Evaluation of genotoxic effects of lead in pottery-glaze workers using micronucleus assay, alkaline comet assay and DNA diffusion assay.

PURPOSE: We investigated genotoxic effects of occupational exposure to lead acetate in pottery-glaze ceramic workers. METHODS: The study was carried out in 30 exposed workers and 30 matched controls, to whom several biochemical parameters-the blood lead (B-Pb; range: exposed, 41.68-404.77; controls, 12-52) and cadmium (B-Cd) level, the activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), the level of vitamin B(12) and folate in serum-were measured. The genotoxic effects were evaluated by the alkaline comet assay, the DNA diffusion assay and micronucleus test in peripheral blood lymphocytes. RESULTS: Subjects exposed to lead had significantly higher B-Pb level and, consequently, increased values of tail intensity (TI), frequency of apoptotic and necrotic cells, and frequency of micronuclei (MN). In contrast, their activity of ALAD, the level of vitamin B(12) and folate in serum were significantly lower compared to controls. Poisson regression analysis showed a significant correlation of profession, duration of exposure, smoking, level of cadmium in blood, ALAD and EP with primary DNA damage. A majority of primary damage repairs in a short period after exposure to a genotoxic agent. In addition, the influence of gender and level of vitamin B(12) and folate in serum MN frequency in exposed group was observed. CONCLUSIONS: In this study, DNA diffusion and micronucleus test showed higher influence of tested parameters to DNA damage. The results indicate a need for concomitant use of at least two different biomarkers of exposure when estimating a genetic risk of lead exposure.

Antioxidants, trace elements and metabolic syndrome in elderly subjects.

OBJECTIVE: To examine whether concentrations of several trace elements and activities of several antioxidant enzymes are modified in metabolic syndrome, and to evaluate their possible association with metabolic syndrome components. Additionally, concentration of CRP, as a marker of inflammation, was measured. DESIGN: Cross-sectional study. PARTICIPANTS: The study group consisted of 100 subjects, aged 71-88 years. MEASUREMENTS: Anthropometric measurements and biochemical analyses of fasting blood samples were performed by standardized methods. According to the International Diabetic Federation (IDF) criteria, metabolic syndrome was diagnosed in 64 subjects. Whole blood glutathione peroxidase (GPx), erythrocyte superoxide dismutase (SOD) and catalase (CAT), serum selenium (SSe), copper (SCu) and zinc (SZn), glucose, lipoprotein profile and C-reactive protein (CRP) were determined in all subjects. RESULTS: No clear influence of metabolic syndrome on SSe, SZn and SCu concentration and SOD and CAT activity was found. However, significantly higher GPx was found in subjects with metabolic syndrome than in subjects without metabolic syndrome (p=0.029), as well as in subjects with hypertriglyceridemia than in control subjects (p=0.038). After adjusting for potentially confounding variables by multiple regression, significant positive relationship between SCu and CRP was found, indicating that elevated levels of Cu could have influence on inflammatory mechanisms. CONCLUSION: Our results suggest that GPx and CRP, as biomarkers of oxidative stress and chronic inflammation, respectively, have significant role in the pathogenesis of metabolic syndrome.

Blood pressure in relation to biomarkers of lead, cadmium, copper, zinc, and selenium in men without occupational exposure to metals.

The interrelationship of blood lead (BPb), activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), blood cadmium (BCd), serum copper (SCu), serum zinc (SZn), serum selenium (SSe), hematocrit (Hct), body mass index (BMI), age, smoking habits, and alcohol consumption to blood pressure was examined in 154 Croatian male subjects 19-53 years of age. None of the subjects had been occupationally exposed to metals, or used any medication that could influence blood pressure or metal metabolism. The median and range values were: BPb, 57 (25-254) microg/L; ALAD, 51.9 (22.8-96.4) European units; EP, 0.68 (0.38-1.68) micromol/L erythrocytes; BCd, 0.83 (0.21-11.93) microg/L; SCu, 1113 (763-1662) microg/L; SZn, 961 (734-1213) microg/L; SSe, 73.6 (44.2-106.9) microg/L; systolic blood pressure, 131 (105-165) mm Hg; and diastolic blood pressure, 94 (71-112) mm Hg. Systolic and diastolic blood pressure significantly correlated with an increase in BMI (P<0.0005 and P<0.0001, respectively), EP (P<0.0002 and P<0.002, respectively), and BPb (P<0.005 and P<0.01, respectively). After adjusting for potential confounding variables by multiple regression, an increase in systolic blood pressure was significantly predictive by BMI (P<0.0005) and log BPb (P<0.02) and inversely by log BPb*SSe interaction term (P<0.007), or alternatively by EP (P<0.0001), BMI (P<0.001), alcohol (P<0.02), and Hct (P<0.05). An increase in diastolic blood pressure was significantly predictive by BMI (P<10(-5)), log BPb (P<0.04), and alcohol (P=0.05) and inversely by log BPb*SSe interaction term (P<0.0007), or alternatively by BMI (P<0.0001), EP (P<0.002), alcohol (P<0.004), and Hct (P<0.04) and inversely by smoking (P<0.04). With respect to the EP range in the study population, an increase of 27 mm Hg in systolic and 14 mm Hg in diastolic blood pressure was found. The study results indicate a significant Pb-related increase in blood pressure, particularly within the low-level Pb exposure range (BPb of 25-75 microg/L, and/or EP of 0.4-1.0 micromol/L erythrocytes).

