RESUMO
BACKGROUND: Caregivers in healthcare settings are exposed to a risk of antineoplastic drug contamination which can lead to adverse health effects. Biological monitoring is necessary to estimate the actual level of exposure of these workers. This study was conducted with the aim of assessing blood contamination levels by irinotecan and its metabolites of pharmaceutical staff operating inside and outside a compounding unit. METHODS: The study took place within the pharmaceutical unit of a French comprehensive cancer centre. Blood samples were collected from the pharmacy workers operating inside and outside the compounding unit, and analysed by UHPLC-MS/MS. Plasma and red blood cell irinotecan and its metabolites (SN-38; APC) were determined with a validated analytical method detection test. RESULTS: A total of 17/78 (21.8%) plasma and red blood cell-based assays were found to be contaminated among staff. Overall, the total number of positive assays was significantly higher for staff members working outside the compounding unit than for workers working inside it (P = 0.022), with respectively 5/42 (11.9%) and 12/36 (33.3%) positive assays. For plasma dosages, the "outside" group had a significantly higher number of positive assays (P = 0.014). For red blood cell-based assays, no significant difference was found (P = 0.309). CONCLUSIONS: This study reveals that pharmaceutical staff serving in health care settings are exposed to a risk of antineoplastic drug contamination, not only inside the compounding room but also in adjacent rooms. The results would help to raise awareness and potentially establish protective measures for caregivers working in areas close to the compounding room as well.
Assuntos
Antineoplásicos , Exposição Ocupacional , Farmácia , Composição de Medicamentos , Contaminação de Medicamentos , Monitoramento Ambiental/métodos , Humanos , Irinotecano/análise , Exposição Ocupacional/análise , Preparações Farmacêuticas , Espectrometria de Massas em TandemRESUMO
Epidemiological studies have demonstrated the link between chronic exposure to particulate matter (PM), especially particles with an aerodynamic diameter lesser than 2.5 µm (PM(2.5) ), and lung cancer. Mechanistic investigations focus on the contribution of the various genotoxicants adsorbed onto the particles, and more particularly on polycyclic aromatic hydrocarbons or nitroaromatics. Most of the previous studies dealing with genotoxic and/or mutagenic measurements were performed on organic extracts obtained from PM(2.5) collected in polluted areas. In contrast, we have evaluated genotoxic and mutagenic properties of urbano-industrial PM(2.5) (PM) collected in Dunkerque (France). Thermally desorbed PM(2.5) (dPM) was also comparatively studied. Suspensions of PM and dPM (5-50 µg per plate) were tested in Salmonella tester strains TA98, TA102 and YG1041 ± S9mix. Significant mutagenicity was observed for PM in YG1041 ± S9 mix. In strain TA102 - S9mix, a slight, but not significant dose-response increase was observed, for both PM and dPM. Genotoxic properties of PM and dPM were evaluated by the measurement of (1) 8-OHdG in A549 cells and (2) bulky DNA adducts on A549 cells and on human alveolar macrophages (AMs) in primary culture. A dose-dependant formation of 8-OHdG adducts was observed on A549 cells for PM and dPM, probably mainly attributed to the core of the particles. Bulky DNA adducts were observed only in AMs after exposure to PM and dPM. In conclusion, using relevant exposure models, suspension of PM(2.5) induces a combination of DNA-interaction mechanisms, which could contribute to the induction of lung cancer in exposed populations.
