A survey of EPA/OPP and open literature on selected pesticide chemicals. II. Mutagenicity and carcinogenicity of selected chloroacetanilides and related compounds.

With this effort, we continue our examination of data on selected pesticide chemicals and their related analogues that have been presented to the U.S. Environmental Protection Agency's (USEPA's) Office of Pesticide Programs (OPP). This report focuses on a group of selected chloroacetanilides and a few related compounds. As part of the registration process for pesticidal chemicals, interested parties (registrants) must submit toxicity information to support the registration including both mutagenicity and carcinogenicity data. Although this information is available to the public via Freedom of Information (FOI) requests to the OPP, publication in the scientific literature allows greater dissemination and examination of the data. For this Special Issue, graphic profiles have been prepared of the mutagenicity and carcinogenicity data available in the submissions to OPP. Also, a discussion is presented about how toxicity data are used to help establish tolerances (limits of pesticide residues in foods). The mutagenicity results submitted by registrants are supplemented by data on these chemicals from the open literature to provide a full perspective of their genetic toxicology. The group of chloroacetanilides reviewed here display a consistent pattern of mutagenic activity, probably mediated via metabolites. This mutagenic activity is a mechanistically plausible factor in the development of tumors seen in experimental animals exposed to this class of chemicals.     

Enabling Policy Planning and Innovation Management through Patent Information and Co-Authorship Network Analyses: A Study of Tuberculosis in Brazil

Introduction

New tools and approaches are necessary to facilitate public policy planning and foster the management of innovation in countries'' public health systems. To this end, an understanding of the integrated way in which the various actors who produce scientific knowledge and inventions in technological areas of interest operate, where they are located and how they relate to one another is of great relevance. Tuberculosis has been chosen as a model for the present study as it is a current challenge for Brazilian research and innovation.

Methodology

Publications about tuberculosis written by Brazilian authors were accessed from international databases, analyzed, processed with text searching tools and networks of coauthors were constructed and visualized. Patent applications about tuberculosis in Brazil were retrieved from the Brazilian National Institute of Industrial Property (INPI) and the European Patent Office databases, through the use of International Patent Classification and keywords and then categorized and analyzed.

Results/Conclusions

Brazilian authorship of articles about tuberculosis jumped from 1% in 1995 to 5% in 2010. Article production and patent filings of national origin have been concentrated in public universities and research institutions while the participation of private industry in the filing of Brazilian patents has remained limited. The goals of national patenting efforts have still not been reached, as up to the present none of the applications filed have been granted a patent. The analysis of all this data about TB publishing and patents clearly demonstrates the importance of maintaining the continuity of Brazil''s production development policies as well as government support for infrastructure projects to be employed in transforming the potential of research. This policy, which already exists for the promotion of new products and processes that, in addition to bringing diverse economic benefits to the country, will also contribute to effective dealing with public health problems affecting Brazil and the World.     

Considerations in the U.S. Environmental Protection Agency's testing approach for mutagenicity.

OPP: This paper provides the rationale and support for the decisions the OPP will make in requiring and reviewing mutagenicity information. The regulatory requirement for mutagenicity testing to support a pesticide registration is found in the 40 CFR Part 158. The guidance as to the specific mutagenicity testing to be performed is found in the OPP's Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic Animals (referred to as the Subdivision F guideline). A revised Subdivision F guideline has been presented that becomes the current guidance for submitters of mutagenicity data to the OPP. The decision to revise the guideline was the result of close examination of the version published in 1982 and the desire to update the guidance based on developments since then and current state-of-the-science. After undergoing Agency and public scrutiny, the revised guideline is to be published in 1991. The revised guideline consists of an initial battery of tests (the Salmonella assay, an in vitro mammalian gene mutation assay and an in vivo cytogenetics assay which may be either a bone marrow assay for chromosomal aberrations or for micronuclei formation) that should provide an adequate initial assessment of the potential mutagenicity of a chemical. Follow-up testing to clarify results from the initial testing may be necessary. After this information as well as all other relevant information is obtained, a weight-of-evidence decision will be made about the possible mutagenicity concern a chemical may present. Testing to pursue qualitative and/or quantitative evidence for assessing heritable risk in relation to human beings will then be considered if a mutagenicity concern exists. This testing may range from tests for evidence of gonadal exposure to dominant lethal testing to quantitative tests such as the specific locus and heritable translocation assays. The mutagenicity assessment will be performed in accordance with the Agency's Mutagenicity Risk Assessment Guidelines. The mutagenicity data would also be used in the weight-of-evidence consideration for the potential carcinogenicity of a chemical in accordance with the Agency's Carcinogen Risk Assessment Guidelines. In instances where there are triggers for carcinogenicity testing, mutagenicity data may be used as one of the triggers after a consideration of available information. It is felt that the revised Subdivision F guideline will provide appropriate, and more specific, guidance concerning the OPP approach to mutagenicity testing for the registration of a pesticide. It also provides a clearer understanding of how the OPP will proceed with its evaluation and decision making concerning the potential heritable effects of a test chemical.(ABSTRACT TRUNCATED AT 400 WORDS)     

