Lung permeability, antioxidant status, and NO2 inhalation: a selenium supplementation study in rats.

Little is known about antioxidant status, selenium status in particular, and lung response to NO2, which acts as a proinflammatory air pollutant. The effects of a low selenium diet (1.3 microg Se/d) with or without selenium supplementation were therefore studied in 128 Wistar rats, 2 mo old, male exposed to either acute (50 ppm, 30 min), intermittent subacute (5 ppm, 6 h/d, 5 d), intermittent long-term NO2 (1 ppm, 10 ppm, 6 h/d, 5 d/wk, 28 d), or normal atmospheric air (controls). Following sacrifice, measurements of lipid peroxidation (thiobarbituric acid-reactive substances, chemiluminescence), antioxidative protective enzymes (glutathione peroxidase [GPx], superoxide dismutase [SOD], glutathione S-transferase [GST], ceruloplasmin), lung damage (lactate dehydrogenase, alkaline and acid phosphatases), lung permeability (total protein, albumin), and inflammation (cell populations), along with the determination of new biomarkers such as CC16 (Clara-cell protein), were performed in serum and bronchoalveolar lavage fluid (BALF). While selenium-supplemented animals had increased GPx activity in serum prior to inhalation experiments, they also had decreased BALF CC16, blood SOD, and GST levels. Nevertheless, the protective role of normal selenium status with respect to NO2 lung toxicity was evident both for long-term and acute exposures, as the increase in BALF total proteins and corresponding decrease in serum (indicating increased lung permeability) was significantly more pronounced in selenium-deficient animals. During the various inhalation experiments, serum CC16 demonstrated its key role as an early marker of increased lung permeability. These findings corroborate the important role of selenium status in NO2 oxidative damage modulation, but also indicate, in view of its negative impact on CC16, a natural anti-inflammatory and immunosuppressor, that caution should be used prior to advocating selenium supplementation.

Micronucleus frequency in peripheral blood lymphocytes and buccal mucosa cells of copper smelter workers, with special regard to arsenic exposure.

Occupational exposure in copper smelters may produce various adverse health effects including cancer which, according to available epidemiologic data, is associated mainly with exposure to arsenic. Despite a number of well-documented studies reporting an increased risk of cancer among copper smelters workers, the data on genotoxic effects in this industry are scarce. In view of the above, an assessment of micronuclei (MN) frequency in peripheral blood lymphocytes and buccal epithelial cells from copper smelter workers was undertaken. Additionally, the clastogenic/aneugenic effect in lymphocytes was assessed with the fluorescence in situ hybridization (FISH). The study was conducted in three copper smelters in southwestern Poland. The subjects (n = 72) were enrolled among male workers at departments where As concentration in the air was up to at 80 microg/m(3). Exposure was assessed by measurement of arsenic concentration in urine and toenail samples. The control group (n = 83) was recruited from healthy male individuals living in central Poland who did not report any exposure to known genotoxins. The results of our study showed a significant increase in MN frequency in peripheral blood lymphocytes and in buccal epithelial cells of smelter workers, compared to the controls (7.96 +/- 4.28 vs. 3.47 +/- 1.70 and 0.98 +/- 0.76 vs. 0.50 +/- 0.52, respectively). The FISH technique revealed the presence of clastogenic and aneugenic effects in peripheral blood lymphocytes in both groups. The clastogenic effect was slightly more pronounced in the smelter workers; however, the difference was not statistically significant. The mean arsenic concentrations in urine (total arsenic species) and in toenail samples in the exposed group were 54.04 +/- 42.26 microg/l and 7.63 +/- 7.24 microg/g, respectively, being significantly different from control group 11.01 +/- 10.84 microg/l and 0.51 +/- 0.05 microg/g. No correlation between As content in urine or toenail samples and the genotoxic effect was found under study.

Pulmonary response to methylcyclopentadienyl manganese tricarbonyl treatment in rats: injury and repair evaluation.

