DNA damage and antioxidant capacity in COPD patients with and without lung cancer.

BACKGROUND AND OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is characterized by chronic inflammation of the lower airways, and COPD patients show two to five times higher risk of lung cancer than smokers with normal lung function. COPD is associated with increased oxidative stress, which may cause DNA damage and lung carcinogenesis. Our aim was to evaluate DNA damage and oxidative stress (lipid peroxidation and antioxidant status) and their relationship in patients with COPD with and without lung cancer. METHODS: We evaluated 18 patients with COPD, 18 with COPD with lung cancer, and 18 controls (former or current smokers). DNA damage was evaluated in peripheral blood lymphocytes using a comet assay; the concentration of malondialdehyde (MDA) and hydrophilic antioxidant performance (HAP) were measured in the plasma. RESULTS: DNA damage was higher in patients with COPD with cancer than in the controls (p = 0.003). HAP was significantly lower in patients with COPD with cancer than in those without cancer and controls. The presence of lung cancer and COPD showed a positive association with DNA strand breaks and the concentration of MDA. CONCLUSION: COPD with lung cancer was associated with elevated DNA damage in peripheral lymphocytes, and cancer and COPD showed a positive correlation with DNA damage. The antioxidant capacity showed a negative association with the interaction COPD and cancer and presence of COPD. The mechanisms underlying the increased incidence of lung cancer in COPD are unknown; DNA damage may be involved. Further research may provide insights into their development and treatment.

Oxidative stress, DNA damage, inflammation and gene expression in occupationally exposed university hospital anesthesia providers.

Considering the importance and lack of data of toxicogenomic approaches on occupational exposure to anesthetics, we evaluated possible associations between waste anesthetic gases (WAGs) exposure and biological effects including oxidative stress, DNA damage, inflammation, and transcriptional modulation. The exposed group was constituted by anesthesia providers who were mainly exposed to the anesthetics sevoflurane and isoflurane (10 ppm) and to a lesser degree to nitrous oxide (150 ppm), and the control group was constituted by physicians who had no exposure to WAGs. The oxidative stress markers included oxidized DNA bases (comet assay), malondialdehyde (high-performance liquid chromatography [HPLC]), nitric oxide metabolites (ozone-chemiluminescence), and antioxidative markers, including individual antioxidants (HPLC) and antioxidant defense marker (ferric reducing antioxidant power by spectrophotometry). The inflammatory markers included high-sensitivity C-reactive protein (chemiluminescent immunoassay) and the proinflammatory interleukins IL-6, IL-8 and IL-17A (flow cytometry). Telomere length and gene expression related to DNA repair (hOGG1 and XRCC1), antioxidant defense (NRF2) and inflammation (IL6, IL8 and IL17A) were evaluated by real-time quantitative polymerase chain reaction. No significant differences (p > .0025) between the groups were observed for any parameter evaluated. Thus, under the conditions of the study, the findings suggest that occupational exposure to WAGs is not associated with oxidative stress or inflammation when evaluated in serum/plasma, with DNA damage evaluated in lymphocytes and leucocytes or with molecular modulation assessed in peripheral blood cells in university anesthesia providers. However, it is prudent to reduce WAGs exposure and to increase biomonitoring of all occupationally exposed professionals.

[Epidemiology of perioperative cardiac arrest and mortality in Brazil: a systematic review]. / Epidemiologia de parada cardíaca e de mortalidade perioperatória no Brasil: revisão sistemática.

