Conventional Toxicity Tests Research Articles

Analysis of the ToxCast Chemical-Assay Space Using the Comparative Toxicogenomics Database.

Many studies have attempted to predict in vivo hazards based on the ToxCast in vitro assay results with the goal of using these predictions to prioritize compounds for conventional toxicity testing. Most of these conventional studies rely on in vivo end points observed using preclinical species (e.g., mice and rats). Although the preclinical animal studies provide valuable insights, there can often be significant disconnects between these studies and safety concerns in humans. One way to address these concerns, for an admittedly more limited set of compounds, is to explore relationships between the in vitro data from human cell lines and observations from human related studies. The Comparative Toxicogenomics Database (CTD; http://ctdbase.org ) is a rich source of data linking chemicals to human diseases/adverse events and pathways. In this study we explored the relationships between ToxCast chemicals, their ToxCast in vitro test results, and their annotations of human disease/adverse event end points as captured in the CTD database. We mined these associations to identify potentially interesting, statistically significant in vitro assay and in vivo toxicity correlations. To the best of our knowledge, this is one of the first studies analyzing the relationships between the ToxCast in vitro assays results and the CTD disease/adverse event end point annotations. The in vitro profiles identified in this analysis may prove useful for prioritizing compounds for toxicity testing, suggesting mechanisms of toxicity, and forecasting potential in vivo human drug induced injury.

Organ-on-a-chip: development and clinical prospects toward toxicity assessment with an emphasis on bone marrow.

Conventional approaches for toxicity evaluation of drugs and chemicals, such as animal tests, can be impractical due to the large experimental scale and the immunological differences between species. Organ-on-a-chip models have recently been recognized as a prominent alternative to conventional toxicity tests aiming to simulate the human in vivo physiology. This review focuses on the organ-on-a-chip applications for high-throughput screening of candidate drugs against toxicity, with a particular emphasis on bone-marrow-on-a-chip. Studies in which organ-on-a-chip models have been developed and utilized to maximize the efficiency and predictability in toxicity assessment are introduced. The potential of these devices to replace tests of acute systemic toxicity in animals, and the challenges that are inherent in simulating the human immune system are also discussed. As a promising approach to overcome the limitations, we further focus on an in-depth analysis of the development of bone-marrow-on-a-chip that is capable of simulating human immune responses against external stimuli due to the key roles of marrow in immune systems with hematopoietic activities. Owing to the complex interactions between hematopoietic stem cells and marrow microenvironments, precise control of both biochemical and physical niches that are critical in maintenance of hematopoiesis remains a key challenge. Thus, recently developed bone-marrow-on-a-chip models support immunogenicity and immunotoxicity testing in long-term cultivation with repeated antigen stimulation. In this review, we provide an overview of clinical studies that have been carried out on bone marrow transplants in patients with immune-related diseases and future aspects of clinical and pharmaceutical application of bone-marrow-on-a-chip.

Screening the toxicity and biodegradability of petroleum hydrocarbons by a rapid colorimetric method.

Crude oil and petroleum products have a wide variety of hazardous components with high toxicity and low biodegradability. Certain dyes change their colors by intercepting electron transfer reactions during the transformation processes. This study applied resazurin and 2,6-dichlorophenol-indophenol indicators for a rapid screening biodegradation capability and toxicity response to various petroleum products such as motor oil, diesel, gasoline, and phenol. Colorimetry tests were performed in test tubes, and the absorbance values were measured over time. We observed different discoloration profiles after degradation tests using Bacillus subtilis inoculum. Phytotoxicity assays were also performed to compare colorimetric screening assays with a conventional toxicity testing with plants (seed germination). The results indicated that biotransformation of oils can increase its overall toxicity. Intermediate byproducts can be formed through biodegradation and thereby increase the toxicity of oils. The assessment of acute toxicity has shown that phenol is extremely toxic to petroleum-biodegrading microbial communities. Low molecular-weight gasoline was considered biodegradable, but it also exhibited a high acute toxic effect, mainly due to its high solubility and the presence of more volatile compounds that can penetrate cells and potentially damage cellular structures.