Assuntos
Carcinógenos/toxicidade , Indústrias , Mutagênicos/toxicidade , Material Particulado/toxicidade , Urbanização , 8-Hidroxi-2'-Desoxiguanosina , Testes de Carcinogenicidade , Carcinógenos/química , Linhagem Celular , Células Cultivadas , Adutos de DNA/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , França , Humanos , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/metabolismo , Testes de Mutagenicidade , Mutagênicos/química , Concentração Osmolar , Tamanho da Partícula , Material Particulado/química , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/metabolismo , Especificidade da Espécie , Fatores de TempoRESUMO
Using an air-liquid interface (ALI) device in dynamic conditions, we evaluated the efficiency of fuel after-treatment strategies (diesel oxidation catalysis, DOC, and diesel particulate filter, DPF, devices) and the impact of 7% and 30% rapeseed methyl esters (RME) blending on oxidative stress and genotoxicity induced in A549 lung cells after 3h exposure to whole Diesel exhausts. Oxidative stress was studied using assays of ROS production, glutathione level, catalase and superoxide-dismutase (SOD) activities. No oxidative stress and no clear differences on cytotoxicity patterns between biodiesel and standard Diesel exhausts were found. A weak but significant genotoxicity (8-oxodGuo adducts) and, for standard Diesel only, a DNA damage response (DDR) as evidenced by ÆH2AX foci, remained after DOC+DPF flowing. All together, these data could contribute to the improvement of the after treatment strategies and to health risk assessment of current diesel exhausts.
Assuntos
Poluentes Atmosféricos/toxicidade , Biocombustíveis , Mutagênicos/toxicidade , Testes de Toxicidade/instrumentação , Emissões de Veículos/toxicidade , Células A549 , Poluentes Atmosféricos/análise , Catalase/metabolismo , Dano ao DNA , Glutationa/metabolismo , Humanos , Testes de Mutagenicidade , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Testes de Toxicidade/métodos , Emissões de Veículos/análiseRESUMO
Diesel exhausts are partly responsible for the deleterious effects on human health associated with urban pollution, including cardiovascular diseases, asthma, COPD, and possibly lung cancer. Particulate fraction has been incriminated and thus largely investigated for its genotoxic properties, based on exposure conditions that are, however, not relevant for human risk assessment. In this paper, original and more realistic protocols were used to investigate the hazards induced by exhausts emitted by the combustion of standard (DF0) vs. bio-diesel fuels (DF7 and DF30) and to assess the impact of exhaust treatment devices (DOC and DPF). Mutagenicity and genotoxicity were evaluated for (1) resuspended particles ("off line" exposure that takes into account the bioavailability of adsorbed chemicals) and for (2) the whole aerosols (particles+gas phase components) under continuous flow exposure ("on line" exposure). Native particles displayed mutagenic properties associated with nitroaromatic profiles (YG1041), whereas PAHs did not seem to be involved. After DOC treatment, the mutagenicity of particles was fully abolished. In contrast, the level of particle deposition was low under continuous flow exposure, and the observed mutagenicity in TA98 and TA102 was thus attributable to the gas phase. A bactericidal effect was also observed in TA102 after DOC treatment, and a weak but significant mutagenicity persisted after DPF treatment for bio-diesel fuels. No formation of bulky DNA-adducts was observed on A549 cells exposed to diesel exhaust, even in very drastic conditions (organic extracts corresponding to 500 µg equivalent particule/mL, 48 h exposure). Taken together, these data indicate that the exhausts issued from the bio-diesel fuels supplemented with rapseed methyl ester (RME), and generated by current diesel engines equipped with after treatment devices are less mutagenic than older ones. The residual mutagenicity is linked to the gas phase and could be due to pro-oxydants, mainly for RME-supplemented fuels.
Assuntos
Biocombustíveis/toxicidade , Brassica rapa/química , Mutagênicos/toxicidade , Nitrobenzenos/toxicidade , Material Particulado/toxicidade , Salmonella typhimurium/efeitos dos fármacos , Emissões de Veículos/toxicidade , Aerossóis , Brônquios/citologia , Brônquios/efeitos dos fármacos , Catálise , Linhagem Celular Tumoral , Dano ao DNA , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Ésteres , Filtração/métodos , Gasolina , Humanos , Testes de Mutagenicidade , Oxirredução , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimentoRESUMO
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.
Assuntos
Agroquímicos/efeitos adversos , Dano ao DNA/genética , Monitoramento Ambiental/métodos , Leucócitos Mononucleares/efeitos dos fármacos , Exposição Ocupacional/efeitos adversos , Praguicidas/efeitos adversos , Estações do Ano , Adulto , Análise de Variância , Eosinófilos/efeitos dos fármacos , França , Humanos , Contagem de Leucócitos/efeitos dos fármacos , Contagem de Linfócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de Risco , Fumar/efeitos adversosRESUMO
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.
