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Determination of perchlorate in drinking water by ion chromatography using macrocycle-based concentration and separation methods.

Macrocycle-based ion chromatography provides a convenient, reliable method for the determination of perchlorate ion, which is currently of great interest to the environmental community. This study shows that effective perchlorate determinations can be made using standard conductimetric detection by combining an 18-crown-6-based mobile phase with an underivatized reversed-phase mobile phase ion chromatography (MPIC) column. One unique feature of this method is the flexibility in column capacity that is achieved through simple variations in eluent concentrations of 18-crown-6 and KOH, facilitating the separation of target analyte anions such as perchlorate. Using a standard anion exchange column as concentrator makes possible the determination of perchlorate as low as 0.2 ug/L in low ionic strength matrices. Determination of perchlorate at the sub-ug/L level in pure water and in spiked local city hard water samples with high background ion concentrations can be achieved this way. However, like other IC techniques, this method is challenged to achieve analyses at the ug/L level in the demanding high ionic strength matrix described by the United States Environmental Protection Agency (EPA) (1,000 mg/L chloride, sulfate and carbonate). We approached this challenge by use of the Cryptand C1 concentrator column, provided by Dionex Corporation, to effectively preconcentrate perchlorate while reducing background ion concentrations in the high ionic strength matrix. The retention characteristics of the concentrator column were studied in order to maximize its effectiveness for perchlorate determinations. The method makes possible the determination of perchlorate at the 5 ug/L level in the highest ionic strength matrix described by the EPA.

Chromatography, Ion Exchange↗

Use of a physiologically based pharmacokinetic model to identify exposures consistent with human biomonitoring data for chloroform.

Biomonitoring data provide evidence of human exposure to environmental chemicals by quantifying the chemical or its metabolite in a biological matrix. To better understand the correlation between biomonitoring data and environmental exposure, physiologically based pharmacokinetic (PBPK) modeling can be of use. The objective of this study was to use a combined PBPK model with an exposure model for showering to estimate the intake concentrations of chloroform based on measured blood and exhaled breath concentrations of chloroform. First, the predictive ability of the combined model was evaluated with three published studies describing exhaled breath and blood concentrations in people exposed to chloroform under controlled showering events. Following that, a plausible exposure regimen was defined combining inhalation, ingestion, and dermal exposures associated with residential use of water containing typical concentrations of chloroform to simulate blood and exhaled breath concentrations of chloroform. Simulation results showed that inhalation and dermal exposure could contribute substantially to total chloroform exposure. Next, sensitivity analysis and Monte Carlo analysis were performed to investigate the sources of variability in model output. The variability in exposure conditions (e.g., shower duration) was shown to contribute more than the variability in pharmacokinetics (e.g., body weight) to the predicted variability in blood and exhaled breath concentrations of chloroform. Lastly, the model was used in a reverse dosimetry approach to estimate distributions of exposure consistent with concentrations of chloroform measured in human blood and exhaled breath.

Chloroform↗

The effects of environmental complexity on the hippocampal formation of the adult rat.

In an effort to determine the plasticity of the proliferative matrix of the hippocampus in an adult rat to environmental manipulation, the following study was conducted. Fifteen adult male Long-Evans rats (6 months old at the start of the experiment) were placed into two different environments for 3 months. Seven animals represented the enriched condition and 8 represented the condition of isolation. At the conclusion of exposure to the different environments, the animals were sacrificed by transcardial perfusion and the dorsal hippocampi examined in matched sections for all animals. Results indicated heavier total brain weights and larger numbers of granule cells in the dorsal hippocampus for animals exposed to the enriched environment. Implications for CNS plasticity were discussed.

Animals↗

Environmental regulation of curli production in Escherichia coli.

Curli are novel surface organelles on E. coli that mediate binding to soluble matrix proteins. The expression of curli is affected by environmental factors, such as temperature, osmolarity, and growth conditions. Curli formation is regulated at the level of transcription, in that the csgA gene can be transcriptionally activated by the cytosolic Crl protein or transcriptionally relieved by a mutation in hns. The expression of curli is also dependent on functional RpoS. E. coli--expressing curli bind to human skin tissue, provided they are precoated with soluble fibronectin, suggesting that curli may act as a colonization factor in host-microbe interactions. Fibronectin is a multifunctional extracellular matrix and plasma protein involved in cell adhesion and cell spreading. It also interacts with a variety of microorganisms, and thus the role of fibronectin in mediating binding of curliated E. coli is of great interest. An investigation of the epitopes of both the fibronectin molecule and the curlin subunit protein involved in the binding of E. coli to tissue will give us more insight into the initial colonization of host surfaces by bacteria.