BACKGROUND AND OBJECTIVES: The perioperative cardiac arrest (CA) and mortality rates in Brazil, a developing country, are higher than in developed countries. The hypothesis of this review was that knowledge of the epidemiology of perioperative CA and mortality in Brazil enables the comparison with developed countries. The systematic review aimed to verify, in studies conducted in Brazil, the epidemiology of perioperative CA and mortality. METHOD AND RESULTS: A search strategy was carried out on different databases (PubMed, EMBASE, SciELO and LILACS) to identify observational studies that reported perioperative CA and/or mortality up to 48 hours postoperatively in Brazil. The primary outcomes were data on epidemiology of perioperative CA and mortality. In 8 Brazilian studies, there was a higher occurrence of perioperative CA and mortality in males; in extremes of age; in patients in worse physical status according to the American Society of Anesthesiologists (ASA); in emergency surgeries; in general anesthesia; and in cardiac, thoracic, vascular, abdominal and neurological surgeries. The patient's disease/condition was the main triggering factor, with sepsis and trauma as the main causes. CONCLUSIONS: The epidemiology of both perioperative CA and mortality events reported in Brazilian studies does not show important differences and, in general, is similar to studies in developed countries. However, sepsis represents one of the major causes of perioperative CA and mortality in Brazilian studies, contrasting with studies in developed countries in which sepsis is a secondary cause.

Epidemiology of perioperative cardiac arrest and mortality in Brazil: a systematic review / Epidemiologia de parada cardíaca e de mortalidade perioperatória no Brasil: revisão sistemática

Abstract Background and objectives: The perioperative cardiac arrest (CA) and mortality rates in Brazil, a developing country, are higher than in developed countries. The hypothesis of this review was that knowledge of the epidemiology of perioperative CA and mortality in Brazil enables the comparison with developed countries. The systematic review aimed to verify, in studies conducted in Brazil, the epidemiology of perioperative CA and mortality. Method and results: A search strategy was carried out on different databases (PubMed, EMBASE, SciELO and LILACS) to identify observational studies that reported perioperative CA and/or mortality up to 48 hours postoperatively in Brazil. The primary outcomes were data on epidemiology of perioperative CA and mortality. In 8 Brazilian studies, there was a higher occurrence of perioperative CA and mortality in males; in extremes of age; in patients in worse physical status according to the American Society of Anesthesiologists (ASA); in emergency surgeries; in general anesthesia; and in cardiac, thoracic, vascular, abdominal and neurological surgeries. The patient's disease/condition was the main triggering factor, with sepsis and trauma as the main causes. Conclusions: The epidemiology of both perioperative CA and mortality events reported in Brazilian studies does not show important differences and, in general, is similar to studies in developed countries. However, sepsis represents one of the major causes of perioperative CA and mortality in Brazilian studies, contrasting with studies in developed countries in which sepsis is a secondary cause.

Resumo Justificativa e objetivos: As incidências de parada cardíaca (PC) e de mortalidade perioperatória no Brasil, um país em desenvolvimento, são mais elevadas em relação às dos países desenvolvidos. A hipótese desta revisão é que o conhecimento da epidemiologia de PC e de mortalidade perioperatória no Brasil possibilita sua comparação com a dos países desenvolvidos. A revisão sistemática teve como objetivo verificar, em estudos realizados no Brasil, a epidemiologia de PC e de mortalidade perioperatória. Conteúdo: Realizou-se estratégia de busca em diferentes bases de dados (PubMed, EMBASE, SciELO e LILACS) para a identificação de estudos observacionais que reportaram PC e/ou mortalidade perioperatória até 48 horas pós-operatório no Brasil. Os desfechos primários foram dados de epidemiologia de PC e de mortalidade perioperatória. Em 8 estudos nacionais, identificou- se maior ocorrência de PC e de mortalidade perioperatória no sexo masculino, em extremos de idade, em pacientes em pior estado físico segundo a American Society of Anesthesiologists (ASA), em cirurgias de emergência, em anestesia geral, e em cirurgias cardíaca, torácica, vascular, abdominal e neurológica. A doença/condição do paciente foi o principal fator desencadeante, tendo como causas principais a sepse e o trauma. Conclusões: Nos estudos nacionais, a epidemiologia dos eventos tanto de PC como de mortalidade perioperatória não apresenta diferenças importantes, e de maneira geral, é semelhante à de estudos de países desenvolvidos. Entretanto, a sepse, nos estudos nacionais, representa uma das principais causas de PC e de mortalidade perioperatória, diferenciando-se dos estudos de países desenvolvidos nos quais a sepse é causa secundária.

Lipid damage is the best marker of oxidative injury during the cardiac remodeling process induced by tobacco smoke.

