Applicability Domains Enhance Application of PPARγ Agonist Classifiers Trained by Drug-like Compounds to Environmental Chemicals.

Peroxisome proliferator activator receptor gamma (PPARγ) agonist activity of chemicals is an indicator of concerned health conditions such as fatty liver and obesity. In silico screening PPARγ agonists based on quantitative structure-activity relationship (QSAR) models could serve as an efficient and pragmatic strategy. Owing to the broad research interests in discovery of PPARγ-targeted drugs, a large amount of PPARγ agonist activity data has been produced in the field of medicinal chemistry, facilitating development of robust QSAR models. In this study, random forest classifiers were developed based on the binary-category data transformed from the heterogeneous PPARγ agonist activity data of drug-like compounds. Coupling with applicability domains, capability of the established classifiers for predicting environmental chemicals was evaluated using two external data sets. Our results demonstrated that applicability domains could enhance application of the developed classifiers to predict environmental PPARγ agonists.

Interspecies differences in perfluoroalkyl substances (PFAS) toxicokinetics and application to health-based criteria.

Toxicokinetics are important for extrapolating health effects and effect levels observed in laboratory animals to humans for purposes of establishing health-based criteria. We conducted a comprehensive review of key absorption, distribution, metabolism, and excretion (ADME) parameters across different mammalian species for five perfluoroalkyl substances (PFAS) and discussed how these data can be used to inform human health risk assessment of these substances. Our analysis revealed several notable differences among the different PFAS regarding species- and substance-specific tissue partitioning, half-life, and transfer to developing offspring via the placenta or lactation, as well as highlighted data gaps for certain substances. We incorporated these observations in an analysis of whether health-based values for specific PFAS can be applied to other PFAS of differing chain length or toxicological mode of action. Overall, our analysis provides one of the first syntheses of available empirical PFAS toxicokinetic data to facilitate interpreting human relevance of animal study findings and developing health-based criteria for PFAS from such studies.

Plastic pollution and potential solutions.

A review is presented of the manufacture and use of different types of plastic, and the effects of pollution by these materials on animal, human and environmental health, insofar as this is known. Since 2004, the world has made as much plastic as it did in the previous half century, and it has been reckoned that the total mass of virgin plastics ever made amounts to 8.3 billion tonnes, mainly derived from natural gas and crude oil, used as chemical feedstocks and fuel sources. Between 1950 and 2015, a total of 6.3 billion tonnes of primary and secondary (recycled) plastic waste was generated, of which around 9% has been recycled, and 12% incinerated, with the remaining 79% either being stored in landfills or having been released directly into the natural environment. In 2015, 407 million tonnes (Mt) of plastic was produced, of which 164 Mt was consumed by packaging (36% of the total). Although quoted values vary, packaging probably accounts for around one third of all plastics used, of which approximately 40% goes to landfill, while 32% escapes the collection system. It has been deduced that around 9 Mt of plastic entered the oceans in 2010, as a result of mismanaged waste, along with up to 0.5 Mt each of microplastics from washing synthetic textiles, and from the abrasion of tyres on road surfaces. However, the amount of plastics actually measured in the oceans represents less than 1% of the (at least) 150 Mt reckoned to have been released into the oceans over time. Plastic accounts for around 10% by mass of municipal waste, but up to 85% of marine debris items - most of which arrive from land-based sources. Geographically, the five heaviest plastic polluters are P. R. China, Indonesia, Philippines, Vietnam and Sri Lanka, which between them contribute 56% of global plastic waste. Larger, primary plastic items can undergo progressive fragmentation to yield a greater number of increasingly smaller 'secondary' microplastic particles, thus increasing the overall surface area of the plastic material, which enhances its ability to absorb, and concentrate, persistent organic pollutants (POPs) such as dichlorodiphenyltrichloroethane (DDT) and polychlorinated biphenyls (PCBs), with the potential to transfer them to the tissues of animals that ingest the microplastic particles, particularly in marine environments. Although fears that such microparticles and their toxins may be passed via food webs to humans are not as yet substantiated, the direct ingestion of microplastics by humans via drinking water is a distinct possibility - since 92% of samples taken in the USA and 72% in Europe showed their presence - although any consequent health effects are as yet unclear. Foodstuffs may also become contaminated by microplastics from the air, although any consequent health effects are also unknown. In regard to such airborne sources, it is noteworthy that small plastic particles have been found in human lung tissue, which might prove an adverse health issue under given circumstances. It is also very striking that microplastics have been detected in mountain soils in Switzerland, which are most likely windborne in origin. Arctic ice core samples too have revealed the presence of microplastics, which were most likely carried on ocean currents from the Pacific garbage patch, and from local pollution from shipping and fishing. Thus, sea ice traps large amounts of microplastics and transports them across the Arctic Ocean, but these particles will be released into the global environment when the ice melts, particularly under the influence of a rising mean global temperature. While there is a growing emphasis toward the substitution of petrochemically derived plastics by bioplastics, controversy has arisen in regard to how biodegradable the latter actually are in the open environment, and they presently only account for 0.5% of the total mass of plastics manufactured globally. Since the majority of bioplastics are made from sugar and starch materials, to expand their use significantly raises the prospect of competition between growing crops to supply food or plastics, similarly to the diversion of food crops for the manufacture of primary biofuels. The use of oxo-plastics, which contain additives that assist the material to degrade, is also a matter of concern, since it is claimed that they merely fragment and add to the environmental burden of microplastics; hence, the European Union has moved to restrict their use. Since 6% of the current global oil (including natural gas liquids, NGLs) production is used to manufacture plastic commodities - predicted to rise to 20% by 2050 - the current approaches for the manufacture and use of plastics (including their end-use) demand immediate revision. More extensive collection and recycling of plastic items at the end of their life, for re-use in new production, to offset the use of virgin plastic, is a critical aspect both for reducing the amount of plastic waste entering the environment, and in improving the efficiency of fossil resource use. This is central to the ideology underpinning the circular economy, which has common elements with permaculture, the latter being a regenerative design system based on 'nature as teacher', which could help optimise the use of resources in town and city environments, while minimising and repurposing 'waste'. Thus, food might be produced more on the local than the global scale, with smaller inputs of fuels (including transportation fuels for importing and distributing food), water and fertilisers, and with a marked reduction in the use of plastic packaging. Such an approach, adopted by billions of individuals, could prove of immense significance in ensuring future food security, and in reducing waste and pollution - of all kinds.