Blood pressure in relation to dietary calcium intake, alcohol consumption, blood lead, and blood cadmium in female nonsmokers.

The interrelationship of dietary calcium (Ca) intake, alcohol consumption, blood lead (BPb), blood cadmium (BCd), age, and body mass index (BMI) to blood pressure was examined in 267 peasant women 40-85 years of age. They were residents of two rural areas in Croatia and differed with regard to dietary Ca intake: 100 women with low Ca intake (approximately 450 mg/day) and 167 women with relatively high Ca intake (approximately 940 mg/day). All of the women were nonsmokers and consumed very little or no alcohol. Median and range BPb values were 74 (29-251) microg/L in women with low Ca intake and 59 (21-263) microg/L in women with high Ca intake (p < 0.0002), whereas corresponding BCd values were 0.6 (0.2-3.6) microg/L and 0.6 (0.3-4.5) microg/L (p > 0.10). Results of multiple regression showed a significant (p < 0.05) increase in systolic blood pressure with age, BMI, and BCd, and marginally with alcohol consumption (multiple r = 0.48, p < 10(- 6)). An increase in diastolic blood pressure was significantly (p < 0.05) associated with BMI, age, and residence area (i.e., it was higher in women with low Ca intake), and marginally with BCd, and alcohol consumption (multiple r = 0.38, p < 10(-6)) When the two groups of women with different Ca intake were subdivided into consumers and nonconsumers of alcohol, BPb was related positively to alcohol consumption and inversely to Ca intake. The highest BPb was found in the subgroup of alcohol consumers with low Ca intake, and the lowest BPb in the subgroup of nonconsumers with high Ca intake: 78 (42-251) microg/L and 51 (22-192) microg/L, respectively (p < 10(-8)). Diastolic blood pressure was significantly higher in the former subgroup as compared to the latter: 95 (72-130) mm Hg and 90 (60-120) mm Hg, respectively (p < 0.05). This cannot be explained by age, BMI, or BCd, which were comparable in the two subgroups. The results indicate that alcohol consumption and low Ca intake can increase BPb, which may significantly contribute to an increase in diastolic blood pressure in female nonsmokers even at relatively low-level Pb exposure.

Lead absorption and psychological function in Zagreb (Croatia) school children.

A cross-sectional study was performed on 275 pupils from the third and fourth grade of three elementary schools (three urban areas with different traffic conditions) in Zagreb. Lead exposure was environmental, mostly through leaded gasoline. The difference in traffic density around the schools was consistent with biological indicators of lead absorption. The aim of the study was to clarify the relationship between characteristic biological indicators of lead absorption including indicators of hematological status with some psychological functions. Lead absorption in pupils was relatively low (mean blood lead: 70.8 +/- 17.88 microgram/L). Pupils' socio-economic status was evaluated by parents' education. The results obtained indicate that gender and school were associated with both biological and psychological variables. After adjusting for age, parental education, and gender, lead appears to have no association with cognitive or psycho-motor measures. The nonstandardized regression coefficients for blood lead-as a measure of the size of lead effect on VIQ, NIQ, and IQ-were -0.016, -0.031, and -0.025, respectively, all nonsignificant.

Semen quality and reproductive endocrine function in relation to biomarkers of lead, cadmium, zinc, and copper in men.

