Metal determinations in biological specimens of diseased and non-diseased hard metal workers

Biological monitoring of Co, Ta and W, using mainly neutron activation analysis, was carried out on the urine, blood, pubic hair and toe nails of 251 subjects occupationally exposed to hard metal dusts (23 individuals were diagnosed as ‘diseased subjects’, affected by asthma and/or lung fibrosis). Airborne dust at hard metal workplaces were also analyzed for Co and W content. Cobalt and tungsten exposure at workplaces varied widely and frequently exceeded the TLV (0.05 mg Co/m³). Cobalt in urine (CoU), which is better than Co in blood (CoB), could represent a valuable indicator in discriminating between exposed groups, but is of little value as an indicator of exposure for single individuals. The high concentrations of cobalt in pubic hair (CoH) and toe nails (CoN) could also be valuable indicators of exposure although they are not useful, however, in establishing quantitatively the levels of cobalt accumulation. Tungsten and tantalum in pubic hair (WH and TaH) and toe nails (WN and TaN) also seem to be useful indicators in proving hard metal exposure qualitatively. The determination of these two elements, rather than cobalt, in the bronchoalveolar lavage (BAL) is very useful in complementing the diagnosis of hard metal disease proving hard metal exposure. The analysis of the BAL subfractions showed that W and Ta were firmly incorporated into the macrophage fraction while Co was distributed between cellular fraction and supernatant, which suggests a different mobility of hard metals in pulmonary tissue. The metal concentrations determined when submitted to statistical analysis indicated a positive correlation with P < 0.001 for the pairs (CoB---CoU), (Co---W) in urine, and (Co---W) in toe nails. Multielement analysis of biological specimens from diseased subjects suggests that hard metal disease does not relate to Co, W and Ta levels in the specimens considered. The disease does not depend on sex, age, working age and length of hard metal exposure. These findings support the theory on the possible haptenic properties of Co which may induce hypersensitivity and immuno-related toxic effects.     

Absorption and excretion of cobalt in the hard metal industry

Absorption and excretion of cobalt in the hard metal industry was investigated by means of ambient air and urine measurements in three factories with high levels of environmental cobalt pollution. In the presence of poor hygiene conditions and permission to smoking during work, there was no relationship between cobalt ambient air and cobalt urine concentrations. Such a finding was therefore attributed to a substantial skin contact. A simple experiment of skin exposure to freshly mixed or waste powder on volunteers identified a ten-fold increase of cobalt in urine in the post-exposure samples, thus confirming the contribution of dermal exposure as a route of entry. An improvement in the hygiene of the working conditions helped to investigate the relationship between exposure and excretion level. Cobalt uptake through the different routes of entry may be substantial, and requires a more prolonged exposure-free period so that the excretion rate can be reduced to the reference population level.     

Chromium in urine, serum and red blood cells in the biological monitoring of workers exposed to different chromium valency states

Using personal air sampling exposure to hexavalent and trivalent chromium was measured in 22 workers mainly exposed to Cr(VI) and in 15 workers mainly exposed to Cr(III) as basic chromium sulphate. Determination of Cr(VI) in the urine of all the subjects using a selective technique by ETA-AAS and liquid anion exchangers failed to show detectable amounts of the hexavalent form, the detection limit of the technique being 0.05 micrograms/L. A clear relationship between exposure and postshift urinary total chromium was found in subjects exposed to Cr(VI), while urinary levels in workers exposed to chromic sulphate high concentration proved lower. Determination of total chromium in serum and red blood cells showed a significant increase of chromium levels in erythrocytes of workers exposed to Cr(VI) while in subjects mainly exposed to Cr(III) an increase of the serum fraction was observed. The results demonstrate that Cr(III) is absorbed through the respiratory tract, but its kinetics and distribution in the body are not the same as for Cr(VI), and are not adequately monitored by short-term urinary determinations. Oxidation states of chromium largely influence uptake, mechanism of absorption, transport and organ distribution as well as toxicity of chromium-containing compounds. In particular, hexavalent derivatives are known to induce adverse effects, both acute and chronic, in occupationally exposed subjects, while there is little conclusive evidence for toxic effects caused by trivalent chromium compounds. Biological monitoring of exposure to chromium(VI) has usually been performed by determining total chromium levels in urine, whereas biological monitoring data in subjects occupationally exposed to Cr(III) are still scanty.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bronchoalveolar lavage and its role in diagnosing cobalt lung disease