水体污染物"三致"效应的生物监测研究进展

由于大量且多种多样化学物质进入到水环境中 ,使得水体不同程度地表现出致突变性、致癌性、致畸性。传统分析技术与综合指标不能客观、及时反映水的这种特性。而短期生物试验以其具有快速的、综合的响应水中这方面变化的特点 ,越来越受到重视与应用。本文主要论述与评价“三致”物作用机理以及几种常用短期生物试验 (Ames试验、微核试验、非程序DNA合成、SOS显色试验、姊妹单体交换测定、单细胞凝胶电泳技术 )。并分类评述了可用于此领域的生物材料的来源、特性 ,便于同类科研工作参考     

Experience with mutagenicity testing of new drugs: viewpoint of a regulatory agency

Quality and quantity of mutagenicity testing were analyzed for drugs with new active compounds which were submitted for registration in the Federal Republic of Germany from mid 1982 to mid 1986. A large variety of deficiencies was found, applying to selection and number of mutagenicity tests as well as to test performances. Only 65 out of the 144 drugs submitted for registration were tested sufficiently in the initial phase of registration. From 1982 to 1986 this situation has not been changed markedly. Inadequate test performance still remains the main reason for insufficient testing, leading in some cases to artificially positive results. For in vivo tests the selection of test species was mainly motivated by technical reasons and not by characteristics of the test compound. Most of the insufficiencies were eliminated during the second phase of registration. In some cases insufficient mutagenicity testing led to consequences concerning risk-benefit assessment of the drug and its regulation.     

Optimizing French scientific and economic performance: the Cifre system of public-private partnership in doctoral research and Servier's contribution

The European Union has set itself the daunting target of becoming the world's most competitive and dynamic knowledge-based economy by 2010. Any hope of success against the United States and the Asian tiger economies lies in the quality of scientific and technological research. In France, postgraduate training has long labored under a deep academia/industry divide. Although the universities have introduced supervised 3 year doctoral courses along the lines of the English-speaking countries, they still produce too many postdocs with little experience or understanding of, and little taste for, the private sector. This ignores career realities: the public sector can offer employment to only half the postdocs it produces. The rest must fall back on positions in the private sector, in some cases with a sense of failure, ill suiting them to drive the intellectual economy forwards compared to their international competitors. To combat the divide and emphasize the quality of the research training available within industry, the public/private National Association for Technical Research (ANRT), acting on behalf of the Ministry of Research, created the Industrial Research Training Agreement (Cifre) scheme in 1981. Higher education laboratories and private companies combine to offer doctoral students the opportunity to undertake their 3 year course in a mixed public/private environment (the exact ratio is not defined but in the case of the Servier Research Group, an early and active participant in the scheme, at least one third of the course is spent in the private sector). The doctoral thesis is thereby transformed into a meaningful career qualification. Funded by the Ministry, with maintenance grants to the students and compensatory payments to the companies, the Cifre scheme, which is currently being expanded, has produced 12,000 postdocs personally and intellectually equipped for careers transiting seamlessly between the public and private sectors, to the enrichment of each.     

Legal and regulatory concerns about transgenic plants in Brazil

Brazil has a biosafety law that was approved in 1995. This law provides for a horizontal type of regulation that coordinates other existing regulatory frameworks in the areas of agriculture, health and environment. Various federal government departments are responsible for implementing the law. The National Technical Biosafety Commission is the national competent authority on biosafety with overall responsibility. In the case of Bt plants or any insecticidal organism, the Agrochemical Law also applies and authorization for laboratory, greenhouse and field studies must be obtained from the Plant Protection Secretariat, the Brazilian Institute of Environment and the National Agency of Health. Furthermore, the National Environmental Council must issue a license for commercialization of any GMO. There is pressure needed for capacity building and to harmonize the regulatory and administrative frameworks among the different federal departments involved. Some perspectives and challenges for the commercial registration of transgenic crops are discussed.     