Methylcyclopentadienyl manganese tricarbonyl (MMT), an organometallic compound, used as an antiknock additive in fuels, may produce alveolar inflammation and bronchiolar cell injury. The aim of the experimental study on female rats was to determine by morphological examination and sensitive biomarkers, the course of the injury and repair process following a single i.p. injection of 5 mg/kg MMT. The animals were sacrificed 12, 24, 48 hours or 7 days post-exposure (PE). The first biochemical changes 12 h PE showed an increase in GSH-S-transferase (GST) activity in the lung parallel to the earliest observed morphological changes -vacuolation and swollen cytoplasm in type I pneumocytes. Alterations in type I pneumocytes were most prevalent in rat lung 24 h PE. Clara cells with dilated smooth endoplasmic reticulum membranes and cytoplasmic vacuolation could be observed. Compared to the values found for controls, Clara cell protein (CC16) in the bronchoalveolar lavage fluid (BALF) at 24 and 48 h PE decreased by 58% and 55%, respectively. At the same time (at 24 and 48 h), the total protein concentration in BALF increased 5 and 7 times, respectively. A significant rise in hyaluronic acid (HA) level was observed 24 and 48 h PE. Divided type II pneumocyte cells and Clara cells in their mitotic phase were observed in immunocytochemistry (detecting BrdU binding into DNA) 48 h PE. Seven days after MMT administration, fibroblasts, macrophages, collagen and elastin fibres could be seen in the alveolar walls as well as neutrophils, lymphocytes, and alveoli macrophages in the alveolar lumen. We conclude that injury and repair of bronchial epithelium cells, especially of Clara cells and type II pneumocyte cells, play an important part in MMT toxicity, probably depending on the antioxidant status of these cells. The sensitive biomarkers of CC16 and hyaluronic acid in BALF and serum reflect lung injury and indicate the time course of pulmonary damage and repair processes.

Assessment of usefulness of J774A.1 macrophages for the assay of IL-1beta promoter activity.

Existing data indicate that the increase of il-1beta gene expression can be a promising marker of Langerhans cells activation after exposure to contact sensitizers. In this study, we were interested in development of an alternative in vitro screening test detecting such sensitizers. Two IL-1beta reporter constructs containing the enhanced green fluorescent protein (GFP) gene and mouse IL-1beta promoter fragments of varying lengths (-500 bp and -4093 bp) were used for transient transfections of J771A.1 murine monocyte-macrophage cells. As a result of the transfections performed using Lipofectamine reagent we did not observe any GFP fluorescence after stimulation of the cells with LPS as well as known sensitizers (potassium tetrachloroplatinate, dinitrochlorobenzene and nickel sulfate). Low transfection efficiency of J774A.1 cells (less than 0.1%) was confirmed using control plasmid containing GFP gene under the control of cytomegalovirus promoter. The fact that, using the same conditions, we were able to transfect murine fibroblasts 3T3-L1 with the control plasmid very efficiently, may support the theory of high metabolic activity of macrophages being responsible for the extremely low transfection efficiency. These data suggest limited suitability of J774A.1 cell line for transient transfections using cationic liposomes.

Prognostic significance of low serum levels of Clara cell phospholipid-binding protein in occupational aluminium neurotoxicity.

The relationship between respiratory and neurological effects of exposure to aluminium (Al) was investigated in a group of foundry workers exposed to Al at concentrations below the threshold limit value (TLV) binding in Poland (2.0 mg Al2O3 m(-3)). Neurological and neurophysiological parameters indicated subclinical effects of Al exposure on the nervous system. The measurement of serum anti-inflammatory Clara cell protein (CC16) was employed as a peripheral marker of the lung epithelium function. There was a strong inverse relationship between serum Al (Al-S) and CC16 concentrations (p = 0.006). The lowest CC16 concentrations were found in serum of workers characterised by subjective symptoms of the central nervous system (CNS) and abnormal results of neurophysiological examinations (EEG and VEP). Low serum CC16 concentrations and enhanced Al and iron (Fe) levels were also observed in the younger age group of workers with the subjective CNS symptoms and abnormal VEP results, which suggests that Fe is implicated in strengthening of the neurotoxic Al potential. The results of our study support the hypothesis that subclinical neurological symptoms (especially abnormal VEP) are most likely associated with internalisation of Al ions with lipid fractions of the lung epithelium, which in turn may help Al ions overcome the blood-brain barrier. Low serum CC16 concentrations (<10 microg L(-1)) were noted in workers with the abnormal results of neurological (CNS) and neurophysiological (EEG and VEP) examinations as well as with Al body burden manifested by urinary excretion (Al-U) below 60 microg L(-1) and Al-S concentration of 2 microg L(-1). This concentration may be considered as a threshold allowable biological concentration of aluminium.