Toxicity screenings of nanomaterials: challenges due to interference with assay processes and components of classic in vitro tests

Given the multiplicity of nanoparticles (NPs), there is a requirement to develop screening strategies to evaluate their toxicity. Within the EU-funded FP7 NanoTEST project, a panel of medically relevant NPs has been used to develop alternative testing strategies of NPs used in medical diagnostics. As conventional toxicity tests cannot necessarily be directly applied to NPs in the same manner as for soluble chemicals and drugs, we determined the extent of interference of NPs with each assay process and components. In this study, we fully characterized the panel of NP suspensions used in this project (poly(lactic-co-glycolic acid)–polyethylene oxide [PLGA–PEO], TiO2, SiO2, and uncoated and oleic-acid coated Fe3O4) and showed that many NP characteristics (composition, size, coatings, and agglomeration) interfere with a range of in vitro cytotoxicity assays (WST-1, MTT, lactate dehydrogenase, neutral red, propidium iodide, 3H-thymidine incorporation, and cell counting), pro-inflammatory response evaluation (ELISA for GM-CSF, IL-6, and IL-8), and oxidative stress detection (monoBromoBimane, dichlorofluorescein, and NO assays). Interferences were assay specific as well as NP specific. We propose how to integrate and avoid interference with testing systems as a first step of a screening strategy for biomedical NPs.

Tissue-based environmental quality benchmarks and standards

Although the use of tissue concentrations (residues) of chemical contaminants as the dose metric to characterize chemical toxicity to aquatic organisms has been gaining acceptance over the past 20years, tissue concentrations are less commonly used in water quality management and have yet to be formally adopted as benchmarks or environmental quality standards (EQS). This synthesis paper addresses advantages and disadvantages for the development and application of tissue-based EQS as an alternative and supplement to exposure-based EQS determined with water and sediment concentration data. Tissue-based EQS can be readily developed in parallel with conventional toxicity tests, and achieved by quantification of chemical concentrations in tissue alongside traditional concentration-response toxicity testing. Tissue-residue toxicity metrics can be used as benchmarks for screening and monitoring water and sediment quality, to derive equivalent water or sediment EQS, and for ecological risk assessments and weight of evidence approaches for assessing ecosystem impairment. Tissue-based toxicity metrics and associated EQS provide several advantages; however, there are some limitations to consider and key knowledge gaps to fill.

Using additive modelling to quantify the effect of chemicals on phytoplankton diversity and biomass

Environmental authorities require the protection of biodiversity and other ecosystem properties such as biomass production. However, the endpoints listed in available ecotoxicological datasets generally do not contain these two ecosystem descriptors. Inferring the effects of chemicals on such descriptors from micro- or mesocosm experiments is often hampered by inherent differences in the initial biodiversity levels between experimental units or by delayed community responses. Here we introduce additive modelling to establish the effects of a chronic application of the herbicide linuron on 10 biodiversity indices and phytoplankton biomass in microcosms. We found that communities with a low (high) initial biodiversity subsequently became more (less) diverse, indicating an equilibrium biodiversity status in the communities considered here. Linuron adversely affected richness and evenness while dominance increased but no biodiversity indices were different from the control treatment at linuron concentrations below 2.4μg/L. Richness-related indices changed at lower linuron concentrations (effects noticeable from 2.4μg/L) than other biodiversity indices (effects noticeable from 14.4μg/L) and, in contrast to the other indices, showed no signs of recovery following chronic exposure. Phytoplankton biomass was unaffected by linuron due to functional redundancy within the phytoplankton community. Comparing thresholds for biodiversity with conventional toxicity test results showed that standard ecological risk assessments also protect biodiversity in the case of linuron.

Rapid assessment of the toxicity of oil sands process-affected waters using fish cell lines

Rapid and reliable toxicity assessment of oil sands process-affected waters (OSPW) is needed to support oil sands reclamation projects. Conventional toxicity tests using whole animals are relatively slow, costly, and often subjective, while at the same time requiring the sacrifice of test organisms as is the case with lethal dosage/concentration assays. A nonlethal alternative, using fish cell lines, has been developed for its potential use in supporting oil sands reclamation planning and to help predict the viability of aquatic reclamation models such as end-pit lakes. This study employed six fish cell lines (WF-2, GFSk-S1, RTL-W1, RTgill-W1, FHML, FHMT) in 24 h viability assays for rapid fluorometric assessment of cellular integrity and functionality. Forty-nine test water samples collected from the surface of oil sands developments in the Athabasca Oil Sands deposit, north of Fort McMurray, Alberta, Canada, were evaluated in blind. Small subsample volumes (8 ml) were mixed with 2 ml of 5× concentrated exposure media and used for direct cell exposures. All cell line responses in terms of viability as measured by Alamar blue assay, correlated well with the naphthenic acids (NA) content in the samples (R (2) between 0.4519 and 0.6171; p<0.0001) when data comparisons were performed after the bioassays. NA or total acid-extractable organics group has been shown to be responsible for most of the acute toxicity of OSPW and our results further corroborate this. The multifish cell line bioassay provides a strong degree of reproducibility among tested cell lines and good relative sensitivity of the cell line bioassay as compared to available in vivo data that could lead to cost effective, high-throughput screening assays.

