effects , Male , Middle Aged , Prospective Studies , Risk Factors , Smoking/adverse effects
20.
Cancer Epidemiol Biomarkers Prev ; 7(10): 929-40, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9796639

ABSTRACT

The alkaline comet assay was used to assess DNA damage in mononuclear leukocytes of farmers before and after a 1-day spraying period with selected pesticides under usual conditions. Two blood samples were collected, one in the morning of the day of spraying (S0) and the second in the morning of the day after (S1). Here, we assessed variations in DNA damage levels between these two sampling times. Four groups of farmers were formed, according to exposure to: (a) various fungicide-insecticide mixtures (including chlorothalonil; group 1, n = 8), (b) the herbicide isoproturon (group 2, n = 11), (c) fungicide triazoles (group 3, n = 14), and (d) a fungicide (chlorothalonil)-insecticide mixture (group 4, n = 8). An increase in DNA damage levels was observed at S1 for groups 1 and 4, who were exposed to similar pesticides. This increase was correlated with area sprayed between S0 and S1 and with the number of spraying tanks used over this 1-day period. No effect was observed on cell viability or on hematological parameters for these two groups. No statistically significant modification of DNA damage level was observed the day after spraying for groups 2 and 3, when each was observed as a whole. However, some farmers presented significantly more DNA damage after exposure, and others presented less damage. In these two groups, a significant decrease of neutrophils was observed at S1, and a decrease of red blood cells was observed in group 3. In parallel, a significant loss of lymphocyte viability was observed in these two groups. A 1-day spraying period seems to be sufficient to significantly modify DNA damage levels in mononuclear leukocytes, but the correlation of this change with pesticide-related exposure parameters depends on the kind of pesticide concerned.


Subject(s)
Agrochemicals/adverse effects , DNA Damage/genetics , Environmental Monitoring/methods , Leukocytes, Mononuclear/drug effects , Occupational Exposure/adverse effects , Pesticides/adverse effects , Adult , Agrochemicals/chemistry , Erythrocyte Count/drug effects , Humans , Leukocyte Count/drug effects , Lymphocytes/drug effects , Male , Middle Aged , Neutrophils/drug effects , Pesticides/chemistry , Prospective Studies , Surveys and Questionnaires , Time Factors
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