Development of a classification model for the antigenotoxic activity of flavonoids.

Genotoxic agents are capable of causing damage to genetic material and the cumulative DNA damage causes mutations, involved in the development of various pathological conditions, including cancer. Antigenotoxic agents possess the potential to counteract these detrimental cellular modifications and may aid in preventing, delaying, or decreasing the severity of these pathological conditions. An important class of natural products for which promising antigenotoxic activities have already been shown, are the flavonoids. In this research, we investigated the quantitative structure-activity relationship (QSAR) of flavonoids and their antigenotoxic activity against benzo[a]pyrene (B[a]P) and its mutagenic metabolite B[a]P-7,8-diol-9,10-epoxide-2. Random Forest classification models were developed, which could be useful as a preliminary in silico evaluation tool, before performing in vitro or in vivo experiments. The descriptors G2S and R8s. were the most significant for predicting the antigenotoxic potential.

Insects as an Alternative Source for the Production of Fats for Cosmetics.

Insects may provide an environmentally friendly way of producing high-quality bio-based materials that can be implemented for cosmetic applications. Insects can be bred on organic waste, in high numbers, and on small surfaces, therefore, making large scale industrial breeding possible. Fats from three insect species: the black soldier fly (BSF) (Hermetia illucens), the locust (Locusta migratoria), and the house cricket (Acheta domesticus) were evaluated for potential use in skin care. Insects were dried and fats were extracted using petroleum ether. The fats were further refined, and the fatty acid composition and the acid value were determined. The fats were used in a hand cream formulation and compared with the currently used mink-and plant-derived oils. Fatty acid analysis indicates that BSF contains > 60% of lauric acid, which makes it less suitable for application in a skin-care product, whereas locust and cricket fats are rich in C16 and C18 fatty acids which makes them more suitable. Phospholipids and free fatty acid levels in the three insect species are relatively high compared with commercial, refined oils, and need to be removed by appropriate refining protocols. Odor and color also need to be removed by physical refinement to improve the applicability.

Identification of in vitro and in vivo disconnects using transcriptomic data.

BACKGROUND: Integrating transcriptomic experiments within drug development is increasingly advocated for the early detection of toxicity. This is partly to reduce costs related to drug failures in the late, and expensive phases of clinical trials. Such an approach has proven useful both in the study of toxicology and carcinogenicity. However, general lack of translation of in vitro findings to in vivo systems remains one of the bottle necks in drug development. This paper proposes a method for identifying disconnected genes between in vitro and in vivo toxicogenomic rat experiments. The analytical framework is based on the joint modeling of dose-dependent in vitro and in vivo data using a fractional polynomial framework and biclustering algorithm. RESULTS: Most disconnected genes identified belonged to known pathways, such as drug metabolism and oxidative stress due to reactive metabolites, bilirubin increase, glutathion depletion and phospholipidosis. We also identified compounds that were likely to induce disconnect in gene expression between in vitro and in vivo toxicogenomic rat experiments. These compounds include: sulindac and diclofenac (both linked to liver damage), naphtyl isothiocyanate (linked to hepatoxocity), indomethacin and naproxen (linked to gastrointestinal problem and damage of intestines). CONCLUSION: The results confirmed that there are important discrepancies between in vitro and in vivo toxicogenomic experiments. However, the contribution of this paper is to provide a tool to identify genes that are disconnected between the two systems. Pathway analysis of disconnected genes may improve our understanding of uncertainties in the mechanism of actions of drug candidates in humans, especially concerning the early detection of toxicity.

Integrating High-Dimensional Transcriptomics and Image Analysis Tools into Early Safety Screening: Proof of Concept for a New Early Drug Development Strategy.

During drug discovery and development, the early identification of adverse effects is expected to reduce costly late-stage failures of candidate drugs. As risk/safety assessment takes place rather late during the development process and due to the limited ability of animal models to predict the human situation, modern unbiased high-dimensional biology readouts are sought, such as molecular signatures predictive for in vivo response using high-throughput cell-based assays. In this theoretical proof of concept, we provide findings of an in-depth exploration of a single chemical core structure. Via transcriptional profiling, we identified a subset of close analogues that commonly downregulate multiple tubulin genes across cellular contexts, suggesting possible spindle poison effects. Confirmation via a qualified toxicity assay (in vitro micronucleus test) and the identification of a characteristic aggregate-formation phenotype via exploratory high-content imaging validated the initial findings. SAR analysis triggered the synthesis of a new set of compounds and allowed us to extend the series showing the genotoxic effect. We demonstrate the potential to flag toxicity issues by utilizing data from exploratory experiments that are typically generated for target evaluation purposes during early drug discovery. We share our thoughts on how this approach may be incorporated into drug development strategies.