The bronchoalveolar lavage (BAL) of two groups of hard metal workers (11 subjects) were examined and compared with BALs of 14 unexposed individuals as controls. The first group of workers included five asymptomatic subjects (2 females and 3 males) with normal lung function tests and chest X-rays. All the workers had been exposed daily to hard metal dust for 5 years before fiberoptic bronchoscopy. The second group of workers included six individuals (1 female and 5 males) with cobalt lung disease at biopsy. The BAL's cytology at the deep lung level is in agreement with a possible immunologic pathogenesis of the lung disease, similar to hypersensitivity pneumonitis (lymphocytosis with helper-suppressor ratio reduction). In order to identify diagnostic key-parameters, the data presented here are compared with data reported in literature for workers with a history of hard metal or cobalt (alone) exposure. Although the BAL can be useful, for a better definition of the reaction at the deep lung level, it seems insufficient per se for the diagnosis of hard metal lung disease.     

The rate of urinary excretion of phosalone residues in occupationally exposed persons

The absorption of malathion and phosalone was followed in occupationally exposed workers by determination of residues excreted in the urine. Because of the high concentrations found in the morning urine samples, the rates of excretion of phosalone metabolites in the urine of a volunteer experimentally exposed to phosalone during one and then again during three subsequent working days were investigated. The urinary excretion of phosalone metabolites was most intense 4–5 hours after exposure. At the beginning of the next day, the metabolites were still well measureable in the urine. Blood and plasma cholinesterase activities were only slightly reduced during exposure. The analyses of 24-hour urine samples, or of urine samples taken 4–5 hours after exposure, are not suitable for the routine control of occupationally exposed persons because of sampling difficulties. Instead, analyses of samples taken immediately before and after work hours have to be performed. A systematic increase in the concentrations of pesticide residues in the morning urine should initiate more efficient and well-timed protection measures.     

Biomonitoring of the adverse effects induced by the chronic exposure to lead and cadmium on kidney function: usefulness of alpha-glutathione S-transferase

A successful prevention of renal diseases induced by occupational exposure to lead (Pb) and/or cadmium (Cd) largely relies on the capability to detect nephrotoxic effects at a stage when they are still reversible or at least not yet compromising renal function. Hence, the aim of this cross-sectional study was to evaluate the usefulness of a set of early biological markers of oxidative stress or nephrotoxicity for the biomonitoring of workers occupationally exposed to Pb and/or Cd in a non-ferrous metal smelter, and gender, age, socioeconomic status, smoking habits, and drug use-matched control individuals. In exposed subjects, mean levels of Pb in blood and urine were also 387.1+/-99.1 microg Pb/L (1.868+/-0.478 micromol Pb/L) and 217.7+/-117.7 microg Pb/g creatinine (1.051+/-0.568 micromol Pb/g creatinine), and mean levels of Cd in blood and urine were 3.26+/-2.11 microg Cd/L (0.029+/-0.019 micromol Cd/L) and 2.51+/-1.89 microg Cd/g creatinine (0.022+/-0.017 micromol Cd/g creatinine), suggesting thereby relatively low occupational exposure levels. Statistically significant variations in zinc protoporphyrin, malondialdehyde, retinol binding protein, alpha-glutathione S-transferase, and urinary protein levels were reported between the two groups, and were closely correlated with Pb and/or Cd exposure levels. Variations in alphaGST levels were closely associated with Pb exposure. Taken together, these results suggest the use of alpha-glutathione S-transferase excretion in urine as a hallmark of early changes in the proximal tubular integrity.     