Serum clara-cell protein and beta2-microglobulin as early markers of occupational exposure to nitric oxides.

Biochemical effects of NOx on 60 workers (both genders) of nitric acid production were studied. The control group consisted of 61 nonexposed people employed elsewhere in the plant. Although the actual threshold limit valuetime weighted averages (TLV-TWA) were not exceeded in the specific conditions of our study, the subjects were exposed to NO2 and NO during several exposure episodes with peak maximal concentrations of 140 ppm and 515 ppm, respectively. Additional cross-week evaluation of several biochemical biomarkers in 15 NOx-exposed workers from one shift was performed. The objective of the study was to evaluate the value of serum Clara-cell protein (CC16) as a marker of bronchoalveolar epithelium activity. Antioxidant status was assessed by measuring activity of enzymes: glutathione peroxidase (GSH-Px), ceruloplasmin (Cp) in plasma, or superoxide dismutase (SOD), gluthatione S-transferase (GST), and nonenzymatic alpha-tocopherol in erythrocytes and thiobarbituric acid-reactive substances (TBARS) in plasma. Serum hyaluronic acid (HA) determining the connective tissue matrix status of airways, and beta2-microglobulin in serum (beta2M-S) and urine (beta2M-U) as a marker of renal function in occupational exposure to NOx were also employed. Exposure to NOx initiates peroxidative chain depleting of lipoprotein pool (alpha-tocopherol) in blood. Serum CC16 levels in NOx-exposed workers were found to be closely connected with alpha-tocopherol content. In NOx-exposed workers, the beta2M-S level was significantly higher than in the nonexposed ones, with the exception of smokers. Results of the cross-week study confirm cumulative systemic effects of NOx on several examined biomarkers. SOD and GST were found to be depleted. A transient higher level of HA after a 5-d shift significantly inversely correlated with CC16 level. The data imply that NOx-depleted levels of CC16 are detectable already after an 8-h shift. Our results demonstrate that even low NOx human exposure can cause characteristic changes in bronchiolar epithelium cells and renal effects. Serum CC16 level, although a nonspecific marker, was lowest in NOx-exposed subjects. The most sensitive parameters in exposed workers were beta2M-S and a-tocopherol. Spirometric assessment was not useful to describe low occupational exposure to NOx. In studying the effects of NOx on biomarkers, it is essential to carefully select suitable time of sampling. Screening of CC16, beta2M-S, and a-tocopherol can be successfully employed for biological monitoring of exposure to NOx.

Glutathione S-transferase M1 and P1 metabolic polymorphism and lung cancer predisposition.

Individual susceptibility to different environmental agents is expected to be associated with alterations in metabolism of xenobiotics. Thus, genetic polymorphism of glutathione S-transferase (GST) can be recognized as a potential risk modifier in lung cancer development. The distribution of GSTM1 and GSTP1 genotypes was studied in a group of 138 diagnosed lung cancer patients and in 165 controls living in central Poland and RFLP-PCR technique was applied. The frequency of GSTM1 null genotype and GSTP1 Val single and duplicated alleles was similar among patients and controls. GSTM1 homozygous deletion was most prevalent in small-cell carcinoma groups (adjusted odds ratio (OR): 2.32, 95% confidence interval (CI): 0.98-5.52). In patients and controls, GSTM1A genotype was most frequent (34.1% vs. 37.0%). The estimated lung cancer risk for GSTM1 null, GSTP1 Ile/Val and GSTP1 Val/Val combined genotype was 1.44 (95% CI: 0.73-2.83), suggesting the absence of modifying effect of defective GSTM1 and GSTP1 alleles on lung cancer predisposition.