Use of transgenic seeds in Brazilian agriculture and concentration of agricultural production to large agribusinesses

We identified the commercial releases of genetically modified organisms (GMOs) in Brazil, their characteristics, the types of genetic transformation used, and the companies responsible for the development of these GMOs, classifying them into two categories: private companies, subdivided into multinational and national, and public institutions. The data came from the data bank of the national registration of cultivars and the service of national protection of cultivars of the Ministry of Agriculture, Fishing and Supply (MAPA). This survey was carried out from 1998 to February 12, 2011. Until this date, 27 GMOs had been approved, including five for soybean, 15 for maize and seven for cotton cultivars. These GMOs have been used for the development of 766 cultivars, of which, 305 are soybean, 445 are maize, and 13 are cotton cultivars. The Monsato Company controls 73.2% of the transgenic cultivars certified by the MAPA; a partnership between Dow AgroSciences and DuPont accounts for 21.4%, and Syngenta controls 4.96%. Seed supply by these companies is almost a monopoly supported by law, giving no choice for producers and leading to the fast replacement of conventional cultivars by transgenic cultivars, which are expensive and exclude small producers from the market, since seeds cannot be kept for later use. This situation concentrates production in the hands of a few large national agribusiness entrepreneurs.     

Public policy and partnership in research on biological control in France

Most public institutions and companies of plant protection industry have a favourable opinion of biological control; they feel concerned by the issue, but weakly engaged in its solution. Biological plant protection is however poorly developed in agriculture, the success of biological control being insufficiently known due to severe competition with chemical pesticides that are reliable, efficient, specific and safer for the environment. In this context, and to improve the implementation of biological control, the following actions are proposed:

1. more interdisciplinarity in research between agronomists, economists and biologists who have to take better advantage of the various concepts of evolutionary biology and to carry out more field experiments on a large scale;
2. wider and more integrated objectives: the successful control of a pest using biological means is not a sufficient aim. We absolutely need to consider all aspects of IPM, in particular resistance to pesticides, cultural means, plant breeding and all alternative biotechnical means;
3. larger partnership: during too many years, only researchers of public institutions were involved in biological control. Today, many partners, including public institutions (Plant Protection Service, Technical Institutes) and private companies (large and small) are engaged in alternative plant protection. To facilitate cooperation between them, a specific committee has been set up in France in order to coordinate experimentations for registration and quality control of biopesticides.

With such improved relationships between all partners, specific arguments and lobbying pressure should be stronger on the distribution and retailers of agricultural goods with a view to promoting plant products obtained via biological or integrated control methods and to seeking public understanding and support.     

Can synthetic pesticides be replaced with biologically-based alternatives?—an industry perspective

Agricultural chemical companies have invested in the discovery and development of biological pesticides to complement synthetic pesticides for the control of insects, diseases, and weeds on agronomic and horticultural crops. For plant disease control, companies envisage biological fungicides entering markets where they have the best chance of performing and which are most receptive to using biological control methods. Fewer regulatory requirements can mean faster registration for a biological than a synthetic pesticide. However, industry’s requirements for competitive performance, effective formulations, and economic production can mean significant investments in time and money for a biological pesticide, although total investment may be less than for a synthetic pesticide. One biocontrol project in which industry has invested is baculoviruses for insect control. Insect baculoviruses, genetically modified to kill insects faster than wild-type viruses, are attractive biocontrol agents because their selectivity to insect pests and safety to beneficial insects and mammals enable them to compete with synthetic insecticides. Industry is looking for similar biocontrol opportunities in disease control. Biocontrol agents for seedling disease, root rot, and postharvest disease control have been registered by the EPA and are trying to compete with synthetic fungicides for market share. To date, effective biocontrol agents have not been identified for the control of serious foliar diseases, such as grape downy mildew, potato late blight, wheat powdery mildew, and apple scab. Farmers must rely on synthetic fungicides and agronomic methods to control these diseases for the foreseeable future. Received 06 February 1997/ Accepted in revised form 01 June 1997     

Evaluation of Ecotoxicological Field Studies for Authorization of Plant Protection Products in Europe

An increasing number of ecotoxicological field studies are being submitted in the European Union procedure for authorization of pesticides. Although there is some guidance on how these studies can be used for risk assessment, not all aspects of field tests are covered and the guidance differs per type of test and per non-target group. To facilitate a more uniform approach by the regulatory authorities in the EU, a basic scheme is proposed with qualitative tools to: (i) assess the scientific reliability of individual field tests, and (ii) to assess the usefulness of field tests for regulatory risk assessment of the pesticide under registration. In this way, the treatment, evaluation, and the mutual comparability of field data for regulatory purposes is harmonized. It thereby provides a more consistent foundation for further risk assessment.     