Blood lead (BPb), activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), blood cadmium (BCd), serum zinc (SZn), seminal fluid zinc (SfZn), serum copper (SCu), and parameters of semen quality and of reproductive endocrine function were measured in 149 healthy male industrial workers 20-43 years of age. The group contained 98 subjects with slight to moderate occupational exposure to Pb and 51 reference subjects. All of the subjects lived in Zagreb, Croatia. Significant (p < 0.05) correlations of BPb, ALAD, and/or EP with reproductive parameters indicated a Pb-related decrease in sperm density, in counts of total, motile, and viable sperm, in the percentage and count of progressively motile sperm, in parameters of prostate secretory function (SfZn, acid phosphatase, and citric acid in seminal fluid), and an increase in abnormal sperm head morphology, serum testosterone, and estradiol. These associations were confirmed by results of multiple regression, which also showed significant (p < 0. 05) influence of BCd, SZn, SCu, smoking habits, alcohol consumption, or age on certain reproductive parameters. These effects were mainly of lower rank and intensity as compared to Pb-related reproductive effects, whereas BCd contributed to a decrease in sperm motility and an increase in abnormal sperm morphology and serum testosterone. No significant Pb- or Cd-related influence was found on levels of the lactate dehydrogenase isoenzyme LDH-C(4) and fructose in seminal fluid or on follicle-stimulating hormone, luteinizing hormone, and prolactin in serum. The seminal fluid concentrations of Pb (SfPb) and Cd (SfCd) were measured in 118 of the 149 subjects, and a highly significant (p < 0.0001) correlation was found between BPb and SfPb levels (r = 0.571) and between BCd and SfCd levels (r = 0.490). The overall study results indicate that even moderate exposures to Pb (BPb < 400 microg/L) and Cd (BCd < 10 microg/L) can significantly reduce human semen quality without conclusive evidence of impairment of male reproductive endocrine function.

Serum copper, zinc and selenium levels with regard to psychological stress in men.

Serum concentrations of copper (S-Cu), zinc (S-Zn) and selenium (S-Se) were measured in 34 apparently healthy male prisoners of war immediately on release from a detention camp, and 85 healthy male subjects of comparable age and body mass index who had not been in a war combat zone. The results expressed as median and range were: 1138 (877-1337) micrograms/L of S-Cu, 1087 (514-1260) micrograms/L of S-Zn and 53 (30-100) micrograms/L of S-Se in the former prisoners, and 1149 (869-1487) micrograms/L of S-Cu, 1131 (874-1351) micrograms/L of S-Zn and 65 (45-109) micrograms/L of S-Se in the reference subjects. Significantly lower S-Zn (p < 0.02) and S-Se (p < 10(-5)) were found in the former prisoners compared to the reference subjects, whereas no significant difference between the groups was found for S-Cu (p > 0.80). In the group of former prisoners, a significant positive correlation was found between the S-Zn and S-Se levels (r = 0.40, p < 0.05) and an inverse correlation between S-Zn and body mass index (r = -0.34, p < 0.05), whereas no significant correlation was found of S-Cu, S-Zn or S-Se with age (38 (19-54) years) or duration of imprisonment (130 (126-270) days). As the body mass index of 23.4 (19.7-28.1) kg/m2 and the body mass relative deviation from nomogram of 105 (89-125)% in the group of former prisoners showed no indication of malnutrition, lowered S-Zn and S-Se levels may be ascribed to increased psychological stress induced by conditions during imprisonment.

Meso-2,3-dimercaptosuccinic acid in the treatment of occupationally exposed lead workers.

The aim of this study was to evaluate the efficacy of meso-2,3-dimercaptosuccinic acid (DMSA) treatment in workers with increased lead absorption and no overt symptoms of lead poisoning. Seven occupationally lead exposed male workers with blood lead concentrations (PbB) exceeding 50 micrograms/100 ml and a positive calcium disodium ethylenediaminetetraacetate (EDTA) lead mobilization test were treated with DMSA for 19 days. Individual doses were 700 mg DMSA, three times a day from day one to five, and twice a day from day six to 19. The treatment intensified urinary lead excretion, most rapidly during the first five days. The increased elimination was followed by a decline of mean PbB to 15% of the pretreatment values. However, 15 days after the treatment, the PbB concentrations rebounded, yet kept below the baseline values and did not exceed 40 micrograms/100 ml. After repeated EDTA lead mobilization test, urine lead was 23-68% of that before DMSA treatment. It can be concluded that DMSA can effectively reduce chelatable lead in occupationally exposed workers.

[Copper levels in the tissues of patients with Wilson's disease]. / Koncentracije bakra u tkivima bolesnika s Wilsonovom bolescu.

