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Purification and immunological characterization of pigeon serum butyrylcholinesterase. Implications on environmental monitoring and toxicological testing of birds.

Butyrylcholinesterase (EC 3.1.1.8) (BChE) was purified from pigeon serum to electrophoretic homogeneity by a four-step procedure involving blue sepharose CL-6B chromatography, ion exchange chromatography, procainamide affinity chromatography and gel filtration. An overall 2789-fold purification was achieved, with a final specific activity of 61.35 mumol/min/mg. The purified enzyme separated into two peaks when filtered through a column of Sephacryl S-300, a smaller peak containing the tetrameric form of BChE (C4) and a larger peak containing the monomeric form of BChE (C1). Native polyacrylamide gel electrophoresis (PAGE) of both peaks revealed single protein bands which coincided with esterase activity, with approximate M(r) values of 84,000 and 340,000, respectively. The C1 monomer represented 85-90% of the activity found in the pigeon serum. It is not clear whether this polymorphism of BChE in vertebrates contributes to the wider inter-individual variations observed in xenobiotics elimination kinetics and in the response to the pharmacological and toxic effects of pesticides. PAGE of the monomeric form of the enzyme in the presence of sodium dodecyl sulphate showed only one protein band with a M(r) of 84,000, while that of the tetrameric form revealed two bands, a major protein band (84,000) and a minor band (170,000), representing the monomer and the dimer of the dissociated tetrameric BChE enzyme under reducing conditions. Highly specific polyclonal antibodies were raised in rabbits against the purified enzyme. These antibodies cross-reacted with other avian BChEs, a criterion which make them useful for the immunopurification of other BChEs from different species as well as for biomonitoring and toxicological studies on the role of esterases as an indicator of avian exposure to organophosphorous pesticides.

Animals↗

Development and validation of methods for environmental monitoring of cyclophosphamide in workplaces.

Methods to monitor contamination of workplaces with antineoplastic drugs have been developed and validated. Cyclophosphamide (CP) was used as a model compound as it is one of the most commonly used antineoplastic drugs. A wipe sampling method to detect contamination with CP at surfaces was developed. A personal air sampling method to sample gas and vapour on solid sorbent tubes and particles with filters was also developed. Wipe and filter samples were extracted and sorbent samples were eluted, all with ethyl acetate. The samples were analysed with liquid chromatography tandem mass spectrometry. (2)H(6)-labelled cyclophosphamide was used as an internal standard. The between-day precision was 2-5% for wipe samples, 4-6% for sorbent samples and 3-8% for filter samples. The limit of detection was 0.02 ng CP per sample for the wipe and filter methods and 0.03 ng CP per sample for the solid sorbent method. Wipe sampling on surfaces made of different materials resulted in mean recoveries between 78-106%. The desorption recovery was between 97-102% for the wipe samples, 97% for the sorbent samples and 101% for the filter samples. Samples were stable for up to 2 months at 5 degrees C and -20 degrees C and for about 2 d at room temperature. The developed methods were applied to the measurement of contamination with CP in a hospital pharmacy. Trace amounts of CP, 1.3 and 1.4 ng, were detected on surfaces in the pharmacy.

Air Pollutants, Occupational↗

Environmental monitoring of sevoflurane and nitrous oxide using the cuffed oropharyngeal airway.

UNLABELLED: We compared exposure to sevoflurane (SEV) and nitrous oxide (N(2)O) during ventilation using the cuffed oropharyngeal airway (COPA) with waste gas exposure using a conventional face mask (FM) without any additional airways or face straps and with the laryngeal mask airway (LMA). Trace concentrations of SEV and N(2)O were assessed by using a direct reading spectrometer during 33 surgical procedures under general anesthesia. Measurements were made at the patients' mouths and in the anesthesiologists' breathing zones. Mean +/- SD concentrations of SEV and N(2)O measured at the patients' mouths were comparable in the COPA (SEV, 8.1 +/- 12.2 ppm; N(2)O, 213.3 +/- 289.2 ppm) and LMA (SEV, 18.5 +/- 25.8 ppm; N(2)O, 283.4 +/- 361.0 ppm) groups but differed significantly from the FM group (SEV, 46.5 +/- 19.6 ppm; N(2)O, 750.7 +/- 308.3 ppm). These values resulted in a comparable contamination of the anesthesiologists' breathing zones (SEV, 0.5 +/- 0.2 ppm; N(2)O, 5.7 +/- 4.8 ppm) for the COPA group, compared with the LMA group (SEV, 1.0 +/- 0.9 ppm; N(2)O, 12.2 +/- 14.3 ppm). This differed significantly from the FM group (SEV, 2.2 +/- 0.9 ppm; N(2)O, 37.5 +/- 14.3 ppm). We conclude that the use of the COPA during short surgical interventions has an occupational safety comparable to that of the LMA and that both resulted in less contamination through waste anesthetic gases. Therefore, the COPA may be a valuable alternative to the conventional FM. IMPLICATIONS: In this study, we have shown that the occupational exposure to waste anesthetic gases is comparable when using the cuffed oropharyngeal airway (COPA) and the laryngeal mask airway and is increased when using the face mask. Therefore, the COPA may be a valuable alternative to the conventional face mask during short surgical procedures.