Assuntos
Agroquímicos/efeitos adversos , Dano ao DNA/genética , Monitoramento Ambiental/métodos , Leucócitos Mononucleares/efeitos dos fármacos , Exposição Ocupacional/efeitos adversos , Praguicidas/efeitos adversos , Adulto , Agroquímicos/química , Contagem de Eritrócitos/efeitos dos fármacos , Humanos , Contagem de Leucócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Neutrófilos/efeitos dos fármacos , Praguicidas/química , Estudos Prospectivos , Inquéritos e Questionários , Fatores de TempoRESUMO
The effect of melatonin, an indole hormone of the pineal gland, on the initiation of N-nitroso-N-methylurea (NMU)-induced carcinogenesis in rats and mutagenesis in vitro has been investigated. Two-month-old female LIO rats (groups 1 and 2) were exposed to a single injection of NMU (50 mg/kg of body weight, i.v.). Rats from group 2 were given melatonin orally (20 mg/l) from 18:00 to 09:00 h over 3 days (2 days before and 1 day after NMU injection). Animals from group 1 (control) were administered the solvent (ethanol/water, 1:1000). Rats were followed up to natural death or were sacrificed when moribund. Tumors developed both in rats treated with NMU alone (50.0%) and in rats exposed to NMU plus melatonin (34.8%). The percentage of malignant tumor-bearing rats in group 2 (21.7%) was lower (P < 0.02) than that in the other group (41.7%). Melatonin also decreased the multiplicity of malignant tumors 1.3-fold and reduced the incidence of malignancies in some organs. Two in vitro tests were used for mutagenesis studies: the Ames test (strains TA 100 and TA 102 of Salmonella typhimurium) and the Single Cell Gel Electrophoresis assay (SCGE assay or COMET assay) performed on CHOK1 cells. Melatonin itself revealed no genotoxic effect in either of the tests. No protective action of melatonin (at doses of up to 2 micromol/plate) towards NMU was found in the Ames test. In contrast, in the SCGE assay a slight, but statistically significant (P < 0.001), dose-related anticlastogenic effect of melatonin (10(-10)-10(-7) M) was observed. Thus, our data indicate that melatonin may act as an anti-initiating hormone in NMU-induced carcinogenesis and possess anticlastogenic activity towards NMU in CHOK1 cells.
Assuntos
Antimutagênicos/farmacologia , Melatonina/farmacologia , Metilnitrosoureia/toxicidade , Neoplasias Experimentais/prevenção & controle , Animais , Dano ao DNA , Feminino , Neoplasias Experimentais/induzido quimicamente , Ratos , Salmonella typhimurium/efeitos dos fármacosRESUMO
Forty female CBA mice aged 3-4 months were exposed twice a week during 2 months to intravaginal applications of polyurethane sponges impregnated with 0.1% solution of 7,12-dimethylbenz[a]anthracene (DMBA) in triethyleneglycol. Three hours after each application the sponges were taken out. Starting from the day of the 1st DMBA application a part of mice was exposed five times a week during 4 months with melatonin in tap water (20 mg/l) given at night time (from 18:00 to 09:00 h). Additional 20 female CBA mice were intact and served as a control. All mice were sacrificed in 6 months after start of the experiment. Seven of 20 mice exposed to DMBA alone developed malignancies in the vagina and cervix uteri and two mice developed benign cervical tumors. No malignancies in vagina and uterine cervix and three vaginal papillomas were observed in mice exposed to DMBA+melatonin. There were no any tumors in intact controls. Two in vitro tests were used for mutagenicity studies: the Ames test (strains TA 97 and TA 98 of Salmonella typhimurium) and the single cell gel electrophoresis assay (SCGE assay or COMET assay) performed on CHOK1 cells. In tested strains melatonin significantly reduced the mutagenicity of DMBA. In the SCGE assay preincubation with melatonin led to a strong inhibition of clastogenic activities of DMBA. Thus, our data indicate that pineal indole hormone melatonin inhibits cervical and vaginal carcinogenesis induced by DMBA in mice and possess antimutagenic and anticlastogenic effect in vitro.