Oxidative stress/reactive metabolite gene expression signature in rat liver detects idiosyncratic hepatotoxicants.

Previously we reported a gene expression signature in rat liver for detecting a specific type of oxidative stress (OS) related to reactive metabolites (RM). High doses of the drugs disulfiram, ethinyl estradiol and nimesulide were used with another dozen paradigm OS/RM compounds, and three other drugs flutamide, phenacetin and sulindac were identified by this signature. In a second study, antiepileptic drugs were compared for covalent binding and their effects on OS/RM; felbamate, carbamazepine, and phenobarbital produced robust OS/RM gene expression. In the present study, liver RNA samples from drug-treated rats from more recent experiments were examined for statistical fit to the OS/RM signature. Of all 97 drugs examined, in addition to the nine drugs noted above, 19 more were identified as OS/RM-producing compounds-chlorpromazine, clozapine, cyproterone acetate, dantrolene, dipyridamole, glibenclamide, isoniazid, ketoconazole, methapyrilene, naltrexone, nifedipine, sulfamethoxazole, tamoxifen, coumarin, ritonavir, amitriptyline, valproic acid, enalapril, and chloramphenicol. Importantly, all of the OS/RM drugs listed above have been linked to idiosyncratic hepatotoxicity, excepting chloramphenicol, which does not have a package label for hepatotoxicity, but does have a black box warning for idiosyncratic bone marrow suppression. Most of these drugs are not acutely toxic in the rat. The OS/RM signature should be useful to avoid idiosyncratic hepatotoxicity of drug candidates.

Overview of in vitro-in vivo extrapolation approaches for the risk assessment of nanomaterial toxicity.

Nanomaterials are increasingly used in many applications due to their enhanced properties. To ensure their safety for humans and the environment, nanomaterials need to be evaluated for their potential risk. The risk assessment analysis on the nanomaterials based on animal or in vivo studies is accompanied by several concerns, including animal welfare, time and cost needed for the studies. Therefore, incorporating in vitro studies in the risk assessment process is increasingly considered. To be able to analyze the potential risk of nanomaterial to human health, there are factors to take into account. Utilizing in vitro data in the risk assessment analysis requires methods that can be used to translate in vitro data to predict in vivo phenomena (in vitro-in vivo extrapolation (IVIVE) methods) to be incorporated, to obtain a more accurate result. Apart from the experiments and species conversion (for example, translation between the cell culture, animal and human), the challenge also includes the unique properties of nanomaterials that might cause them to behave differently compared to the same materials in a bulk form. This overview presents the IVIVE techniques that are developed to extrapolate pharmacokinetics data or doses. A brief example of the IVIVE methods for chemicals is provided, followed by a more detailed summary of available IVIVE methods applied to nanomaterials. The IVIVE techniques discussed include the comparison between in vitro and in vivo studies, methods to rene the dose metric or the in vitro models, allometric approach, mechanistic modeling, Multiple-Path Particle Dosimetry (MPPD), methods using organ burden data and also approaches that are currently being developed.

Evaluation of miR-122 and other biomarkers in distinct acute liver injury in rats.

The detection of drug-induced hepatotoxicity remains an important safety issue in drug development. A liver-specific microRNA species, microRNA-122 (miR-122), has recently shown potential for predicting liver injury in addition to the standard hepatic injury biomarkers. The objective of this study was to measure miR-122 together with several other liver markers in distinct settings of acute liver toxicity in rats to determine the value of miR-122 as a biomarker for liver injury in this species. Rats were exposed to 3 well-established liver toxicants (acetaminophen, allyl alcohol, and α-naphthyl isothiocyanate), a liver-enzyme inducer (phenobarbital), or a cardiotoxicant (doxorubicin). There was a clear increase in plasma miR-122 following administration of acetaminophen, allyl alcohol, and α-naphthyl isothiocyanate. The response of miR-122 paralleled that of other markers and was consistent with liver injury as indicated by histopathological evaluation. Furthermore, the changes in miR-122 were detected earlier than standard liver injury markers and exhibited a wide dynamic range. In contrast, miR-122 responses to phenobarbital and doxorubicin were low. Based on these findings, miR-122 shows significant promise and may provide added value for assessing liver toxicity in drug development.