The speciation of the chemical forms of arsenic in the biological monitoring of exposure to inorganic arsenic

Total As content may be determined in blood and urine by means of an AAS method that involves reduction of As to its volatile hydride and ashing at 600°C with MgO and Mg (NO₃)₂. Separation of inorganic As (InAs), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMAA) by ion-exchange chromatography, followed by direct AAS analysis, allows the determination of each As species in the urine.In a reference population of 148 subjects with only normal environmental exposure to As, total As concentration in the urine averages 17.2 ± 11.1 μg/l. Urinary As consists of 10% each of InAs, MMAA and DMAA, the remaining 70% consisting of other forms of organic As.Blood As concentration averages 5.1 ± 6.9 μg/l and correlates significantly with the urinary concentration of InAs and the sum of its metabolites (InAs + MMAA + DMAA).Inorganic arsenic undergoes methylation in the organism. After ingestion of high quantities of As₂O₃, the time course of excretion of its metabolites indicates that As methylation occurs by a saturable mechanism.In workers exposed to As₂O₃, InAs, MMAA and DMAA are the only chemical forms of As excreted in the urine that are relevant to a study of occupational exposure. Blood As concentration is proportional to exposure and correlates only with urinary DMAA excretion; DMAA seems to be the most appropriate single indicator of exposure. At high levels of exposure (total As excretion above 200 μg/l), As accumulates in the organism and DMAA excretion reflects its accumulation. At low levels of exposure (total As excretion below 50 μg/l) a short-term accumulation does not occur and the best biological indicator of exposure is InAs excretion.Seafood ingestion brings about a marked increase in urinary excretion of total As that lasts for 24–48 h and is not accompanied by any increase in InAs, MMAA or DMAA excretion. Organic As from seafood does not mix with the pool of inorganic As in the organism and may be separately detected in urine. In the biological monitoring of human exposure to As, particularly in the case of high urinary values, the speciation of the chemical forms of As in urine is necessary in order to establish with certainty the source, industrial or alimentary, of exposure.     

Effects of genetic polymorphisms CYP1A1, GSTM1, GSTT1 and GSTP1 on urinary 1-hydroxypyrene levels in sugarcane workers

Sugarcane workers in Brazil are exposed to various genotoxic compounds, including polycyclic aromatic hydrocarbons (PAHs), derived from an incomplete combustion process of burnt sugarcane fields. The effects of the occupational exposure to sugarcane fields burning were measured in urine samples of sugarcane workers from the northwest of the State of S?o Paulo when exposed (harvesting) and when non-exposed (non-harvesting). The urinary levels of 1-hydroxypyrene (1-OHP) and the influence of the genetic polymorphisms CYP1A1, GSTM1, GSTT1 and GSTP1 were evaluated. Our results showed that the 1-OHP levels were significantly higher (P<0.0000) in the exposed sugarcane workers (0.318 mumol mol(-1) creatinine) than in the non-exposed workers (0.035 mumol mol(-1) creatinine). In an unvaried analysis, no influence regarding the polymorphisms was observed. However, multivariate regression analysis showed that the CYP1A1()4 polymorphism in the exposed group, and age and the GSTP1 polymorphism in the non-exposed group significantly influenced urinary 1-OHP excretion levels (P<0.10). The same group of sugarcane workers was significantly more exposed to PAHs during the harvesting period than during the non-harvesting period.     

Observations on renal function in workers exposed to lead

Renal function was studied in 40 workers chronically exposed to lead at a smelter. The geometric means of lead were 40 (range 24.0 - 63.0) micrograms/dl in blood and 47 (range 19 - 115) micrograms/g creatinine or 71 (range 20 - 250) micrograms/l in urine, respectively. There were no significant differences between the lead-exposed group and a control group in the concentrations of beta 2-microglobulin in serum, urinary total protein and IgG, whereas the level of beta 2-microglobulin in urine was significantly higher in the lead-exposed group (p less than 0.01). No individual lead worker showed abnormally increased level of urinary beta 2-microglobulin. The significance of increased excretion of beta 2-microglobulin on the basis of the group remains to be elucidated.     