Assuntos
9,10-Dimetil-1,2-benzantraceno/antagonistas & inibidores , Anticarcinógenos/uso terapêutico , Antimutagênicos/uso terapêutico , Melatonina/uso terapêutico , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias Vaginais/prevenção & controle , 9,10-Dimetil-1,2-benzantraceno/toxicidade , Animais , Células CHO , Carcinógenos/antagonistas & inibidores , Carcinógenos/toxicidade , Ensaio Cometa , Cricetinae , Relação Dose-Resposta a Droga , Interações Medicamentosas , Feminino , Camundongos , Camundongos Endogâmicos CBA , Mutagênese/efeitos dos fármacos , Mutagênicos/toxicidade , Lesões Pré-Cancerosas/induzido quimicamente , Lesões Pré-Cancerosas/patologia , Lesões Pré-Cancerosas/prevenção & controle , Ratos , Neoplasias do Colo do Útero/induzido quimicamente , Neoplasias do Colo do Útero/patologia , Neoplasias Vaginais/induzido quimicamente , Neoplasias Vaginais/patologiaRESUMO
An in vitro study of the combined cytotoxicity of either cisplatin (CDDP) or carboplatin and amphotericin B (AmB) was undertaken on a set of different ovarian carcinoma (IGROVI, IGROVI-C10, OAW42) and peritoneal malignant mesothelioma (CFB-CARP1) cell lines and ascitic cells freshly obtained from ovarian cancer patients so as to investigate the possibility of overcoming their resistance to platinum compounds. Growth-inhibition curves obtained 6 days after a 2-h period of exposure to the drugs showed that AmB at 5-10 mg/l allowed a 5- to 10-fold decrease in the 50% growth-inhibitory concentrations (IC50) of CDDP and carboplatin on either sensitive or resistant cells. Intracellular platinum assays with IGROVI cells showed that AmB acted by increasing dramatically the platinum uptake at a proportion that accounted for the increase in cytotoxicity. In the subline IGROVI-C10, a 10-fold resistant subline of IGROVI, AmB at 10 mg/l allowed recovery to the level of sensitivity seen in the parental cell line in the absence of AmB but not to the level observed in the presence of AmB. Acquisition of resistance mechanisms that are independent of the regulation of platinum uptake might be involved in this cell line. Thus, AmB might act by increasing the intracellular concentration of platinum without modifying the resistance mechanism involved downstream. However, in our models an increase in the intracellular level of platinum was always sufficient for the recovery of chemosensitivity in vitro. We also show that the phosphodiesterase inhibiting methylxanthines act synergistically with AmB. The latter drugs are weakly toxic and could also attenuate the nephrotoxicity of AmB.
Assuntos
Adenocarcinoma/tratamento farmacológico , Anfotericina B/farmacologia , Antineoplásicos/farmacologia , Carboplatina/farmacologia , Cisplatino/farmacologia , Mesotelioma/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Peritoneais/tratamento farmacológico , Sinergismo Farmacológico , Feminino , Humanos , Resultado do Tratamento , Células Tumorais CultivadasRESUMO
UDP-Galactose: N-acetylglucosaminyl glycoprotein beta 1-4 galactosyltransferase (GT) catalyzes the transfer of galactose to N-acetylglucosamine from UDP-[3H]Gal. The uncharged reaction product (tritiated N-acetyllactosamine) is separated from the unreacted UDP-[3H]Gal by ion-exchange chromatography. The major advantage of this method is its rapidity compared to other isotopic techniques.