Adult↗

Multi-component assessment of worker exposures in a copper refinery. Part 1. Environmental monitoring.

The exposure characterisation described in this paper for 135 copper refinery workers (45 females, 90 males) focuses on the concentrations of copper, nickel and other trace elements in the inhalable aerosol fractions, as well as in the water-soluble and water-insoluble subfractions. Some information is also provided on the thoracic and respirable aerosol fractions. Further, results are presented for volatile hydrides of arsenic and selenium released in the copper purification steps of the electrorefining process. For the pyrometallurgical operations, a comparison of the geometric means for the inhalable aerosol fraction indicated that water-soluble copper levels were on average 19-fold higher compared to nickel (p < 0.001) and a significant association was evident between them (r = 0.87, p < 0.001); for the insoluble subfraction, the copper : nickel ratio was 12.5 (p < 0.001) and the inter-element correlation had r = 0.98 and p < 0.001. Although for the electrorefinery workers the relative inhalable concentrations of copper and nickel were not significantly different (p > 0.05), the corresponding inter-element associations were: slope of 7.7, r= 0.54, p < or =0.001 for the water-soluble subfraction and slope of 1.3, r = 0.71 and p < or =0.001 for the water-insoluble subfraction. On average, a good proportion of the inhalable copper and nickel were found in the thoracic (40%) and respirable (20%) aerosol fractions. Cobalt air concentrations were generally low with geometric means and 95% confidence intervals of 3.1 (2.4-4.2)microg m(-3) (pyrometallurgical workers) and 0.3 (0.4-0.5) microg m(-3)(electrorefinery workers). Similarly, the maximum concentrations of cadmium and lead were low, respectively 4 and 25 microg m(-3). Of the hydrides, tellurium and antimony could not be detected, but for the arsenic (arsine) and selenium hydrides measurable exposure occurred for almost all electrorefinery workers, although the levels were generally low at 0.2 microg m(-3).

Adult↗

Field uptake rates of hydrophobic organic contaminants by semipermeable membrane devices: environmental monitoring considerations.

The uptake rates of selected hydrophobic organic contaminants (HOCs) by semipermeable membrane devices (SPMDs)--a polyethylene layflat containing the lipid triolein--were investigated under natural conditions. SPMDs were exposed in three sampling sites (industrial, urban, and agricultural areas) in the Tajo River (Toledo, Spain) for 5, 11 and 20 d. The organochlorine compounds 4,4'-DDT, 4,4'-DDE, alpha-HCH, gamma-HCH, pentachlorobenzene, hexachlorobenzene, and polychlorinated biphenyls (PCBs), and the 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) were detected in the SPMDs deployed in the three sampling sites. A linear uptake rate was found for DDTs and for 4-Cl- and 5-Cl-substituted PCB congeners in all sampling sites. Concentrations of HCHs (80.3 ng g(-1) SPMD for alpha-HCH and 109 ng g(-1) SPMD for gamma-HCH after 20 d of exposure) increased according to a linear uptake rate in the SPMDs deployed in the sampling site located in the agricultural area. Likewise, a marked increase of total PAH concentration (up to 300 ng g(-1) SPMD after 20 d of exposure) was solely found in the sampling site situated near a thermoelectric power station. Examination of individual PAHs revealed that PAHs with log K(OW) between 4.2 and 5.7 displayed a linear uptake rate over the 20 d of exposure. Water concentrations (ng L(-1)) of HCB (0.80-2.48), lindane (1.30-11.5), 4,4'-DDT (0.61-2.02), 4,4'-DDE (6.89-11.6) and total PAHs (12.0-26.7) estimated by a linear uptake kinetic model were found to be high in comparison with other polluted aquatic systems, and similar to concentrations in other Spanish rivers. Our results suggest that SPMD kinetic uptake studies in the natural environment are recommended for identifying point-pollution sources, and that shorter times of SPMD exposure (approximately 1 week) are desirable to minimize one of the main problems of field SPMD deployment, i.e., the biofouling, which negatively affects the estimation of the dissolved HOC concentrations.

