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The toxicity of brominated and mixed-halogenated dibenzo-p-dioxins and dibenzofurans: an overview.

Brominated dibenzo-p-dioxins and dibenzofurans can be formed under laboratory conditions by pyrolysis of flame retardants based on polybrominated biphenyls and biphenyl ethers. Their occurrence in the environment, however, is due to combustion processes such as municipal waste incineration and internal combustion engines. As these processes generally take place in the presence of an excess of chlorine, predominantly mixed brominated and chlorinated compounds have been identified so far in environmental samples. Brominated dibenzo-p-dioxins or dibenzofurans bind to the cytosolic Ah receptor about as avidly as their chlorinated congeners and induce hepatic microsomal enzymes with comparable potency. The same holds true for mixed brominated-chlorinated compounds. Gross pathologic symptoms-hypothyroidism, thymic atrophy, wasting of body mass, lethality-also occur at doses that, on a molar concentration basis, are virtually identical to those seen with the chlorinated compounds. Their potency to induce malformations in mice following prenatal exposure is equivalent to that of chlorinated dibenzo-p-dioxins and dibenzofurans. Possible activities as (co)carcinogens and endocrine disrupters have not been evaluated, but are likely to exist. Considering the overall similarity in action of chlorinated and brominated dibenzo-p-dioxins and dibenzofurans, environmental and health assessments should be based on molar body burdens without discrimination for the nature of the halogen.

Animals↗

Distribution of volatile halogenated organic compounds between rat blood serum and adipose tissue.

A 31-day rat-dosing sequence was used to study some aspects of the physiological distribution, metabolism, storage and rate of elimination of chloroform, trichloroethylene, carbon tetrachloride and bromodichloromethane from rat adipose tissue and blood serum. Data points were collected nine times during the dosing period and twice after dosing had ceased. Purge/trap/desorb methodology was employed using a liquid sample concentrator, a gas-liquid chromatograph and a Hall electrolytic conductivity detector in series. Compound identities were confirmed using a GC/MS analytical system. For these volatile compounds, tissue levels fluctuated but did not indicate increased storage with time. Adipose tissue to blood serum levels never differed by more than a factor of three. Within 3-6 days after dosing was terminated, practically all of the halogenated compounds had left the examined tissues. Metabolic conversion of trichloroethylene and carbon tetrachloride into chloroform and/or a chloroform precursor was observed; bromodichloromethane-dosed animals did not have serum chloroform levels exceeding those observed for the control animals.

Adipose Tissue↗

Formation of halide-ions on one-electron reduction of halogenated nitroimidazoles in aqueous solution. A radiolytic study.

Reactions of e-aq, (CH3)2COH and CO.2- with a series of halogenated nitroimidazoles (ArNO2) results predominantly in the formation of their one-electron reduced species (ArNO.2-), the properties of which are consistent with those of other nitroimidazole radical anions. Further, one-electron reduction of ArNO2 ultimately leads to production of halide ion (X-); G(X-) = 1.2 to 2.2. In the presence of either the nitroxyl free radical, norpseudopelletierine-N-oxyl (NPPN), or oxygen the yield of X- is reduced and 'residual' yields of X- were observed, G(X-) = 0.08 to 0.43. From the results it is suggested that less than or equal to 10 per cent of the yield of reducing species results in the production of X- and an imidazolyl-type radical and that the majority of the yield of X- results from subsequent reactions of ArNO.2- (e.g. bimolecular reactions).

Halogens↗

Halogenated secondary metabolites from Laurencia obtusa.

A new sesquiterpene (1), and a halogenated C15 acetogenin (2), a stereoisomer of neoisoprelaurefucin were isolated from Laurencia obtusa. Four known compounds laurencienyne (3), rogiolenyne B (4), obtusenol (5), and (3E)-dactomelyne (6) were also isolated from this alga. Rogiolenyne B (4) and (3E)-dactomelyne (6) were found for the first from this species. The structures of these compounds were elucidated by spectroscopic methods. The unambiguous assignments of the 1H and 13C NMR spectral data of (5) and 13C NMR data of (6) were also reported for the first time.