Genotoxic effects of occupational exposure to lead and cadmium.

This study was designed to assess genotoxic damage in somatic cells of workers in a Polish battery plant after high-level occupational exposure to lead (Pb) and cadmium (Cd), by use of the following techniques: the micronucleus (MN) assay, combined with in situ fluorescence hybridization (FISH) with pan-centromeric probes, analysis of sister chromatid exchanges (SCEs), and the comet assay. Blood samples from 44 workers exposed to lead, 22 exposed to cadmium, and 52 unexposed persons were used for SCE and MN analysis with 5'-bromodeoxyuridine (BrdU) or cytokinesis block, respectively. In parallel, the comet assay was performed with blood samples from the same persons for detection of DNA damage, including single-strand breaks (SSB) and alkali-labile sites (ALS). In workers exposed mostly to lead, blood Pb concentrations ranged from 282 to 655 microg/l, while the range in the controls was from 17 to 180 microg/l. Cd concentration in lead-exposed workers fell in the same range as for the controls. In workers exposed mainly to cadmium, blood Cd levels varied from 5.4 to 30.8 microg/l, with respective values for controls within the range of 0.2-5.7 microg/l. Pb concentrations were similar as for the controls. The incidence of MN in peripheral lymphocytes from workers exposed to Pb and Cd was over twice as high as in the controls (P<0.01). Using a combination of conventional scoring of MN and FISH with pan-centromeric probes, we assessed that this increase may have been due to clastogenic as well as aneugenic effects. In Cd- and Pb-exposed workers, the frequency of SCEs as well as the incidence of leukocytes with DNA fragmentation in lymphocytes were slightly, but significantly increased ( P<0.05) as compared with controls. After a 3h incubation of the cells to allow for DNA repair, a clear decrease was found in the level of DNA damage in the controls as well as in the exposed workers. No significant influence of smoking on genotoxic damage could be detected in metal-exposed cohorts. Our findings indicate that lead and cadmium induce clastogenic as well as aneugenic effects in peripheral lymphocytes, indicating a potential health risk for working populations with significant exposures to these heavy metals.

Immunohistochemical evaluation of P21ras and P53 proteins expression in human non-small-cell lung cancers.

Human non-small-cell lung cancers (NSCLCs) of 48 patients were analyzed immunohistochemically to detect P21 ras and P53 proteins expression. The relationship between P21 ras and P53 proteins expression and clinicopathologic findings was also assessed. DAKO EnVision TM detection system was employed in the study. The P21 ras and P53 proteins expression was shown in 75% (36/48) and 33.3% (16/48) studied NSCLCs, respectively. In both cases the difference was significant when compared with adequate negative control. Simultaneous expression of both studied proteins was observed in all cases in which P53 expression was noticed. No significant association of P21 ras and P53 expression was found with age, histologic type, histologic grade, tumor size or lymph node metastasis of the studied NSCLCs. Therefore, our study suggests that P21 ras and P53 protein play a role in the pathogenesis of NSCLCs but they have no value as a prognostic markers in the case of lung cancers.

Oxidative-stress markers in blood of lung cancer patients occupationally exposed to carcinogens.

The study covered 152 lung cancer patients and 210 controls. The results of the study indicated decreased selenium (Se) concentrations and lowered activity of erythrocyte antioxidant enzymes (glutathione peroxidase, superoxide dismutase, glutathione-S-transferase) in the blood of lung cancer patients, as well as significantly increased concentrations of vitamin E in erythrocytes and thiobarbituric acid reactive substances in the plasma of the study population. Low plasma Se concentrations (< 45.7 microg/L) enhance the estimated risk of lung cancer (odds ratio = 3.047, p < 0.001). A more precise exposure assessment is required to identify the association between lung cancer incidence and occupational exposure to carcinogens.