Carcinogenic risk of emerging persistent organic pollutant perfluorooctane sulfonate (PFOS): A proposal of classification.

Perfluoroalkyls are stable synthetic chemicals, able to repel oils, fats and water. These compounds have been used in the manufacturing of products such as Teflon®, lubricants, paints, fire-fighting foams, coatings for pans, carpets, clothes, and paperboard for packaging, among others. It is believed that populations are exposed constantly to them. Its regulation in the world is under development and several controversies are in the course of litigation. One occupational study shows bladder cancer risk. This paper intends to review scientific information on the most critical perfluoroalkyl compound and proposes a procedure to get a cancer-risk categorization which PFOS can cause to populations. METHODS: As a guiding axis, we used the IARC process for developing monographs of carcinogenic risks. We used the SIGN guides for evaluating the quality of studies in human populations; and finally, we used the Squire method for evaluating studies in laboratory animals. Inadequate evidence of carcinogenicity was found in human studies mainly due to chance, threshold effect and confounders. In experimental animal studies, inadequate evidence of carcinogenicity was found in view of the number of affected species, different types of neoplasms, dose-response relationship and genotoxicity found in in-vivo and in-vitro studies. In this proposal, we concluded that cancer risk for PFOS, according to the IARC method, is not classifiable as carcinogenic to humans (group 3).

Conformal Prediction Classification of a Large Data Set of Environmental Chemicals from ToxCast and Tox21 Estrogen Receptor Assays.

Quantitative structure-activity relationships (QSAR) are critical to exploitation of the chemical information in toxicology databases. Exploitation can be extraction of chemical knowledge from the data but also making predictions of new chemicals based on quantitative analysis of past findings. In this study, we analyzed the ToxCast and Tox21 estrogen receptor data sets using Conformal Prediction to enhance the full exploitation of the information in these data sets. We applied aggregated conformal prediction (ACP) to the ToxCast and Tox21 estrogen receptor data sets using support vector machine classifiers to compare overall performance of the models but, more importantly, to explore the performance of ACP on data sets that are significantly enriched in one class without employing sampling strategies of the training set. ACP was also used to investigate the problem of applicability domain using both data sets. Comparison of ACP to previous results obtained on the same data sets using traditional QSAR approaches indicated similar overall balanced performance to methods in which careful training set selections were made, e.g., sensitivity and specificity for the external Tox21 data set of 70-75% and far superior results to those obtained using traditional methods without training set sampling where the corresponding results showed a clear imbalance of 50 and 96%, respectively. Application of conformal prediction to imbalanced data sets facilitates an unambiguous analysis of all data, allows accurate predictive models to be built which display similar accuracy in external validation to external validation, and, most importantly, allows an unambiguous treatment of the applicability domain.