Plasma cells are not restricted to the CD27+ phenotype: characterization of CD27-CD43+ antibody-secreting cells.

Circulating antibody-secreting cells are present in the peripheral blood of healthy individuals reflecting the continued activity of the humoral immune system. Antibody-secreting cells typically express CD27. Here we describe and characterize a small population of antibody-secreting class switched CD19+CD43+ B cells that lack expression of CD27 in the peripheral blood of healthy subjects. In this study, we characterized CD27-CD43+ cells. We demonstrate that class-switched CD27-CD43+ B cells possess characteristics of conventional plasmablasts as they spontaneously secrete antibodies, are morphologically similar to antibody-secreting cells, show downregulation of B cell differentiation markers, and have a gene expression profile related to conventional plasmablasts. Despite these similarities, we observed differences in IgA and IgG subclass distribution, expression of homing markers, replication history, frequency of somatic hypermutation, immunoglobulin repertoire, gene expression related to Toll-like receptors, cytokines, and cytokine receptors, and antibody response to vaccination. Their frequency is altered in immune-mediated disorders. Conclusion: we characterized CD27-CD43+ cells as antibody-secreting cells with differences in function and homing potential as compared to conventional CD27+ antibody-secreting cells.

Phospholipidosis in rats treated with amiodarone: serum biochemistry and whole genome micro-array analysis supporting the lipid traffic jam hypothesis and the subsequent rise of the biomarker BMP.

To provide mechanistic insight in the induction of phospholipidosis and the appearance of the proposed biomarker di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate (BMP), rats were treated with 150 mg/kg amiodarone for 12 consecutive days and analyzed at three different time points (day 4, 9, and 12). Biochemical analysis of the serum revealed a significant increase in cholesterol and phospholipids at the three time points. Bio-analysis on the serum and urine detected a time-dependent increase in BMP, as high as 10-fold compared to vehicle-treated animals on day 12. Paralleling these increases, micro-array analysis on the liver of treated rats identified cholesterol biosynthesis and glycerophospholipid metabolism as highly modulated pathways. This modulation indicates that during phospholipidosis-induction interactions take place between the cationic amphiphilic drug and phospholipids at the level of BMP-rich internal membranes of endosomes, impeding cholesterol sorting and leading to an accumulation of internal membranes, converting into multilamellar bodies. This process shows analogy to Niemann-Pick disease type C (NPC). Whereas the NPC-induced lipid traffic jam is situated at the cholesterol sorting proteins NPC1 and NPC2, the amiodarone-induced traffic jam is thought to be located at the BMP level, demonstrating its role in the mechanism of phospholipidosis-induction and its significance for use as a biomarker.

Impact of Particle Size on Toxicity, Tissue Distribution and Excretion Kinetics of Subchronic Intratracheal Instilled Silver Nanoparticles in Mice.

The unique physicochemical properties of silver nanoparticles (AgNPs) make them useful in a wide range of sectors, increasing their propensity for human exposure, as well as the need for thorough toxicological assessment. The biodistribution of silver, hematological parameters and GSH/GSSG levels in the lung and liver were studied in mice that were intratracheally instilled with AgNP (5 and 50 nm) and AgNO3 once a week for 5 weeks, followed by a recovery period of up to 28 days (dpi). Data was gathered to build a PBPK model after the entry of AgNPs into the lungs. AgNPs could be absorbed into the blood and might cross the physiological barriers and be distributed extensively in mice. Similar to AgNO3, AgNP5 induced longer-lasting toxicity toward blood cells and increased GSH levels in the lung. The exposure to AgNP50 increased the GSH from 1 dpi onward in the liver and silver was distributed to the organs after exposure, but its concentration decreased over time. In AgNP5 treated mice, silver levels were highest in the spleen, kidney, liver and blood, persisting for at least 28 days, suggesting accumulation. The major route for excretion seemed to be through the urine, despite a high concentration of AgNP5 also being found in feces. The modeled silver concentration was in line with the in vivo data for the heart and liver.

Insects as Diet and Therapy: Perspectives on Their Use for Combating Diabetes Mellitus in Tanzania.