Amino Acid Sequence↗

Serum stimulation, cell-cell interactions, and extracellular matrix independently influence smooth muscle cell phenotype in vitro.

Vascular injury profoundly alters the vessel wall microenvironment, and smooth muscle cells respond with cell cycle re-entry, loss of contractile elements, extracellular matrix remodeling, and altered signaling by endogenous growth factors and their receptors. Environmental cues include stimulation by exogenous mitogens and both cell-cell and cell-matrix interactions. Modeling this process in smooth muscle cells in vitro, these environmental determinants were varied independently and the phenotypic consequences assessed. Mitogenic stimulation with serum promoted the synthesis of collagen and fibronectin and the expression of fibroblast growth factor receptor-1 and suppressed the content of smooth muscle alpha-actin, myosin heavy chain, and basic fibroblast growth factor. Low cell density (reduced cell-cell contact) was also associated with enhanced extracellular matrix protein production, increased fibroblast growth factor receptor-1 expression, and reduced contractile protein and basic fibroblast growth factor content. The influence of serum stimulation and reduced cell-cell contact were independent and additive. Provision of a type I collagen matrix blunted the influence of serum and cell-cell contact on collagen synthesis but had minor effects on other measures of phenotype. Environmental factors thus independently influence smooth muscle cell phenotype, including endogenous growth factor expression and responsiveness, which can in turn influence the microenvironment of the vessel wall after injury.

Actins↗

Sampling of water, soil and sediment to trace organic pollutants at a river-basin scale.

Sampling is considered a crucial step in the analysis of organic compounds in the environment. This review describes field sampling techniques and provides detailed step-by-step procedures for collection and preservation of all major environmental matrices (water, sediment and soil) integrated as part of the river-basin water cycle. Attention is given to the prerequisites for obtaining reliable samples, and the practical issues of sample collection (planning, field sampling, sampling strategies and equipment and data quality assessment) are considered. Considering the heterogeneity of environmental matrices, special considerations for each matrix are given to solve typical problems and to find the most appropriate solutions to ensure the quality of the sample. The procedures described in the next sections are commonly used protocols that reflect true field conditions and current state-of-the-art techniques used in the sampling of organic compounds. The aim is to signify the importance of sampling to the overall analytical procedure. Finally, quality control issues to be considered in environmental sampling are given.

Geologic Sediments↗

A new luciferase reporter gene assay for the detection of androgenic and antiandrogenic effects based on a human prostate specific antigen promoter and PC3/AR human prostate cancer cells.

We developed a new mammalian cell-based luciferase reporter gene assay for androgenic and antiandrogenic activities of chemicals and environmental samples. Environmental samples usually have a complex matrix that may contain the constituents acting as androgen receptor (AR) agonists, AR antagonists or aryl hydrocarbon receptor (AhR) agonists. AhR agonists are known to elicit the antiandrogenic effect through cross-talk between AR and AhR signal transduction pathways. In this study, PC3/AR human prostate carcinoma cells were transiently transfected with a prostate-specific antigen (PSA) promoter-driven luciferase expression plasmid. The cells were treated with a test compound or an environmental sample for 24 h at 37 degrees C and then measured for luciferase activity. The luciferase activity was induced by dihydrotestosterone (DHT) in a concentration-dependent manner in a concentration range from 10 fM to 1 nM. R1881, a synthetic androgen receptor agonist, induced luciferase activity and its inductive effects was additive to that of DHT. The luciferase activity was not induced by cortisol, a glucocorticoid, progesterone, a progestin, and 17beta-estradiol, an estrogen in a concentration range of up to 1 microM. DHT-induced luciferase activity was reduced by bicalutamide and cyproterone acetate, AR antagonists, and also by benzo[a]pyrene, an aryl hydrocarbon receptor agonist, through AhR-mediated pathways. All of these findings indicate that the present assay system correctly responds to AR agonists, AR antagonists and AhR agonist and, therefore, it is a powerful tool for the sensitive and selective screening of chemicals and environmental samples for their androgenic and antiandrogenic activities. We developed the first assay system, in which the expression of luciferase was driven by the promoter of a prostate-specific antigen gene, a typical human androgen-regulated gene.

Androgen Antagonists↗

Regulation of expression of the chondrocytic phenotype in a skeletal cell line (CFK2) in vitro.