Acetogenins↗

Quantum descriptors for predictive toxicology of halogenated aliphatic hydrocarbons.

In order to improve Quantitative Structure-Activity Relationships (QSARs) for halogenated aliphatics (HA) and to better understand the biophysical mechanism of toxic response to these ubiquitous chemicals, we employ improved quantum-mechanical descriptors to account for HA electrophilicity. We demonstrate that, unlike the lowest unoccupied molecular orbital energy, ELUMO, which was previously used as a descriptor, the electron affinity can be systematically improved by application of higher levels of theory. We also show that employing the reciprocal of ELUMO, which is more consistent with frontier molecular orbital (FMO) theory, improves the correlations with in vitro toxicity data. We offer explanations based on FMO theory for a result from our previous work, in which the LUMO energies of HA anions correlated surprisingly well with in vitro toxicity data. Additional descriptors are also suggested and interpreted in terms of the accepted biophysical mechanism of toxic response to HAs and new QSARs are derived for various chemical categories that compose the data set employed. These alternate descriptors provide important insight and could benefit other classes of compounds where the biophysical mechanism of toxic response involves dissociative attachment.

Chemical Phenomena↗

Photocatalytic degradation of p-halophenols in TiO2 aqueous suspensions: halogen effect on removal rate, aromatic intermediates and toxicity variations.

The influence of the halogen upon the TiO(2) photocatalytic degradation of p-halophenols in water has been investigated. Phenol was used as the reference compound. Compared with its value for phenol, the apparent first-order rate constant of removal, k, was slightly but significantly higher for p-fluorophenol and p-chlorophenol, and slightly but significantly lower for p-bromophenol. For p-iodophenol, k was about half that of phenol. The relative values confirm that k is roughly correlated to the Hammett constant; this constant reflects the electron density on the aromatic ring and, accordingly, the reactivity towards electrophilic species generated by UV-irradiation of TiO(2). All compounds were found to be poorly adsorbed on TiO(2). Accordingly, k was not related to the differences observed in the very low adsorbed amounts. The detected aromatic intermediate products included hydroquinone (HQ), benzoquinone (BQ) and various halodihydroxybenzenes. HQ, BQ, 4-chloro-1,2 (and 1,3)-dihydroxybenzenes and 4-bromo-1,3-dihydroxybenzene were quantified. Mechanisms are tentatively suggested to interpret the differences in the degradation pathways of the p-halophenols. The organic intermediate products accounted for only a few percents of the total carbon during the degradation. The toxicity (1/EC50) measured by the Microtox test almost did not vary in the course of the degradation of phenol, p-chlorophenol and p-bromophenol until complete removal of these compounds. By contrast, the value of 1/EC50 was multiplied by ca. 2.5 when ca. 45% of p-iodophenol had been removed; concentrations of BQ higher than with the other p-halophenols are tentatively suggested to be at the origin of this increase. Interpretation of a surprising substantial increase in the 1/EC50 value when the removal of p-fluorophenol increased from 80 to 95% requires further investigation.

Adsorption↗

Molecular determinants of hormone mimicry: halogenated aromatic hydrocarbon environmental agents.