Agriculture↗

Environmental monitoring for genotoxicity with plant systems. Results and recommendations.

In the first phase of a collaborative study by the International Programme on Chemical Safety (IPCS), four coded chemicals, i.e. azidoglycerol (AG, 3-azido-1,2-propanediol), methyl nitrosourea (MNU), sodium azide (NaN3) and maleic hydrazide (MH), and ethyl methanesulfonate (EMS) as a positive control were tested in four plant bioassays, namely the Arabidopsis embryo and chlorophyll mutation assay, the Tradescantia stamen hair assay (Trad-SH assay), the Tradescantia micronucleus assay (Trad-MCN), and the Vicia faba root tip assay. Seventeen laboratories from diverse regions of the world participated with four to six laboratories each using one plant assay. For the Arabidopsis assay, laboratories were in agreement with MNU and AG giving positive responses and NaN3 giving a negative response. With the exception of one laboratory which reported MH as weakly mutagenic, no mutagenic response was reported for MH by the other laboratories. For the Vicia faba assay, all laboratories reported a positive response for MNU, AG, and MH, whereas two of the six laboratories reported a negative response for NaN3. For the Trad-SH assay, MH was reported as giving a positive response and a positive response was also observed for MNU with the exception of one laboratory. NaN3, which exhibited a relatively high degree of toxicity, elicited a positive response in three of the five laboratories. AG was found positive in only one of the two laboratories which tested this chemical. For the Trad-MCN assay, MNU and MH were reported as positive by all laboratories, while four out of five laboratories reported NaN3 to be positive. Only one of three laboratories reported AG to be positive. The major sources of variability were identified and considered to be in the same range as found in similar studies on other test systems. Recommendations were made for minor changes in methodology and for initiating the second phase of this study.

Arabidopsis↗

Environmental monitoring of occupational exposure to N,N-dimethylformamide: comparison between active and diffusive sampling.

OBJECTIVES: The objective of this study is to optimize the evaluation of the exposure to N,N-dimethylformamide (DMF) in synthetic leather factories by diffusive samplers. The DMF exposure was monitored in synthetic leather factories by two sampler types: active and diffusive. METHODS: Air measurements were carried out using two different personal air samplers, a diffusive and an active one. The diffusive sampling method, TK200 with charcoal filters, was examined in comparison with pumping through NIOSH silica gel tubes workplace air as with the currently available "gold standard". The evaluation was carried out, in two different years but in the same season, for all the duration of the shift, i.e. 8 h on workers employed in five different factories in the district of Florence and Prato (Italy). RESULTS: The statistical and graphical analysis of data show a good correlation between active and passive samplers (r = 0.96, P < 0.001, n = 91), a good linear regression (DMF(diffusive )= 0.95 DMF(active) + 0.15, R (2) = 0.92), a not statistically significant difference between data (tested by paired t test and non-parametric Wilcoxon test). Moreover, all these results are confirmed for data lower and higher than TLV-TWA, in particular we found a significant Pearson correlation (r = 0.92, P < 0.001, n = 83; r = 0.92, P < 0.05, n = 8, respectively) and a significant linear regression (DMF(diffusive )= 0.88 DMF(active) + 0.73, R (2 )= 0.86; DMF(diffusive )= 0.90 DMF(active) + 3.76, R (2 )= 0.85). Besides, the analysis of graphical representations confirmed the previous evidences. Finally, we can not find a significant difference between different types of job. CONCLUSIONS: Due to the good agreement between the two groups of data, the TK200 samplers can be considered as a simpler approach than the pump for screening worker exposures to DMF.

Air Pollutants, Occupational↗

Measurement and application of uranium isotopes for human and environmental monitoring.