Mapping the Dawn of Nanoecotoxicological Research

Some researchers consider nanotechnology the next industrial revolution, and consumer products and a variety of industries increasingly use synthetic nanoparticles. In this Account, we review the initial accomplishments of nanoecotoxicology, a discipline that is just a decade old. This new subdiscipline of ecotoxicology faces two important and challenging problems: the analysis of the safety of nanotechnologies in the natural environment and the promotion of sustainable development while mitigating the potential pitfalls of innovative nanotechnologies. In this Account, we provide a snapshot of the publicly available scientific information regarding the ecotoxicity of engineered nanoparticles. We pay special attention to information relevant to aquatic freshwater species commonly used for risk assessment and regulation. Just as the development of ecotoxicology has lagged behind that of toxicology, nanoecotoxicological research has developed much more slowly than nanotoxicology. Although the first nanotoxicolology papers were published in 1990s, the first nanoecotoxicology papers came out in 2006. A meta-analysis of scientific publications covering different environmental impacts of nanomaterials showed that the importance of research into the environmental impact of nanotechnology has gradually increased since 2005. Now the most frequently cited papers in the environmental disciplines are often those that focus on synthetic nanoparticles. The first nanoecotoxicology studies focused on adverse effects of nanoparticles on fish, algae and daphnids, which are ecotoxicological model organisms for classification and labeling of chemicals (these model organisms are also used in the EU chemical safety policy adopted in 2007: Registration, Evaluation, Authorization, and Restriction of Chemicals (REACH)). Based on our experience, we propose a multitrophic battery of nanoecotoxicological testing that includes particle-feeding and a priori particle-"proof" prokaryotic and eukaryotic organisms at different food-chain levels. Using this battery of selected test organisms, we demonstrated that TiO₂ nanoparticles were toxic to algae and that ZnO and CuO nanoparticles were toxic to several aquatic invertebrate test species. Thus, one single biotest cannot predict the ecotoxicological effects of chemicals/nanoparticles, and researchers should use several tests instead. Moreover, produced nanoparticles usually vary in features such as size, shape, and coating; therefore, a single nanoparticle species may actually include many entities with different physicochemical properties. An ecotoxicity analysis of all these variants would require a huge number of laboratory tests. To address these issues, high throughput bioassays and computational (QSAR) models that serve as powerful alternatives to conventional (eco)toxicity testing must be implemented to handle both the diversity of nanomaterials and the complexity of ecosystems.

An evaluation of the toxicity of 95 ingredients added individually to experimental cigarettes: approach and methods

Context: Ingredients have been used in modern cigarette manufacturing to facilitate tobacco processing, provide flavor, and preserve tobacco. Concern has been raised regarding the use of ingredients in cigarette manufacturing due to the possible generation of toxic chemicals resulting from their combustion when added to tobacco.Objective: Investigate the impact of individual ingredients on cigarette smoke toxicity.Materials and methods: A total of 95 ingredients were tested individually through addition at different concentrations to the tobacco of experimental cigarettes. Mainstream cigarette smoke chemistry analysis, bacterial mutagenicity testing, and cytotoxicity testing were conducted. Additionally, 31 of the ingredients were tested in 90-day nose-only rat inhalation studies using mainstream cigarette smoke. Studies were designed following conventional toxicity testing methods employed for food additives and other consumer products.Results: The studies reported here demonstrate that high levels of some ingredients can change the quantity of some smoke constituents, altering the smoke chemistry profile. From the panel of biological endpoints monitored, these added ingredients produced minimal changes in the overall toxicity profile of mainstream cigarette smoke. In some cases, the addition of high levels of an ingredient caused a small reduction in toxicity findings, probably due to displacement of burning tobacco.Conclusion: The battery of testing results presented here is a useful addition to the available scientific information for cigarette ingredients and extends the dataset which can be used for evaluating their appropriate use.