American Cancer Society perspectives on environmental factors and cancer.

Cancer prevention is central to the mission of the American Cancer Society (ACS). The ACS's prevention activities take many forms, but are primarily focused on modifiable risk factors that have been demonstrated to have the largest impact on cancer risk in the general population (with particular emphasis on tobacco use because of its large impact on cancer), and well-proven policy and program interventions. The ACS addresses nutrition, physical inactivity and obesity, alcohol consumption, excessive sun exposure, prevention of certain chronic infections, and selected other environmental factors through a variety of venues, including consensus guidelines (eg, nutrition and physical activity, human papillomavirus vaccination) and developing educational materials for health care providers and the general public. In contrast to the broad definition of environmental factors used by the ACS and most other public health agencies, some members of the general public associate the term "environmental" only with toxic air and water pollutants and other, predominantly manmade, hazards that people encounter, often involuntarily, in their daily life. This article will provide an overview of the ACS's approach to the prevention of cancer associated with such toxic pollutants in the context of its mission and priorities with respect to cancer prevention.

[APPLICATION OF BIOMONITORING METHODOLOGY FOR THE ASSESSMENT OF EXPOSURE TO ENVIRONMENTAL POLLUTANTS].

In this article there are presented the current views on the technology for the assessment of the exposure to chemical pollutants with the use of the methodology of human biomonitoring and the main advantages of the latter are highlighted. There are presented main problems of the implementation of biomonitoring studies in Russia such as: beginning with the lack of the national system of the biomonitoring, and accomplishing with the inconsistency in the data within the country, the inability to assess the trend according to levels of exposure in the regional or national context. Due to the inconvenience of the Russian regulatory basis on human biomonitoring, there is persisted technological backwardness in terms of the delivery of the design, presentation and evaluation of research results, which results in the decline of the significance of biomonitoring for public health in the country. There is preserved a need for standardization and harmonization of methods and procedures of human biomonitoring (HBM) in Russia with international requirements. A serious concern is the lack of Russian programs on standardization of procedures and interlaboratcy comparison of results according to biomarkers of the exposure, the insufficient involvement of national laboratories in international programs of the comparison, the difficulties with the acquisition of standard samples of the compositionfor different environmental pollutants in biological tissues. The restraint ofthe development of HBM in the Russian Federation is caused by a complex of reasons. The most urgent task is the development of the national concept of the system with subsequent formation of technological, institutional and organizational framework of biomonitoring, as well as the improvement of Russian normative-methodical base.

High-throughput models for exposure-based chemical prioritization in the ExpoCast project.

The United States Environmental Protection Agency (U.S. EPA) must characterize potential risks to human health and the environment associated with manufacture and use of thousands of chemicals. High-throughput screening (HTS) for biological activity allows the ToxCast research program to prioritize chemical inventories for potential hazard. Similar capabilities for estimating exposure potential would support rapid risk-based prioritization for chemicals with limited information; here, we propose a framework for high-throughput exposure assessment. To demonstrate application, an analysis was conducted that predicts human exposure potential for chemicals and estimates uncertainty in these predictions by comparison to biomonitoring data. We evaluated 1936 chemicals using far-field mass balance human exposure models (USEtox and RAIDAR) and an indicator for indoor and/or consumer use. These predictions were compared to exposures inferred by Bayesian analysis from urine concentrations for 82 chemicals reported in the National Health and Nutrition Examination Survey (NHANES). Joint regression on all factors provided a calibrated consensus prediction, the variance of which serves as an empirical determination of uncertainty for prioritization on absolute exposure potential. Information on use was found to be most predictive; generally, chemicals above the limit of detection in NHANES had consumer/indoor use. Coupled with hazard HTS, exposure HTS can place risk earlier in decision processes. High-priority chemicals become targets for further data collection.

Classification of polycyclic aromatic hydrocarbons based on mutagenicity in lung tissue through DNA microarray.