More than 450 million people worldwide are suffering from diabetes and this number is expected to increase. In developing countries, such as Tanzania, the number of patients suffering from diabetes and associated diseases is increasing as well. Up to 80% of the Tanzanian people rely on traditional medicines for their health care services. The nature of Tanzanian is very rich in different plant and insect species, and this could be exploited through their implementation in preventive and/or curative approaches in the battle against diabetes. The implementation of healthy insects in the diets of people may help in the prevention of obesity, which is a risk factor in the etiology of diabetes, while the identification of small molecules in insects may help in the discovery of potential new drugs that can be used in the treatment of diabetes. In this paper, an overview on the potential implementation of insects against diabetes is presented.

Valorisation Potential of Using Organic Side Streams as Feed for Tenebrio molitor, Acheta domesticus and Locusta migratoria.

Due to increasing welfare and population, the demand for alternative protein sources, obtained with minimal use of natural resources, is rising in today's society. Insects have the potential to be used as an alternative protein source since they are considered to be able to convert low-value biomass into high-value components, resulting in opportunities for valorisation of organic side streams. Moreover, insects are suggested to be a sustainable protein source, referring to the efficient "feed to body" mass conversion potential. The aim of this review was to explore the potential to rear the yellow mealworm (Tenebrio molitor), the house cricket (Acheta domesticus) and the migratory locust (Locusta migratoria) on low or not yet valorised organic side streams within the food supply chain. This was performed by collecting research information focusing on the rearing of the insects in scope on organic biomass. In addition, the nutritional composition of the produced insects as well as their dietary requirements will be reviewed. Finally, the availability of side streams in the EU will be discussed as well as their potential to be used as insects feed.

Automated analysis of contractility in the embryonic stem cell test, a novel approach to assess embryotoxicity.

The embryonic stem cell test (EST) is an ECVAM-validated assay to detect embryotoxicity. The output of the assay is the effect of test compounds on the differentiation of murine-derived embryonic stem cells (D3 cells), recorded by visual analysis of contracting cardiomyocyte-like cells. Incorporation of a system to assess the contractility in an automated manner is proposed, to increase the throughput in the EST independent of observer bias. The automated system is based on image recording of each well, resulting in the area (pixels) and frequency of contractility (Hz). Four test compounds were assessed for their embryotoxic potency in the 96-well version of the EST, with both manual and automated analysis: 6-Aminonicotinamide, Valproic Acid, Boric Acid, and Penicillin G. There was no statistically significant difference in the outcome of both methods in the fraction of contractility (p<0.05), resulting in the same rank-order of Relative Embryotoxic Potency (REP) values: 6-aminonicotinamide (1)>valproic acid (0.007-0.013)>Boric Acid (0.002-0.005)>Penicillin G (0.00001). The automated image recording of contractile cardiomyocyte-like cells in the EST allows for an unbiased high throughput method to assess the embryotoxic potency of test compounds, resulting in an outcome comparable to manual analysis.

Microarray analyses in dendritic cells reveal potential biomarkers for chemical-induced skin sensitization.

The assessment of the skin sensitising capacity of chemicals is up to now investigated using in vivo animal tests. However there has been an increasing public and governmental concern regarding the use of animals for chemical screening. This has raised the need for the development of validated in vitro alternatives. Langerhans cells are potent antigen-presenting cells that play a crucial role in the development of allergic contact dermatitis. We used CD34(+) progenitor-derived dendritic cells from cord blood as an in vitro alternative for Langerhans cells. The cells were exposed to four contact allergens (nickel sulphate, dinitrochlorobenzene, oxazolone and eugenol) and two irritants (sodium dodecyl sulphate and benzalkonium chloride) for 3, 6, 12 and 24h. Using microarray analyses we revealed a set of 25 genes with an altered gene expression pattern after exposure to allergens and not to irritants. Five out of these 25 genes were selected and their gene expression changes were confirmed with real-time reverse transcriptase polymerase chain reaction. The list of 25 genes represent valuable candidates to be further evaluated for their capacity to predict the sensitizing potential of different classes of chemicals in studies using a more extended set of (non) allergic substances.

Evaluation of existing (Q)SAR models for skin and eye irritation and corrosion to use for REACH registration.