The potential of ostensibly structurally diverse environmental chemicals to modulate endocrine processes in biological systems has been recognized. Difficulty in classifying endocrine system modulators by chemical structure may in large part be due to lack of understanding of mechanisms of action. New developments in understanding nuclear receptor mechanisms of hormone action support a more complex mechanism, possibly involving dimerization/aggregation events leading to multimeric receptor complexes in agonist action. Because of the requirement for high structural specificity in agonist action, it is suggested that most environmental chemicals of concern are likely to function as imperfect hormones with partial agonist-antagonist properties, especially at environmentally realistic concentrations. In the absence of having appropriately placed molecular recognition domains to affect agonist action, partial agonism-antagonism may be associated with favorable low-energy conformational flexibility and complementary receptor protein flexibility. The halogenated aromatic hydrocarbons are of particular concern as hormone mimics since they often have (1) similar molecular recognition factors but in many cases relatively more flexible structures, (2) similar bulk physico-chemical properties controlling uptake and distribution in biological systems, and (3) are relatively more resistant to metabolism and elimination. Some important molecular reactivity properties underlying thyromimetic and estrogenic actions of some of these chemicals are identified and described in terms of structure-activity relationships (SARs). It is proposed that specificity of hormone action in the nucleus could be associated with differential interaction of ligand-bound receptor dimeric forms with other transcription factors specific to the target cell. The small-molecule ligand can be viewed as playing a central, multifunctional role in nuclear receptor action as an organic unmasking and reclustering agent for critical macromolecules. Evidence is discussed in support of a nuclear heterodimerization model for dioxin and related compound action involving a structural transition mechanism. These models with some molecular detail also have utility in understanding the different structural properties of agonists and antagonists. There would appear to be ample opportunities for environmental chemicals to act as antagonists for multiple receptor systems with little more than anchor-ring similarities in structure. The application of three-dimensional quantitative structure-activity (3D QSAR) models incorporating such structural information should be a useful adjunct for identifying endocrine system modulating chemicals. This data has implications for (1) improved drug design, (2) understanding of chemical interaction toxicity, (3) removing undesirable chemicals from our environment, and (4) reducing their chemical release.

Environmental Pollutants↗

Mutagenicity of halogenated and oxygenated three-carbon compounds.

Four structurally related three-carbon compounds, known for their antifertility activity in the male, and the brominated derivatives of two of these compounds were tested for mutagenic activity by the Salmonella typhimurium test of Ames et al. In the presence of strain TA-100, a base-pair substituion detector strain, 1,2-dibromo-3-chloropropane (DBCP), was the most active compound tested but required enzymatic conversion by 59 microsomal preparation to an active mutagen. Three of these compounds containing an epoxide group-epichlorohydrin, epibromohydrin, and glycidol-were highly active direct mutagens, not requiring 59 for activation, alpha-Chlorohydrin was the least active compound tested; alpha-bromohydrin was 40 times more active than its chlorinated analog. Epibromohydrin was only slightly more active than epichlorohydrin, but both were highly active. With both of the halogenated epoxides, 59 preparation caused a substantial decrease in mutagenic activity at every concentration tested. All six compounds showed dose-related responsiveness for the base-pair substitution detector strains used. However, they were relatively inactive against the frameshift detector strain of S. typhimurium, TA-98. Glycerol, propylene glycol, and n-propanol, which are also three-carbon compounds containing one or more hydroxy groups, were inactive when trested at high concentrations with strain TA-100.

Alcohols↗

Immunomodulation in C57Bl/6 mice following consumption of halogenated aromatic hydrocarbon-contaminated coho salmon (Oncorhynchus kisutch) from Lake Ontario.

This report describes studies designed to assess the immunomodulatory effects associated with the consumption of coho salmon containing halogenated aromatic hydrocarbons (HAHs) and other compounds naturally bioaccumulated from Lake Ontario. Diets containing 33% coho salmon from Lake Ontario or the Pacific Ocean were fed to juvenile C57Bl/6 mice for 2-4 mo. Following 60 d, the mice that consumed Lake Ontario salmon had reduced IgM, IgG, and IgA plaque-forming cell responses to sheep erythrocytes. No changes were observed in total numbers of spleen lymphocytes, total T-lymphocytes or T-lymphocyte subsets as determined by flow cytometry. Cellular immunity, assessed by the cytotoxic T-lymphocyte response to allogeneic tumor target cells, was not altered following dietary exposure to Lake Ontario coho salmon for 4 mo. The observed humoral immunomodulation correlated with elevated PCB levels in the Lake Ontario salmon diets. The levels of pollutants such as mercury, tin compounds and other metals, PCDDs, and PCDFs were not examined.

Animals↗

Relationships between the electrostatic potential, epoxide hydrase inhibition and carcinogenicity for some hydrocarbon and halogenated hydrocarbon epoxides.