Revisiting the Brazilian scenario of registry and protection of cultivars: an analysis of the period from 1998 to 2010, its dynamics and legal observations

During the last 20 years, the national production of grains has increased 156.1%; productivity increased 93.8% and there has been an increase of 29.1% in cultivated area. Currently, agribusiness is responsible for 40% of Brazilian exports. Nevertheless, there is little quantitative information on the main plant species of economic interest that have been registered and protected in the Agriculture, Fisheries and Food Supply Ministry (MAPA) by public and private companies, as well as by public-private partnerships. Consequently, we investigated the registry and protection of 27 species of economic interest, including the 15 that are the basis of the Brazilian diet, based on the information available on the site CultivarWeb, of MAPA, for the period from 1998 to August 30, 2010. We also examined the legislation that regulates registration and protection procedures and its implications for plant breeding and plant product development. It was found that the private sector controls 73.1% of the registrations and 53.56% of the protections, while 10.73% of the protections were of material developed overseas. Public-private partnerships contributed little to the development of new cultivars, with 0.5% of the registries and 3.61% of the protections. We conclude that plant protection directed private investment to development of wheat and rice varieties, with the greatest public investments directed to corn and sorghum. After the Cultivar Protection Law was implemented, there was restriction of access to germplasm banks, which could inhibit advances in Brazilian plant breeding programs, indicating a need for revision of this legal barrier.     

Scientific Integrity in Brazil

This article focuses on scientific integrity and the identification of predisposing factors to scientific misconduct in Brazil. Brazilian scientific production has increased in the last ten years, but the quality of the articles has decreased. Pressure on researchers and students for increasing scientific production may contribute to scientific misconduct. Cases of misconduct in science have been recently denounced in the country. Brazil has important institutions for controlling ethical and safety aspects of human research, but there is a lack of specific offices to investigate suspected cases of misconduct and policies to deal with scientific dishonesty.     

Market Assessment of Tuberculosis Diagnostics in Brazil in 2012

Background

Improved diagnostics for the diagnosis of tuberculosis (TB) are urgently needed. However, test developers and investors require market size data to support new product development. This study assessed the served available market for TB diagnostics in Brazil in 2012 and the market segmentation in the public and private sectors.

Methods

Data were collected on test volumes done in the public and private sectors for the diagnosis of latent and active TB, drug susceptibility testing and treatment follow-up. Tests included were tuberculin skin tests, interferon-gamma releases assays, smear microscopy, solid and liquid cultures, nucleic acid amplification tests and phenotypic drug susceptibility tests. The data were collected by means of an electronic survey via the Brazilian State laboratories and from sales information provided by manufacturers. Test costs for the public sector were calculated using a components approach, while costs for the private sector were based on prices paid by patients. The overall market value (expenditure) for the entire country was calculated using the public sector test costs.

Results

During 2012, an estimated total of 2.4 million TB diagnostic tests were done in Brazil, resulting in an estimated overall market value of USD 17.2 million. The public sector accounted for 91% of the test volumes and 88% of the market value. Smear microscopy was the most commonly test (n = 1.3 million; 55% of total) at an estimated value of USD 3.7 million. Culture overall (n = 302,761) represented 13% of test volumes and 40% (USD 6.9 million) of the market value. On average, USD 208 was spent on TB diagnostics for every notified TB patient in Brazil, in 2012.

Conclusion

The TB diagnostics market value in Brazil in 2012 was over USD 17 million. These study results will help test developers to understand the current and potential market for replacement or add-on diagnostic technologies.     

A Pragmatic Approach to Assess the Exposure of the Honey Bee (Apis mellifera) When Subjected to Pesticide Spray

Plant protection spray treatments may expose non-target organisms to pesticides. In the pesticide registration procedure, the honey bee represents one of the non-target model species for which the risk posed by pesticides must be assessed on the basis of the hazard quotient (HQ). The HQ is defined as the ratio between environmental exposure and toxicity. For the honey bee, the HQ calculation is not consistent because it corresponds to the ratio between the pesticide field rate (in mass of pesticide/ha) and LD₅₀ (in mass of pesticide/bee). Thus, in contrast to all other species, the HQ can only be interpreted empirically because it corresponds to a number of bees/ha. This type of HQ calculation is due to the difficulty in transforming pesticide field rates into doses to which bees are exposed. In this study, we used a pragmatic approach to determine the apparent exposure surface area of honey bees submitted to pesticide treatments by spraying with a Potter-type tower. The doses received by the bees were quantified by very efficient chemical analyses, which enabled us to determine an apparent surface area of 1.05 cm²/bee. The apparent surface area was used to calculate the exposure levels of bees submitted to pesticide sprays and then to revisit the HQ ratios with a calculation mode similar to that used for all other living species. X-tomography was used to assess the physical surface area of a bee, which was 3.27 cm²/bee, and showed that the apparent exposure surface was not overestimated. The control experiments showed that the toxicity induced by doses calculated with the exposure surface area was similar to that induced by treatments according to the European testing procedure. This new approach to measure risk is more accurate and could become a tool to aid the decision-making process in the risk assessment of pesticides.     