BACKGROUND: Oxidative stress is one potential mechanism that explain the direct effects of smoking on cardiac remodeling process. However, no study has compared different myocardial products of macromolecule oxidation after tobacco smoke exposure. Thus, the aim of this study was to investigate the lipid hydroperoxide (LH) levels, protein carbonyl concentrations and DNA damage in cardiac tissue of rats exposed to tobacco smoke. METHODS: Male Wistar rats were divided into two groups: group C (control, n = 14) composed of animals not exposed to cigarette smoke; group ETS (exposed to tobacco smoke, n = 14) composed by animals exposed to cigarette smoke. The animals were exposed to 2 month of ETS and morphological, biochemical and functional analyses were performed. RESULTS: Cardiac cotinine levels were elevated in the ETS group. In addition, the myocyte cross-sectional area was higher in the ETS group. (C = 266.6 ± 23.2 µm2 and ETS = 347.5 ± 15.1 µm2, p <  0.001). Cardiac LH was higher in the ETS group than in group C (C = 196.4 ± 51.5 nmol/g and ETS = 331.9 ± 52.9 nmol/g, p <  0.001). However, there were no between-group differences in cardiac protein carbonyl concentration or DNA damage. CONCLUSIONS: Therefore, our results suggest that, in this model, lipid damage is a good marker of oxidative damage during the cardiac remodeling process induced by 2 months of exposure to tobacco smoke.

Spondias mombin supplementation attenuated cardiac remodelling process induced by tobacco smoke.

The objective of this study was to investigate the influence of Spondias mombin (SM) supplementation on the cardiac remodelling process induced by exposure to tobacco smoke (ETS) in rats. Male Wistar rats were divided into 4 groups: group C (control, n = 20) comprised animals not exposed to cigarette smoke and received standard chow; group ETS (n = 20) comprised animals exposed to cigarette smoke and received standard chow; group ETS100 (n = 20) received standard chow supplemented with 100 mg/kg body weight/d of SM; and group ETS250 (n = 20) received standard chow supplemented with 250 mg/kg body weight/d of SM. The observation period was 2 months. The ETS animals had higher values of left cardiac chamber diameters and of left ventricular mass index. SM supplementation attenuated these changes. In addition, the myocyte cross-sectional area (CSA) was lower in group C compared with the ETS groups; however, the ETS250 group had lower values of CSA compared with the ETS group. The ETS group also showed higher cardiac levels of lipid hydroperoxide (LH) compared with group C; and, groups ETS100 and ETS250 had lower concentrations of LH compared with the ETS group. Regarding energy metabolism, SM supplementation decreased glycolysis and increased the ß-oxidation and the oxidative phosphorylation. There were no differences in the expression of Nrf-2, SIRT-1, NF-κB, interferon-gamma and interleukin 10. In conclusion, our results suggest that ETS induced the cardiac remodelling process. In addition, SM supplementation attenuated this process, along with oxidative stress reduction and energy metabolism modulation.

Comparison of waste anesthetic gases in operating rooms with or without an scavenging system in a Brazilian University Hospital / Comparação de resíduos de gases anestésicos em salas de operação com ou sem sistema de exaustão em hospital universitário brasileiro

Abstract Background and objectives Occupational exposure to waste anesthetic gases in operating room without active scavenging system has been associated with adverse health effects. Thus, this study aimed to compare the trace concentrations of the inhalational anesthetics isoflurane and sevoflurane in operating room with and without central scavenging system. Method Waste concentrations of isoflurane and sevoflurane were measured by infrared analyzer at different locations (near the respiratory area of the assistant nurse and anesthesiologist and near the anesthesia station) and at two times (30 and 120 min after the start of surgery) in both operating room types. Results All isoflurane and sevoflurane concentrations in unscavenged operating room were higher than the US recommended limit (2 parts per million), regardless of the location and time evaluated. In scavenged operating room, the average concentrations of isoflurane were within the limit of exposure, except for the measurements near the anesthesia station, regardless of the measurement times. For sevoflurane, concentrations exceeded the limit value at all measurement locations and at both times. Conclusions The exposure to both anesthetics exceeded the international limit in unscavenged operating room. In scavenged operating room, the concentrations of sevoflurane, and to a lesser extent those of isoflurane, exceeded the recommended limit value. Thus, the operating room scavenging system analyzed in the present study decreased the anesthetic concentrations, although not to the internationally recommended values.