Hard metal disease: clinical aspects

On the basis of the data available in the literature and of our experience, the clinical patterns of respiratory troubles which can be observed in workers exposed to inhalation of hard metal dusts can be schematized as follows: Irritation forms, mild and transient, or severe up to pulmonary oedema, dose-correlated, which occur in all subjects exposed to sufficiently high atmospheric concentrations; Asthmatic factors, either reversible after cessation of exposure or persistent after stopping the exposure, which occur in a relatively low percentage of exposed subjects and also apply to the states quoted below; Dyspnoic patterns due to alveolitis (lymphocytic alveolitis with inverted helper/suppressor ratio, or giant cell-eosinophilic alveolitis, with or without fibrotic changes of pulmonary interstitium); Interstitial sclerotics lung disease, associated with or without an alveolitic component. The present diagnostic potentialities, particularly bronchoalveolar lavage (BAL), have helped in defining the clinical patterns and have confirmed the fundamental role of individual susceptibility in the occurrence of clinical manifestations (with the exception of the irritation forms), but so far have not yet enabled us to clarify whether the different clinical patterns are the results of a single pathogenetic mechanism or constitute pathogenetically distinct entities.     

Cobalt speciation in urine of hard metal workers. A study carried out by nuclear and radioanalytical techniques

A method for the determination of inorganic and organically-bound cobalt in human urine has been developed and applied to the urine of hard metal workers. The development was based on the use of the radionuclides ⁵⁷Co, ⁵⁸Co and ⁶⁰Co-labelled Co compounds such as Co-Vitamin B12 and CO²⁺ ions which allowed the study of their biotransformations in human and rat urine. The proposed procedure is based on the use of Chelex 100 resin which retains quantitatively the inorganic Co from the urine while the organic complexed form of the element is eluted. Cobalt is detected in both column and eluate by neutron activation analysis (NAA). The method has been applied to speciate inorganic and organically-bound Co in the urine of hard metal workers. There is a significant increase (P < 0.02) of the ratio inorganic/organic Co (2.3) in the urine of workers compared with controls (1.01), showing an increase of the inorganic fraction of Co in the urine of workers. The ratio was constant for the wide range of urinary Co analyzed (from 180 μg to 1254 μg Co/l). Therefore, the discrimination between inorganic and organic Co in urine should not represent progress in the biological monitoring of Co compared with the determination of total urinary Co. However, a large amount of organically-complexed Co is formed in the body of hard metal workers and excreted in urine, thus, investigationsof the nature of the organocobalt compounds are of fundamental importance in establishing their possible clinical significance.     

Assessment through environmental and biological measurements of total daily exposure to volatile organic compounds of office workers in Milan, Italy

Personal exposure to total volatile organic compounds (TVOCs), benzene and toluene of 100 Milan office workers was assessed through personal air monitoring at home, in the office, and during commuting. Biological monitoring was performed by measuring blood benzene and toluene concentrations together with urinary trans-trans-muconic acid (t,t-MA) and cotinine at the end of the monitoring period. The geometric means of the total 24-h personal exposure were 514 micrograms/m3 for TVOCs, 21.2 micrograms/m3 for benzene and 35.2 micrograms/m3 for toluene. Daily exposure to the volatile organic compounds was almost totally determined by indoor exposure at home and in the office, with a minor contribution in the transport means. An important factor determining exposure to benzene was found to be tobacco smoke, both for active smokers and for non-smokers exposed to environmental tobacco smoke (ETS). All the mean levels of the biological indicators were significantly higher in active smokers than in non-smoking subjects non-exposed to ETS; urine cotinine and t,t-MA levels were also significantly higher in non-smokers exposed to ETS than in non-smokers non-exposed to ETS.     

Levels of selenium and antioxidative enzymes following occupational exposure to inorganic mercury

Levels of selenium and mercury in blood and urine were analysed in 37 male workers exposed to elemental mercury vapour in a chloralkali plant and in 39 unexposed controls of the same age. Mean urinary Hg was 223 nmol l-1 (15 nmol/mmol creatinine) in the exposed group and 26 nmol l-1 (2.0 nmol/mmol creatinine) in the controls. Mean blood and plasma Hg levels were 46 and 36 nmol l-1, respectively, in the exposed group, as compared with 17 and 7 nmol l-1 in the controls. The concentrations of Se in plasma and erythrocytes did not differ between the two groups. Urinary Se levels were, however, slightly but significantly lower in the exposed group (median values 23 vs 29 nmol/mmol creatinine), and there was a negative correlation between urinary Se and plasma Hg in the exposed group. This may be due to a retention of Se in the kidneys. In a subgroup of exposed workers and controls, glutathione peroxidase, superoxide dismutase and catalase were also analysed. No differences were found between the groups with respect to these antioxidative enzymes. The effect on Se status of moderate Hg exposure seems to be of minor clinical importance.     