For a group of nine hydrocarbon and halogenated hydrocarbon epoxides, there is shown to be a good correlation between their abilities to inhibit epoxide hydrase and the quantity Vmin /Es, where Vmin is the most negative value of the molecule's electrostatic potential in the neighborhood of the epoxide oxygen and Es is a factor that takes account of steric effects. It is also demonstrated that carcinogenicity, for thirteen epoxides, appears to be associated with Vmin having a relatively large negative value. On this basis, several other epoxides are predicted to be carcinogenic. The electrostatic potentials used in developing these relationships have been computed by an ab initio self-consistent-field molecular orbital procedure, using optimized molecular geometries.

Animals↗

A comparison of tungsten-quartz-halogen, plasma arc and light-emitting diode light sources for the polymerization of an orthodontic adhesive.

This study investigated whether there were differences between the debond stress and adhesive remnant index (ARI) of an adhesive cured with three different orthodontic light sources. Sixty sound premolar teeth were divided into three groups of 20. A standard pre-adjusted edgewise premolar bracket (Victory Series) was bonded to each tooth using a light-cured orthodontic adhesive, Transbond X. Group 1 (control) specimens were cured with an Ortholux XT (tungsten-quartz-halogen bulb) light for 20 seconds, group 2 with an Ortho lite (plasma arc) for 6 seconds and group 3 with an Ortholux LED light-emitting diode for 10 seconds. The specimens were debonded 24 hours later using a universal mechanical testing machine, operating at a crosshead speed of 0.5 mm minute(-1). The Weibull modulus and a Logrank test showed no statistically significant differences between the three groups for debond stress. The ARI was assessed at x10 magnification. The ARI scores for group 2 were significantly different (P < 0.01) from those of groups 1 and 3 (between which there was no significant difference). For group 2 there was a greater tendency for failure to occur at the adhesive/tooth interface than for the other two groups. There appears to be no reason why any of the three types of light source cannot be used in orthodontics. Polymerization, as effective as that produced by conventional bulb light sources, was obtained with the short exposure times recommended for the plasma arc or light-emitting diode sources.

Bicuspid↗

Quantitative structure-activity relationship models correctly predict the toxic and aneuploidizing properties of six halogenated methanes in Aspergillus nidulans.

In a previous study, the relationships between the chemical structure and the ability of 35 chlorinated aliphatic hydrocarbons to induce aneuploidy and toxicity in Aspergillus nidulans were analyzed. Quantitative structure-activity relationships (QSAR) were defined for each of the biological activities under study: ARR (the dose able to block mitotic growth), D37 (the dose with 37% of survival) and LEC (the lowest efficient concentration in aneuploidy induction). In this study, these QSAR equations were used to predict the toxic and genetic activity of a further six chemicals, not included in the previous data base: bromotrichloromethane, bromoform, bromochloromethane, bromodichloromethane, dibromochloromethane and dibromochlorofluoromethane. Their ARR, D37 and LEC values were measured, and were in agreement with the predicted values, with correlation coefficients around 0.99. Furthermore, the QSAR model, which had previously been developed to discriminate between aneugenic and inactive halogenated hydrocarbons, correctly predicted the aneugenic activity of five out of six methanes. These correct predictions confirmed the validity of our QSAR model, according to which the induction of aneuploidy in A. nidulans depends on both the electrophilic and steric properties of the chemicals, whereas toxicity mainly depends on steric factors.

Aneuploidy↗

Predicting vehicle effects on the dermal absorption of halogenated methanes using physiologically based modeling.

Occupational and environmental settings present opportunities for humans to come into contact with a variety of chemicals via the dermal route. The chemicals contacting the skin are likely to be diluted with a vehicle or present as a component of a mixture. In order to support risk assessment activities, we evaluated the vehicle effects on dermal penetration of two halogenated hydrocarbons, dibromomethane (DBM) and bromochloromethane (BCM). In vivo exposures to 15 combinations of of these in water, mineral oil, and corn oil vehicles were conducted, and blood was sampled for dibromomethane and bromochloromethane during the exposure at 0.5, 1, 2, 4, 8, 12, and 24 h. A physiologically based pharmacokinetic (PBPK) model was used to estimate the total amounts of dibromomethane or bromochloromethane that were absorbed during the exposure, and the dermal permeability coefficients were determined. While the permeability coefficients for dibromomethane and bromochloromethane were approximately 73- and 40-fold higher, respectively, in the water vehicle than in the corn oil, the permeability coefficient, when normalized for the skin:vehicle matrix partition coefficient, varied by less than a factor of 2. The permeability in an aqueous vehicle was then successfully used to predict the permeability coefficient for dibromomethane in a nonpolar vehicle, peanut oil.