Resumo Justificativa e objetivos A exposição ocupacional aos resíduos de gases anestésicos em salas de operação (SO) sem sistema ativo de exaustão tem sido associada a efeitos adversos à saúde. Assim, o objetivo do estudo foi comparar os resíduos dos anestésicos inalatórios isoflurano e sevoflurano em SO com e sem sistema de exaustão. Método Concentrações residuais de isoflurano e sevoflurano foram mensuradas por analisador infravermelho em diferentes locais (próximo à área respiratória do auxiliar de enfermagem e do anestesiologista e próximo à estação de anestesia) e em dois momentos (30 e 120 min após o início da cirurgia) em ambos os tipos de SO. Resultados Todas as concentrações de isoflurano e sevoflurano nas SO sem sistema de exaustão foram mais elevadas em relação ao valor limite recomendado pelos EUA (2 partes por milhão), independentemente do local e momento avaliados. Nas SO com sistema de exaustão, as concentrações médias de isoflurano ficaram dentro do limite de exposição, exceto para as mensurações próximas à estação de anestesia, independentemente dos momentos avaliados. Para o sevoflurano, as concentrações excederam o valor limite em todos locais de medição e nos dois momentos. Conclusões A exposição a ambos os anestésicos excedeu o limite internacional nas SO sem sistema de exaustão. Nas SO com sistema de exaustão, as concentrações de sevoflurano, e em menor extensão, as de isoflurano excederam o valor limite recomendado. Dessa forma, o sistema de exaustão das SO analisado no presente estudo diminuiu as concentrações dos anestésicos, embora não tenha reduzido a valores internacionalmente recomendados.

Yacon (Smallanthus sonchifolius) Leaf Extract Attenuates Hyperglycemia and Skeletal Muscle Oxidative Stress and Inflammation in Diabetic Rats.

The effects of hydroethanolic extract of Yacon leaves (HEYL) on antioxidant, glycemic, and inflammatory biomarkers were tested in diabetic rats. Outcome parameters included glucose, insulin, interleukin-6 (IL-6), and hydrophilic antioxidant capacity (HAC) in serum and IL-6, HAC, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in soleus. The rats (10/group) were divided as follows: C, controls; C + Y, HEYL treated; DM, diabetic controls; and DM + Y, diabetic rats treated with HEYL. Diabetes mellitus was induced by administration of streptozotocin. C + Y and DM + Y groups received 100 mg/kg HEYL daily via gavage for 30 d. Hyperglycemia was improved in the DM + Y versus DM group. Insulin was reduced in DM versus C group. DM rats had higher IL-6 and MDA and lower HAC in the soleus muscle. HEYL treatment decreased IL-6 and MDA and increased HAC in DM rats. DM + Y rats had the highest CAT activity versus the other groups; GPx was higher in C + Y and DM + Y versus their respective controls. The apparent benefit of HEYL may be mediated via improving glucoregulation and ameliorating oxidative stress and inflammation, particularly in diabetic rats.

[Comparison of waste anesthetic gases in operating rooms with or without an scavenging system in a Brazilian University Hospital]. / Comparação de resíduos de gases anestésicos em salas de operação com ou sem sistema de exaustão em hospital universitário brasileiro.