An improved method is described utilizing extraction chromatography, anion exchange, and alpha spectroscopy for measurement of uranium isotopes in human and environmental surveillance studies. These methods provide a sensitivity of approximately 0.7 mBq per isotope per sample and are generally accurate within the precision of the measurements. The extraction chromatography methods greatly simplify separation of uranium from iron in silicate matrices and provide increased sample throughput and data quality for water, soil, and air filter samples. For bioassay samples, the coprecipitation/anion exchange/alpha spectrometric methods provide rapid throughput and sufficient sensitivity to meet new analytical performance standards in human monitoring studies. In addition, the 234U:238U data can be used as a fingerprint of natural vs. anthropogenic sources of uranium. For 1995 data from our laboratory, a large percentage (79-94% by matrix) of samples appear to be of natural 234U:238U isotopic composition. For all matrices, samples with higher uranium concentration generally have more depleted isotopic composition (smaller 234U:238U). A small percentage of soil (11%), air filter (3%), urine (3%), and water (3%) samples have depleted isotopic signatures at the 95% confidence interval, indicating anthropogenic contributions of uranium to these samples.

Air Pollutants, Radioactive↗

Mutagenic/carcinogenic hazards in a cold-rolling steel plant exposed to mineral oils: environmental monitoring phase.

A survey was carried out in a cold-rolling steel plant exposed to mineral oils to study the mutagenic/carcinogenic hazards, following a coupled environmental/biological monitoring. The present paper deals only with the environmental phase by determining polycyclic aromatic hydrocarbons (PAH) and applying a mutagenicity test (Ames test), and includes the following steps: (a) work process and environment study; (b) sampling and analysing mineral oils before and after use (PAH determination and mutagenicity analysis); (c) sampling and analysing oil mist (TWA determination, PAH determination and mutagenicity analysis). The results showed that: (1) both unused and used (recycled) mineral oils contained only trace amounts of PAH and were not mutagenic; (2) the TWA concentrations of oil mists were lower than the TLV (less than 5 mg/m3); (3) oil mists contained only trace amounts of PAH and were not mutagenic. The authors suggest that these results could be due to the moderate temperature of the oil during the work process and that there is a relationship between low PAH content and absence of mutagenicity in the oils and air samples.

Air Pollutants, Occupational↗

A comparison of alcohol ethoxylate environmental monitoring data using different analytical procedures.

Several analytical methods have been developed for analyzing alcohol ethoxylates (AE) in aqueous environmental samples. These methods differ widely in their selectivity and sensitivity for measuring the AE components; that is, they vary in their resolution of alkyl chain length homologs and ethoxymer distributions (degree of ethoxylation for each homolog). Given these differences, AE monitoring results from different studies often are inconsistent and, sometimes, are deemed to be incomparable. To address these differences, three currently available methods for determining AE concentrations in environmental matrices were compared among a common set of wastewater treatment plant samples. These methods included the detection of hydrogen bromide-derivatized homologs by gas chromatography/mass spectrometry, the detection of aqueous homologs by high-pressure liquid chromatography/electrospray mass spectrometry, and the detection of pyridinium-derivatized homologs by high-pressure liquid chromatography/electrospray mass spectrometry. Results from the present study showed that all three methods responded differently in determining the complex suite of chemical species that comprise AE in the environment. The collective information, however, allowed a consistent comparison among the methods. This comparison was then used to reevaluate results from previous AE monitoring studies. Results from this reevaluation provided a more realistic profile of both historical AE removal during wastewater treatment as well as the occurrence of AE in U.S. surface waters.

Alcohols↗

Long-term environmental monitoring and application of low-level 3H, 7Be, 137Cs and 210Pb activity concentrations in the non-biotic compartments of the Danube in Austria.

The aim, methods, and results of long-term low-level measurements of 3H, 7Be, 137Cs and 210Pb in water and sediment of the Austrian part of the Danube are presented. The results are discussed focused on recent and obvious future applications in applied radioecology and environmental research. The long-term radiometric data supported by the low-level monitoring program act as basis for ecological modelling and environmental assessment.

Austria↗

Functional analysis of a metal response element in the regulatory region of flounder cytochrome P450 1A and implications for environmental monitoring of pollutants.