Administration, Cutaneous↗

Development and modification of a recombinant cell bioassay to directly detect halogenated and polycyclic aromatic hydrocarbons in serum.

Polycyclic and halogenated aromatic hydrocarbons (PAHs/HAHs) are a diverse group of widespread and persistent environmental contaminants that can cause a variety of detrimental effects in vertebrates. As most available methods to detect these contaminants are expensive, labor and time intensive, and require large amounts of tissue for extraction and analysis, several rapid mechanistically based bioassay systems have been developed to detect these chemicals. Here we describe application and optimization of a recently developed recombinant mouse cell bioassay system that responds to both PAHs and HAHs with the rapid induction of firefly luciferase for the detection of these chemicals in whole serum samples. This chemically activated luciferase expression (CALUX) bioassay has been modified to allow rapid (4-h) and direct analysis of small volumes (25-50 microl) of whole serum in a 96-well microtiter plate format without the need for solvent extraction. This bioassay can detect as little as 10 parts per trillion of the most potent HAH, 2,3,7,8-TCDD, and is also sensitive to other HAHs and PAHs. The use of simple procedures corrects for interplate and intraplate variability and the Ah receptor dependence of the induction response is accounted for by use of the antagonist 4-amino-3-methoxyflavone.

Algorithms↗

PBPK modeling of canine inhalation exposures to halogenated hydrocarbons.

Human exposure guidelines for halogenated hydrocarbons (halons) and halon replacement chemicals have been established using dose-response data obtained from canine cardiac sensitization studies. In order to provide a tool for decision makers and regulators tasked with setting guidelines for egress from exposure to halon replacement chemicals, a quantitative approach, using a physiologically based pharmacokinetic model, was established that allowed exposures to be assessed in terms of the chemical concentrations in blood during the exposure. This model, which includes a respiratory tract compartment containing a dead-space region, a pulmonary exchange area, and a breath-by-breath description of respiratory tract uptake, allows successful simulation of exhaled breath concentrations of humans during the first minute of exposure to the anesthetics halothane, isoflurane, and desflurane. In the current study, the human model was modified with canine parameters and validated with data obtained from dog studies with halothane, isoflurane, desflurane, and CFC-11. With consideration of appropriate values for ventilation and cardiac output, the model successfully simulated data collected under a variety of exposure scenarios. The canine model can be used for simulating blood concentrations associated with the potential for cardiac sensitization. These target blood concentrations can then be used with the human model for establishing safe human exposure duration. Development of the canine model stresses the need for appropriate data collection for model validation.

Administration, Inhalation↗

Toxicokinetics and oral bioavailability of halogenated acetic acids mixtures in naïve and GSTzeta-depleted rats.