We have examined in vitro the spontaneous and regulated expression of phenotypic characteristics associated with differentiated chondrocytes in an established skeletal cell line (CFK2) derived from fetal rat calvariae. Extended culture of CFK2 cells resulted in the appearance of glycosaminoglycans and type II collagen in the cell layer in association with the formation of focal nodes of cells. In addition, induction of mRNA-encoding link protein, cartilage-specific proteoglycan core protein, and thrombospondin was observed in the differentiated population (dCFK2 cells). The expression of these mRNAs was present for at least two passages after subculturing the dCFK2 cells. The dCFK2 cells also demonstrated enhanced parathyroid hormone (PTH)-stimulated adenylate cyclase activity. Proliferation of CFK2 cells was stimulated by the peptide regulatory factors EGF and PTH and inhibited by the steroidal agents dexamethasone and retinoic acid. EGF and retinoic acid inhibited the formation of cell foci and glycosaminoglycan deposition and the expression of mRNA-encoding link protein. In contrast, PTH and dexamethasone enhanced the formation of focal cellular nodes and augmented matrix deposition and link protein mRNA expression. These studies therefore show that the CFK2 cell line can serve as a nontransformed model of rat chondrocytic cells in which both induction and regulation of the expression of cartilaginous matrix components can be observed. This line thereby provides a unique renewable source of chondrocytic precursor cells and an excellent in vitro model for evaluating temporal and environmental control of chondrocyte differentiation and cartilage matrix production.

Adenylyl Cyclases↗

Guidelines for analysis of human milk for environmental chemicals.

When analyzing human milk for environmental chemicals, in either a monitoring or research project, researchers must ensure that the technique used for measuring the target analytes complies with certain analytical criteria and that a quality control/quality assurance program is in place. In this report, these analytical criteria and the accompanying quality assurance program are elaborated in more detail. Examples of representative methods are also given for the determination of a number of classes of chemicals of environmental concern, such as polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, polychlorinated biphenyls, volatile organics, and halogenated flame retardants. Most of these techniques are physical/chemical in nature, but the possibility of the use of bioassays for screening of analytes in human milk is noted. The described methods are not exclusive, but any laboratory undertaking the analysis of human milk should demonstrate proficiency in their execution. This is usually accomplished by producing reliable data in interlaboratory studies. Human milk is an ideal matrix for estimating exposure to certain environmental chemicals, and researchers must ensure that the techniques used to determine these compounds adhere to known and accepted analytical requirements.

Environmental Monitoring↗

Optical biosensing of nitrite ions using cytochrome cd1 nitrite reductase encapsulated in a sol-gel matrix.

Nitrite is an important human health and environmental analyte. As such, the European Union (EU) has imposed a limit for nitrite in potable water of 0.1 mg l-1 (2.18 microM). In order to develop an optical biosensing system for the determination of nitrite ions in environmental waters, cytochrome cd1 nitrite reductase has been extracted and purified from the bacterium Paracoccus pantotrophus. The protein has been spectroscopically characterised in solution and important kinetic parameters of nitrite reduction of the cytochrome cd1 enzyme, i.e., Km, Vmax and kcat have been determined. The influence of pH on the activity of the cytochrome cd1 has been investigated and the results suggest that this enzyme can be used for the determination of nitrite in the pH range 6-9. Biosensing experiments with the cytochrome cd1 in solution suggested that the decrease in intensity of the absorption band associated with the d1 haem (which is the nitrite binding site), at 460 nm, with increasing nitrite concentrations would enable the measurement of this analyte with the optimum limit of detection. The cytochrome cd1 has been encapsulated in a bulk sol-gel monolith with no structural changes observed and retention of enzymatic activity. The detection of nitrite ions in the range 0.075-1.250 microM was achieved, with a limit of detection of 0.075 microM. In order to increase the speed of response, a sol-gel sandwich thin film structure was formulated with the cytochrome cd1. This structure enabled the determination of nitrite concentrations within ca. 5 min. The sol-gel sandwich entrapped cytochrome cd1 enzyme was found to be stable for several months when the films were stored at 4 degrees C.

Biosensing Techniques↗

Adaptation of an amphibian mucociliary clearance model to evaluate early effects of tobacco smoke exposure.