The performance of the (Q)SAR models Derek Nexus, Toxtree and Case Ultra for the prediction of skin and eye irritation/corrosion is investigated. For irritation and corrosion of the skin, 117 compounds and for the eye, 125 compounds were listed. The balance between the groups positive and negative for irritation and corrosion was maintained. The obtained predictions were compared with experimental data and the numbers of true and false positives and negatives were determined. Based on these results several performance parameters of the tested (Q)SAR models were calculated. Despite all the efforts to make good and valid models, the results indicate a poor predictivity of the current models: a lot of compounds were not predicted, were out of the applicability domain or were predicted wrong. Considering our results, it can be concluded that the tested models are not yet sufficiently powerful for implementation. Possibly the training-sets used within the current models are not yet comprehensive enough or the incorporated data are not of enough quality. Although the use of these models as stand-alone evaluation is not recommended, these models can be of value as weight-of-evidence in the context of expert knowledge in an Integrated Approach to Testing and Assessment.

Interactions between dendritic cells and epithelial cells in allergic disease.

Dendritic cells (DCs) play a crucial role in the sensitisation process. Upon encounter with an allergen, DCs require interactions with other cells and factors for triggering a primary or secondary immune response. Epithelial cells (ECs) express features of accessory cells, such as expression of HLA-DR, co-stimulatory molecules, functional FcgammaR, molecules of the antigen-processing machinery, and display an ability to internalise antigen. These features may authorize them to function as immunomodulators (e.g. amplification of memory T cells during secondary immune responses). ECs may increase chemokine (e.g. CCL20) secretion thereby attracting DCs. Epithelial human TSLP activates DC, which allow them to prime naive T cells for the production of pro-inflammatory cytokines, while down-regulating IFN-gamma and IL-10. ECs may also influence the local polarization of types l and 2 antigen-presenting cells via PGE(2) by impairing the ability of maturing DC to produce bioactive IL-12 p70. PGE(2) is synergistic with IL-1beta and TNF-alpha in the induction of functional and phenotypic maturation of DC and induce IL12 p40 production. Sensitisation via the respiratory route may be Th(2) skewed, possibly because the antigen recognition by DC occurs in an environment rich of airway EC-product such as PGE(2).

Respiratory immunotoxicity: an in vitro assessment.

As yet, in vitro assessment of the immunotoxic potency of respiratory agents is not possible. The complexity of the endpoint and the respiratory tract, and the limited availability of well-documented respiratory agents are the main reasons. The evidence that epithelial cells (ECs) are triggered by compounds to express in vitro surface proteins and soluble mediators, has stimulated their use for developing tests for respiratory immunotoxicity. A variety of airway ECs and EC-lines have been assessed, but the available information seems to point at human alveolar cells (e.g., A549) as the most convenient cell type. EC-based test formats with various degrees of complexity have been assessed. Sofar, promising results were obtained using a 3D model using the human A549 lung cell line. Dendritic cells (DCs) have been subjected to intensive research. However, currently available tests are not well suited to discern among the potency of sensitizers. Potential explanations include the lack of standardised protocols for the generation of DCs, no good standards for estimating the quality of in vitro derived DC-cultures, and limited dynamics of the currently used end-points. Alveolar macrophages (AMs) have so far received less attention. This may proof unjustified as macrophages may link innate responses to adaptive immunity. The observation that ECs, DCs and AMs affect each other, suggests that test formats are required combining at least two of these cell types if ranking of compounds according to their sensitising potency is the aim. In addition, the capacity of compounds to cross a cellular membrane is an important property of an immunotoxic compound, which can be assessed only in 3D reconstituted human tissue models. While promising data have been reported for the skin, immunocompetent 3D reconstituted human lung remains to be evaluated for respiratory immunotoxicity. Obviously, the success of any of these simplified test (as compared to the complexity of the immune response) is highly dependent on the availability of early stage biomarkers (expressed at mucosal barrier level) that are predictive for relevant immunotoxicity mechanisms occurring down-stream of the immune response. As yet, such biomarkers are not yet available.

Cytokine transcript profiling in CD34+-progenitor derived dendritic cells exposed to contact allergens and irritants.