Disinfection of drinking water typically produces a mixture of mono-, di-, and tri-halogenated acetic acids (HAAs). In this study, we investigated the toxicokinetics of HAA mixtures in naive and glutathione transferase zeta 1 (GSTzeta)-depleted male F344 rats administered orally or iv to Mixture-1 (monobromo [MBAA]- dichloro- [DCAA], chlorodibromo- [CDBAA], tribromo- [TBAA] acetic acids) or Mixture-2 (bromochloro- [BCAA], dibromo- [DBAA], trichloro- [TCAA] bromodichloro- [BDCAA] acetic acids) at a dose of 25 micromol/kg HAA. Serial blood samples were collected at various times up to 36 h, and the plasma concentrations of each HAA quantified by GC-ECD. Rats were pretreated for 7 d with drinking water containing 0.2 g/l DCAA to deplete the GSTzeta (GSTZ1-1) activity in the liver. An additional group of GSTzeta-depleted rats were orally dosed with each mixture and euthanized at 0.25, 0.5, 1, 2, and 4 h to determine tissue distribution of mixture components. In both mixtures, GSTzeta depletion primarily affected the toxicokinetics of di-HAAs (DCAA, BCAA, and DBAA), with the total body clearance (Cl b) decreasing 3- to 10-fold. Interestingly, DCAA pretreatment appeared to increase the elimination of Mixture-2 tri-HAAs (TCAA and BDCAA). After oral administration, DCAA exhibited a complex time-course plasma profile with secondary peaks appearing long after completion of the initial absorption phase. This phenomenon coincided with elevated DCA levels in the lower portion of the GI tract compared to CDBAA and TBAA. Comparison of the results with previous studies employing similar or higher doses of individual HAAs indicated the primary difference in HAA toxicokinetics when administered as mixture was a reduction in Cl b. These results suggest competitive interactions between tri- and di-HAAs beyond what would be predicted from individual HAA studies. For di-HAAs, the total dose is important, as clearance is dose dependent due to competition for GSTzeta. When considering HAA dosimetry, importance should be placed on both the components of the mixture and prior exposure history to di-HAAs.

Acetates↗

Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation.

Five ORFs were detected in a fragment from the Streptomyces venezuelae ISP5230 genomic DNA library by hybridization with a PCR product amplified from primers representing a consensus of known halogenase sequences. Sequencing and functional analyses demonstrated that ORFs 11 and 12 (but not ORFs 13-15) extended the partially characterized gene cluster for chloramphenicol (Cm) biosynthesis in the chromosome. Disruption of ORF11 (cmlK) or ORF12 (cmlS) and conjugal transfer of the insertionally inactivated genes to S. venezuelae gave mutant strains VS1111 and VS1112, each producing a similar series of Cm analogues in which unhalogenated acyl groups replaced the dichloroacetyl substituent of Cm. 1H-NMR established that the principal metabolite in the disrupted strains was the alpha-N-propionyl analogue. The sequence of CmlK implicated the protein in adenylation, and involvement in halogenation was inferred from biosynthesis of analogues by the cmlK-disrupted mutant. A role in generating the dichloroacetyl substituent was supported by partial restoration of Cm biosynthesis when a cloned copy of cmlK was introduced in trans into VS1111. Complementation of the mutant also indicated that inactivation of cmlK rather than a polar effect of the disruption on cmlS expression had interfered with dichloroacetyl biosynthesis. The deduced CmlS sequence resembled sequences of FADH2-dependent halogenases. Conjugal transfer of cmlK or cmlS into S. venezuelae cml-2, a chlorination-deficient strain with a mutation mapped genetically to the Cm biosynthesis gene cluster, did not complement the cml-2 lesion, suggesting that one or more genes in addition to cmlK and cmlS is needed to assemble the dichloroacetyl substituent. Insertional inactivation of ORF13 did not affect Cm production, and the products of ORF14 and ORF15 matched Streptomyces coelicolor A3(2) proteins lacking plausible functions in Cm biosynthesis. Thus cmlS appears to mark the downstream end of the gene cluster.

Amino Acid Sequence↗

Modification of halogen specificity of a vanadium-dependent bromoperoxidase.

The halide specificity of vanadium-dependent bromoperoxidase (BPO) from the marine algae, Corallina pilulifera, has been changed by a single amino acid substitution. The residue R397 has been substituted by the other 19 amino acids. The mutant enzymes R397W and R397F showed significant chloroperoxidase (CPO) activity as well as BPO activity. These mutant enzymes were purified and their properties were investigated. The maximal velocities of CPO activities of the R397W and R397F enzymes were 31.2 and 39.2 units/mg, and the K(m) values for Cl(-) were 780 mM and 670 mM, respectively. Unlike the native enzyme, both mutant enzymes were inhibited by NaN(3). In the case of the R397W enzyme, the incorporation rate of vanadate into the active site was low, compared with the R397F and the wild-type enzyme. These results supported the existence of a specific halogen binding site within the catalytic cleft of vanadium haloperoxidases.

Binding Sites↗