RATIONALE: Inhaled side-stream tobacco smoke brings in all of its harmful components impairing mechanisms that protect the airways and lungs. Chronic respiratory health consequences are a complex multi-step silent process. By the time clinical manifestations require medical attention, several structural and functional changes have already occurred. The respiratory system has to undergo an iterative process of injury, healing and remodeling with every exposure. METHODS: To have a better understanding of the initial changes that take place when first exposed to environmental tobacco smoke, we have developed an exposure model, using the frog palate that closely represents the features of obstructive airways where ciliary dysfunction and mucus hypersecretion occur. RESULTS: Mucus transport was significantly reduced, even after exposure to the smoke of one cigarette (p < 0.05) and even further with 4-cigarettes exposure (p < 0.001). Morphometric and ultrastructural studies by SEM show extensive areas of tissue disruption. Gelatinase zymography shows activation of MMP9 in mucus from palates exposed to tobacco smoke. CONCLUSIONS: The clearance of mucus on the frog palate is significantly reduced after exposure to environmental tobacco smoke. Cilia and the extracellular matrix are anatomically disrupted. Tobacco smoke triggers an increased activity of matrix metalloproteinases associated with a substantial defoliation of ciliated epithelium. These studies enhance the knowledge of the changes in the mucociliary apparatus that occur initially after exposure to environmental tobacco smoke, with the goal of understanding how these changes relate to the genesis of chronic airway pathologies in humans.

Animals↗

Microbe-surface interactions in biofouling and biocorrosion processes.

The presence of microorganisms on material surfaces can have a profound effect on materials performance. Surface-associated microbial growth, i.e. a biofilm, is known to instigate biofouling. The presence of biofilms may promote interfacial physico-chemical reactions that are not favored under abiotic conditions. In the case of metallic materials, undesirable changes in material properties due to a biofilm (or a biofouling layer) are referred to as biocorrosion or microbially influenced corrosion (MIC). Biofouling and biocorrosion occur in aquatic and terrestrial habitats varying in nutrient content, temperature, pressure and pH. Interfacial chemistry in such systems reflects a wide variety of physiological activities carried out by diverse microbial populations thriving within biofilms. Biocorrosion can be viewed as a consequence of coupled biological and abiotic electron-transfer reactions, i.e. redox reactions of metals, enabled by microbial ecology. Microbially produced extracellular polymeric substances (EPS), which comprise different macromolecules, mediate initial cell adhesion to the material surface and constitute a biofilm matrix. Despite their unquestionable importance in biofilm development, the extent to which EPS contribute to biocorrosion is not well-understood. This review offers a current perspective on material/microbe interactions pertinent to biocorrosion and biofouling, with EPS as a focal point, while emphasizing the role atomic force spectroscopy and mass spectrometry techniques can play in elucidating such interactions.

Bacterial Physiological Phenomena↗

Food web dynamics in correlated and autocorrelated environments.

The densities of populations in a community or food web vary as a consequence of both population interactions and environmental (e.g. weather) fluctuations. Populations often respond to the same kinds of environmental fluctuations, and therefore experience correlated environments. Furthermore, some environmental factors change slowly over time, thereby producing positive environmental autocorrelation. We show that the effects of environmental correlation and autocorrelation on the dynamics of the populations in a food web can be large and unintuitive, but can be understood by analyzing the eigenvectors of the community (system) matrix of interactions among populations. For example, environmental correlation and autocorrelation may either obscure or enhance the cyclic dynamics that generally characterize predator-prey interactions even when there is no direct effect of the environment on how species interact. Thus, understanding the population dynamics of species in a food web requires explicit attention to the correlation structure of environmental factors affecting all species.

Algorithms↗

Interaction between the aryl hydrocarbon receptor and retinoic acid pathways increases matrix metalloproteinase-1 expression in keratinocytes.

Exposure to the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) results in a variety of pathological lesions in humans via activation of the aryl hydrocarbon receptor (AhR) pathway. It has become apparent that this pathway interacts with a variety of signaling pathways that are believed to be involved in mediating TCDD/AhR biological effects. Our hypothesis is that TCDD mediates these pathological lesions by directly altering the expression of genes involved in matrix deposition and remodeling and that the retinoic acid signaling pathway is involved in modulating TCDD-induced effects. Therefore, we examined the effect of TCDD and all-trans retinoic acid (atRA) on the expression of matrix metalloproteinase-1 (MMP-1, interstitial collagenase), one of the proteolytic enzymes that degrade type I collagen, in normal human keratinocytes. The data show that TCDD exposure results in increased MMP-1 expression in keratinocytes that is further enhanced by co-treatment with all-trans retinoic acid. TCDD-induced expression of MMP-1 appears to be mediated through two AP-1 elements in the proximal promoter of the MMP-1 gene. However, retinoic acid-mediated induction of keratinocyte MMP-1 is a result of both promoter activation and increased mRNA stability. These findings are the first to demonstrate TCDD-induced expression of MMP-1 and to demonstrate interactions between the TCDD/AhR and retinoic acid pathways on MMP-1 expression.