Selenium status of low-selenium area residents: Polish experience.

The concentration of selenium (Se) in human organism varies widely between geographical areas depending on its content in soil and plants, dietary Se intake, bioavailability and retention, mineral interactions and other factors. The study includes healthy inhabitants of different regions of Poland; pregnant women, lactating women, children from 0 to 15 years of age and adults. Systematic determinations allow us to observe changes of the concentration of Se in time, which may be significant for developing preventive action. The results obtained confirm our thesis that Se concentration in the blood of the inhabitants of Poland depends on the region of the country. In recent years, in a considerable number of Polish inhabitants, the concentration of Se in blood plasma has been relatively low-about 50-55 microg/l, and the calculated daily dietary intake about 30-40 microg/day. The low levels of the element in the blood and urine are probably due to its deficiency in the diet.

Glutaraldehyde inhalation exposure of rats: effects on lung morphology, Clara-cell protein, and hyaluronic acid levels in BAL.

Glutaraldehyde (GA) is a biocide widely used in hospital and laboratory practice. GA is a volatile substance and, under certain circumstances, significant airborne concentrations may be generated at room temperature. Occupational exposure to GA by inhalation is suspected of causing delayed irritating effects. In recent years, GA has emerged as the main cause of occupational asthma among health-care workers. The aim of the present study was to evaluate effects of GA inhalatory exposure (0.025 ppm or 0.1 ppm, for 28 days) in rats exposed corresponding to the occupational shift cycle, at time point 24 h, 48 h, and 7 days postexposure (PE). Numerous vacuoles and dilated spaces in epithelial cells in bronchioles showing a destructive effect of GA on the cellular membrane were observed at 24 h PE in 0.1 ppm exposed rats. Lipid vacuoles observed after 48 h PE in higher GA exposure, in the Clara cells of the bronchial epithelium, and in endothelial cells of the alveolar capillaries are probably attributable to disturbed lipid metabolism. Many foci of collagen fibers were observed already after 7 days postexposure. Monitoring of inflammatory response and repair was made possible by using two biomarkers: Clara-cell protein (CC16) and hyaluronic acid (HA). Our results show that the inflammatory repair response contributed to progenitor Clara cells and HA plays a role in the development of fibrotic changes in the lung of rats. Glutaraldehyde in rats causes fibrotic effects at the actual threshold limit value-time weighted average (TLV-TWA) level for GA as specified by current Polish and other national regulations.

Selenium, zinc, and copper concentrations in the blood and milk of lactating women.

The aim of the study was to determine Se, Zn, and Cu concentrations in blood plasma and milk of lactating women from central Poland who were in different stages of lactation and to investigate the relationship between the content of trace elements in mothers' blood and concentrations of microelements in their milk. Se and Zn concentrations in blood plasma of mothers were the lowest and Cu was the highest on the first 4 d of lactation (colostrum, n = 43) and were found to be 34.9 +/- 11.8 microg/L, 0.51 +/- 0.13 mg/L, and 1.70 +/- 0.55 mg/L, respectively. The highest plasma level of Se and Zn and the lowest content of Cu could be observed between d 10 and 30 of lactation (mature milk, n = 41), and were found to be 54.3 +/- 14.6 microg/L for Se (p < 0.001), 0.76 +/- 0.20 mg/L for Zn (p < 0.001), and 1.03 +/- 0.30 mg/L (p < 0.001) for Cu. The results of Se, Zn, and Cu determination in breast milk samples demonstrate a pattern of decline in their concentration with advancing stages of lactation. We found out that Se, Zn, and Cu concentrations were the highest in colostrum (n = 43) and amounted to 24.8 +/- 10.1 microg/L, 8.2 +/- 2.8 mg/L, and 0.45 +/- 0.11 mg/L, respectively. The content of all determined microelements declined significantly during the time of lactation. Statistically significant linear correlation was found between concentrations of Zn in blood plasma and milk in the first stage of lactation. Weak but statistically significant linear correlations were also found between plasma Se content in plasma and in transitional and mature milk of breast-feeding women.