Acute Cardiorespiratory Effects of PS1, an Antineoplastic Glucan, in Rats

PS1 is a high molecular weight glucan extracted from Bacillus Calmette–Guerin (BCG) vaccine. It has demonstrated considerable in-vivo antineoplastic activity, apparently without associated toxicity. Toxicity of the BCG vaccine used in the clinical treatment of human superficial bladder cancer has been associated with the use of a viable mycobacterial preparation; an active extract would be anticipated to have less toxicity but has to be tested. Most toxicity tests require considerable quantities of the test material which in this case are currently not available. Accordingly, an in-vivo study of the effects produced on blood gases, haemoglobin and pulse and breathing rates after intravenous administration 5 μg kg−1 PS1 was carried out using cannulated rats and a sensitive methodology developed for other purposes. Following the various parameters for up to 1 h after administration showed no significant deviation from baseline values obtained before injection. PS1 at a dose of 5 μg kg−1 had no observable acute toxicity in rats. Nevertheless, chronic effects at this and higher doses have to be determined before more conventional toxicity testing is carried out.

Toxicity evaluation of verapamil and tramadol based on toxicity assay and expression patterns of Dhb, Vtg, Arnt, CYP4, and CYP314 in Daphnia magna

In this study, the toxicities of two pharmaceuticals, verapamil and tramadol were evaluated in Daphnia magna using the conventional toxicity tests (acute and chronic test) and the expression patterns of five stress responsive genes. In the chronic toxicity test, several parameters, such as the survival percentage, the body length of D. magna, the time of first reproduction, and the number of offspring per female, were adversely affected during the exposure to 4.2 mg L(-1) verapamil and 34 mg L(-1) tramadol. During the 24-h short-term exposure, verapamil particularly caused a downregulated expression of the CYP4 and CYP314 genes, whereas tramadol upregulated the expression of the CYP314 gene. Neither pharmaceutical affected the expression of Dhb, Arnt, and Vtg. However, during the 21-day long-term exposure, both verapamil and tramadol significantly reduced the expression level of the Vtg gene, a biomarker of the reproduction ability in an oviparous animal, whereas neither affected the other genes.

REP-LECOTOX: an example of FP 6 INCO project to strengthen ecotoxicological research in WBC (Western Balkan countries)

REP-LECOTOX project - INCO-CT-2006-043559-REP-LECOTOX UNSPMF - Reinforcement of research potential of Laboratory for Ecotoxicology (LECOTOX) of University of Novi Sad Faculty of Sciences funded by EC FP6 (2007-2009) presented a crucial milestone for LECOTOX, both in terms of overall research performance and its visibility and presentation to international scientific community. The project enabled upgrading and renewal of ST School of Bioscience, University of Birmingham, UK and RECETOX, Masaryk University, Brno, Czech Republic. The project enabled LECOTOX, focusing mainly on two research topics, i.e. endocrine disruption/reproductive toxicity, and identification and characterisation of aquatic toxicity, to form a functional competitive research team ready for combining transcriptomics with the established conventional toxicity tests and traditional function-based biomarkers. LECOTOX became well-equipped and experienced research group confident enough to join any FP or otherwise funded ecotoxicological research project and network.

Behavioural ecotoxicology, an “early warning” signal to assess environmental quality