Assuntos
Lactose Sintase/sangue , N-Acetil-Lactosamina Sintase/sangue , Cromatografia por Troca Iônica , Feminino , Humanos , Masculino , Valores de ReferênciaRESUMO
We present a comparative study of several biological markers (galactosyltransferase, CA 125, isoenzymes of amylase and alkaline phosphatase) with a view to ovarian carcinoma follow-up. Serum samples were obtained from a population of 75 patients under clinical observation. After a minimum 18-months period, we assessed the prognostic value of the markers. No marker permits the detection of discrete, evolving carcinomas. CA 125 is the marker that gives the best results, particularly in terms of sensitivity. Galactosyltransferase has a lower sensitivity except in the case of endometrioid carcinomas. Simultaneous analysis with CA 125 and galactosyltransferase results in no decisive improvement, other than greater precision in unfavourable prognoses. Isoenzymes of amylase and alkaline phosphatase are of no interest in the follow-up of such carcinomas.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma/sangue , Neoplasias Ovarianas/sangue , Fosfatase Alcalina/sangue , Amilases/sangue , Antígeno Ca-125/sangue , Carcinoma/enzimologia , Feminino , Seguimentos , Galactosiltransferases/sangue , Humanos , Isoenzimas/sangue , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/secundário , Neoplasias Ovarianas/enzimologia , Prognóstico , Sensibilidade e Especificidade , gama-Glutamiltransferase/sangueRESUMO
Isoelectric focusing on agarose gel was used to separate the isoenzymes of serum galactosyltransferase (uridine diphosphogalactose: N-acetylglucosaminyl galactosyltransferase, EC 2.4.1.22) from 8 healthy women, and 11 ovarian cancer patients of whom 4 were in clinical remission. In all cases, we found 7 major peaks with isoelectric points ranging from 4.0-5.4. The most acidic peaks were preferentially elevated in the tumor-bearing patients, particularly the peak with pI 4.44.
Assuntos
Carcinoma/enzimologia , Galactosiltransferases/metabolismo , Isoenzimas/metabolismo , Neoplasias Ovarianas/enzimologia , Idoso , Feminino , Galactosiltransferases/sangue , Humanos , Focalização Isoelétrica/métodos , Isoenzimas/sangue , Pessoa de Meia-IdadeRESUMO
The effect of a potent endogenous antioxidant, the pineal gland indole melatonin (MLT) on the mutagenicity of twelve well-known mutagens and carcinogens has been investigated using two in vitro tests the Ames test and the single cell gel electrophoresis assay (SCGE assay or COMET assay). The 12 mutagens used were 7, 12-dimethylbenz(a)anthracene (DMBA), benzo(a)pyrene (BP), 2-aminofluorene (AF), 1,2-dimethylhydrazine (DMH), bleomycin, cyclophosphamide (CP), 4-nitroquinoline-N-oxide (NQO), 2,4, 7-trinitro-9-fluorenone (TNF), 9-aminoacridine (AA), N-nitrosomethylurea (NMU), mitomycin C and sodium azide tested in the absence or in the presence of S9 mix. MLT alone turned out neither toxic nor mutagenic in the Ames test and revealed clastogenic activity at the highest concentration tested (100 microM) in the SCGE assay. In four Salmonella typhimurium tester strains TA 97, TA 98, TA 100 and TA 102 MLT significantly reduced the mutagenicity of chemicals which require S9 activation. In the SCGE assay performed on CHO cells, preincubation with MLT led to a strong inhibition of clastogenic activities of DMBA and CP, and in a lesser extent with BP and NMU. With mitomycin C, MLT exacerbated responses in both tests. The possible mechanisms of MLT's inhibitory action are discussed.