The disposition and metabolism of durene (1,2,4,5-tetramethylbenzene) in rats.

The organ and tissue distribution, excretion and metabolism of [3H]1,2,4,5-tetramethylbenzene ([3H]durene) in male Wistar albino rats were investigated following a single i.p. administration (40 mg/kg) and within 9 days after five daily repeated administrations. Urine proved to be the main route of tritium excretion. Within the first 24 h after a single administration 69% of the radioactivity was excreted in the urine and only 9% in the feces. The highest level of tritium binding was found in the fat tissue, liver, kidneys and adrenal glands. The accumulation of tritium in the plasma proceeded with a kinetic constant of 0.49 h(-1), whereas the half-life of radioactivity decay amounted to about 6.3 h. In erythrocytes, the tritium level was found to be about three times lower than in blood plasma. The total amount eliminated during the 9 days following repeated administration was about 94% of the five doses given. The highest level of tritium was found in fat tissue and adrenal glands, followed by the liver, kidneys, sciatic nerve and muscle. A gradual decline in tritium levels was observed during the following 4 days in most tissues to reach about 2% of the dose given. The main urinary metabolites resulting from the administration of durene were 2,4,5-trimethylbenzyl alcohol (about 22%), 4,5-dimethyl-1,2-benzdialdehyde (about 19%), 2,4,5-trimethylbenzaldehyde (about 19%) and 2,4,5-trimethylbenzoic acid (about 16%). The oxygen-containing metabolites accounted for almost 80%, whereas sulphur-containing metabolites accounted for approximately 10% of the products of biotransformation. In conclusion, most of the durene administered has a relatively rapid turnover rate, with minor levels retained in the tissues for longer time periods.

Blood concentration of essential trace elements and heavy metals in workers exposed to lead and cadmium.

The aim of the study was to determine blood concentration of essential trace elements (Se, Zn, Cu) and toxic metals (Pb, Cd), markers of antioxidant (activities of glutathione peroxidase (GPx), superoxidase dismutase and ceruloplasmin) and prooxidant processes (thiobarbituric acid reactive substances (TBARS)) in workers exposed to Pb and Cd. Forty three male workers of the lead-acid batteries department, aged 25-52 years, and twenty two workers, including 15 women, aged 36-51 years, exposed to Cd in the alkaline batteries department were examined. The reference group consisted of 52 healthy inhabitants of the same region. It was found that Se concentration and GPx activity in both erythrocytes and plasma of Cd exposed workers were significantly lower (p < 0.001) than in the reference group. We found an inverse linear correlation between blood Se and Cd concentrations in the workers exposed to Cd (r = -0.449; p < 0.01). Moreover, the activity of erythrocyte and plasma GPx was shown to be significantly lower in the study group of workers (p < 0.001). It was observed that TBARS concentration in plasma was significantly higher (p < 0.05) in the lead exposed workers than in the group without contact with Pb. Our results indicate that exposure to Pb and Cd affects the antioxidant potential of blood in workers exposed to heavy metals.

Subchronic inhalation toxicity of 1,2,3-trimethylbenzene (hemimellitene) in rats.

Toxic effects of exposure to 1,2,3-trimethylbenzene (hemimellitene) in the condition of subchronic inhalation experiment were examined. Rats were exposed to vapours of hemimellitene at concentrations of 123 mg/m3, 492 mg/m3 and 1230 mg/m3, 6 h/day, 5 days/week for 3 months. After termination of a 3-month inhalation, animals were necropsied. Blood samples were obtained and selected organs were weighed and prepared for histological examinations. Subchronic inhalation exposure to hemimellitene resulted in an overall, low systemic toxicity. There were no changes in body weight gain and food consumption. At a concentration of 1230 mg/m3, the increase in relative liver weight was observed in male rats. It was accompanied by slight increase in sorbitol dehydrogenase activity. The increase in alkaline phosphatase activity was found in females only. Some disturbances in haematological parameters, characterised by the decrease in red blood cells and slight increase in white blood cells, segmented neutrophils and lymphocytes were observed in rats at high exposure concentration of 1230 mg/m3. The pulmonary lesions as well as the increased number of goblet cells and interstitial lung parenchyma infiltration were noted in male and female rats from the highest exposure groups.