BackgroundIn this review, the position of behavioural ecotoxicology within the available means to assess the status of marine environments is described as filling the gap for the needed “early warning” signals. A few examples of studies performed since the 1960s are discussed to highlight the sensitivity of these approaches in investigating the effects of chemicals, including priority pollutants and emerging contaminants, relative to conventional toxicity tests measuring survival.DiscussionThe advantage of the behavioural response is due to the integration of biochemical and physiological processes that reflect changes at higher levels of organisation with ecological relevance. Avoidance often represents a behavioural symptom easily detected in many animals exposed to contaminants and would be a useful test to explore more widely. This rapid response would reflect a defence mechanism protective against further exposure and the potential development of more pronounced deleterious effects, whilst in some cases, escape could lead to the relocation of a species with negative consequences. An investigation of the avoidance behaviour of mud shrimp, Corophium volutator, along with the chemical analyses of sediments and amphipods to assess the quality of harbour sediments is summarised. The body burden of the amphipods was 1,000 times lower than the one associated with narcosis, emphasizing the sensitivity of this endpoint. The application of this acute toxicity test is briefly compared to additional work that involved intertidal mussels collected in the field.ConclusionsRecent research undertaken with mud snails, Ilyanassa obsoleta, and harbour sediments confirmed the usefulness of the escape behaviour as an assessment tool. However, the limits of the state of knowledge regarding the fate of contaminants in species with the ability to metabolise contaminants is further discussed along with directions to be pursued to address questions arising from the reviewed literature.

Disrupting Conventional Policy: The Three Faces of Nanotechnology

Technological innovation almost always comes at a price in terms, of environmental health. In large part, this equation can be traced to one of the fundamental differences between technological innovation and regulatory innovation. The former can often occur swiftly, coming from numerous diffuse sources within the market. The latter, with rare exceptions, moves slowly, flowing from centralized state, national or international sources. (1) Thus, regulatory response typically lags well beyond the introduction of new products and technology. Nanotechnology, defined for these purposes as the use, manipulation or control of materials at the nanometer scale, is yet another case in point. Nanomaterials already are present in hundreds of consumer and industrial applications, yet are subject to minimal regulation in the United States and elsewhere. Despite the paucity of comprehensive testing of nanomaterials, there is a growing body of evidence that exposure to some types of nanoparticles may pose significant health risks to workers and the general public. (2) The very properties that make engineered nanomaterials valuable--small size, large surface area and highly variable physical and chemical characteristics--both complicate our ability to monitor exposures to nanomaterials and increase the potential that some categories of nanomaterials will prove harmful. Notwithstanding the expected benefits of emerging nanotechnologies and nanomaterials, the hazards associated with them have lead to a cacophony of appeals for various types of regulatory response, ranging from a moratorium to industry self-regulation and virtually everything in between. With this background in mind, the UCLA Working Conference on Nanotechnology Regulatory Policy commissioned a series of papers to form the centerpiece of pragmatic discussion regarding the way forward. Commentators often characterize nanotechnology as disruptive, meaning it is a form of radical technological innovation that fundamentally challenges the existing product/technology market and leads to new competitive opportunities. (3) Yet this new technology is disruptive in three other significant ways as well. It presents difficult technical issues regarding toxicity testing and hazard assessment so central to conventional regulatory policy. Likewise, it highlights the limits of existing risk governance approaches in use in the United States and abroad, particularly with respect to the role of third parties and the general public in policymaking. Lastly, it strains an already overburdened--and some might say obsolete--regulatory system, raising calls for innovative approaches more suited to managing this strange new technology. The papers that follow deal thoughtfully with these three themes, providing useful background and provocative prescriptions for policymakers, industry and academia alike. THE NEED FOR A NEW TOXICOLOGY In The Scientific Basis for the Regulation of Nanoparticles: Challenging Paracelsus and Pare, Dr. Bernard Goldstein describes how the unique attributes of nanomaterials can turn commonly accepted laws of toxicology on their head. Consequently, he argues, conventional toxicity testing and assessment are neither protective nor cost-effective. He appeals for substantial investment in new approaches to toxicology, including new toxicological testing modalities, improved exposure assessment, and post-marketing surveillance of nanomaterials in products. Dr. Goldstein's emphasis on developing a new toxicology for nanotechnology is consistent with the growth of predictive toxicology more generally, (4) and with work currently underway at universities and other research centers. (5) Dr. Goldstein sees a direct link between the new toxicological science and regulatory policy, arguing that better science is required for appropriate risk management. He cautions against embracing the precautionary principle, concerned about the likelihood of incurring unreasonably high social costs. …

Stage‐dependent differences in effects of carbaryl on population growth rate in Japanese medaka (Oryzias latipes)