Cytochrome P450 1A (CYP 1A) is a member of a multigene family of xenobiotic metabolizing enzymes. CYP 1A is highly inducible by numerous environmental contaminants including polycyclic aromatic hydrocarbons (PAHs) and is widely used in biomonitoring studies. Therefore, understanding the regulation of this gene is important for accurate interpretation of biomarker data. We describe here the functional role of a metal response element (MRE) in the European flounder CYP 1A promoter region. To help elucidate the potential role of this MRE, reporter gene constructs, with or without site-directed mutagenesis, were used in conjunction with a dual-luciferase assay. The electrophoretic mobility shift assay (EMSA) was also used to investigate potential protein binding at this MRE site. Treatment with the prototypical PAH 3-methylcholanthrene (3MC) (1.0 microM) produced a dose-dependent response at the CYP 1A promoter, whereas treatment with cadmium (0-1.0 microM) produced little transcriptional activity at either the wild-type or mutated promoter. Cotreatment with cadmium (1.0 microM) and 3MC (1.0 microM) reduced induction at this promoter to 1.83-fold compared to 3MC treatment alone (4.0-fold induction). Mutation of the MRE site resulted in abolishment of this cadmium-related loss of 3MC-dependent activity. Furthermore, a retarded band was observed in the EMSA when the MRE was used as a probe and incubated with liver nuclear protein from flounder treated with cadmium. The results not only add to knowledge of the diversity in vertebrate CYP 1A regulation but also raise the complexity of interpretation of CYP 1A induction in monitoring studies that involve mixtures of PAHs and metals.

Animals↗

Applications of electrochemical immunosensors to environmental monitoring.

This paper discusses basic electrochemical immunoassay technology. Factors limiting the practical application of antibodies to analytical problems are also presented. It addresses the potential use of immunoassay methods based on electrochemical detection for the analysis of environmental samples. It provides examples for the detection and quantitation of environmental samples using conducting electroactive polymers (CEPs). CEP-based immunosensing systems are compared with conventional environmental immunoassay procedures. The advantages of using these types of sensors for rapid, sensitive, and cost-effective analysis of pesticides and toxic chemicals are analysed and discussed. CEP-based immunosensing technology might eventually be used for continuous monitoring of effluents such as waste streams to determine compliance with regulations. CEP-based sensors are suitable for monitoring ground-water, waste stream effluents, agricultural run-offs and for monitoring the effectiveness of remediation, or for other situations where a real-time monitoring capability is desired.

Biosensing Techniques↗

EC intercomparisons for laboratories monitoring environmental radioactivity.

International measurement comparisons are organised regularly for EU laboratories involved in monitoring radioactivity, with emphasis on meeting routine measurement conditions. Using the recent comparison of 137Cs in air filters as an example, the whole cycle is described: establishment of traceable reference values, spiking of individual filters for the comparison and their quality assurance, treatment and measurement of filters in the participating laboratories and evaluation of comparison results. The treatment of an individual result, deviating widely from the reference value, is discussed. Monte-Carlo simulations allow to estimate the maximum errors possibly made due to a non-suitable measurement geometry.

Air Pollutants, Radioactive↗

Environmental monitoring in a laboratory animal facility.

A study was made of the microbial environmental status of an animal facility. Cultures were made of animal and surgical room floors; the germicidal effectiveness of the phenolic disinfectant-detergent employed in the facility was tested against standard test organisms as well as against other microorganisms isolated from the facility, and killing power of the disinfectant-detergent was evaluated during various steps of the usual cleaning procedures, ie, mops and mop bucket solutions were tested before, during, and after mopping a room. It was found that colony counts for animal rooms cleaned with a chlorhexidine disinfectant were much lower than those cleaned with a phenolic disinfectant. The phenolic disinfectant killed some organisms after 10 min exposure, but no others. Pseudomonads were the most resistant organisms. Contaminated mops and mop bucket solutions appeared responsible for the high counts on floors cleaned with the phenolic disinfectant. Guidelines for achievable levels of cleanliness were suggested.

Animals↗

[Biological and environmental monitoring of exposure to chlorinated aromatic hydrocarbons].

Polychlorinated hydrocarbons due to their lipophilic properties can cumulate in animal organisms by entering of various food chains. Food, especially of animal origin, is regarded as a major source of these compounds for man. For several years the Department of Environmental Toxicology of the National Institute of Hygiene jointly with sanitary epidemiological stations perform the monitoring of organochlorine compounds in food, including infant formula and breast milk. The results of this monitoring are presented in this paper. In order to assess the environmental exposure of man to organochlorine insecticides (DDT and HCH) and other contaminants i.e. PCBs and HCB were chosen. Samples were taken in 18 voivodships in Poland. The results show that the exposure to above compounds from food of plant origin is negligible, and that the food of animal origin is the predominant source of intake of these compounds. The results of biological monitoring show that infants fed with breast milk are of greatest risk.

Animals↗