We here investigated wether genes encoding the interleukins IL-1beta, IL-6 and IL-8, and the chemokines CCL2, CCL3, CCL3L1 and CCL4 are useful markers for sensitization testing in CD34+-progenitor derived dendritic cells (CD34-DC). CD34-DC from at least three donors were exposed during 0.5 up to 24h to the chemical sensitizers nickel sulphate, oxazolone, 2,4-dinitrochlorobenzene (DNCB) and eugenol, and to the irritants sodium dodecyl sulphate (SDS) and benzalkonium sulphate (BC). mRNA expression was evaluated using real-time RT-PCR. We observed a large inter-individual variation in mRNA expression in CD34-DC exposed to the chemicals. No or limited effects on expression were observed for the irritant BC and the weak sensitizer eugenol. All other chemicals modulated the transcript levels of most cytokines that were investigated. Most of the time, no clear-cut distinctions could be made between the sensitizers and SDS. After 24 h, consistent upregulatory effects of all sensitizing compounds on transcript expression of CCL2, CCL3 and CCL4 were observed, whereas SDS (and BC) had no effect. Our findings suggest that the CCL2, CCL3 and CCL4 genes may be selective end-point markers in the CD34-DC model to discern chemical sensitizers from irritants.

Release of (and lessons learned from mining) a pioneering large toxicogenomics database.

AIM: We release the Janssen Toxicogenomics database. This rat liver gene-expression database was generated using Codelink microarrays, and has been used over the past years within Janssen to derive signatures for multiple end points and to classify proprietary compounds. MATERIALS & METHODS: The release consists of gene-expression responses to 124 compounds, selected to give a broad coverage of liver-active compounds. A selection of the compounds were also analyzed on Affymetrix microarrays. RESULTS: The release includes results of an in-house reannotation pipeline to Entrez gene annotations, to classify probes into different confidence classes. High confidence unambiguously annotated probes were used to create gene-level data which served as starting point for cross-platform comparisons. Connectivity map-based similarity methods show excellent agreement between Codelink and Affymetrix runs of the same samples. We also compared our dataset with the Japanese Toxicogenomics Project and observed reasonable agreement, especially for compounds with stronger gene signatures. We describe an R-package containing the gene-level data and show how it can be used for expression-based similarity searches. CONCLUSION: Comparing the same biological samples run on the Affymetrix and the Codelink platform, good correspondence is observed using connectivity mapping approaches. As expected, this correspondence is smaller when the data are compared with an independent dataset such as TG-GATE. We hope that this collection of gene-expression profiles will be incorporated in toxicogenomics pipelines of users.

Immunologic biomarkers in relation to exposure markers of PCBs and dioxins in Flemish adolescents (Belgium).

In this study, we investigated 17- to 18-year-old boys and girls to determine whether changes in humoral or cellular immunity or respiratory complaints were related to blood serum levels of polychlorinated biphenyls (PCBs) and dioxin-like compounds after lifetime exposure in Flanders (Belgium). We obtained blood samples from and administered questionnaires to 200 adolescents recruited from a rural area and two urban suburbs. Physicians recorded medical history and respiratory diseases. We measured immunologic biomarkers such as differential blood cell counts, lymphocyte phenotypes, and serum immunoglobulins. As biomarkers of exposure, we determined the serum concentrations of PCBs (PCB 138, PCB 153, and PCB 180) and dioxin-like compounds [chemical-activated luciferase expression (CALUX) bioassay]. The percentages of eosinophils and natural killer cells in blood were negatively correlated with CALUX toxic equivalents (TEQs) in serum (p = 0.009 and p = 0.05, respectively). Increased serum CALUX TEQs resulted in an increase in serum IgA levels (p = 0.05). Furthermore, levels of specific IgEs (measured by radioallergosorbent tests) of cat dander, house dust mite, and grass pollen were also significantly and negatively associated with the CALUX TEQ, with odds ratios (ORs) equal to 0.63 [95% confidence interval (CI), 0.42-0.96], 0.68 (0.5-0.93), and 0.70 (0.52-0.95), respectively. In addition, reported allergies of the upper airways and past use of antiallergic drugs were negatively associated with CALUX TEQs, with ORs equal to 0.66 (0.47-0.93) and 0.58 (0.39-0.85), respectively. We found a negative association between IgGs and marker PCBs in serum (p = 0.009). This study shows that immunologic measurements and respiratory complaints in adolescents were associated with environmental exposure to polyhalogenated aromatic hydrocarbons (PHAHs). The negative correlation between PHAHs and allergic responses in adolescents suggested that exposure may entail alterations in the immune status.