Fish embryo toxicology is important because embryos are considered more susceptible than adult fish to the effects of toxic chemicals. Recently, fish embryo bioassay was proposed to replace the conventional fish acute toxicity chemical test of the Organization for Economic Cooperation and Development guidelines because it offers the advantages of fewer reagents, easy handling, and efficient data production. To accelerate the establishment of a chemical toxicity database for the protection of environmental and human health, we need to determine whether the conventional toxicity test can safely be replaced by such fish embryo toxicity tests. For instance, it is unclear how the presence of the chorion moderates the toxic effects of some chemicals. If such chemical toxicities do differ between embryos and, for example, the larval stage, then different toxic effects should appear in later life. We tested the later-life effects of the neurotoxic insecticide carbaryl at sublethal concentrations (0 [control] and 5 and 10 mg/L) in embryos and posthatch larvae of the freshwater fish medaka, Oryzias latipes. Although embryos exposed until hatching showed multiple developmental malformations and reductions in subsequent survival rates over three months, no significant reduction was observed in tolerance to starvation for 7 d and in intrinsic population growth rate (r). Exposure of larvae for 96 h resulted in dose-responsive vertebral fracture, significant reduction in tolerance to starvation for 7 d, and reduced three-month survival rate; r was reduced significantly and consistently. These results suggested that posthatch larvae were more susceptible than embryos to carbaryl exposure and that the toxic cascades may differ between larvae and embryos. The influences of carbaryl exposure on population growth rate differed significantly with developmental stage.

Using field data and weight of evidence to develop water quality criteria

In the United States, ambient aquatic life water quality criteria are derived using guidelines developed in 1985 that include a clear and consistent methodology using data from standard toxicity tests. The methodology from these guidelines has been successful, but a broader methodology is needed because some effects of pollutants do not lend themselves to conventional toxicity testing. Criterion assessment is proposed as that methodology. In criterion assessment, a specific environmental goal is translated into a measurable benchmark of effect that is used together with a modeled exposure-response relationship to estimate a range of exposures that will achieve the specific goal. The model of the exposure-response relationships and the benchmark effect are developed from field data and laboratory data using multiple analytical methods. Then the model is solved for the effect, thereby estimating the criterion, an upper threshold for acceptable exposures. The resulting candidate criteria are synthesized to select criteria and other benchmark values, such as remedial goals. The criterion assessment process is illustrated using the US Environmental Protection Agency Framework for Developing for Suspended and Bedded Sediments Water Quality Criteria, which recommends developing alternative candidate criterion values and then evaluating them to select a final criterion. Candidate criteria may be derived from models of field observations, field manipulations, laboratory tests, or empirical and theoretical models. Final selection of a criterion uses a weight-of-evidence comparison that engenders confidence because causal associations are confirmed on the basis of different assumptions, independent data sets, and varied statistical methods, thereby compensating for the concerns raised by individual studies and methods. Thus, it becomes possible to specify criteria for agents with biological or physical modes of action, as well as those with chemical modes of action, to best achieve environmental goals.

Predicting survival of grass shrimp (Palaemonetes pugio) during ethylnaphthalene, dimethylnaphthalene, and phenanthrene exposures differing in concentration and duration

Both exposure duration and concentration determine the lethal consequences of polycyclic aromatic hydrocarbons (PAHs) released during oil spills. Many factors, such as weathering, tidal transport, and addition of surfactants, can change the composition of individual dissolved compounds and the duration over which an individual is exposed. Conventional toxicity testing methods produce effect metrics, such as the median lethal concentration (LC50), that are not applicable to predicting mortality at all toxicant exposure durations that are likely to occur during a spill. In the present study, survival time models were developed that explicitly include toxicant exposure duration and concentration to predict time-to-death for grass shrimp (Palaemonetes pugio) exposed to three PAHs (1-ethylnaphthalene, 2,6-dimethylnaphthalene, and phenanthrene) commonly found in the water-soluble fraction derived from oil. Conventional 48-h LC50s also were calculated for the compounds (ethylnaphthalene, 295 microg/L; dimethylnaphthalene, 500 microg/L; and phenanthrene, 360 microg/L). In contrast to LC50s, survival models and associated response surfaces can be used to predict the proportions of shrimp that will die at various times throughout the exposure period.