Hair can be a good biomarker of occupational exposure to mercury vapor: simulated experiments and field data analysis

Generally, urine mercury (U-Hg) is widely used for assessment of inorganic mercury (I-Hg) exposure in humans. The reliability of scalp hair as an indicator of mercury vapor exposure is contentious. However, significant correlations were found between hair total mercury (T-Hg) and U-Hg and between hair I-Hg and total gaseous mercury (TGM) in ambient air in our previous studies. Simulated experiments were designed to assess the contribution of direct absorption/adsorption of mercury vapor in the hair. Results indicated that the increases of hair T-Hg concentrations were less than 1 μg/g, which was negligible compared with hair T-Hg concentrations in occupationally exposed workers. The β-mercaptoethanol washing can remove 30% of mercury (Hg) in the exposed hair samples. The inhaled Hg constituted the major fraction (97.4%) of I-Hg exposure for the artisanal Hg mining workers. From the simulated experiments and field data analysis, we can conclude that hair I-Hg can be a useful tool for monitoring occupational exposure to Hg vapor.     

Cobalt powdersintering industry (stone cutting diamond wheels): a study of environmental-biological monitoring, workplace improvement and health surveillance

This communication deals with an aspect of occupational hygiene in a factory producing granite cutting diamond wheels by sintering, in moulds, of fine cobalt powder. The factory has been studied between 1988 and 1991; the Department of Preventive and Occupational Medicine of the Local Sanitary Unit of Reggio Emilia has followed the evolution of the local exhaust ventilation equipment supplied by the employer in that period. At the same time, the following measurements and observations were carried out: (a) cobalt exposure by personal sampling, (b) airborne cobalt measurements by area sampling, (c) biological monitoring of cobalt in urine, (d) health surveillance.     

Extremely low frequency-magnetic fields (ELF-EMF) occupational exposure and natural killer activity in peripheral blood lymphocytes

Extremely Low Frequency-Magnetic Fields (ELF-MF) are possible carcinogens to humans and some data suggest that they can act as promoters or progressors. Since NK cells play a major role in the control of cancer development, an adverse effect on ELF-MF on NK function has been hypothesized. We examined NK activity in 52 workers exposed to different levels of ELF-MF in various activities. Individual exposure was monitored during 3 complete work-shifts using personal dosimeters. Environmental exposure was also monitored. ELF-MF levels in the workers were expressed as Time-Weighted Average (TWA) values. NK activity was measured in peripheral blood lymphocytes (PBL). In the whole group the median occupational TWA was 0.21 microT. According to the TWA levels, workers were classified as low exposed (26 subjects, TWA < or =0.2 microT) and higher exposed workers (26 subjects; TWA >0.2 microT). In higher exposed workers, we observed a trend to reduce NK activity compared to low exposed, but the difference was not significant. Then we selected a subgroup of highest exposed workers (12 subjects; TWA >1 microT); no difference was observed between low and highest exposed subjects in the main personal variables. Considering both E:T ratios from 12:1 to 50:1 and Lytic Units, a significant reduction in NK activity was observed in the highest exposed workers compared to the low exposed. Multivariate analysis showed a significant negative correlation between exposure and LU, while no correlation was evidenced with other personal characteristics. ELF-MF are considered possible carcinogens, and existing data suggest that they can act as promoters. Due to the role of NK activity in host defence against cancer, the results obtained in this study in workers exposed to ELF-MF levels exceeding 1 microT are in agreement with this hypothesis, and support the need for further investigation in this field.     