BACKGROUND AND OBJECTIVES: Occupational exposure to waste anesthetic gases in operating room (OR) without active scavenging system has been associated with adverse health effects. Thus, this study aimed to compare the trace concentrations of the inhaled anesthetics isoflurane and sevoflurane in OR with and without central scavenging system. METHOD: Waste concentrations of isoflurane and sevoflurane were measured by infrared analyzer at different locations (near the respiratory area of the assistant nurse and anesthesiologist and near the anesthesia station) and at two times (30 and 120minutes after the start of surgery) in both OR types. RESULTS: All isoflurane and sevoflurane concentrations in unscavenged OR were higher than the US recommended limit (2 parts per million), regardless of the location and time evaluated. In scavenged OR, the average concentrations of isoflurane were within the limit of exposure, except for the measurements near the anesthesia station, regardless of the measurement times. For sevoflurane, concentrations exceeded the limit value at all measurement locations and at both times. CONCLUSIONS: The exposure to both anesthetics exceeded the international limit in unscavenged OR. In scavenged OR, the concentrations of sevoflurane, and to a lesser extent those of isoflurane, exceeded the recommended limit value. Thus, the OR scavenging system analyzed in the present study decreased the anesthetic concentrations, although not to the internationally recommended values.

Does occupational exposure to anesthetic gases lead to increase of pro-inflammatory cytokines?

INTRODUCTION: There is great concern about the possible harmful effects of exposure to volatile anesthetics. The current study aimed at evaluating, for the first time, the effects of occupational exposure to anesthetic gases on physicians who work in operating rooms, by determining several inflammatory cytokines. MATERIALS AND METHODS: Plasma inflammatory cytokines (IL-1ß, -6, -8, -10, -12, TNF-α) were investigated in 30 individuals who were allocated into two groups of 15: the exposed group, consisting of operating room medical personnel exposed to a mixture of anesthetic gases for 3 years, and a control group composed of medical personnel not exposed to anesthetic gases. The concentrations of volatile anesthetics were measured in the operating room by means of an infrared portable analyzer RESULTS AND CONCLUSIONS: Our findings suggest an increase of the pro-inflammatory IL-8 (p<0.05) in medical personnel exposed to high concentrations of anesthetic gases, even for a relatively short period.

DNA damage and antioxidant status in medical residents occupationally exposed to waste anesthetic gases.

PURPOSE: To investigate the effects of occupational exposure to waste anesthetic gases on genetic material and antioxidant status in professionals during their medical residency. METHODS: The study group consisted of 15 medical residents from Anesthesiology and Surgery areas, of both genders, mainly exposed to isoflurane and to a lesser degree to sevoflurane and nitrous oxide; the control group consisted of 15 young adults not exposed to anesthetics. Blood samples were drawn from professionals during medical residency (eight, 16 and 22 months of exposure to waste anesthetic gases). DNA damage was evaluated by comet assay, and antioxidant defense was assessed by total thiols and the enzymes glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT). RESULTS: When comparing the two groups, DNA damage was significantly increased at all time points evaluated in the exposed group; plasma thiols increased at 22 months of exposure and GPX was higher at 16 and 22 months of exposure. CONCLUSION: Young professionals exposed to waste anesthetic gases in operating rooms without adequate scavenging system have increased DNA damage and changes in redox status during medical residency. There is a need to minimize exposure to inhalation anesthetics and to provide better work conditions.

DNA damage and antioxidant status in medical residents occupationally exposed to waste anesthetic gases

To investigate the effects of occupational exposure to waste anesthetic gases on genetic material and antioxidant status in professionals during their medical residency. The study group consisted of 15 medical residents from Anesthesiology and Surgery areas, of both genders, mainly exposed to isoflurane and to a lesser degree to sevoflurane and nitrous oxide; the control group consisted of 15 young adults not exposed to anesthetics. Blood samples were drawn from professionals during medical residency (eight, 16 and 22 months of exposure to waste anesthetic gases). DNA damage was evaluated by comet assay, and antioxidant defense was assessed by total thiols and the enzymes glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT). When comparing the two groups, DNA damage was significantly increased at all time points evaluated in the exposed group; plasma thiols increased at 22 months of exposure and GPX was higher at 16 and 22 months of exposure. Young professionals exposed to waste anesthetic gases in operating rooms without adequate scavenging system have increased DNA damage and changes in redox status during medical residency. There is a need to minimize exposure to inhalation anesthetics and to provide better work conditions.

Comparison of inflammatory cytokine profiles in plasma of patients undergoing otorhinological surgery with propofol or isoflurane anesthesia.