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Concern about the toxicity of chemicals released into the environment has been increasing recently. Many chemicals are suspected to have hazardous effects, but evaluation of their toxicity is still difficult and challenging. One of these difficulties is the presence of chemicals that are reported to have an adverse effect on organisms despite negative results in conventional toxicity tests. Thus, a new technique has been desired in order to evaluate the effects of chemicals, Recent advances in molecular biology have provided a technique for better understanding the responses of organisms to chemicals; this emerging field is known as toxicogenomics. Toxicogenomics is defined as an integration of genomics (transcriptomics, proteomics, metabolomics) and toxicology. For example, the DNA microarray can be used to explore the gene expression profiles (transcriptomics) of organisms in response to chemicals. Exposure to chemicals results in characteristic gene expression profiles, suggesting that the DNA microarray can be used to evaluate chemical effects. Toxicogenomics is also expected to be useful in gaining a mechanistic understanding of these effects. Although it still has some limitations, this technique can be developed to assess chemicals.

The enhancement of the subacute repeat dose toxicity test OECD TG 407 for the detection of endocrine active chemicals: comparison with toxicity tests of longer duration

The OECD conventional 28-day repeat dose toxicity test (OECD TG 407) is widely employed in the initial hazard identification and characterization for commercial chemicals. The OECD has recently undertaken an international effort to "enhance" the conventional 28-day repeat dose toxicity test (OECD TG 407) in order to ensure that chemicals acting through (anti)estrogenic, (anti)androgenic, and (anti)thyroid mechanisms are identified. The enhancements include additional parameters based on the respective target organs from the male and female reproductive tracts, the thyroid, and circulating hormone levels. Ten chemicals with known endocrine modes of action and different potencies were administered using the "enhanced TG 407" test protocol to investigate the performance of this procedure. In the present evaluation, these "enhanced TG 407" protocol results, drawn from a report of the OECD validation studies, are compared to studies of the same or similar chemicals with longer and/or in utero exposures in order to evaluate the capability of the this "enhanced TG 407" in identifying the chemicals' mode of action. The major conclusions that can be drawn from these comparisons are: 1. The "enhanced TG 407" will reliably identify chemicals with a strong to moderate potential to act through endocrine modes of action on the gonads and the thyroid. In addition, this test method gives a first indication for the dose-related potency. 2. Substances with a low potency for an endocrine mode of action, i.e., having only marginal effects in the most comprehensive in vivo studies such as multi-generation studies, may not elicit clear endocrine-related effects in the "enhanced TG 407". In these cases, the primary or principal effects observed will be driven by other toxic actions of the test materials in the "enhanced TG 407". 3. It may be concluded from the present database that prolongation of exposure from 28 days up to 90 days is unlikely to improve the chance of detecting an endocrine-mediated effect 4. A number of higher tier studies with in utero and pubertal exposure show that prenatally exposed rats may be more sensitive to exposures to compounds with very low estrogenic or antiandrogenic potential in some cases than young adult rats as used in the "enhanced TG 407". 5. Overall, these comparisons support the use of the "enhanced TG407" for the detection of endocrine active chemicals. It is therefore recommended to fully accept the enhancements and include them in the test method for toxicological and regulatory use.

The Acute and Chronic Toxicity of Hexadecyl and Heptadecyl Sulfate to Aquatic Organisms

HSAS (high-solubility alkyl sulfate) is a new anionic surfactant composed predominantly of methyl and ethyl branched hexadecyl and heptadecyl sulfate. Effects of HSAS on a wide range of fish, algae, and invertebrates were investigated in conventional laboratory toxicity tests as well as in exposures conducted as part of an experimental stream model ecosystem study. For invertebrates and fish, C(16.7)HSAS (average alkyl chain length 16.7) acute LC(50) values ranged from 0.23 (channel catfish) to 2.9 (Asiatic clam, Corbicula) mg/L in well and river waters. LC(50) values for those species tested in both waters were typically within a factor of 1.5 and all were within a factor of 2 of each other, suggesting bioavailability is similar in these waters. Chronic toxicity values ranged from 0.070 (fathead minnow) to 0.42 (amphipod, Hyalella) mg/L across fish and invertebrates with algal chronic toxicity values ranging from 0.5 (blue-green algae, Anabaena flos-aquae) to 7.8 (green algae, Scenedesmus) mg/L. The order of sensitivity to HSAS acute and chronic toxicity was fish = invertebrate > algae. Based on the chronic single species sensitivity distribution, the concentrations protective of 90 and 95% of species were estimated to be 0.058 and 0.036 mg/L, respectively. These compare well with the model ecosystem NOEC of 0.064 mg/L.