Assuntos
Antioxidantes/farmacologia , Melatonina/farmacologia , Mutagênicos/toxicidade , Animais , Células CHO/efeitos dos fármacos , Cricetinae , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genéticaRESUMO
The mutagenic potency of nine methylnitrocarbazoles, four methylaminocarbazoles and the methylcarbazole parent compounds was evaluated in Salmonella typhimurium TA98 and TA100, in the absence and presence of S9 isolated from Aroclor-induced rat liver. Nitro derivatives were additionally tested in TA98NR and TA98/1,8DNP6, and mutagenicity of nitrocarbazoles bearing methyl groups in positions 1 and 4 was also determined in TA1537 and TA1977, with and without S9. The addition of methyl groups on non-mutagenic carbazole can induce a mutagenic response where the intensity and nature of the effect depends on the position of the substitution: base-pair substitutions were only observed for N-methylated carbazoles, whereas 1,4-dimethylated compounds exhibited frameshift mutagenicity. All these activities depended on the presence of S9. From its dependence on classical nitroreductases, direct mutagenicity of methylnitro derivatives should be attributed to bacterial reduction of nitro groups. The influence of the methyl groups (and other additional substituents) on mutagenicity of these derivatives is discussed through their effects on life-time and reactivity of the intermediates (i.e., hydroxylamines and nitrenium ions), taking into account the nature, the position and the number of substituted sites Mutagenic activity of methylnitrocarbazoles was also tentatively correlated with various molecular descriptors. Among them hydrophobicity was found to be strongly correlated with the mutagenicity of the 1,4diMe3NC isomers. On the other hand, mutagenic potency of the nitrated and aminated methylcarbazoles varied independently of parameters linked to their oxidoreduction properties (i.e., reduction and oxidation potentials, LUMO and HOMO energies).
Assuntos
Aminas/farmacologia , Carbazóis/farmacologia , Mutagênicos/farmacologia , Nitrocompostos/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Aminas/química , Animais , Biotransformação , Carbazóis/química , Microssomos Hepáticos/metabolismo , Estrutura Molecular , Testes de Mutagenicidade , Nitrocompostos/química , Ratos , Salmonella typhimurium/genética , Relação Estrutura-AtividadeRESUMO
The effects of two fungicides (carbendazim and chlorothalonil) on the induction of DNA damage in human peripheral blood lymphocytes (human PBL) have been investigated using the single cell gel electrophoresis assay (SCGE assay or comet assay) immediately after a 1-h treatment and after a 24-h post-treatment incubation. The assessment of etoposide (an effective antitumour agent) effects on human PBL in terms of cell viability and dose-DNA damage relationships was made and etoposide selected as a positive control. The results indicate that etoposide induces significant (p < 0.01) dose-dependent DNA damages for concentrations at which the loss of cell viability is low. After a 24-h recuperation period, all observed DNA damages has disappeared. With SCGE assay performed after a 1-h treatment, similar positive results were observed with chlorothalonil alone or in association with carbendazim, without any loss of cell viability. However, a dramatic loss of cell viability was measured after 24 h and was associated with a large proportion of highly damaged cells. In contrast, carbendazim was not cytotoxic on human PBL and did not induced DNA damage using the SCGE assay either immediately after treatment or after a 24-h post-treatment incubation. These results point to the necessity of an adequate evaluation of immediate and long-term cytotoxicity of compounds that are to be assessed by the SCGE assay.
Assuntos
Benzimidazóis/farmacologia , Carbamatos , Dano ao DNA , Etoposídeo/farmacologia , Linfócitos/efeitos dos fármacos , Nitrilas/farmacologia , Adulto , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Fungicidas Industriais/farmacologia , Humanos , Masculino , Fatores de TempoRESUMO
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.