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental pollutants produced in the combustion of organic matter. Exposure to PAHs raises the risk of lung cancer and inflammatory and allergic disorders such as asthma. DNA microarray technologies have been applied to research on toxicogenomics in the recent years. To evaluate the mutagenicity of PAHs and constituents of environmental pollutants in lung tissue, including metabolic activation, human alveolar epithelial type II cells (A549) were treated with nonmutagenic PAH pyrene and with the mutagenic PAHs benzo-[a]-pyrene, 1-nitropyrene, or 1,8-dinitropyrene. Comparison of genome-wide microarray expression profiles between a nonmutagenic and a mutagenic PAH-treated group revealed that xenobiotic response genes such as CYP1B1 were commonly upregulated in two groups and that DNA damage induced genes, especially p53-downstream genes such as p21 (CDKN1A) were upregulated only in the mutagenic PAH-treated group. Pretreatment with cytochrome P450 inhibitor α-naphthoflavone or p53 inhibitor pifithrin-α inhibited the benzo-[a]-pyrene-induced p21 expression. These data suggest that when PAHs enter the cells, lung epithelium induces PAH metabolic activating enzymes, and then the DNA damages-recognition signal is converged with p53 downstream genes. This metabolic activation and DNA damage is induced in lung epithelium, and the mutagenicity of PAHs can be classified by DNA microarray expression profiles.

A docking-based receptor library of antibiotics and its novel application in predicting chronic mixture toxicity for environmental risk assessment.

As organisms are typically exposed to chemical mixtures over long periods of time, chronic mixture toxicity is the best way to perform an environmental risk assessment (ERA). However, it is difficult to obtain the chronic mixture toxicity data due to the high expense and the complexity of the data acquisition method. Therefore, an approach was proposed in this study to predict chronic mixture toxicity. The acute (15 min exposure) and chronic (24 h exposure) toxicity of eight antibiotics and trimethoprim to Vibrio fischeri were determined in both single and binary mixtures. The results indicated that the risk quotients (RQs) of antibiotics should be based on the chronic mixture toxicity. To predict the chronic mixture toxicity, a docking-based receptor library of antibiotics and the receptor-library-based quantitative structure-activity relationship (QSAR) model were developed. Application of the developed QSAR model to the ERA of antibiotic mixtures demonstrated that there was a close affinity between RQs based on the observed chronic toxicity and the corresponding RQs based on the predicted data. The average coefficients of variations were 46.26 and 34.93 % and the determination coefficients (R (2)) were 0.999 and 0.998 for the low concentration group and the high concentration group, respectively. This result convinced us that the receptor library would be a promising tool for predicting the chronic mixture toxicity of antibiotics and that it can be further applied in ERA.

[The concept of the development of the state of chemical-analytical environmental monitoring].

Chemical and analytical monitoring of the quality of environment is based on the accounting of the trace amount of substances. Considering the multicomponent composition of the environment and running processes of transformation of substances in it, in determination of the danger of the exposure to the chemical pollution of environment on population health there is necessary evaluation based on the simultaneous account of complex of substances really contained in the environment and supplying from different sources. Therefore, in the analytical monitoring of the quality and safety of the environment there is a necessary conversion from the orientation, based on the investigation of specific target substances, to estimation of real complex of compounds.

[Harmonization with international approaches to guidance documents on methods of assessment of the mutagenic properties of chemical environmental factors].

Evaluation of mutagenic activity--an indispensable element of the toxicological characteristics of chemicals in their hygienic regulation. In the article there is performed an analysis of the adopted in the Organization for Economic Cooperation and Development (OECD) and in Russia guidance documents on methods of assessment of the mutagenic properties of chemicals. In the OECD there are 17 manuals, each of which describes a single method, in Russia there were approved five guidance documents for specific groups of chemicals (drugs, pesticides, nanomaterials, substances normalized in the water and the air), which represent the basic and auxiliary methods of testing mutagens. Basic methods aimed to the evaluation of gene, chromosomal and genomic mutations include: assessment of gene mutations in bacteria, methods for estimating mutations in cell cultures of human and mammals in vitro, methods for inspecting mutations in somatic and germ cells of mammals in vivo. Analysis of Russian documents shows that the protocols of basic tests of assessment of the mutagenic activity are close to the protocols of the OECD. It is necessary to publish guidance documents on tests of assessment of the mutagenic activity of chemicals harmonized with the OECD documents.