Transgenerational effects from exposure to environmental toxic substances

Exposure of mouse germ cells to radiation and chemicals results in mutation, malformation, cancer and other adverse effects (e.g., functional disorders) in the offspring, though these findings have not been proven in human studies. Environmental toxic substances such as urethane (ethyl carbamate) which had been injected subcutaneously to 50 million people as a co-solvent of analgesics and dioxin (an endocrine disruptor) have been found to be associated with adverse effects in the progeny of mice after parental exposures. There are some reports on congenital malformations in the progeny of fathers who had been exposed to dioxin. However, these substances have not shown mutagenicity in in vitro assay systems such as bacterial systems even with S9, cell transformation assays, etc., in spite of their potent teratogenicity and carcinogenicity in in vivo systems. Urethane was negative in the mouse specific locus test for germ cell mutations, but elicited a significant response at the same loci in the offspring of mice treated during pregnancy. Further, urethane is a mutagen in Drosophila germ cell tests, specifically inducing point mutations. Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin) does not induce in vivo somatic mutations in mice and rats. It does not induce chromosomal aberrations when the mouse and/or human sperm are treated, but induces mutations at ESTR (expanded simple tandem repeat) loci in mice at low frequencies and also congenital malformations. In this paper, we first present an overview of the results of our studies on transgenerational effects of these toxic substances, compare the results with those obtained after radiation exposure, and then discuss our subsequent studies to reconcile the problems underlying their mutagenicity, teratogenicity and carcinogenicity.     

Adjustments of the Pesticide Risk Index Used in Environmental Policy in Flanders

Indicators are used to quantify the pressure of pesticides on the environment. Pesticide risk indicators typically require weighting environmental exposure by a no effect concentration. An indicator based on spread equivalents (ΣSeq) is used in environmental policy in Flanders (Belgium). The pesticide risk for aquatic life is estimated by weighting active ingredient usage by the ratio of their maximum allowable concentration and their soil halflife. Accurate estimates of total pesticide usage in the region are essential in such calculations. Up to 2012, the environmental impact of pesticides was estimated on sales figures provided by the Federal Government. Since 2013, pesticide use is calculated based on results from the Farm Accountancy Data Network (FADN). The estimation of pesticide use was supplemented with data for non-agricultural use based on sales figures of amateur use provided by industry and data obtained from public services. The Seq-indicator was modified to better reflect reality. This method was applied for the period 2009-2012 and showed differences between estimated use and sales figures of pesticides. The estimated use of pesticides based on accountancy data is more accurate compared to sales figures. This approach resulted in a better view on pesticide use and its respective environmental impact in Flanders.     

Alleged misconceptions' distort perceptions of environmental cancer risks.

In a series of papers, Ames and colleagues allege that the scientific and public health communities have perpetuated a series of 'misconceptions' that resulted in inaccurate identification of chemicals that pose potential human cancer risks, and misguided cancer prevention strategies and regulatory policies. They conclude that exposures to industrial and synthetic chemicals represent negligible cancer risks and that animal studies have little or no scientific value for assessing human risks. Their conclusions are based on flawed and untested assumptions. For instance, they claim that synthetic residues on food can be ignored because 99.99% of pesticides humans eat are natural, chemicals in plants are pesticides, and their potential to cause cancer equals that of synthetic pesticides. Similarly, Ames does not offer any convincing scientific evidence to justify discrediting bioassays for identifying human carcinogens. Ironically, their arguments center on a ranking procedure that relies on the same experimental data and extrapolation methods they criticize as being unreliable for evaluating cancer risks. We address their inconsistencies and flaws, and present scientific facts and our perspectives surrounding Ames' nine alleged misconceptions. Our conclusions agree with the International Agency for Research on Cancer, the National Toxicology Program, and other respected scientific organizations: in the absence of human data, animal studies are the most definitive for assessing human cancer risks. Animal data should not be ignored, and precautions should be taken to lessen human exposures. Dismissing animal carcinogenicity findings would lead to human cancer cases as the only means of demonstrating carcinogenicity of environmental agents. This is unacceptable public health policy.