The evaluation of tissue copper concentration is essential for the diagnosis of Wilson's disease. It is characterized by symptoms of the damages to parenchymatous organs, primarily liver and brain, due to chronic copper intoxication. The paper presents the autopsy tissue analysis of a 39-year-old patient diagnosed to suffer from the Wilson's disease while still alive. The patient died from sepsis due to burns caused by direct flame exposure. The standard histochemical staining of tissue samples failed to demonstrate the presence of copper but it was qualitatively proved by line spectrum-based mass spectrography. The copper concentrations in the liver, brain, and kidney (240, 73.8, and 30 micrograms/g wet tissue weight, respectively), measured by flame-atomic absorption spectrometry, were significantly elevated. In this study, the Wilson's disease was verified by a postmortem determination of increased copper concentration in the tissues. The results obtained contribute to the understanding of this rare disease.

Cadmium in the blood and seminal fluid of nonoccupationally exposed adult male subjects with regard to smoking habits.

Blood cadmium (B-Cd) and seminal fluid cadmium (Sf-Cd) were measured in 120 adult male subjects not occupationally exposed to cadmium (Cd), comprising 42 nonsmokers (including nine former smokers) and 78 smokers. The respective median and range values were: 0.46 (0.19-1.49) microgram/l of B-Cd and 0.54 (0.17-1.67) microgram/l of Sf-Cd in nonsmokers, and 4.33 (0.49-13.33) micrograms/l of B-Cd and 0.85 (0.29-3.56) microgram/l of Sf-Cd in smokers. Both indicators showed a highly significant difference in Cd exposure between the groups (P < 0.0001), although the increase in B-Cd was considerably more pronounced than that of Sf-Cd in smokers compared with nonsmokers. The results suggest a nonlinear relationship (log Sf-Cd/log B-Cd: r = 0.501, P < 0.0001), rather than linear relationship (Sf-Cd/B-Cd: r = 0.430, P < 0.0001), between the indicators. Significant correlations were found between smoking habits, i.e., the number of cigarettes per day, and an increase in B-Cd in smokers (r = 0.296, P < 0.01) and in all 120 subjects (r = 0.685, P < 0.0001), as well as between smoking habits and an increase in Sf-Cd in smokers (r = 0.378, P < 0.001) and in all 120 subjects (r = 0.488, P < 0.0001). Both indicators are necessary for evaluation of individual internal Cd dose, since they appear to differ in reflecting recent and long-term cumulative Cd exposure and/or the amount of Cd at the site(s) of its effect(s) in the body.

Lead poisoning associated with the use of Ayurvedic metal-mineral tonics.

OBJECTIVE: The aim of this study was to confirm the connection between lead poisoning and the use of traditional Ayurvedic metal mineral tonics. METHODS: The study group comprised 29 subjects (26 adults and three children) who had previously taken Ayurvedic metal mineral tonics. All subjects were tested for lead absorption by blood lead, erythrocyte delta-aminolevulinic acid dehydratase activity and erythrocyte protoporphyrin. RESULTS: Eighteen samples of Ayurvedic preparations were obtained from 15 subjects and analyzed for lead content. Five adult subjects with established lead poisoning received chelation therapy. In Ayurvedic preparations a wide range of lead content was found (0.9-72,990 micrograms Pb/g; 0.35-29,900 micrograms Pb/capsule or tablet). The blood lead, erythrocyte delta-aminolevulinic acid dehydratase and erythrocyte protoporphyrin of the subjects, grouped according to the remedies used, correlated with the lead content in the preparations (p < 0.001). Chelation therapy was successful in normalization of laboratory findings and clinical recovery. CONCLUSION: Ayurvedic metal-mineral tonics are again identified as a potential source of high lead. The import of such tonics should be strictly controlled.

A follow-up study in a lead smelter community following the introduction of an effective pollution control system.

A follow-up study using biological and environmental monitoring was performed in a lead smelter community to assess the health risk of inhabitants and to establish how long it takes after the introduction of an effective emission control system to declare a lead-contaminated area safe for normal living. In specific population groups (mothers and children) biological indicators of lead exposure and lead effect on hemoglobin synthesis were determined 2 years before and 11 consecutive years after the installation of bag filters. At the same time lead was monitored in the airborne and deposited particulates, in household dust and in soil. The results clearly show that the process of normalization is rather slow. The difference in blood lead and erythrocyte delta-aminolevulinic acid dehydratase between the exposed and control groups, even 12 years after bag filters installation, was still remarkable. It appears that soil, contaminated with lead from past exposure, is a major source of increased lead absorption in a population for many years.