PAH exposure in a Ghanaian population at high risk for aflatoxicosis

It was postulated that a population in sub-Saharan Africa, known to be at high risk for aflatoxicosis due to frequent ingestion of aflatoxin (AF)-contaminated foods could also be exposed to polycyclic aromatic hydrocarbons (PAHs) from a variety of environmental sources. Previously, participants in this population were shown to be highly exposed to AFs, and this exposure was significantly reduced by intervention with NovaSil clay (NS). Objectives of this study were 1) to assess PAH exposure in participants from the AF study using urinary biomarker 1-hydroxypyrene (1-OHP); 2) examine the effect of NS clay and placebo (cellulose) treatment on 1-OHP levels; and 3) determine potential association(s) between AF and PAH exposures. A clinical trial was conducted in 177 Ghanaians who received either NS capsules as high dose or low dose, or placebo (cellulose) for a period of 3 months. At the start and end of the study, urine samples were analyzed for 1-OHP. Of the 279 total samples, 98.9% had detectable levels of 1-OHP. Median 1-OHP excretion in nonsmokers was 0.64 µmol/mol creatinine at baseline and 0.69 µmol/mol creatinine after 3 months. Samples collected at both time points did not show significant differences between placebo and NS-treated groups. There was no linear correlation between 1-OHP and AF-albumin adduct levels. Results show that this population is highly exposed to PAHs (and AFs), that NS and cellulose treatment had no statistically significant effect on 1-OHP levels, and that this urinary biomarker was not linearly related with AF exposure.     

Benzene in blood as a biomarker of low level occupational exposure.

The occupational airborne exposure to benzene of 150 workers employed in petrol stations and a refinery plant was assessed using personal sampling pumps. All workers provided blood samples after the end of work and on the following morning before resuming work. Benzene concentrations in the blood of 243 non-occupationally-exposed subjects were also measured. The median occupational benzene exposure for all 150 workers studied was 80 micrograms/m3. Overall median blood benzene of all workers was 251 ng/l at the end of the shift, and 174 ng/l the following morning. The benzene concentrations measured in blood collected the following morning proved to be significantly lower than those measured at the end of the shift. Median blood benzene for the 243 'normal' subjects was 128 ng/l, which was significantly lower than that measured in the workers before a new work shift. The median blood benzene concentration was significantly higher in smokers than in non-smokers, both in the general population (210 ng/l vs. 110 ng/l) and in the exposed workers at the end of the shift (476 ng/l vs. 132 ng/l) and the following morning (360 ng/l vs. 99 ng/l). End-of-shift blood benzene correlated significantly with environmental exposure; this correlation was better in the 83 non-smokers than in the 67 smokers. In non-smokers with the median benzene occupational exposure of 50 micrograms/m3, no difference was found in blood benzene concentration in exposed and non-exposed subjects.     

Relationship between "chelatable lead" and the indicator of exposure and effect in current and past occupational exposure

The behaviour of blood (PbB) and two biological indicators of effect: erythrocyte protoporphyrin (EP) and urinary aminolaevulinic acid (ALAU), were studied in subjects who had ceased working with inorganic lead for at least one year. The relationships between these indices and chelatable lead (PbU-EDTA) were also analysed. As a comparison, a group of subjects currently exposed to inorganic lead was studied. At a given PbU-EDTA level, the PbB values were significantly lower in the workers with past exposure than those found in currently exposed subjects. The relationship between EP and PbU-EDTA illustrates that, for corresponding levels of chelatable lead, the values of the erythrocyte metabolite are identical in the two groups. Considering the EP-PbB relationship, however, at a given PbB level the protoporphyrin values appear distinctly more altered in the subjects with past exposure. The relationship between ALAU and PbU-EDTA shows that, at a given PbU-EDTA level the urinary metabolite in past exposed subjects is distinctly lower than in currently exposed workers, while the relationship between ALAU and PbB shows that, at similar blood lead values, the ALAU levels are identical in both groups. On the basis of these results, it is concluded that in subjects with past exposure, erythrocyte protoporphyrin can be used in establishing the persistence and extent of an "active deposit" of lead in the organism, while PbB and ALAU are of very limited value.