[Results of analysis of ratios of indices of toxic and sensory effect of chemicals].

In the article there are presented results of the analysis of the quantitative relations between the parameters of toxicometry and indices of sensory effect of chemicals permitting to improve the accuracy of prediction of reference concentrations.

Combinatorial × computational × cheminformatics (C3) approach to characterization of congeneric libraries of organic pollutants.

Congeners are molecules based on the same carbon skeleton but are different by the number of substituents and/or a substitution pattern. Examples are 1-chloronaphthalene, 1,4-dichloronaphthalene, and 1,3,8-trichloronaphthalene. Various persistent organic pollutants (POPs) exist in the environment as families of congeners. Very large numbers of possible congeners make their experimental characterization and risk assessment unfeasible. Computational high-throughput and quantitative structure-property relationship (QSPR) modeling has been limited by the lack of tools and approaches facilitating analysis of such POP families. We present a comprehensive approach that enables modeling of extremely large congeneric libraries. The approach involves three steps: (1) combinatorial generation of a library of congeners, (2) quantum chemical characterization of each structure at the PM6 semiempirical level to obtain molecular descriptors, and (3) analysis of the information generated in step 2. In steps 1-3, we employ combinatorial, computational, and cheminformatics techniques, respectively. Therefore, this hybrid approach is named "Combinatorial × Computational × Cheminformatics", or just abbreviated as C(3) (or C-cubed) approach. We demonstrate the usefulness of this approach by generating and characterizing Br- and Cl-substituted congeneric families of 23 typical POPs. The analysis of the resulting set of 1 840 951 congeners that includes Cl-, Br-, and mixed Br/Cl-substituted species, proves that, based on structural similarities defined by the molecular descriptors' values, the existing QSPR models developed originally for Cl- and Br-substituted congeners can be applied also to mixed Br/Cl-substituted ones. Thus, the C(3) approach may serve as a tool for exploring structural applicability domains of the existing QSPR models for congeneric sets.

Low-dose effects and nonmonotonic dose-responses of endocrine disrupting chemicals: has the case been made?

Vandenberg et al. (2012) claim that "most if not all [endocrine-disrupting chemicals (EDCs)] are likely to have low-dose effects" and "nonmonotonicity is a common occurrence after exposures to hormones and EDCs in cell culture and animals and across human populations." They present examples as anecdotes without attempting to review all available pertinent data, selectively citing studies without evaluating most of them or examining whether their putative examples are consistent and coherent with other relevant information. They assume that any statistically significant association indicates causation of an adverse effect, and their limited evaluation of specific studies is not done uniformly (i.e., studies with positive results are evaluated differently than those with null results). They also do not evaluate whether exposures in studies are truly "low-dose" and relevant to humans. They propose a number of different nonmonotonic dose-response curves, but do not consider reasons for why they should be expected to apply generally across species. Many of their examples would be - and indeed have been - questioned by many scientists. Overall, Vandenberg et al. put forth many asserted illustrations of their two conclusions without providing sufficient evidence to make the case for either and while overlooking evidence that suggests the contrary.

The Matthew effect in environmental science publication: a bibliometric analysis of chemical substances in journal articles.

BACKGROUND: While environmental research addresses scientific questions of possible societal relevance, it is unclear to what degree research focuses on environmental chemicals in need of documentation for risk assessment purposes. METHODS: In a bibliometric analysis, we used SciFinder to extract Chemical Abstract Service (CAS) numbers for chemicals addressed by publications in the 78 major environmental science journals during 2000-2009. The Web of Science was used to conduct title searches to determine long-term trends for prominent substances and substances considered in need of research attention. RESULTS: The 119,636 journal articles found had 760,056 CAS number links during 2000-2009. The top-20 environmental chemicals consisted of metals, (chlorinated) biphenyls, polyaromatic hydrocarbons, benzene, and ethanol and contributed 12% toward the total number of links- Each of the top-20 substances was covered by 2,000-10,000 articles during the decade. The numbers for the 10-year period were similar to the total numbers of pre-2000 articles on the same chemicals. However, substances considered a high priority from a regulatory viewpoint, due to lack of documentation, showed very low publication rates. The persistence in the scientific literature of the top-20 chemicals was only weakly related to their publication in journals with a high impact factor, but some substances achieved high citation rates. CONCLUSIONS: The persistence of some environmental chemicals in the scientific literature may be due to a 'Matthew' principle of maintaining prominence for the very reason of having been well researched. Such bias detracts from the societal needs for documentation on less well known environmental hazards, and it may also impact negatively on the potentials for innovation and discovery in research.