Assuntos
Carbamatos , Aberrações Cromossômicas , Dano ao DNA , Testes de Mutagenicidade/métodos , Praguicidas/toxicidade , Compostos de Fenilureia , Animais , Benzimidazóis/toxicidade , Células CHO , Cricetinae , Eletroforese em Gel de Ágar , Fungicidas Industriais/toxicidade , Herbicidas/toxicidade , Compostos de Metilureia/toxicidade , Nitrilas/toxicidadeRESUMO
The alkaline single cell gel electrophoresis (comet) assay was used to assess in vitro and in vivo genotoxicity of etoposide, a topoisomerase II inhibitor known to induce DNA strand breaks, and chlorothalonil, a fungicide widely used in agriculture. For in vivo studies, rats were sacrificed at various times after treatment and the induction of DNA strand breaks was assessed in whole blood, bone marrow, thymus, liver, kidney cortex and in the distal part of the intestine. One hour after injection, etoposide induced DNA damage in all organs studied except kidney, especially in bone marrow, thymus (presence of HDC) and whole blood. As observed during in vitro comet assay on Chinese hamster ovary (CHO) cells, dose- and time-dependent DNA effects occurred in vivo with a complete disappearance of damage 24 h after administration. Even though apoptotic cells were detected in vitro 48 h after cell exposure to etoposide, such a result was not found in vivo. After chlorothalonil treatment, no DNA strand breaks were observed in rat organs whereas a clear dose-related DNA damage was observed in vitro. The discrepancy between in vivo and in vitro models could be explained by metabolic and mechanistic reasons. Our results show that the in vivo comet assay is able to detect the target organs of etoposide and suggest that chlorothalonil is devoid of appreciable in vivo genotoxic activity under the protocol used.
Assuntos
Dano ao DNA , Etoposídeo/toxicidade , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Nitrilas/toxicidade , Animais , Células Sanguíneas/efeitos dos fármacos , Células da Medula Óssea/efeitos dos fármacos , Células CHO , Cricetinae , Intestinos/efeitos dos fármacos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Masculino , Especificidade de Órgãos , Ratos , Ratos Sprague-Dawley , Timo/efeitos dos fármacos , Fatores de TempoRESUMO
We have compared serum galactosyltransferase activity in pregnant women and in newborn children. Subjects in this study were 75 women at different stages of pregnancy and 60 newborns, including premature infants. Activity ratios in pregnant women are based on the period of gestation, expressed in weeks of amenorrhea. Average values were 265 nmol/h/ml for up to 12 weeks of amenorrhea, 369 from 13 to 28 weeks and 483 over 28 weeks versus 232 nmol/h/ml for non-pregnant women. In infants, galactosyltransferase activity decreased with increasing age from conception, but the activity level was always much higher in newborns than in women at the ninth month of pregnancy. We discuss the origin of the enzyme in these various samples.
Assuntos
Galactosiltransferases/sangue , Feminino , Humanos , Recém-Nascido , GravidezRESUMO
We have compared three assays for serum galactosyltransferase activity, which use ovomucoïd, asialo agalactofetuin and free N-acetylglucosamine respectively as exogenous acceptors. A very good correlation between the three assays is obtained, for the whole range of GT activity. When the methods are compared to one another, the slopes of the regression lines are similar whether the sera are collected from healthy controls, pregnant women, or women suffering from benign gynecologic diseases or ovarian neoplasms. The methods which uses free N-acetylglucosamine as acceptor is rapid, cheaper and easier to perform.
Assuntos
Galactosiltransferases/sangue , Feminino , Doenças dos Genitais Femininos/sangue , Humanos , Métodos , Neoplasias Ovarianas/sangue , GravidezRESUMO
Airborne particulate matter has recently been classified by the IARC as carcinogenic to humans (group 1). However, the link between PM chemical composition and its carcinogenicity is still unclear. The aim of the present study was to evaluate and to compare genotoxic potencies of 6 native PM samples collected in spring-summer or autumn-winter, either in industrial, urban or rural area. We evaluated their mutagenicity through Ames test on YG1041, TA98, and TA102 tester strains, and their clastogenicity on human bronchial epithelial BEAS-2B cells using comet assay, γ-H2AX quantification, and micronucleus assay. Ames test results showed a strong positive response, presumably associated with nitro-aromatics content. In addition, at least 2 positive responses were observed out of the 3 genotoxicity assays for each of the 6 samples, demonstrating their clastogenicity. Our data suggest that PM samples collected in autumn-winter season are more genotoxic than those collected in spring-summer, potentially because of higher concentrations of adsorbed organic compounds. Taken together, our results showed the mutagenicity and clastogenicity of native PM2.5 samples from different origins, and bring additional elements to explain the newly recognized carcinogenicity of outdoor air pollution.