OBJECTIVE AND DESIGN: The effects of anesthetics on cytokine release in patients without comorbidities who undergo minor surgery are not well defined. We compared inflammatory cytokine profiles in adult patients undergoing minimally invasive surgery who received isoflurane or propofol anesthesia. METHODS: Thirty-four patients without comorbidities undergoing minor surgery were randomly assigned to receive an inhaled anesthetic (isoflurane; n = 16) or an intravenous anesthetic (propofol; n = 18). Blood samples were drawn before premedication and anesthesia (T1), 120 min after anesthesia induction (T2), and on the first post-operative day (T3). Plasma concentrations of interleukins (IL-) 1ß, 6, 8, 10 and 12 and tumor necrosis factor (TNF)-α were measured using flow cytometry. RESULTS: The pro-inflammatory cytokine IL-6 was increased in the isoflurane group at T2 and T3 compared to T1 (P < 0.01). In the propofol group, IL-6 and IL-8 were significantly increased at T3 compared to T1. However, there were no significant differences in cytokine concentrations between the isoflurane and propofol groups. CONCLUSION: An inflammatory response occurred earlier in patients who received an inhaled agent compared with an intravenous anesthetic, but no differences in plasma cytokine profiles were evident between isoflurane and propofol anesthesia in patients without comorbidities undergoing minimally invasive surgeries.

Genotoxicity, cytotoxicity and gene expression in patients undergoing elective surgery under isoflurane anaesthesia.

There are numerous studies reporting on the effects of inhalation anaesthesia in cells of exposed individuals but not much is known about the ability of isoflurane (ISF) to induce oxidative DNA damage. However, surgery is often associated with a temporary perioperative immunological alteration, and some volatile anaesthetics seem to contribute to a transient lymphocytopenia after surgery. We conducted a study to evaluate a possible genotoxic effect, including oxidative DNA damage, and apoptosis in peripheral lymphocytes of 20 patients American Society of Anaesthesiologists physical status I undergoing minor elective surgery lasting at least 120 min, under anaesthesia with ISF. We also investigated the expression of several genes in blood cells. Blood samples were collected at three time points: before anaesthesia (T(1)), 2 h after the beginning of anaesthesia (T(2)) and on the first post-operative day (T(3)). General DNA damage and oxidised bases (Fpg and endo III-sites) in blood lymphocytes were evaluated using the comet assay. Lymphocytes were phenotyped and apoptosis was evaluated by flow cytometry. In addition, expressions of hOGG1 and XRCC1, genes involved in DNA repair, and BCL2, a gene related to apoptosis, were assessed by quantitative real-time polymerase chain reaction. Results showed no statistically significant difference in the level of DNA damage and oxidised bases among the three sampling times. Anaesthesia with ISF did not increase the percentage of cells in early or late apoptosis in cytotoxic or helper T lymphocytes. Lower hOGG1 and BCL2 expressions were detected at T(3) in comparison to the other two previous time points, and there was significantly lower expression of XRCC1 at T(3) in relation to T(2). In conclusion, the exposure to ISF did not result in genotoxicity and cytotoxicity in lymphocytes and in toxicogenomic effect in leukocytes, although DNA repair and apoptosis-related genes were down-regulated on the first post-operative day.

Evaluation of DNA damage and lipoperoxidation of propofol in patients undergoing elective surgery.

BACKGROUND AND OBJECTIVES: Inhaled anaesthetics have been studied regarding their genotoxic and mutagenic potential in vivo. Propofol differs from volatile anaesthetics because it does not show mutagenic effects and it has been reported to be an antioxidant. However, there are no studies with propofol and genotoxicity in vivo. The study aimed to evaluate the hypothesis that propofol is not genotoxic and it inhibits lipid peroxidation [malondialdehyde (MDA)] in patients undergoing propofol anaesthesia. METHODS: ASA physical status I patients scheduled for elective surgery, lasting at least 90 min, were enrolled in this study. Initially, the estimated plasma concentration of propofol was targeted at 4 microg ml(-1) and then maintained at 2-4 microg ml(-1) until the end of surgery. Haemodynamic data were determined at baseline (before premedication) and in conjunction with target-controlled infusion of propofol: after tracheal intubation, 30, 60 and 90 min after anaesthesia induction and at the end of the surgery. Venous blood samples were collected at baseline, after tracheal intubation, at the end of the surgery and on the postoperative first day for evaluating DNA damage in white blood cells (WBCs), by comet assay, and MDA levels. RESULTS: Haemodynamic data did not differ among times. No statistically significant differences were observed for the levels of DNA damage in WBCs, nor in plasma MDA, among the four times. CONCLUSION: Propofol does not induce DNA damage in WBCs and does not alter MDA in plasma of patients.