Exposure to phthalates among premenstrual girls from rural and urban Gharbiah, Egypt: a pilot exposure assessment study.

BACKGROUND: Phthalates have been identified as endocrine active compounds associated with developmental and reproductive toxicity. The exposure to phthalates in premenstrual Egyptian females remains unknown. The objective of this study was to characterize phthalate exposure of a potentially vulnerable population of premenstrual girls from urban and rural Egypt. MATERIALS AND METHODS: We collected one spot urine sample from 60 10-13 year old females, 30 from rural Egypt, and 30 from urban Egypt from July to October 2009. Samples were analyzed for 11 phthalate metabolites. Additionally, we collected anthropometrics as well as questionnaire data concerning food storage behaviors, cooking practices, and cosmetic use. Phthalate metabolite concentrations were compared between urban and rural Egyptians as well as to age and gender matched Americans. RESULTS: Monoethyl phthalate (MEP), was detected at the highest concentration in urine of Egyptian girls (median: 43.2 ng/mL in rural, 98.8 ng/mL in urban). Concentrations of urinary metabolites of di-(2-ethylhexyl) phthalate and dibutyl phthalate were comparable between Egyptians and age matched US girls. Storage of food in plastic containers was a statistically significant predictor of urinary mono-isobutyl phthalate (MiBP) concentrations when comparing covariate adjusted means. CONCLUSIONS: Urinary concentrations of phthalate metabolites were similar in Egyptian and US populations, suggesting that phthalate exposure also occurs in developing nations. Dietary intake is likely an important route of exposure to phthalates in both urban and rural populations.

Estimating persistence of brominated and chlorinated organic pollutants in air, water, soil, and sediments with the QSPR-based classification scheme.

We have estimated degradation half-lives of both brominated and chlorinated dibenzo-p-dioxins (PBDDs and PCDDs), furans (PBDFs and PCDFs), biphenyls (PBBs and PCBs), naphthalenes (PBNs and PCNs), diphenyl ethers (PBDEs and PCDEs) as well as selected unsubstituted polycyclic aromatic hydrocarbons (PAHs) in air, surface water, surface soil, and sediments (in total of 1,431 compounds in four compartments). Next, we compared the persistence between chloro- (relatively well-studied) and bromo- (less studied) analogs. The predictions have been performed based on the quantitative structure-property relationship (QSPR) scheme with use of k-nearest neighbors (kNN) classifier and the semi-quantitative system of persistence classes. The classification models utilized principal components derived from the principal component analysis of a set of 24 constitutional and quantum mechanical descriptors as input variables. Accuracies of classification (based on an external validation) were 86, 85, 87, and 75% for air, surface water, surface soil, and sediments, respectively. The persistence of all chlorinated species increased with increasing halogenation degree. In the case of brominated organic pollutants (Br-OPs), the trend was the same for air and sediments. However, we noticed that the opposite trend for persistence in surface water and soil. The results suggest that, due to high photoreactivity of C-Br chemical bonds, photolytic processes occurring in surface water and soil are able to play significant role in transforming and removing Br-OPs from these compartments. This contribution is the first attempt of classifying together Br-OPs and Cl-OPs according to their persistence, in particular, environmental compartments.

Time- and dose-dependent induction of HSP70 in Lemna minor exposed to different environmental stressors.

The objective of this study was to examine the influence of different stressors, including cadmium (heavy metal), anthracene (polycyclic aromatic hydrocarbon-PAH) and chloridazon (herbicide), on population growth and biosynthesis of cytoplasmic HSP70 in Lemna minor (duckweed) in short (4 h)- and long (7 days)-term tests. A heat shock response was confirmed in Lemna exposed to high temperature: 35, 37.5, 40, or 42.5°C in short-term (4 h) treatments. The chemicals tested stimulated the biosynthesis of the cytoplasmic HSP70 protein in a concentration-dependent way (0.5-5 µM), higher in fronds exposed to lower doses of stressors. Additionally, production of HSP70 was greater after 4 h of incubation than after 7 days. The results suggest that HSP70 could be applied as a non-specific and sensitive detector of stress induced by different chemicals at concentrations below those that produce the type of response observed in classical cytotoxicity tests, such as growth inhibition.