Sub-chronic inhalation toxicity of 1,2,4-trimethylbenzene (pseudocumene) in rats.

Toxic effects of exposure of 1,2,4-trimethylbenzene (pseudocumene) in the condition of sub-chronic inhalation experiment were examined. Rats were exposed to vapours of pseudocumene at concentrations of 123 mg/m3, 492 mg/m3 and 1230 mg/m3, 6 h/day, 5 days/week for 3 months. After 3 months of inhalation exposure animals were necropsied. Blood samples were obtained and selected organs were weighted and prepared for histological examinations. Sub-chronic inhalation exposure to pseudocumene resulted in an overall low degree of systemic toxicity. There were no changes in body weight gain, food consumption and absolute and relative organ weights. Slightly higher activity of sorbitol dehydrogenase was observed in male rats exposed to all concentrations applied. Some disturbances in hematological parameters characterised by decrease in red and increase in white blood cells were observed in male rats exposed to high concentration of 1230 mg/m3. The pulmonary lesions observed in male and female rats were statistically significant at mid and high concentrations of pseudocumene.

Studies on dermal, ocular and respiratory effects of 4-ethyltoluene in experimental animals.

The toxicity of 4-ethyltoluene to experimental animals was studied after single and repeated exposures. It was found that 4-ethyltoluene can be classified as a very mild skin and eye irritant. Sensory respiratory irritation of 4-ethyltoluene was studied in Balb/C male mice using the plethysmographic method. The concentration at which the respiratory rate decreased to 50% (RD50 value) was determined to be 4216 mg/m3 (2795-5850 mg/m3 for 95% confidence interval). To study repeated-dose inhalation toxicity, male and female outbred Wistar rats were exposed in a dynamic inhalation chamber to 4-ethyltoluene vapours at concentrations of 477 or 2337 mg/m3, 6 h/day, 5 days/week for 4 weeks (20 exposure days). No significant changes were observed in food consumption and body weight gain. Statistically significant, concentration-dependent changes in the number of total cells, as well as of macrophages, polymorphonuclear leucocytes and lymphocytes were found in bronchoalveolar lavage. In the fluid of bronchoalveolar lavage, a significant, concentration-related increase was noted in total protein and mucoproteins and the activity of beta-glucuronidase, gamma-glutamyl transferase, lactate dehydrogenase and acid phosphatase. Histopathology revealed an increased rate of bronchitis and pneumonia and perivascular lymphoid infiltrations in rats exposed to 2337 mg/m3 of 4-ethyltoluene.

DNA damage detected by the comet assay in the white blood cells of workers in a wooden furniture plant.

The study was aimed at the assessment of genotoxic effects in workers of a wooden furniture manufacture, based on the level of DNA damage in white blood cells (WBC). The alkaline single cell gel electrophoresis assay (known as the comet assay) in individual cells was adapted for detecting damaged DNA in WBC. The level of DNA damage was determined as the percentage of cells with comets. It was assessed in cells before and after incubation in RPMI 1640 medium and CO(2) at 37 degrees C for 1 h to repair DNA breaks. Thirty-five woodworkers and 41 control subjects were studied. In the woodworkers, significantly more cells with DNA damage (21.5%) were observed than in the control persons (9.7%). A slight but significant decrease in the level of DNA damage was found in the WBC of woodworkers after incubation (17.2%). Significantly higher levels of damaged DNA was observed in woodworkers who either smoked (22.1%) or did not smoke cigarettes (20.8%) than in smokers (13.2%) and non-smokers (7.0%) from the control group. After incubation, a slight decrease in the level of DNA damage was found in both smoking and non-smoking woodworkers compared to the respective subjects in the control group. The increased levels of DNA damage observed in the woodworkers could be associated with the occupational exposure to wood dust in the furniture manufacture.