Genotoxicity in primary human peripheral lymphocytes after exposure to radiopacifiers in vitro.

Taking into consideration that DNA damage plays an important role in carcinogenesis, the purpose of this study was to evaluate whether some radiopacifiers widely used in clinical practice are able to induce genetic damage in primary human cells in vitro. Human peripheral lymphocytes obtained from 10 healthy volunteers were exposed to barium sulphate (BaSO(4)), zirconium oxide (ZnO(2)) and bismuth oxide (Bi(2)O(3)) at final concentrations ranging from 1 to 1000 microg/mL for 1 h at 37 degrees C. The negative control group was treated with vehicle control (phosphate buffer solution) for 1 h at 37 degrees C and the positive control group was treated with hydrogen peroxide (at 100 microM) for 5 min on ice. Results were analyzed by the Friedman non-parametric test. The results pointed all compounds tested out did not induce DNA breakage in human peripheral lymphocytes as depicted by the mean tail moment and tail intensity in all concentrations tested. In summary, our results indicate that exposure to these radiopacifiers may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single cell gel (comet) assay.

Influence of endogenous and synthetic female sex hormones on human blood cells in vitro studied with comet assay.

The comet assay has been conducted with numerous cell lines to assess in vitro genotoxicity. In order to use the comet assay as part of an in vitro test for evaluating genotoxicity, however, there are cell-specific factors that need to be better understood. In this present study we have evaluated some factors that may impact upon the DNA damage detected in whole blood (WB) cells and lymphocytes (ILs). Experiments were conducted comparing responses of both cells, and investigating the effects of the female hormonal cycle, and oral contraceptive (OC) use on DNA damage detection in the in vitro comet assay, at three sampling time. No significant differences were detected in the basal levels of DNA damage detected in ILs and WB cells from women OC users and non-users and from men. Basal DNA damage in ILs was unaffected by gender and stage of the menstrual cycle or the stage of the treatment schedule. Our results also indicated that the H2O2 induces DNA damage in human lymphocytes independently of gender, low-dose OC use and hormonal fluctuation. However, data showed that in 3rd sampling of menstrual cycle, lymphocytes were more resistant to H2O2-induced DNA damage than those from OC users and men.

Lack of genotoxicity induced by endogenous and synthetic female sex hormones in peripheral blood cells detected by alkaline comet assay.

The etiology of hormone-induced cancers has been considered to be a combination of genotoxic and epigenetic events. Currently, the Comet assay is widely used for detecting genotoxicity because it is relatively simple, sensitive, and capable of detecting various kinds of DNA damage. The present study evaluates the genotoxic potential of endogenous and synthetic sex hormones, as detected by the Comet assay. Blood cells were obtained from 12 nonsmoking and 12 smoking women with regular menstrual cycles and from 12 nonsmoking women taking low-dose oral contraceptives (OC). Peripheral blood samples were collected at three phases of the menstrual cycle (early follicular, mean follicular, and luteal phases), or at three different moments of oral contraceptive intake. Three blood samples were also collected from 12 healthy nonsmoking men, at the same time as oral contraceptive users. Results showed no significant difference in the level of DNA damage among the three moments of the menstrual cycle either in nonsmoking and smoking women, or between them. No significant difference in DNA damage was also observed among oral contraceptive users, nonusers, and men. Together, these data indicate lack of genotoxicity induced by the physiological level of the female sex hormones and OC as assessed by the alkaline Comet assay. In conclusion, normal fluctuation in endogenous sex hormones and use of low-doses of oral contraceptive should not interfere with Comet assay data when this technique is used for human biomonitoring.