Blotting, Northern↗

2,3,7,8-tetrachlorodibenzo-p-dioxin specifically reduces mRNA for the mineralization-related dentin sialophosphoprotein in cultured mouse embryonic molar teeth.

Previous studies show that the most toxic dioxin congener, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), interferes with mineralization of the dental matrices in developing mouse and rat teeth. Culture of mouse embryonic molar teeth with TCDD leads to the failure of enamel to be deposited and dentin to undergo mineralization. Lactationally exposed rats show defectively matured enamel and retardation of dentin mineralization. To see if the impaired mineralization is associated with changes in the expression of dentin sialophosphoprotein (Dspp), Bono1 and/or matrix metalloproteinase-20 (MMP-20), thought to be involved in mineralization of the dental hard tissues, we cultured mouse (NMRI) E18 mandibular molars for 3, 5 or 7 days and exposed them to 1 microM TCDD after 2 days of culture. As detected by in situ hybridization of tissue sections, localization and intensity of Bono1 and MMP-20 expression showed no definite difference between the control and exposed tooth explants, suggesting that TCDD does not affect their expression. On the contrary, TCDD reduced or prevented the expression of Dspp in secretory odontoblasts and decreased it in presecretory ameloblasts. The results suggest that the retardation of dentin mineralization by TCDD in mouse molar teeth involves specific interference with Dspp expression.

Animals↗

Modelling the structure and function of extracellular polymeric substances in biofilms with new numerical techniques.

The objective of this study is the mathematical description of the structure and function of the extracellular polymeric substances (EPS) in biofilms. The basic assumptions of the EPS biofilm model are: the production of EPS in biofilms is coupled to the growth of micro-organisms the production of EPS is additionally coupled to the substrate conditions the EPS represent a considerable volume fraction of the matrix in biofilms and thus the density of the biofilms is strongly influenced by the EPS sorption of biocides and pollutants in biofilms occurs mainly to EPS the EPS can be used as an energy source during substrate limited phases. The mathematical model has been derived as a system of partial differential equations. The numerical solution of these complex balance equations has been done by a self-adaptive Galerkin-h-p-method. It can be shown, that on the one hand the simulation of substrate conversion and biofilm growth with the EPS-biofilm model yields similar results as the known biofilm models without consideration of the EPS fraction. On the other hand the advantage of the EPS-biofilm model is a better understanding of biofilm structure, which is mainly influenced by the EPS fraction in the biofilm. Furthermore, the sorption of pollutants, such as heavy metals and chlorinated organic substances, can be simulated in more detail.

Adsorption↗

PsB multiprotein complex of Dictyostelium discoideum. Demonstration of cellulose binding activity and order of protein subunit assembly.

The differentiated spores of Dictyostelium are surrounded by an extracellular matrix, the spore coat, which protects them from environmental factors allowing them to remain viable for extended periods of time. This presumably is a major evolutionary advantage. This unique extracellular matrix is composed of cellulose and glycoproteins. Previous work has shown that some of these spore coat glycoproteins exist as a preassembled multiprotein complex (the PsB multiprotein complex) which is stored in the prespore vesicles (Watson, N., McGuire, V., and Alexander, S (1994) J. Cell Sci. 107, 2567-2579). Later in development, the complex is synchronously secreted from the prespore vesicles and incorporated into the spore coat. We now have shown that the PsB complex has a specific in vitro cellulose binding activity. The analysis of mutants lacking individual subunits of the PsB complex revealed the relative order of assembly of the subunit proteins and demonstrated that the protein subunits must be assembled for cellulose binding activity. These results provide a biochemical explanation for the localization of this multiprotein complex in the spore coat.

Animals↗

Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase MMP-9 expression in glial cells.

Inorganic lead (Pb) is a ubiquitous environmental contaminant that produces a variety of deleterious effects in the central nervous system (CNS). Matrix metalloproteinases (MMPs), specifically MMP-9, induced by inflammatory cytokines, are increasingly being implicated in CNS pathology. The present study demonstrates that low concentrations of either pro-inflammatory cytokines (TNF-alpha and IL-1beta) or Pb did not influence the MMP-9 expression in a glial cell line (C6) when added separately. However, combined administration of Pb and cytokines induced a marked synergized elevation of MMP-9 expression in spite of a reduction in the number of glial cells. These results demonstrate a possible new mechanism by which Pb may induce neuropathological processes.

Animals↗